Time to Stop Holding the Elevator: A New Piece of the Transport Protein Mechanism Puzzle.
In this issue of Structure, McCoy et al. (2016) describe the 2.55-Å X-ray structure of the outward-facing occluded conformation of the Bacillus cereus maltose transporter MalT. This structure represents the penultimate piece needed to complete the picture of the transport cycle of the glucose superfamily of membrane-spanning EIIC components.


Spinal cord compression by extramedullary hematopoiesis in agnogenic myeloid metaplasia.
Extramedullary hematopoiesis presented as a diffuse mass causing almost complete obliteration of the spinal subarachnoid space in a patient with known agnogenic myeloid metaplasia. Pre- and post-radiation therapy films correlate well with significant clinical improvement in his neurological status.


Stressful life events, psychosocial health and general health in preschool children before age 4.
The impact of stressful life events (SLEs) in early childhood is often ignored. We aimed to examine longitudinal associations between SLEs and psychosocial and general health in preschool children. Twelve SLEs occurring before the age of 24 months were assessed and categorized by frequency (no events, 1-2 SLEs, and > 2 SLEs) and overall tension (no events, low, and high) (n = 1431). Psychosocial and general health were measured three times at the age of 24, 36 and 45 months. The associations were examined by logistic regression models using generalized estimating equations to handle repeated measurements. Half (48.4%) of the families experienced SLEs, and 23.8% perceived high-tension SLEs before the children were aged 24 months. Gender differences were observed in the association between SLEs and psychosocial health. Compared to girls without SLEs, girls who experienced > 2 SLEs [OR = 3.31, 95% confidence interval (CI) 2.05-5.35] or high-tension SLEs (OR = 3.01, 95% CI 2.07-4.39) had higher odds of psychosocial problems from 24 to 45 months. The odds ratios in boys were 2.10 (95% CI 1.36-3.24) and 1.47 (95% CI 1.06-2.03), respectively. Moreover, only girls' risk of psychosocial problems increased after experiencing 1-2 SLEs (OR = 2.15, 95% CI 1.54-3.00) or low-tension SLEs (OR = 1.90, 95% CI 1.31-2.74). Regarding general health, children who experienced > 2 SLEs (OR = 1.96, 95% CI 1.21-3.18) and high-tension SLEs (OR = 1.60, 95% CI 1.12-2.28) had higher odds of poor general health from 24 to 45 months. The findings emphasized that young children's psychosocial and general health can be impacted by experiencing SLEs in early childhood. Attention and adequate support for families experiencing SLEs are needed to minimize the potential negative effect of SLEs on child health, particularly in girls.


Alterations in repolarization of cardiac Purkinje fibers recovering from ischemic-like conditions: genesis of early afterdepolarizations.
Triggered activity initiated from delayed after-depolarizations has been proposed as a possible cause of arrhythmias during reperfusion of ischemic myocardium. However, the potential for abnormal repolarization and early afterdepolarizations (EADs) to develop under similar conditions has not been fully explored. Repolarization of the cell membrane during recovery from ischemic-like conditions was analyzed from transmembrane recordings in isolated rabbit Purkinje fibers paced at different basic cycle lengths. Preparations were exposed to conditions of hypoxia (defined as oxygen tension < 30 mmHg, high potassium, and zero substrate) plus lactic acidosis (pH 6.7) for 45 minutes followed by recovery in normal Tyrode's solution. Compared to control, action potentials during recovery at basic cycle length of 3,000 msec (n = 11) were characterized by a: (1) -7.2 +/- 2.1 mV shift in plateau potential (P < 0.01); (2) 126.1 +/- 63.6 msec increase in plateau duration (P < 0.05); and (3) 0.29 +/- 0.07 V/sec slowing of the maximum rate of terminal repolarization (phase 3; P < 0.01). These changes were larger when 10 to 20 microM amiloride was added to the hypoxic, acidotic test solution but were smaller when tissues were conditioned with hypoxia alone (zero lactate, pH 7.4). Following hypoxia plus acidosis, with or without amiloride, repolarization at long basic cycle lengths was often accompanied by EADs predominantly generated from potentials positive to -40 mV. These afterpotentials were blocked by Ca2+ channel antagonists and exhibited an activation threshold of -26.3 +/- 1.8 mV (n = 7). These data are consistent with the hypothesis that the combined negative voltage shift in the plateau and increase in its duration lead to the genesis of low membrane potential EADs by allowing reactivation of Ca2+ channels. Moreover, these results suggest that bradycardia-dependent EADs in Purkinje tissue may underlie arrhythmias in the intact heart during reperfusion of ischemic myocardium by mechanisms that are related in part to the acidosis established during the preceding ischemic conditions.


Mutation of a neutral amino acid in the transit peptide of rat mitochondrial malate dehydrogenase abolishes binding and import.
We have investigated the function of a leucine residue in the transit peptide of the rat mitochondrial malate dehydrogenase precursor using in vitro mutagenesis. Amino acid replacement of leucine 13 with glutamic acid and asparagine abolished import into mitochondria, while substitutions with proline, histidine, and arginine severely diminished uptake. In contrast, glutamine, tyrosine, valine, and alanine replacement resulted in normal levels of import, suggesting that there is a requirement for an uncharged residue at this position. Mutants involving rearrangements of the native sequence at positions 12-14 were imported as efficiently as the wild-type mitochondrial malate dehydrogenase, indicating that there was not an obligatory order of amino acid residues. However, deletion of leucine 13 resulted in diminished import. Binding studies with isolated mitochondria revealed that several position 13 mutants were deficient in binding to the mitochondrial surface, accounting for the reduced import of these proteins. This impairment could be distinguished from the effects due to decreased positive charge. We conclude that while translocation depends on the net positive charge, binding to the mitochondrial surface is mediated by uncharged residues within the transit peptides of mitochondrial precursor proteins.


Factors Governing the Accuracy of Subvisible Particle Counting Methods.
A number of new techniques for subvisible particle characterization in biotechnological products have emerged in the last decade. Although the pharmaceutical community is actively using them, the current knowledge about the analytical performance of some of these tools is still inadequate to support their routine use in the development of biopharmaceuticals (especially in the case of submicron methods). With the aim of increasing this knowledge and our understanding of the most prominent techniques for subvisible particle characterization, this study reports the results of a systematic evaluation of their accuracy. Our results showed a marked overcounting effect especially for low concentrated samples and particles fragile in nature. Furthermore, we established the relative sample size distribution as the most important contributor to an instrument's performance in accuracy counting. The smaller the representation of a particle size within a solution, the more difficulty the instruments had in providing an accurate count. These findings correlate with a recent study examining the principal factors influencing the precision of the subvisible particle measurements. A more thorough understanding of the capabilities of the different particle characterization methods provided here will help guide the application of these methods and the interpretation of results in subvisible particle characterization studies.


What is the minimal dose for resistance exercise effectiveness in prostate cancer patients? Systematic review and meta-analysis on patient-reported outcomes.
Active treatments for prostate cancer are well known to result in several adverse effects such as fatigue, depression and anxiety symptoms, impacting the overall quality of life (QoL) and wellbeing of a considerable proportion of patients. Resistance-based exercise interventions have shown positive effects to reduce or mitigate these treatment-related side effects. However, the minimal dosage required to derive these benefits is unknown. We systematically reviewed the resistance training effects in prostate cancer patients to determine the minimal dosage regarding the exercise components (mode, duration, volume and intensity) on fatigue, QoL, depression and anxiety. Using PRISMA guidelines, MEDLINE, CINAHL, EMBASE, SPORTDiscus and Web of Science databases were searched. Eligible randomised controlled trials examined prostate cancer patients undertaking resistance-based exercise programs during or following treatment. Meta-analysis was undertaken when more than three studies were included. Associations between resistance exercise components and its effects were tested by meta-regression analysis. Eighteen trials involving 1112 men with prostate cancer were included. Resistance-based exercise programs resulted in significant effects on fatigue (effect size = -0.3, 95% CI: -0.4 to -0.2, P < 0.001) and QoL (effect size = 0.2, 95% CI: 0.0 to 0.4, P = 0.018), with significant effects in specific questionnaires and domains of these outcomes. Resistance-based exercise effects on depression (effect size = -0.3, 95% CI: -0.7 to 0.0) and anxiety symptoms (effect size = -0.3, 95% CI: -0.5 to 0.0) were positive but not significant (P = 0.071 to 0.077). Meta-regression indicated no significant association between resistance exercise components with fatigue and QoL outcomes (P = 0.186-0.689). Low volume resistance exercise undertaken at a moderate-to-high intensity is sufficient to achieve significant fatigue and QoL benefits for men with prostate cancer and also mitigate depression and anxiety symptoms. A lower resistance exercise dosage than usually prescribed may help enhance adherence by reducing exercise barriers.


Relation of a coronary artery calcium score higher than 400 to coronary stenoses detected using multidetector computed tomography and to traditional cardiovascular risk factors.
Coronary artery calcium (CAC) is an excellent surrogate for atherosclerosis. However, this calcium is nonspecific for obstructive heart disease. This study sought to determine (1) the frequency of significant computed tomographic (CT) angiographic stenoses as a function of CAC scores, and (2) whether high CAC scores were associated with these stenoses independent of traditional risk factors. Subjects (n = 664) underwent Agatston CAC scoring and multidetector CT angiography using current 64-slice technology. Significant stenoses were defined as >60% diameter compromise. Self-reported risk factors and frequency of stenoses were analyzed as a function of CAC scores. The prevalence of risk factors increased significantly as CAC scores increased. Significant univariate associations included age (p <0.001), male gender (p <0.001), hypertension (p <0.001), and hyperlipidemia (p <0.001). There was also a significant association between CAC scores and the frequency of significant CT angiographic stenoses (p <0.001 for trend). The frequency of CT angiographic stenoses increased as CAC scores increased, with 7.9%, 8.3%, 14.5%, and 27.2% prevalences of significant stenoses in those with CAC scores of 1 to 100, 101 to 400, 401 to 1,000, and >1,000, respectively. Conversely, no significant lesions were found in those with no CAC. Multivariable logistic regression analysis controlling for traditional risk factors showed odds ratios for CAC score of 401 to 1,000 and >1,000 for having significant stenoses of 3.1 (95% confidence interval 1.6 to 6.0) and 6.9 (95% confidence interval 3.5 to 13.5), respectively. In conclusion, a CAC score >400 was significantly associated with multidetector CT angiographic stenoses independently of traditional risk factors.


Verrucous carcinoma, then and now.
The salient clinical and pathological features of verrucous carcinoma, as first described by Ackerman in 1948, are reviewed. Two case reports are presented. The first concerns a personal experience with a patient who had five surgical procedures for multifocal oral verrucous carcinoma and remains alive and well 28 years after her initial treatment. The second report highlights the remarkable story of President Grover Cleveland's secret surgery for verrucous carcinoma of the palate in 1893. Current concepts about the management of this uncommon tumor are reviewed, as well as some recent investigations that deal with its etiology.


Affinity labeling of forskolin-binding proteins. Comparison between glucose carrier and adenylate cyclase.
An [125I]iodoazidosalicylic acid derivative of forskolin was synthesized for identification of the diterpene's binding sites on the catalytic subunit of adenylate cyclase and on glucose transport proteins. The affinity label was selectively incorporated into proteins of Mr 40,000-60,000 in membranes from human erythrocytes and from various other tissues. The iodoazidosalicylic acid derivative also specifically labeled the catalytic moiety of adenylate cyclase from rabbit myocardial membranes. However, the structural requirements of the two forskolin-binding sites must be different, since the affinity of the photolabel for the glucose carriers is much higher than that for the cyclase catalyst. Furthermore, the label is readily competed with by D-glucose and cytochalasin B for its binding site on the glucose carrier but not on adenylate cyclase.


Enhancing the detection sensitivity of a high-resolution β - γ coincidence spectrometer.
A high-resolution β - γ coincidence spectrometry system has been set-up and calibrated at the UK CTBT Radionuclide Laboratory (known as GBL15) at AWE. The system has been configured specifically to measure the signatures of radioxenon isotopes that can be indicative of a nuclear explosion. The high purity germanium (HPGe) and PIPSBox detectors have been placed in an ultra-low-background lead shield to reduce the background count-rate and new software allows the combination of signals from four detectors (two HPGe detectors and two silicon-based detectors) to cover a larger solid angle. Measurements of samples of radioxenon isotopes are used to realise an improved detection efficiency and background acquisitions have demonstrated the achievable detection limits to reach 1.3 mBq for 133Xe and ≤0.3 mBq for the metastable isomers 131mXe and 133mXe. Due to the improved energy resolution in both photon and electron detectors, the detection sensitivity remains high in the presence of interfering signals from other radioxenon isotopes, such as those that may be present due to the significant levels of atmospheric radioxenon in parts of the world. This paper summarises the detector setup, efficiency calibration measurements and determination of the limits of detection. This work demonstrates the benefits of high-resolution coincidence detector systems for re-measurement of samples from the CTBT International Monitoring System (IMS) - improved detectability of metastable isomers 131mXe and 133mXe in the presence of 133Xe, compared to the current laboratory system.


Regulation of in situ to invasive breast carcinoma transition.
The transition of ductal carcinoma in situ (DCIS) to invasive carcinoma is a poorly understood key event in breast tumor progression. Here, we analyzed the role of myoepithelial cells and fibroblasts in the progression of in situ carcinomas using a model of human DCIS and primary breast tumors. Progression to invasion was promoted by fibroblasts and inhibited by normal myoepithelial cells. Molecular profiles of isolated luminal epithelial and myoepithelial cells identified an intricate interaction network involving TGFbeta, Hedgehog, cell adhesion, and p63 required for myoepithelial cell differentiation, the elimination of which resulted in loss of myoepithelial cells and progression to invasion.


Unique patterns of lower respiratory tract microbiota are associated with inflammation and hospital mortality in acute respiratory distress syndrome.
The lung microbiome maintains the homeostasis of the immune system within the lungs. In acute respiratory distress syndrome (ARDS), the lung microbiome is enriched with gut-derived bacteria; however, the specific microbiome associated with morbidity and mortality in patients with ARDS remains unclear. This study investigated the specific patterns of the lung microbiome that are correlated with mortality in ARDS patients. We analyzed the lung microbiome from the bronchoalveolar lavage fluid (BALF) of patients with ARDS and control subjects. We measured the copy numbers of 16S rRNA and the serum and BALF cytokines (interleukin [IL]-6, IL-8, receptor for advanced glycation end products, and angiopoietin-2). We analyzed 47 mechanically ventilated patients diagnosed with (n = 40) or without (n = 7; control) ARDS. The alpha diversity was significantly decreased in ARDS patients compared with that of the controls (6.24 vs. 8.07, P = 0.03). The 16S rRNA gene copy numbers tended to be increased in the ARDS group compared with the controls (3.83 × 106 vs. 1.01 × 105 copies/mL, P = 0.06). ARDS patients were subdivided into the hospital survivor (n = 24) and non-survivor groups (n = 16). Serum IL-6 levels were significantly higher in the non-survivors than in the survivors (567 vs. 214 pg/mL, P = 0.027). The 16S rRNA copy number was significantly correlated with serum IL-6 levels in non-survivors (r = 0.615, P < 0.05). The copy numbers and relative abundance of betaproteobacteria were significantly lower in the non-survivors than in the survivors (713 vs. 7812, P = 0.012; 1.22% vs. 0.08%, P = 0.02, respectively). Conversely, the copy numbers of Staphylococcus, Streptococcus and Enterobacteriaceae were significantly correlated with serum IL-6 levels in the non-survivors (r = 0.579, P < 0.05; r = 0.604, P < 0.05; r = 0.588, P < 0.05, respectively). The lung bacterial burden tended to be increased, and the alpha diversity was significantly decreased in ARDS patients. The decreased Betaproteobacteria and increased Staphylococcus, Streptococcus and Enterobacteriaceae might represent a unique microbial community structure correlated with increased serum IL-6 and hospital mortality. The institutional review boards of Hiroshima University (Trial registration: E-447-4, registered 16 October 2019) and Kyoto Prefectural University of Medicine (Trial registration: ERB-C-973, registered 19 October 2017) approved an opt-out method of informed consent.


Thermal decomposition of acetic anhydride-nitric acid mixtures.
The adoption of Ac2O/HNO3 mixtures for the nitration of organics or the preparation of nitric esters is not free from some risks of explosion. The behaviour of these mixtures at varying Ac2O/HNO3 molar ratios is studied by means of adiabatic and scanning calorimetry. A simplified kinetic model, based on two lumped reactions (one of which leads to the formation of tetranitromethane) is developed to simulate the thermal decomposition of the investigated system under adiabatic conditions. The use of this model allows satisfactory prediction of the temperature profiles in the reactor whereas calculated pressures are generally overestimated due to some model inadequacies to account for the absorption of gaseous decomposition products into the reacting solution.


Acupuncture for migraine prophylaxis: a randomized controlled trial.
Acupuncture is commonly used to treat migraine. We assessed the efficacy of acupuncture at migraine-specific acupuncture points compared with other acupuncture points and sham acupuncture. We performed a multicentre, single-blind randomized controlled trial. In total, 480 patients with migraine were randomly assigned to one of four groups (Shaoyang-specific acupuncture, Shaoyang-nonspecific acupuncture, Yangming-specific acupuncture or sham acupuncture [control]). All groups received 20 treatments, which included electrical stimulation, over a period of four weeks. The primary outcome was the number of days with a migraine experienced during weeks 5-8 after randomization. Our secondary outcomes included the frequency of migraine attack, migraine intensity and migraine-specific quality of life. Compared with patients in the control group, patients in the acupuncture groups reported fewer days with a migraine during weeks 5-8, however the differences between treatments were not significant (p > 0.05). There was a significant reduction in the number of days with a migraine during weeks 13-16 in all acupuncture groups compared with control (Shaoyang-specific acupuncture v. control: difference -1.06 [95% confidence interval (CI) -1.77 to -0.5], p = 0.003; Shaoyang-nonspecific acupuncture v. control: difference -1.22 [95% CI -1.92 to -0.52], p < 0.001; Yangming-specific acupuncture v. control: difference -0.91 [95% CI -1.61 to -0.21], p = 0.011). We found that there was a significant, but not clinically relevant, benefit for almost all secondary outcomes in the three acupuncture groups compared with the control group. We found no relevant differences between the three acupuncture groups. Acupuncture tested appeared to have a clinically minor effect on migraine prophylaxis compared with sham acupuncture. Clinicaltrials.gov NCT00599586.


Identification and characterization of selenate- and selenite-responsive genes in a Se-hyperaccumulator Astragalus racemosus.
Plants with capacity to accumulate high levels of selenium (Se) are desired for phytoremediation and biofortification. Plants of genus Astragalus accumulate and tolerate high levels of Se, but their slow growth, low biomass and non-edible properties limit their direct utilization. Genetic engineering may be an alternative way to produce edible or high biomass Se-accumulating plants. The first step towards this goal is to isolate genes that are responsible for Se accumulation and tolerance. Later, these genes can be introduced into other edible and high biomass plants. In the present study, we applied fluorescent differential display to analyze the transcript profile of Se-hyperaccumulator A. racemosus treated with 20 μM selenate (K(2)SeO(4)) for 2 weeks. Among 125 identified Se-responsive candidate genes, the expression levels of nine were induced or suppressed more than twofold by selenate treatment in two independent experiments while 14 showed such changes when treated with selenite (K(2)SeO(3)). Six of them were found to respond to both selenate and selenite treatments. A novel gene CEJ367 was found to be highly induced by both selenate (1,920-fold) and selenite (579-fold). Root- or shoot-preferential expression of nine genes was further investigated. These identified genes may allow us to create Se-enriched transgenic plants.


Iterative-build OMIT maps: map improvement by iterative model building and refinement without model bias.
A procedure for carrying out iterative model building, density modification and refinement is presented in which the density in an OMIT region is essentially unbiased by an atomic model. Density from a set of overlapping OMIT regions can be combined to create a composite 'iterative-build' OMIT map that is everywhere unbiased by an atomic model but also everywhere benefiting from the model-based information present elsewhere in the unit cell. The procedure may have applications in the validation of specific features in atomic models as well as in overall model validation. The procedure is demonstrated with a molecular-replacement structure and with an experimentally phased structure and a variation on the method is demonstrated by removing model bias from a structure from the Protein Data Bank.


Intrahepatic arterioportal fistulae: role of transcatheter embolization.
Management of 26 arterioportal fistulae (APFs) is reported. Among 13 hepatoma-induced fistulae (group A), conservative treatment was ineffective in 8 patients, and arterial embolization alleviated portal hypertension in the other 5. Of 10 iatrogenic APFs (group B), the 3 largest were successfully embolized, the remaining lesions resolved spontaneously. Three spontaneous nonmalignant APFs (group C) were embolized. Excellent results were obtained in 2 patients, and the other died of severe postembolization hepatic failure. Because long-standing APFs may cause severe portal hypertension with consequent variceal bleeding they should be treated. Arterial embolization is indicated in most patients.


Implant Shell Characteristics: The Science Behind the Surface.
Modern breast implant design emphasizes the host response at the surface. Implant surfaces are characterized by their roughness, surface area, and potential for bacterial attachment. The future of implant design may lie in the ability of bioengineers to transform both the structure and chemical properties of the device surface and therefore affect potential long-term outcomes.


Molecular basis of the JAHK (RH53) antigen.
The JAHK antigen was first described in 1995 as a low-frequency red blood cell antigen. Family studies confirmed the association of the antigen with the rare r(G) phenotype of the Rh blood group system, which is associated with weak expression of C and e, but normal G expression. JAHK was allocated the Rh number RH53. The serologic findings indicated the location of the antigen on the RhCE protein, although the molecular basis for JAHK has not been known. The RHCE gene of eight persons from three unrelated families was analyzed by exon amplification and direct sequencing. Four of the samples were JAHK+ the remaining four were JAHK-. In one JAHK+ sample, the entire RHCE gene was sequenced. The remaining samples were sequenced for exons 1 to 3. A polymerase chain reaction procedure with sequence specific primers was developed for the specific detection of the JAHK allele. Analysis of the entire RHCE gene of one JAHK+ sample showed the expected CcEe-specific nucleotide sequences and revealed an additional nucleotide change (365C>T) in exon 3. This change represented a missense mutation, which led to an amino acid substitution from serine to leucine at position 122 of the RhCE protein. Three JAHK+ samples from two other unrelated families showed the 365C>T mutation and confirmed the association of the Ser122Leu substitution with the JAHK+ phenotype. The molecular basis of the JAHK antigen (RH53) is defined by a 365C>T mutation in exon 3 of the RHCE gene leading to the amino acid substitution Ser122Leu. Because the position 122 is predicted to be located in the transmembrane region adjacent to the second loop, the substitution of the neutral serine by the hydrophobic leucine seems to be the cause of the JAHK antigen by a conformational change of the RhCE protein. This structural change may also cause the weakened expression of the C and e antigens observed in JAHK+ individuals. Based on our results it is concluded that the JAHK-specific mutation is associated with a dCe haplotype.


The effect of anaesthetics on the dynamic heterogeneity of lipid membranes.
The influence of membrane-perturbing drugs such as anaesthetics on the lipid membrane properties is analyzed theoretically on the basis of a general microscopic interaction model of the gel-to-fluid chain melting transition of one-component phospholipid membranes and phospholipid membranes with a low content of cholesterol. Monte Carlo computer simulation of the model shows that the gel-to-fluid transition of the lipid membrane, manifested in the formation of dynamically coexisting domains of gel and fluid lipids, is strongly influenced by the presence of anaesthetics. Macroscopically the effect of anaesthetics on the membrane properties is seen in a depression of the transition temperature and a smearing of thermodynamic response functions like the specific heat. Microscopically the calculations reveal that anaesthetics have a high affinity to the fluctuating domain interfaces that are dominated by kink-like lipid-chain conformations. This leads to formation of more interfaces and to a locally high concentration of anaesthetics in the interfacial regions, which is much larger than the global concentration in the membrane. Important membrane components like cholesterol, which also has been shown to be interfacially active, are found to decrease the absorption of anaesthetics and to squeeze out anaesthetics from the interfaces. The results of the general model study of anaesthetics-membrane interactions are discussed in relation to both general anaesthetics, like halothane, and local anaesthetics like cocaine-derivatives.


Moving towards a new era of genomics in the neuronal ceroid lipofuscinoses.
The neuronal ceroid lipofuscinoses (NCL) are a group of disorders defined by shared clinical and pathological features, including seizures and progressive decline in vision, neurocognition, and motor functioning, as well as accumulation of autofluorescent lysosomal storage material, or 'ceroid lipofuscin'. Research has revealed thirteen distinct genetic subtypes. Precisely how the gene mutations lead to the clinical phenotype is still incompletely understood, but recent research progress is starting to shed light on disease mechanisms, in both gene-specific and shared pathways. As the application of new sequencing technologies to genetic disease diagnosis has grown, so too has the spectrum of clinical phenotypes caused by mutations in the NCL genes. Most genes causing NCL have probably been identified, underscoring the need for a shift towards applying genomics approaches to achieve a deeper understanding of the molecular basis of the NCLs and related disorders. Here, we summarize the current understanding of the thirteen identified NCL genes and the proteins they encode, touching upon the spectrum of clinical manifestations linked to each of the genes, and we highlight recent progress leading to a broader understanding of key pathways involved in NCL disease pathogenesis and commonalities with other neurodegenerative diseases.


Nicotinamide mononucleotide induces lipolysis by regulating ATGL expression via the SIRT1-AMPK axis in adipocytes.
Nicotinamide adenine dinucleotide (NAD+) -dependent protein deacetylase SIRT1 plays an important role in the regulation of metabolism. Although the administration of nicotinamide mononucleotide (NMN), a key NAD+ intermediate, has been shown to ameliorate metabolic disorders, such as insulin resistance and glucose intolerance, the direct effect of NMN on the regulation of lipid metabolism in adipocytes remains unclear. We here investigated the effect of NMN on lipid storage in 3T3-L1 differentiated adipocytes. Oil-red O staining showed that NMN treatment reduced lipid accumulation in these cells. NMN was found to enhance lipolysis in adipocytes since the concentration of glycerol in the media was increased by NMN treatment. Western blotting and real-time RT-PCR analysis revealed that adipose triglyceride lipase (ATGL) expression at both protein and mRNA level was increased with NMN treatment in 3T3-L1 adipocytes. Whereas NMN increased SIRT1 expression and AMPK activation, an AMPK inhibitor compound C restored the NMN-dependent upregulation of ATGL expression in these cells, suggesting that NMN upregulates ATGL expression through the SIRT1-AMPK axis. NMN administration significantly decreased subcutaneous fat mass in mice on a high-fat diet. We also found that adipocyte size in subcutaneous fat was decreased with NMN treatment. Consistent with the alteration of fat mass and adipocyte size, the ATGL expression in subcutaneous fat was slightly, albeit significantly, increased with NMN treatment. These results indicate that NMN suppresses subcutaneous fat mass in diet-induced obese mice, potentially in part via the upregulation of ATGL. Unexpectedly, the reduction in fat mass as well as ATGL upregulation with NMN treatment were not observed in epididymal fat, implying that the effects of NMN are site-specific in adipose tissue. Thus, these findings provide important insights into the mechanism of NMN/NAD+ in the regulation of metabolism.


Flow and aspiration of artificial feeds through nasogastric tubes.
The physical characteristics of 12 nasogastric tubes (Clinifeed, Portex prima, Vygon 6 and 9, Dobbhoff, Nutriflex and 6 Ryles tubes) and 13 feeds (4 Clinifeed Preparations, Nutranel, Nutrauxil, Express, Triosorbon, Fortison, Enteral 400, Elemental 028, Complan and Complan + Caloreen have been assessed under laboratory conditions. The accuracy of the stated internal diameter of the tubes tested varied from 88-119%. The rate of flow was inversely related to the length of the feeding tube and was greatly affected by diameter (an increase from 1 to 2 mm increased flow 10-fold); and was virtually unaffected by the side holes at the end of the tube. Giving sets provide only a small resistance to flow. The viscosities of liquid feeds did not correlate with their osmolarities. The values ranged from 1.9-28 centipois (cps) at 24 degrees C and universally decreased to 1.4-19 cps at 37 degrees C. All enteral feeds except Clinifeed 500, flow satisfactorily through fine bore tubes under a low head of pressure. Undiluted Clinifeed 500 (a high protein feed) requires pumping. Feeds containing casein (e.g. Complan, Nutrauxil, Fortison, Triosorbon) curdle in the presence of acid which hinders aspiration through even widebore tubes. Foods containing whey protein (Express, Enteral 400) or no protein (Elemental 028) do not coagulate with change of pH. Batch to batch variations in liquid feeds are usually small but with one preparation were sufficiently large to cause problems in delivery. The results of these studies provide further data on which to base the choice of liquid feed and giving tubes in clinical practice.


Cellularity and apoptosis after radiofrequency-induced shrinkage of collagenous tissue: assessment of postoperative immobilization using an in vivo rabbit model.
Electrothermally-assisted capsular shrinkage has been gaining increased acceptance in the treatment of shoulder instability. Its indication in ACL-deficient knees has been discussed recently. We examined the influence of immobilization on cell homeostasis of healing collagenous tissue after radiofrequency energy was applied to the patellar tendon in 23 rabbits. The animals were killed immediately after surgery (n = 6) or 3 weeks after surgery (n = 17). 10 rabbits were allowed normal cage activity, whereas the treated hind limb of 7 animals was immobilized for 3 weeks in a cast. Feulgen staining was used to stain the DNA of cell nuclei. Cells undergoing apoptosis were identified by the TUNEL method. Quantitative histological assessment was performed using imaging analysis software. Severe cellular damage in RF-treated collagenous tissue was partly induced by the immediate onset of apoptosis. At 3 weeks after surgery, non-immobilized tendon showed increased cellularity and apoptosis, whereas immobilization prevented the increase in cellularity and apoptosis significantly. The calculated ratio of apoptosis was not influenced by any postoperative treatment. Diminished cellularity and apoptosis during tissue remodeling, due to immobilization, may protect the shortened collagenous scaffold from stretching and further optimize the clinical outcome after radiofrequency shrinkage. To stabilize the shrunken tissue, proliferation during postoperative wound healing should be minimized by careful rehabilitation.


Lyme neuroborreliosis: Progressive cerebral vasculitis responsive to cyclophosphamide. A case report and review of the literature.
Less than 1 % of patients with Lyme Neuroborreliosis (LNB) present with a cerebrovascular event. Ischaemic strokes occur more commonly than parenchymal or subarachnoid haemorrhages. If cerebral vasculitis due to LNB is suspected, antibiotic treatment should be started immediately, which will normally lead to remission. Very rarely progression and recurrent strokes are observed despite sufficient antibiotic therapy, even if steroids are added. Currently there are no guidelines on the adequate treatment of cerebral vasculitis due to LNB which is not responsive to antibiotics and steroids, but in very few reported cases cyclophosphamide led to disease stabilisation. We reviewed the literature regarding cyclophosphamide treatment in these patients and want to share our experience of cyclophosphamide therapy in progressive cerebral vasculitis due to LNB. We report a 71-year-old female patient with cerebral vasculitis and multiple strokes as a complication of LNB. Progression could only be halted by additional immunosuppressive treatment using cyclophosphamide. However, at that point the patient had already suffered severe ischaemic brain damage. Similarly, in existing case reports cyclophosphamide had been administered only at a time when patients already showed serious neurological deficits. Cerebral vasculitis in patients with LNB is very rare and normally responds to antibiotic treatment. A minority of patients show disease progression despite antibiotics and steroids. Our case report strengthens the recommendation that in those patients - even if signs of progressive vasculitis are only detectable on imaging and not clinically - cyclophosphamide should be considered without delay to prevent further cerebrovascular events.


Hydrocephalus caused by conditional ablation of the Pten or beta-catenin gene.
To investigate the roles of Pten and beta-Catenin in the midbrain, either the Pten gene or the beta-catenin gene was conditionally ablated, using Dmbx1 (diencephalon/mesencephalon-expressed brain homeobox gene 1)-Cre mice. Homozygous disruption of the Pten or beta-catenin gene in Dmbx1-expressing cells caused severe hydrocephalus and mortality during the postnatal period. Conditional deletion of Pten resulted in enlargement of midbrain structures. beta-catenin conditional mutant mice showed malformation of the superior and inferior colliculi and stenosis of the midbrain aqueduct. These results demonstrate that both Pten and beta-Catenin are essential for proper midbrain development, and provide the direct evidence that mutations of both Pten and beta-catenin lead to hydrocephalus.


Polymorphisms, genomic imprinting and cancer susceptibility.
Polymorphisms have been identified in proto-oncogenes and tumor suppressor genes that predispose people to cancer. Recent evidence indicates that genomic imprinting, an epigenetic form of gene regulation that results in uniparental gene expression, can also function as a cancer predisposing event. Thus, cancer susceptibility is increased by both Mendelian inherited genetic and non-Mendelian inherited epigenetic events. Consequently, chemical and physical agents cannot only induce cancer through the formation of genetic mutations but also through epigenetic changes that result in the inappropriate expression of imprinted proto-oncogenes and tumor suppressor genes. The role of genomic imprinting in carcinogenesis and cancer susceptibility is examined in this review.


Psychiatric disorders among the Mapuche in Chile.
The Mapuche are the largest indigenous group in Chile; yet almost all data on the mental health of indigenous populations are from North America. The study examines the differential DSM-III-R prevalence rates of psychiatric disorders and service utilization among indigenous and non-indigenous community residence. The Composite International Diagnostic Interview (CIDI) was administered to a stratified random sample of 75 Mapuche and 434 non-Mapuche residents of the province of Cautín. Lifetime prevalence and 12-month prevalence rates were estimated. Approximately 28.4% of the Mapuche population had a lifetime, and 15.7% a 12-month, prevalent psychiatric disorder compared to 38.0% and 25.7%, respectively, of the non-Mapuche. Few significant differences were noted between the two groups; however, generalized anxiety disorder, simple phobia, and drug dependence were less prevalent among the Mapuche. Service utilization among the Mapuche with mental illness was low. This is a preliminary study based on a small sample size. Further research on the mental health of indigenous populations of South America is needed.


Immunohistochemical localization of cytokeratins in normal eccrine glands, with monoclonal antibodies in routinely processed, formalin-fixed, paraffin-embedded sections.
Immunohistochemical detection in paraffin sections would be of value in the retrospective analysis of formalin-fixed, paraffin-embedded tissues. In this study with several monoclonal antibodies directed against fixation-resistant keratin antigens, we performed immunohistochemical staining on routine paraffin-embedded sections to localize specific keratins in the eccrine sweat glands and to determine the tissue distribution of various keratins. We clarified the localization of specific keratins in different segments of normal eccrine glands. Immunohistochemical localization of individual cytokeratins showed different patterns in secretory and ductal portions, although cytokeratins were expressed in all segments of normal sweat glands. Different immunoreactivities were noted in the luminal and basal cells of eccrine ducts. There were also some differences in immunophenotype between the acrosyringium and the dermal ducts. These data provide information for the determination of the histogenesis and differentiation of skin tumors with eccrine differentiation.


Modulation of transcriptional corepressor activity of prospero-related homeobox protein (Prox1) by SUMO modification.
Prospero-related homeobox protein (Prox1) plays essential roles in the development of many tissues and organs. In the present study, we show that Prox1 is modified by the small ubiquitin-like protein SUMO-1 in cultured cells. Mutation analysis identified at least four potential sumoylation sites within the repression domain of Prox1. Our data indicate that sumoylation of Prox1 reduces its interaction with HDAC3 and as a result downregulates its corepressor activity. These findings suggest that sumoylation may serve as a novel mechanism for the regulation of Prox1's corepressor activity.


Experimental realization of hyperbolic dispersion metamaterial for the whole visible spectrum based on liquid crystalline phase soft template.
We experimentally demonstrated a metamaterial composed of hexagonal arrays of silver nanowires that exhibits hyperbolic dispersion and negative refraction in the entire visual wavelength range. The nanowires with extremely small size of 10 nm diameter and 15 nm center-to-center distance were fabricated using the reverse hexagonal liquid crystalline phase template containing AgNO(3) solution. Through the experiments of angle dependent reflectance for s-polarization and p-polarization, the dielectric constants were measured in several wavelengths. Calculations and experiments both show hyperbolic dispersion relations from 370 nm to 750 nm which indicates the presence of all-angle negative refraction. For all the experimental wavelengths, the permittivities of the material are in good agreement with the theoretical calculations.


Chemonucleolysis of lumbar intervertebral disc prolapse with chymopapain: outcome after 1 year.
One hundred and one consecutive patients with lumbar disc prolapse were treated by chymopapain chemonucleolysis and their response and favourable pre-treatment criteria determined. Most improvement occurred within the first month, and one year after treatment outcome was judged satisfactory (excellent or good) in 71%. Individual patient characteristics associated with a satisfactory response were sciatica of greater severity than back pain (p = 0.005) and duration of symptoms less than 18 months (p = 0.03). Patients fulfilling 3 or 4 of four immediate pre-treatment clinical and radiographic criteria (sciatica more severe than back pain, reduced straight leg raising, neurological deficit, radiographic abnormality) had a satisfactory response more often than others (p less than 0.05). Reported success one year after surgery for disc prolapse is similar. Chemonucleolysis, however, requires less resources and does not preclude subsequent operation. Our results therefore suggest that it might be considered an alternative to surgery when conservative treatment has failed.


Influence of thyroid state and improved hypoxia tolerance on noise-induced cochlea damage.
Guinea pigs were exposed to pure tone noise (2.7 kHz, 130 dB, 1 h) and cochlear microphonic potentials were measured 24 h after exposure. There is the possibility to modify the resulting noise-induced cochlea damage by regulating the function of the thyroid gland to alter the rate of metabolism. A hypofunction of the thyroid gland during sound exposure lessens, an over-function aggravates the damage. After gradual adaptation of the animals to a simulated 10,000 m altitude, the electrophysiologically demonstratable noise-induced damage was reduced. This might be explained by the greater hypoxia tolerance and perhaps additional better oxygen supply to the receptor cells.


A decade of community MRSA in New Zealand.
In 1992, isolates with a distinctive phage pattern were identified amongst the 186 MRSA recovered in New Zealand. These unusual isolates were recovered in the Auckland region from individuals who came from or had visited Western Samoa, and were called Western Samoan phage pattern (WSPP) MRSA. They were almost exclusively community based and were mainly responsible for the alarming 15-fold increase in MRSA seen in New Zealand over the next 6 years. Since 2000, the number of infections attributable to WSPP MRSA appears to be declining. WSPP isolates are clonal, possess a unique type IV SCC mec element, and a distinctive multilocus sequence allelic profile (ST30). WSPP isolates are invariably not multiresistant with methicillin MICs generally <or=32 microg/ml. Virulence of the WSPP clone appears to be related to its adhesive and consistent toxin- (e.g. Panton-Valentine leukocidin, alpha- and gamma-haemolysins) producing capabilities. Isolates are most frequently associated with cutaneous lesions in younger age groups. Since 1998, MRSA isolates belonging to the UK-derived EMRSA-15 strain (also type IV SCC mec) have continued to increase in New Zealand, and together with WSPP, these strains now dominate MRSA isolations in New Zealand.


Numerical Analysis of Conventional Air Pollutants and PM10 Concentration Affected to Respiratory Disease Patients in Bangkok, Thailand.
Ambient air pollution is a major cause of death and disease globally. The linear regression between respiratory disease patients, PM10 and other criteria pollutants were used in this study. Among respiratory disease patients and CO was statistically significant with P-value of <0.01 (r=0.198, 95% CI=48.74-288.16). As for respiratory disease patients and NOx was statistically significant with P-value of <0.01 (r=0.190, 95% CI=1379.74-9241.43).


Aberrant expression of NPPB through YAP1 and TAZ activation in mesothelioma with Hippo pathway gene alterations.
Mesothelioma is a neoplastic disease associated with asbestos exposure. It is highly malignant and has a poor prognosis; thus, early detection is desirable. Recent whole-genome analysis has revealed that mesothelioma is characterized by a high frequency of mutations in a set of genes involved in the Hippo pathway, such as NF2 and LATS2. However, a rapid, simple, and precise method for finding mesothelioma with these mutations has not yet been established. Clustering of Hippo pathway gene alteration groups and the differential expression of each gene in mesothelioma patients were analyzed using The Cancer Genome Atlas database. Gene expression levels in various tumors and normal tissues were analyzed using public databases. Knockdown or transient expression of YAP1 or TAZ was performed to evaluate the regulation of gene expression by these genes. NT-proBNP was measured in the pleural effusions of 18 patients and was compared with NF2 expression in five cases where cell lines had been successfully established. NPPB mRNA expression was markedly higher in the group of mesothelioma patients with Hippo pathway gene mutations than in the group without them. NPPB expression was low in all normal tissues except heart, and was highest in mesothelioma. Mesothelioma patients in the high NPPB expression group had a significantly worse prognosis than those in the low NPPB expression group. NPPB expression was suppressed by knockdown of YAP1 or TAZ. NT-proBNP was abundant in the effusions of mesothelioma patients and was particularly high in those with impaired NF2 expression. NPPB, whose levels can be measured in pleural effusions of mesothelioma patients, has the potential to act as a biomarker to detect NF2-Hippo pathway gene alterations and/or predict patient prognosis. Additionally, it may provide useful molecular insights for a better understanding of mesothelioma pathogenesis and for the development of novel therapies.


Small-Molecule MYC Inhibitors Suppress Tumor Growth and Enhance Immunotherapy.
Small molecules that directly target MYC and are also well tolerated in vivo will provide invaluable chemical probes and potential anti-cancer therapeutic agents. We developed a series of small-molecule MYC inhibitors that engage MYC inside cells, disrupt MYC/MAX dimers, and impair MYC-driven gene expression. The compounds enhance MYC phosphorylation on threonine-58, consequently increasing proteasome-mediated MYC degradation. The initial lead, MYC inhibitor 361 (MYCi361), suppressed in vivo tumor growth in mice, increased tumor immune cell infiltration, upregulated PD-L1 on tumors, and sensitized tumors to anti-PD1 immunotherapy. However, 361 demonstrated a narrow therapeutic index. An improved analog, MYCi975 showed better tolerability. These findings suggest the potential of small-molecule MYC inhibitors as chemical probes and possible anti-cancer therapeutic agents.


The culture broker in post-RN education: a view from a distance.
Education is becoming decentralized. The paradigm shift requires us to rethink the ways educational values are delivered. As Connick asserts, education has become a process rather than a place on a campus. In the distance education site at Gallup, N.M., we realized that the context is organized around a new set of relationships. Reality, interpreted in another context, has a culture of its own. Even where facts are known there is often confusion about their meaning. The site coordinator is a valuable force in the aligning and managing of meaning between contexts.


Exercise Reduces the Resumption of Tumor Growth and Proteolytic Pathways in the Skeletal Muscle of Mice Following Chemotherapy.
The pathogenesis of muscle atrophy plays a central role in cancer cachexia, and chemotherapy contributes to this condition. Therefore, the present study aimed to evaluate the effects of endurance exercise on time-dependent muscle atrophy caused by doxorubicin. For this, C57 BL/6 mice were subcutaneously inoculated with Lewis lung carcinoma cells (LLC group). One week after the tumor establishment, a group of these animals initiated the doxorubicin chemotherapy alone (LLC + DOX group) or combined with endurance exercise (LLC + DOX + EXER group). One group of animals was euthanized after the chemotherapy cycle, whereas the remaining animals were euthanized one week after the last administration of doxorubicin. The practice of exercise combined with chemotherapy showed beneficial effects such as a decrease in tumor growth rate after chemotherapy interruption and amelioration of premature death due to doxorubicin toxicity. Moreover, the protein degradation levels in mice undergoing exercise returned to basal levels after chemotherapy; in contrast, the mice treated with doxorubicin alone experienced an increase in the mRNA expression levels of the proteolytic pathways in gastrocnemius muscle (Trim63, Fbxo32, Myostatin, FoxO). Collectively, our results suggest that endurance exercise could be utilized during and after chemotherapy for mitigating muscle atrophy promoted by doxorubicin and avoid the resumption of tumor growth.


Recurrent sixth nerve palsy associated with a hypoplastic Dorello's canal.
Recurrent sixth nerve palsies are usually idiopathic, although they have been postulated to be caused by a postinflammatory insult or abnormal cerebral circulation. We report the case a 9-month-old boy who had 2 episodes of a left abduction deficit that resolved spontaneously. A 3-dimensional high-resolution T2-weighted sequence magnetic resonance imaging scan of the head revealed a hypoplastic left 6th nerve and a small left Dorello's canal.


MicroRNAs involved in the TGF-β signaling pathway in atherosclerosis.
Atherosclerosis (AS) is a chronic inflammatory vascular disease with a multifactorial pathogenesis. It becomes a global health concern, especially causing an array of fatal consequences among the elderly. However, the mechanisms of AS remain unexplained. The transforming growth factor-β (TGF-β) signaling pathway is widely involved in the inflammation, immune function, proliferation, differentiation,and apoptosis in vivo. Based on previous researches, it has not been confirmed whether the TGF-β pathway promotes or inhibits atherosclerosis. Furthermore, more and more studies have found that microRNAs can regulate atherosclerosis through the TGF-β signaling pathway. In this review, we summarize and discuss the role of microRNAs in the pathogenesis of atherosclerosis via the TGF-β signaling pathway.


Theoretical studies of energetic nitrogen-rich ionic salts composed of substituted 5-nitroiminotetrazolate anions and various cations.
We have performed density functional theory and volume-based thermodynamics calculations to study the effects of different combinations of energetic anions and cations on the crystal densities, heats of formation, energetic properties, and thermodynamics of formation for a series of 5-nitroiminotetrazolate-based ionic salts. The results show that the substitution of the -NO2, -NF2, -N3, or -C(NO2)3 group is helpful for increasing the densities of the salts. Incorporating every substituent (-NH2, -NO2, -NF2, -N3, or -C(NO2)3) into the salt is favorable for improving its HOF and detonation performance. Incorporating the cation B1, B2, B10, or B11 into the salts is helpful for improving its detonation properties. Increasing negative charge for the 5-nitroiminotetrazolate-based salts is unfavorable for enhancing the density and detonation performance, but is helpful for increasing the HOFs. Many salts present comparable detonation performance with commonly used explosives RDX or HMX. Among them, 21 salts have near or better properties than HMX. The thermodynamics of formation of the salts show that the majority of the 5-nitroiminotetrazolate salts with the cation B1, B3, B9, B10, B12 could be synthesized by the proposed reactions.


Transient ECM protease activity promotes synaptic plasticity.
Activity-dependent proteolysis at a synapse has been recognized as a pivotal factor in controlling dynamic changes in dendritic spine shape and function; however, excessive proteolytic activity is detrimental to the cells. The exact mechanism of control of these seemingly contradictory outcomes of protease activity remains unknown. Here, we reveal that dendritic spine maturation is strictly controlled by the proteolytic activity, and its inhibition by the endogenous inhibitor (Tissue inhibitor of matrix metalloproteinases-1 - TIMP-1). Excessive proteolytic activity impairs long-term potentiation of the synaptic efficacy (LTP), and this impairment could be rescued by inhibition of protease activity. Moreover LTP is altered persistently when the ability of TIMP-1 to inhibit protease activity is abrogated, further demonstrating the role of such inhibition in the promotion of synaptic plasticity under well-defined conditions. We also show that dendritic spine maturation involves an intermediate formation of elongated spines, followed by their conversion into mushroom shape. The formation of mushroom-shaped spines is accompanied by increase in AMPA/NMDA ratio of glutamate receptors. Altogether, our results identify inhibition of protease activity as a critical regulatory mechanism for dendritic spines maturation.


Retrieval of Migrated Coils From Distal Cerebral Vasculature Using Stent Retriever: A Case Series.
The incidence of coil dislocation during an endovascular embolization of intracranial aneurysm is low but it can lead to serious thrombo-embolic complications. Therefore, coil displacement/migration often requires either retrieval or fixation of the errant coil with a stent. There are no standard recommended methods of coil retrieval. We present a series of three cases in which off-label application of a stent retriever allowed successful retrieval of herniated coils.


Phase II study of everolimus in refractory testicular germ cell tumors.
Testicular germ cell tumors (TGCTs) represent a highly curable disease; however, a small proportion of patients develop disease recurrence. Loss of the tumor-suppressor gene phosphatase and tensin homolog marks the transition from intratubular germ cell neoplasia to invasive GCT and is correlated with disease progression. Inactivation of phosphatase and tensin homolog is associated with deregulation of the PI3K/Akt pathway and increased mammalian target of rapamycin signaling. This study aimed to determine the efficacy and toxicity of a mammalian target of rapamycin inhibitor, everolimus, in patients with refractory TGCTs. From December 2011 to February 2015, 15 patients with refractory GCTs were enrolled in the phase II study. All patients were pretreated with at least 2 cisplatin-based therapies; 4 tumors (26.7%) were absolutely refractory to cisplatin and 9 patients (60.0%) had visceral nonpulmonary metastases. Everolimus was administered at a dose of 10mg daily until progression or unacceptable toxicity. The primary end point was the objective response rate, according to Response Evaluation Criteria in Solid Tumors. No objective response was observed, but 6 patients (40.0%) achieved 12-week progression-free survival. During a median follow-up period of 3.6 months (range: 1-35.1mo), all patients experienced disease progression and 11 patients (80.0%) died. Median progression-free survival was 1.7 months (95% CI: 1.1-4.0mo) and median overall survival was 3.6 months (95% CI: 2.0-11.0mo). This study failed to achieve its primary end point and our data suggest limited efficacy of everolimus against unselected heavily pretreated refractory TGCTs. Everolimus showed limited efficacy in unselected heavily pretreated refractory TGCTs. Prolonged disease stabilization could be achieved in selected patients.


Addressing disparities in speech-language pathology and laryngology services with telehealth.
The COVID-19 pandemic disproportionately affected the health and well-being of marginalized communities, and it brought greater awareness to disparities in health care access and utilization. Addressing these disparities is difficult because of their multidimensional nature. Predisposing factors (demographic information, social structure, and beliefs), enabling factors (family and community) and illness levels (perceived and evaluated illness) are thought to jointly contribute to such disparities. Research has demonstrated that disparities in access and utilization of speech-language pathology and laryngology services are the result of racial and ethnic differences, geographic factors, sex, gender, educational background, income level and insurance status. For example, persons from diverse racial and ethnic backgrounds have been found to be less likely to attend or adhere to voice rehabilitation, and they are more likely to delay health care due to language barriers, longer wait times, a lack of transportation and difficulties contacting their physician. The purpose of this paper is to summarize existing research on telehealth, discuss how telehealth offers the potential to eliminate some disparities in the access and utilization of voice care, review its limitations, and encourage continued research in this area. A clinical perspective from a large volume laryngology clinic in a major city in northeastern United States highlights the use of telehealth in the provision of voice care by a laryngologist and speech-language pathologist during and after the COVID19 pandemic.


A novel strategy to improving Rhodobacter azotoformans denitrification efficiency: Insight into the role of a two-component system NtrX/Y in denitrification regulation.
Transcription factors (TFs) can manage the coordinated expression of genes clusters or multiple genes. TF was used to improve bacterial denitrification ability in this study. During denitrification, the ntrY of R. azotoformans, which encodes the sensor of NtrX/Y system, was significantly upregulated in transcription. Denitrification of the mutant △ntrY was significantly inhibited, and it was recovered after replenishing this gene to the mutant, which indicates the NtrX/Y system plays an important role in regulating bacterial denitrification. According to additional research, the NtrX/Y system regulates bacterial denitrification by directly promoting the expression of the nitrite reductase. ntrY overexpression appears to accelerate bacterial denitrification, and the introduction of a strong promoter tac in conjunction with iron supply optimization increases the rate by 72% further. This study realizes bacterial denitrification enhancement from the perspective of global transcription regulation, which provides a novel strategy for improving microbial ability to degrade pollutants.


Temperament and adolescent suicide attempts: a case-control study with multi-ethnic Asian adolescents.
Suicide is the leading cause of death for adolescents in several parts of Asia, including Singapore. This study examines the relationship between temperament and youth suicide attempts in a sample of multi-ethnic Singaporean adolescents. A case-control design compared 60 adolescents (Mage = 16.40, SDage = 2.00) with a recent suicide attempt (i.e., past 6 months) with 58 adolescents (Mage = 16.00, SDage = 1.68) without any history of suicide attempts. Presence of suicide attempts was established using the semi-structured interviewer-administered Columbia Suicide Severity Rating Scale. Participants also completed self-report measures on temperament traits, psychiatric diagnoses, stressful life events, and perceived parental rejection in an interview-based format. Psychiatric comorbidity, recent stressful life events, perceived parental rejection, and all five "difficult temperament" traits, were significantly overrepresented among adolescent cases relative to healthy controls. Adjusted logistic regression models revealed significant associations between suicide attempt, MDD comorbidity (OR: 10.7, 95% Cl: (2.24-51.39)), "negative mood" trait (OR: 1.12-1.18, 95% Cl: (1.00-1.27)), and the interaction term of "positive mood" and "high adaptability" traits (OR: 0.943 - 0.955, 95% Cl: (0.900 - 0.986)). Specifically, "positive mood" predicted lower likelihood of a suicide attempt when "adaptability" was high (OR: 0.335 - 0.342, 95% Cl: (0.186 - 0.500)) but not low (OR: 0.968 - 0.993, 95% Cl: (0.797 - 1.31)). Temperament screening may be important to identify adolescents at higher or lower risk of suicide at an early stage. More longitudinal and neurobiological research converging on these temperament findings will be helpful in ascertaining temperament screening as an effective suicide prevention methodology for adolescents.


E2Fs up-regulate expression of genes involved in DNA replication, DNA repair and mitosis.
The E2F family of transcription factors plays a pivotal role in the regulation of cell proliferation in higher eukaryotes. We used DNA microarrays and cell lines containing either inducible E2F-1 or inducible E2F-3 to identify novel E2F target genes. Our data indicate that E2F up-regulates the expression of genes not previously described as E2F target genes. A number of these E2F-regulated genes are involved in DNA replication, DNA repair and mitosis. These results suggest that E2F affects cell cycle progression both at S phase and during mitosis. Furthermore, our findings indicate that E2F-dependent gene activation may contribute to the cellular response to DNA damage.


Award-winning articles and posters from the World Federation of Chiropractic's 11th Biennial Congress 2011.
This editorial reviews the scientific sessions and announces the 4 award-winning scientific articles from the World Federation of Chiropractic 11th Biennial Congress held in Rio de Janeiro from April 6, to 9, 2011, that are published in this issue of the Journal of Manipulative and Physiological Therapeutics.


Aortic dissection presenting as spinal cord ischemia with a false-negative aortogram.
This article presents a case of type III aortic dissection presenting as a spinal cord syndrome with a prospectively negative initial aortogram. The patient serendipitously illustrates a multimodality approach to aortic dissection imaging. The advantages and disadvantages of conventional radiography, aortography, computed tomography, echocardiography, and magnetic resonance imaging are discussed.


A potent therapeutics for gallbladder cancer by combinatorial inhibition of the MAPK and mTOR signaling networks.
Gallbladder cancer (GBC) is the most common type of cancer with the worst prognosis among the bile duct cancers. There still remains a clear need for effective mechanism-based novel therapeutic approaches. A crosstalk between mitogen-activated protein kinase (MAPK) and the mammalian target of Rapamycin (mTOR) signaling pathways has been reported in several cancers. We hypothesized that targeting both pathways in combination will be a potent therapeutic for GBC. Expression of phospho-ERK and phospho-S6rp protein were evaluated by immunostaining in surgically resected GBC specimens (n = 30). GBC cell lines and a xenograft model were treated with CI-1040, an inhibitor of MEK (mitogen-activated protein kinase kinase) and RAD001, an inhibitor of mTOR, alone or in combination, and then, we examined the cell proliferation and tumor growth, cell cycle status, and apoptosis. Analysis of human GBC tissues demonstrated that MAPK and mTOR signaling pathways were frequently coordinately dysregulated in one third of them. The combination therapy inhibited both signaling pathways and subsequently inhibited human GBC cell proliferation in vitro and xenograft tumor growth in vivo. Compared to the single treatment, the combination therapy significantly induced cell cycle arrest and apoptosis with decreased cyclin D1 expression. The double blockade of MAPK and mTOR signaling pathways inhibits the signal crosstalk and shows anti-tumor activity, which can be a potent therapeutic for GBC, especially for the patients with hyperactivated signaling of both pathways.


Innovative technique for the direct determination of proteins in calcified aortic valves.
Aortal valve mineralization very frequently causes a genesis of aortic stenosis, which is the most often surgically treated heart disease. Hydroxyapatite deposits have been identified as one of the causes leading to the loss of elasticity of the aortic valves. It is known that phosphates/calcium is accumulated in valve tissues during mineralization, but the mechanism of this process remains unclear. The work is focused mainly on the study of protein composition of mineralized aortic valves by nano-liquid chromatography electrospray ionization in a quadrupole orthogonal acceleration time-of-flight mass spectrometry. New methodological approach based on direct enzymatic digestion of proteins contained in hydroxyapatite deposits was developed for the study of pathological processes connected with osteogenesis. Our objectives were to simplify the traditional analytical protocols of sample preparation and to analyze the organic components of the explanted aortic valves for significant degenerative aortic stenosis. The study of aortic valve mineralization on the molecular level should contribute to understanding this process, which should consequently lead to effective prevention as well as to new ways of treatment of this grave disease.


Ileal interposition surgery for treatment of type 2 diabetes mellitus-pros and cons.
Nowadays, the surgical techniques for treating type 2 diabetes (T2DM) include: Ileal Interposition (II), Ileal Interposition in combination with Sleeve Gastrectomy (IISG) and Diverted Sleeve Gastrectomy (IIDSG). These procedures are not only for obese subjects, but are also used for non-obese subjects. These types of surgical procedures can improve glycemic control, and lead to a significant reduction in oral hypoglycemic agents (OHAs) and insulin therapy. The results of various studies have shown the safety, feasibility, and efficacy of the surgical procedure for the treatment of T2DM. Although it is an effective treatment option, this procedure is not recommended for general use and long-term studies are needed to confirm these findings and potential side effects on a larger number of patients.


Severe hypoalbuminaemia in a systemic lupus erythematosus-like patient.
Diffuse oedema due to hypoalbuminaemia is a common manifestation in patients affected with systemic lupus erythematosus. Hypoalbuminaemia is usually secondary to an ongoing glomerulonephritis leading to proteinuria or, more rarely, to decreased protein synthesis or deficient protein intake. Here, we report the case of a 6-year-old girl with a previous history of Evans' syndrome and mesangial glomerulonephritis who subsequently developed severe anasarca without apparent proteinuria. Faecal oal-antitrypsin showed a persistent severe intestinal protein loss (ranging from 1500 to 6.500 yl/g humid faeces, normal value < 200), consistent with the diagnosis of protein-losing enteropathy. An echographic examination of the abdomen revealed a diffuse thickening of the intestinal wall, particularly at the level of the small bowel, with an almost exclusive involvement of the submucosal layer. Colonoscopy revealed the presence of diffuse purpuric lesions at the level of the submucosa. Aggressive immunosuppressive treatment completely resolved the clinical picture. protein-losing enteropathy is an uncommon cause of hypoalbuminaemia during the course of systemic lupus erythrematosus. It may be considered as a clinical syndrome related to many pathological conditions leading to an excessive intestinal protein loss. Some conditions are related with an altered mucosal permeability, others with primary or secondary intestinal lymphangiectasia. A review of the possible causes of systemic lupus erythrematosus-associated protein-losing enteropathy reported in the paediatric literature is given.


sox4b is a key player of pancreatic alpha cell differentiation in zebrafish.
Pancreas development relies on a network of transcription factors belonging mainly to the Homeodomain and basic Helix-Loop-Helix families. We show in this study that, in zebrafish, sox4, a member of the SRY-like HMG-box (SOX) family, is required for proper endocrine cell differentiation. We found that two genes orthologous to mammalian Sox4 are present in zebrafish and that only one of them, sox4b, is strongly expressed in the pancreatic anlage. Transcripts of sox4b were detected in mid-trunk endoderm from the 5-somite stage, well before the onset of expression of the early pancreatic gene pdx-1. Furthermore, by fluorescent double in situ hybridization, we found that expression of sox4b is mostly restricted to precursors of the endocrine compartment. This expression is not maintained in differentiated cells although transient expression can be detected in alpha cells and some beta cells. That sox4b-expressing cells belong to the endocrine lineage is further illustrated by their absence from the pancreata of slow-muscle-omitted mutant embryos, which specifically lack all early endocrine markers while retaining expression of exocrine markers. The involvement of sox4b in cell differentiation is suggested firstly by its up-regulation in mind bomb mutant embryos displaying accelerated pancreatic cell differentiation. In addition, sox4b knock-down leads to a drastic reduction in glucagon expression, while other pancreatic markers including insulin, somatostatin, and trypsin are not significantly affected. This disruption of alpha cell differentiation is due to down-regulation of the homeobox arx gene specifically in the pancreas. Taken together, these data demonstrate that, in zebrafish, sox4b is expressed transiently during endocrine cell differentiation and plays a crucial role in the generation of alpha endocrine cells.


Age-related reduction of [3H]strychnine binding sites in the cochlear nucleus of the Fischer 344 rat.
The present study used quantitative receptor autoradiography to examine the effects of aging on the binding profile of the strychnine-sensitive glycine receptor in the Fischer 344 rat. Glycine receptor binding sites were localized using [3H]strychnine in two principal subdivisions of the cochlear nucleus; the dorsal and anteroventral cochlear nucleus. These central auditory brainstem structures are known to receive extensive glycinergic inputs. In young rats, single concentrations of [3H]strychnine showed significantly higher binding levels in the dorsal cochlear nucleus than the anteroventral cochlear nucleus (+38%, P < 0.001). Little binding was detected in regions of the posteroventral cochlear nucleus, and no specific binding was apparent in the cerebellum. Saturation analysis in the dorsal cochlear nucleus revealed an affinity constant (Kd) of 16.9 nM and a maximum number of binding sites of 850 fmol/mg protein. A significant age-related decrease in [3H]strychnine (8 nM) binding was observed in the anteroventral cochlear nucleus (-37%, P = 0.003) and dorsal cochlear nucleus (-23%, P = 0.034) of 26-month-old rats compared with three-month-old rats. Saturation analysis indicated that the observed decrease in binding was due to a decrease in the total number of binding sites with no significant change in affinity. In the dorsal cochlear nucleus, the number of binding sites was reduced (-26%) in 26-month-old rats compared with three-month-old adults (P = 0.011). Kd was decreased (-22%) in 26-month-old rats when compared with young adults, but this decrease was not statistically significant (P = 0.377).(ABSTRACT TRUNCATED AT 250 WORDS)


How essentialist reasoning about language acquisition relates to educational myths and policy endorsements.
How people conceptualize learning is related to real-world educational consequences across many domains of education. Despite its centrality to the educational system, we know little about how the public reasons about language acquisition, and the potential consequences for their thinking about real-world issues (e.g., policy endorsements). The current studies examined people's essentialist beliefs about language acquisition (e.g., that language is innate and biologically based), then investigated how individual differences in these beliefs related to the endorsement of educational myths and policies. We probed several dimensions of essentialist beliefs, including that language acquisition is innate, genetically based, and wired in the brain. In two studies, we tested specific hypotheses regarding the extent to which people use essentialist thinking when reasoning about: learning a specific language (e.g., Korean), learning a first language more generally, and learning two or more languages. Across studies, participants were more likely to essentialize the ability to learn multiple languages than one's first language, and more likely to essentialize the learning of multiple languages and one's first language than the learning of a particular language. We also found substantial individual differences in the degree to which participants essentialized language acquisition. In both studies, these individual differences correlated with an endorsement of language-related educational neuromyths (Study 1 and pre-registered Study 2), and rejection of educational policies that promote multilingual education (Study 2). Together, these studies reveal the complexity of how people reason about language acquisition and its corresponding educational consequences.


Value of angiography in the management of abdominal aortic aneurysm.
The value of angiography in the management of abdominal aortic aneurysms (AAA) was assessed in 100 consecutive patients with AAA. Angiographic information influenced management decisons and/or surgery performance in 75: In 23 patients at high risk for surgery because of associated medical problems, it helped in deferring surgery; in 52 patients it resulted in a change of operation from a standard aneurysm resection with conventional grafting to a more conservative procedure (three patients), more extensive grafting (45 patients) and/or the addition of other vascular reconstructions (32 patients). Angiography is considered an integral step in the routine preoperative workup of AAA and is particularly valuable for the determination of important anatomic details about the aneurysm (upper and lower extensions, relation to the renal arteries), the detection of associated vascular disease (of renal, visceral, pelvic and peripheral arteries), and the demonstration of aberrant renal arteries and collateral visceral circulation. Catheter techniques are considered most suitable and safe for examination of AAA.


Positive regulation of the expression of the Escherichia coli pts operon. Identification of the regulatory regions.
The pts operon of Escherichia coli is composed of the ptsH, ptsI and crr genes coding for three proteins central to the phosphoenolpyruvate dependent phosphotransferase system (PTS), the HPr, enzyme I and EIIIGlc proteins, respectively. We previously showed that transcription from the promoter region located upstream from the pts operon is regulated by two control circuits, which can occur independently from each other. Transcription of the pts operon is (1) stimulated by the CAP-cAMP complex and (2) enhanced during growth on glucose, a PTS substrate. The DNA regions involved in regulation of the expression of the pts operon have been identified. Two promoters, P0 and P1, separated by 100 bp are located upstream from the pts operon. In these promoter regions, we identified two sequences showing similarity with the consensus of CAP-binding sites, CAPa located near P0 and CAPb located in the -35 region of P1. In vivo experiments showed that binding of CAP-cAMP at the CAPa site stimulates transcription from the P0 promoter. The binding sites of CAP-cAMP and/or RNA-polymerase on a DNA fragment containing both P0 and P1 promoters as well as both CAPa and CAPb sites were examined by the technique of DNase I footprinting. These in vitro experiments suggested that CAP-cAMP binding at the CAPb site might also play a role in regulation of the pts operon expression. In addition, we showed that the DNA region carrying the CAPa site is important for regulation by glucose. We finally propose that the expression of the pts operon is controlled by two alternative positive regulatory mechanisms, which are designed to allow activation of the pts operon under a great variety of growth conditions.


[Overlooked osteosarcoma in a patient undergoing total knee arthroplasty. A case report].
A 72-year-old woman presented with pain, swelling, and decreased range of motion of the left knee joint after total knee arthroplasty in 2005. We performed standard x-rays, which were highly suspicious for an osteosarcoma of the distal femur; this was proven by open biopsy. Retrospectively, the x-rays taken before implantation of the prosthesis showed suspicious findings. Because the femur was tumor-contaminated, with the intramedullary adjustment far-reaching proximally, a limb salvage procedure was no longer possible. To improve function, we decided to perform a stump-lengthening procedure using a special implant. With regard to the dismal consequences, we recommend that every suspicious finding before an elective surgical procedure be examined with further diagnostics and, if necessary, histological confirmation.


A histofluorescent procedure for identifying marijuana cannabinoids.
A rapid and reliable fluorescence procedure is described as a test for the microscopical identification of the glandular hairs of Cannabis sativa. The proposed method, designated as the IFIM test (induced fluorescence identification for marijuana test), is based on the induction of a red fluorescence in cannabinoids by a hot clearing solution. The results, compared to those obtained by the classical RIM test, offer the possibility of more satisfactory identification of cannabis, hashish or marijuana in suspected samples.


Lectin-binding characteristics of mouse oviduct and uterus associated with pregnancy block by autologous antiprogesterone monoclonal antibody.
Oviducts and uteri were removed from BALB/cJ and F1 (CBA/Ca X BALB/cJ) mice at known stages post coitum and following treatment with an antiprogesterone monoclonal antibody (DB3) or a non-specific immunoglobulin (DNP). Thin sections of tissue were prepared and reacted with fluorescent conjugates of a wide range of lectins to determine if saccharide alterations were associated with the pregnancy-inhibiting effect of the DB3 antibody in BALB/c, but not F1, individuals. The ampullary region of the DB3-treated BALB/c mice showed the most marked changes, with an almost total inhibition of lectin binding, particularly for N-acetylglucosamine residues. There was also a reduced affinity for a lectin reactive with N-acetylgalactosamine in the uteri of DB3-treated BALB/c mice, associated with an extended expression during gestation of this saccharide in the proximal region of the oviduct in such mice. These are the first biochemical alterations in reproductive tract epithelia to be associated with the efficacy of the DB3 antibody in preventing pregnancy.


Effects of leucine supplemented diet on intestinal absorption in tumor bearing pregnant rats.
It is known that amino acid oxidation is increased in tumor-bearing rat muscles and that leucine is an important ketogenic amino acid that provides energy to the skeletal muscle. To evaluate the effects of a leucine supplemented diet on the intestinal absorption alterations produced by Walker 256, growing pregnant rats were distributed into six groups. Three pregnant groups received a normal protein diet (18% protein): pregnant (N), tumor-bearing (WN), pair-fed rats (Np). Three other pregnant groups were fed a diet supplemented with 3% leucine (15% protein plus 3% leucine): leucine (L), tumor-bearing (WL) and pair-fed with leucine (Lp). Non pregnant rats (C), which received a normal protein diet, were used as a control group. After 20 days, the animals were submitted to intestinal perfusion to measure leucine, methionine and glucose absorption. Tumor-bearing pregnant rats showed impairment in food intake, body weight gain and muscle protein content, which were less accentuated in WL than in WN rats. These metabolic changes led to reduction in both fetal and tumor development. Leucine absorption slightly increased in WN group. In spite of having a significant decrease in leucine and methionine absorption compared to L, the WL group has shown a higher absorption rate of methionine than WN group, probably due to the ingestion of the leucine supplemented diet inducing this amino acid uptake. Glucose absorption was reduced in both tumor-bearing groups. Leucine supplementation during pregnancy in tumor-bearing rats promoted high leucine absorption, increasing the availability of the amino acid for neoplasic cells and, mainly, for fetus and host utilization. This may have contributed to the better preservation of body weight gain, food intake and muscle protein observed in the supplemented rats in relation to the non-supplemented ones.


Intracellular recordings from salamander olfactory receptor cells.
Intracellular recordings were obtained from salamander olfactory receptor cells. The occurrence of an intracellular spike in response to the antidromic stimulation of the olfactory fibers was considered as a physiological criterion of a neuronal impalement. The mean resting potential was -56 +/- 9 mV (mean +/- S.D.; n = 70). Fifty-two cells presented a spontaneous spike activity lower than 2 impulses/s. Appropriate olfactory stimulation generally evoked a slow and graded decrease (up to 28 mV) of the intracellular potential. The input resistance of the cell decreased markedly during the response. The slow potential change induced a repetitive firing. Increasing the intensity of the olfactory stimulation increased the instantaneous frequency of firing (up to 25 s-1) and reduced the spike amplitude. The spikes presented an inflexion in the rising phase indicating a two-stage depolarization. With the strongest intensities of stimulation the impulse activity was stopped during the repolarizing phase of the cell response when the membrane potential was still appreciably depolarized.


The nursing costs of urinary incontinence in a nursing home population.
The labor resources and costs of caring for incontinence in a nursing home population are explored. Data from a work measurement study have been applied to patient assessment results to estimate the total costs of labor for three states and the U.S. as a whole. Incontinence care costs an estimated $1.1 billion per year in the US; this represents 3 to 4 per cent of all Medicaid payments made to nursing homes.


Molluscicidal activity of Solanum species of the Northeast of Brazil on Biomphalaria glabrata.
Seven species of Solanum were screened for their molluscicidal properties against Biomphalaria glabrata, the intermediated host of Schistosoma mansoni, Solanum agrarium, S. jabrense, S. melissarum, S. megalonyx, S. paludosum, S. paraibanum and S. stipulaceum. Four extracts showed molluscicidal activity with LC(50) from 22 to 56 microg/ml.


[To screen or systematically treat a population at risk? A method based on the analysis of costs applicable at the level of district health].
With severely limited resources, it is necessary for the health services to select the most efficient strategy for managing health problems in a population at risk. A simple method for cost analysis, which can be used at the district health care level, is proposed so that one can choose between selective treatment on the basis of case identification, or indiscriminate treatment of the entire population at risk. The treatment of anaemia at the antenatal clinic in health centres in southwestern Chad was studied and is presented as an example. The conditions for use and the limitations of the method are discussed.


Tissue Doppler echocardiography before and after the surgical reconstruction of an insufficient mitral valve in a patient with Fontan circulation.
We report a patient with tricuspid atresia after total cavopulmonary connection who developed significant mitral valve regurgitation. Doppler myocardial echocardiography tracings showed "pseudonormalization" of diastolic velocities. After successful surgical reconstruction of the mitral valve, the diastolic myocardial velocities changed to the typical Fontan physiology with predominant late diastolic ventricular filling.


Rapid Detection and Identification of Fasciola spp. and Dicrocoelium spp. Isolated from the Ruminant Livestock of Northwest Iran Using High-Resolution Melting Analysis (HRM).
The liver flukes of the Fasciola species and Dicrocoelium spp. are recognised as parasites of domestic and wild herbivores. Both species of F. hepatica and F. gigantica as well as D. dendriticum are distributed in Iran. The present study aimed to identify Fasciola spp. and Dicrocoelium spp. using mitochondrial Cox1 (cytochrome c oxidase I) gene by HRM method. Totally, thirty infected liver specimens were collected from the sheep (n:23) and cattle (n:7) at the abattoirs of Qazvin Province, northwest Iran in 2022. DNA extraction and PCR amplification of Cox1 gene were conducted by HRM technique. DnaSP v.5.0 was used for compression of diversity indices of ribosomal 28S rDNA and mitochondrial Cox1 markers of Dicrocoelium spp. The taxonomic status of Dicrocoelium spp. was performed by sequencing and phylogenetic analysis. Overall, 26 and 4 isolates were identified as F. hepatica and F. gigantica, respectively. D. dendriticum was the sole infecting species of Dicrocoelium revealed by HRM analysis. Genomic analysis showed a moderate (28S rDNA genes: 0.600±0.215) to high (Cox1: 0.733±0.155) haplotype diversity for D. dendriticum. The parasite-dependent mitochondrial gene (Cox1) could identify a higher genetic diversity of D. dendriticum compared to nuclear 28S rDNA gene. HRM technique in the present study found to be a reliable technique for identification and genetic diversity of liver flukes but more comprehensive and in-depth studies in different parts of the country are needed.


Immunophenotypic characterization of acute leukemia at a public oncology reference center in Maranhão, northeastern Brazil.
The incidence of acute leukemia (AL) subtypes varies according to geographical distribution. The aim here was to determine the incidence of morphological and immunophenotypic AL subtypes in the state of Maranhão, Brazil, and to correlate the expression of aberrant phenotypes in children with acute lymphoblastic leukemia (ALL) with prognostic factors. Single prospective cohort study at a public oncology reference center in Maranhão. Seventy AL cases were diagnosed between September 2008 and January 2010. For the diagnosis, complete blood cell counts, myelograms (at diagnosis and at the end of the induction phase), cytochemical analysis and immunophenotyping were performed. Among adult patients (n = 22), the incidence of AL types was: ALL (22.7%) and acute myeloid leukemia (AML) (77.3%). The subtype AML M0 occurred most frequently (29.4%). In children (n = 48), the types were: AML (18.7%), most frequently subtype AML M4 (33.4%); biphenotypic acute leukemia (BAL) (4.2%); and ALL (77.1%), including the subtypes B-ALL (72.9%) and T-ALL (27.1%). Among the children with ALL, there were no statistically significant differences between patients with and without aberrant phenotypes, in relation to hematological parameters and treatment response. This work demonstrates that the frequencies of AML M0 cases among adults and T-ALL cases among children in Maranhão were high. This suggests that there may be differences in AML subtype incidence, as seen with ALL subtypes, in different regions of Brazil. No association was found between the expression of aberrant phenotypes and prognostic factors, in children with ALL.


Genome editing with CRISPR-Cas nucleases, base editors, transposases and prime editors.
The development of new CRISPR-Cas genome editing tools continues to drive major advances in the life sciences. Four classes of CRISPR-Cas-derived genome editing agents-nucleases, base editors, transposases/recombinases and prime editors-are currently available for modifying genomes in experimental systems. Some of these agents have also moved rapidly into the clinic. Each tool comes with its own capabilities and limitations, and major efforts have broadened their editing capabilities, expanded their targeting scope and improved editing specificity. We analyze key considerations when choosing genome editing agents and identify opportunities for future improvements and applications in basic research and therapeutics.


The protein factor which binds to the upstream activating sequence of Saccharomyces cerevisiae ENO1 gene.
Using a gel retardation assay it was shown that the 87 bp DNA fragment (UAS87) containing the upstream activating sequence (UAS) of S. cerevisiae EN01 gene and a nuclear extract gave rise to three migration-retarded species specific to UAS87. Heat- or proteinase-treatment of the nuclear extract revealed that these species were protein-DNA complexes. The precise binding region of the protein identified by DNaseI protection analysis was found to include a CCAAACA sequence which forms a dyad-symmetrical structure. The amount of one of the three migration-retarded species significantly increased when cells were grown in medium containing a gluconeogenic carbon source. The introduction of pGCR8, a multicopy plasmid containing GCR1 gene, a regulatory gene controlling the expression of several glycolytic enzymes, showed no effect on the amount of three migration-retarded species.


Aberrant dentate gyrus cytoarchitecture and fiber lamination in Lis1 mutant mice.
Mutant mice with a heterozygous deletion of LIS1, show varying degrees of hippocampal abnormality and enhanced excitability. To examine how LIS1 affects cytoarchitecture and fiber lamination in dentate gyrus (DG), we performed a series of immunohistochemistry studies. By using different neuronal- and glial-specific antibodies, we found that the majority of hippocampal cell populations were affected by heterozygous mutation of LIS1; some reelin-positive Cajal-Retzius cells were left undisturbed. Granule cell dispersion was significant in hippocampal sections from Lis1-deficient mice. However, the fiber termination of commissural/associational fibers and mossy fibers appeared relatively compact despite obvious granule cell dispersion and CA1-CA3 pyramidal cell disorganization. vGlut1-immunoreactive axon terminals were found aberrantly traversing the dispersed granule cell layer. Consistent with previous observations, we also found that immature granule cells in Lis1 mutants, here stained with antibodies to doublecortin (DCX) and Mash-1, are aberrantly located and bear an abnormal cellular morphology. Our findings suggest that LIS1 mutants exhibit abnormal cell positioning and aberrant hippocampal neurogenesis, but maintain relatively normal fiber termination patterns. The functional consequences of hippocampal granule cell dispersion could offer critical insight to the epileptic and cognitive disorder associated with LIS1 haploinsufficiency.


Carbamazepine at environmentally relevant concentrations caused DNA damage and apoptosis in the liver of Chinese rare minnows (Gobiocypris rarus) by the Ras/Raf/ERK/p53 signaling pathway.
To assess genetoxicity and the underlying mechanisms of carbamazepine (CBZ) toxicity in fish, adult Chinese rare minnows (Gobiocypris rarus) were exposed to 1, 10, and 100 μg/L CBZ for 28 d. Comet assays indicated that hepatic DNA damage was significantly increased in groups of minnows exposed to CBZ at all concentrations in a dose-dependent manner compared to those of the control groups (p < 0.05). Liver levels of 8-hydroxydeoxyguanosine (8-OHdG) were significantly increased at 10 and 100 μg/L CBZ (p < 0.05). TUNEL assays indicated that the average apoptotic rates of the livers of female and male minnows were significantly increased following exposure to CBZ at all concentrations for 28 d (p < 0.05). Significant increases in caspase 3 and 9 activities after CBZ exposure at all concentrations and caspase 8 at 10 and 100 μg/L CBZ exposure reflected the presence of mitochondrial apoptosis (p < 0.05). The mRNA levels of gadd45a, mdm2, casp3 and casp9 in female and male minnows exposed to CBZ at all concentrations were significantly increased compared with those in the control groups (p < 0.05). Significant increases in the levels of p21 in female minnows exposed to 1 and 100 μg/L CBZ, p53 in female minnows at all CBZ treatments and bcl2 in male minnows exposed to 1 and 100 μg/L CBZ were observed, indicating p53 pathway activation. The inhibition of ras levels in females and males exposed to CBZ at all concentrations and increased levels of raf1 in males exposed to CBZ at all concentrations indicated Ras/Raf1/MAPK (ERK) activation. Therefore, the present study demonstrates that CBZ at environmentally relevant levels induces DNA damage and apoptosis in Chinese rare minnows by the Ras/Raf/ERK/p53 signaling pathway.


On the computation of the magnetic phase shift for magnetic nano-particles of arbitrary shape using a spherical projection model.
The magnetic phase shift of an electron wave traveling through a magnetized object is computed by considering the object to be made up of a collection of uniformly magnetized spheres arranged on the nodes of a cubic grid. In the limit of vanishing grid size, this approach becomes equivalent to other numerical approaches. Update equations are derived for the change of the magnetic phase shift when the magnetization of a single object voxel is modified. Example phase shift calculations are presented for a uniformly magnetized sphere, circular disks with an infinitely sharp vortex core and a smooth core, and an oval disk with a pair of vortices and an antivortex.


Are We Undercounting the True Burden of Mortality Related to Suicide, Alcohol Use, or Drug Use? An Analysis Using Death Certificate Data From Colorado Veterans.
Knowledge regarding deaths due to suicide or alcohol- or drug-related causes may be limited by inconsistent and/or restrictive case definitions, resulting in concerns regarding validity of findings and underestimates of burden. In this proof-of-concept study, we assessed varying case definitions (suicide, alcohol-related, and drug-related mortality using underlying-cause-of-death (UCOD) versus multiple-cause-of-death (MCOD) International Classification of Diseases, Tenth Revision (ICD-10) codes) on the basis of counts and rates among Colorado veterans who died (2009-2020). Suicide, alcohol-related, or drug-related ICD-10 codes were identified, and 2 case definitions were compared: UCOD (qualifying ICD-10 code listed as the UCOD) and MCOD (qualifying ICD-10 code in any cause-of-death field). Of 109,314 decedents, the number of deaths and the age-adjusted mortality rate (per 100,000 persons) significantly increased when MCOD codes were included: n = 4,930 (110.3 deaths/100,000 persons) for UCOD versus n = 6,954 (138.4 deaths/100,000 persons) for MCOD. While rates of suicide mortality did not change, rates of alcohol-related mortality doubled with the more inclusive case definition: 1,752 (27.3 deaths/100,000 persons) for UCOD versus 3,847 (59.8 deaths/100,000 persons) for MCOD. Alcohol-use disorder codes accounted for 71% of additional alcohol-related deaths captured with the MCOD definition. Studies that rely on UCOD codes may be underestimating the burden of deaths, especially alcohol-related deaths. Increased effort is required to reevaluate current classifications of deaths associated with suicide, alcohol use, or drug use.