Insights from biochemical reconstitution into the architecture of human kinetochores.
Chromosomes are carriers of genetic material and their accurate transfer from a mother cell to its two daughters during cell division is of paramount importance for life. Kinetochores are crucial for this process, as they connect chromosomes with microtubules in the mitotic spindle. Kinetochores are multi-subunit complexes that assemble on specialized chromatin domains, the centromeres, that are able to enrich nucleosomes containing the histone H3 variant centromeric protein A (CENP-A). A group of several additional CENPs, collectively known as constitutive centromere associated network (CCAN), establish the inner kinetochore, whereas a ten-subunit assembly known as the KMN network creates a microtubule-binding site in the outer kinetochore. Interactions between CENP-A and two CCAN subunits, CENP-C and CENP-N, have been previously described, but a comprehensive understanding of CCAN organization and of how it contributes to the selective recognition of CENP-A has been missing. Here we use biochemical reconstitution to unveil fundamental principles of kinetochore organization and function. We show that cooperative interactions of a seven-subunit CCAN subcomplex, the CHIKMLN complex, determine binding selectivity for CENP-A over H3-nucleosomes. The CENP-A:CHIKMLN complex binds directly to the KMN network, resulting in a 21-subunit complex that forms a minimal high-affinity linkage between CENP-A nucleosomes and microtubules in vitro. This structural module is related to fungal point kinetochores, which bind a single microtubule. Its convolution with multiple CENP-A proteins may give rise to the regional kinetochores of higher eukaryotes, which bind multiple microtubules. Biochemical reconstitution paves the way for mechanistic and quantitative analyses of kinetochores.


The prometastatic ribosomal S6 kinase 2-cAMP response element-binding protein (RSK2-CREB) signaling pathway up-regulates the actin-binding protein fascin-1 to promote tumor metastasis.
Metastasis is the leading cause of death in patients with breast, lung, and head and neck cancers. However, the molecular mechanisms underlying metastases in these cancers remain unclear. We found that the p90 ribosomal S6 kinase 2 (RSK2)-cAMP response element-binding protein (CREB) pathway is commonly activated in diverse metastatic human cancer cells, leading to up-regulation of a CREB transcription target Fascin-1. We also observed that the protein expression patterns of RSK2 and Fascin-1 correlate in primary human tumor tissue samples from head and neck squamous cell carcinoma patients. Moreover, knockdown of RSK2 disrupts filopodia formation and bundling in highly invasive cancer cells, leading to attenuated cancer cell invasion in vitro and tumor metastasis in vivo, whereas expression of Fascin-1 significantly rescues these phenotypes. Furthermore, targeting RSK2 with the small molecule RSK inhibitor FMK-MEA effectively attenuated the invasive and metastatic potential of cancer cells in vitro and in vivo, respectively. Taken together, our findings for the first time link RSK2-CREB signaling to filopodia formation and bundling through the up-regulation of Fascin-1, providing a proinvasive and prometastatic advantage to human cancers. Therefore, protein effectors of the RSK2-CREB-Fascin-1 pathway represent promising biomarkers and therapeutic targets in the clinical prognosis and treatment of metastatic human cancers.


Urinary outcomes are significantly affected by nerve sparing quality during radical prostatectomy.
To assess the effect of nerve sparing (NS) quality on self-reported patient urinary outcomes after radical prostatectomy. A total of 102 preoperatively potent men underwent laparoscopic or robotic radical prostatectomy; NS was prospectively graded at surgery using a 0-4 scale/neurovascular bundle. Urinary functional outcomes were measured by validated Expanded Prostate Cancer Index Composite questionnaire at baseline and follow-up time points (1, 3, 6, 9, and 12 months) in 99 men who underwent various degrees of NS. Mixed linear regression was used to analyze the effect of NS quality and other clinical factors on urinary outcomes. Patients with at least 1 neurovascular bundle spared completely, along with its supportive tissues (NS grade 4/4), noted significantly improved Expanded Prostate Cancer Index Composite urinary functional and continence outcomes as early as 1 month postoperatively and up to 12 months. Significantly less urinary bother was also noted in these men by 9-12 months postoperatively. Multivariate analysis revealed that bilateral or unilateral excellent NS (at least 1 bundle graded 4/4), increasing time from surgery, young patient age, and lower body mass index positively and significantly affected urinary functional outcomes, including pad use. Men who received excellent unilateral NS recovered urinary function about as well as men who had both neurovascular bundles spared in similar fashion. The quality of NS significantly influences patient-defined urinary functional convalescence. Completely sparing at least 1 neurovascular bundle along with its supportive tissues has a dramatic effect on the recovery of urinary continence and quality of life in preoperatively potent men.


Arctigenin represses TGF-β-induced epithelial mesenchymal transition in human lung cancer cells.
Arctigenin (ARC) is a lignan that is abundant in Asteraceae plants, which show anti-inflammatory and anti-cancer activities. The current study investigated whether ARC affects cancer progression and metastasis, focusing on EMT using invasive human non-small cell lung cancer (NSCLC) cells. No toxicity was observed in the cells treated with different doses of ARC (12-100 μM). The treatment of ARC repressed TGF-β-stimulated changes of metastatic morphology and cell invasion and migration. ARC inhibited TGF-β-induced phosphorylation and transcriptional activity of smad2/3, and expression of snail. ARC also decreased expression of N-cadherin and increased expression of E-cadherin in dose-dependent and time-dependent manners. These changes were accompanied by decreased amount of phospho-smad2/3 in nucleus and nuclear translocation of smad2/3. Moreover, ARC repressed TGF-β-induced phosphorylation of ERK and transcriptional activity of β-catenin. Our data demonstrate anti-metastatic activity of ARC in lung cancer model.


Methemoglobinemie consecutive a l'abus de gel de lidocaïne visqueuse.
Lidocaine use may lead to methemoglobinemia. Lidocaine is clinically widely used as an anaesthesic drug, however its toxic effect is rare. We report a high methemoglobinemia level of 31.1 % following repeated Xylocaine® gel oral applications. New metabolic pathways of lidocaine have been reported leading to dimethylphenylhydroxylamine in humans. This metabolism is very similar to the aniline derivative ones, and may explain lidocaine methemoglobinemia effect.


Recent developments in hereditary nonpolyposis colorectal cancer.
Hereditary non-polyposis colorectal cancer (HNPCC) is characterized by early onset of colorectal carcinoma (CRC), usually located proximally to the splenic flexure and reportedly carrying a better survival as compared to sporadic-type CRC. Depending on the absence or presence of extracolonic tumours, particularly carcinomas of the endometrium, stomach and urinary tract, HNPCC can be divided into Lynch syndromes I and II, respectively. Although first described in 1913, the elucidation of the molecular basis of this disease has only recently started to unfold, and is reviewed in this article. Literature survey of published articles. Comparison of HNPCC tumours with sporadic-type CRC had already revealed that no significant differences were found in APC, Ki-ras and p53 gene alterations. Instead, microsatellite instability was found to be the hallmark of HNPCC being present in 80% of cases compared to only 13% of sporadic type CRC. Since studies in yeast and bacteria had shown that microsatellite instability resulted from mutations in so-called postreplicative DNA mismatch repair genes, it was hypothesized that a similar mechanism might underlie the observed microsatellite instability in HNPCC and, hence, its hereditary character. In due course, four human homologues of yeast and bacterial postreplicative DNA mismatch repair genes were cloned and denoted hMSH2, hMLH1, hPMS1, and hPMS2. Current estimates suggest that mutations in hMSH2 account for 50%, in hMLH1 for 30%, in hPMS1 for 5% and in hPMS2 for 5% of HNPCC. Despite many issues still to be resolved, accurate molecular screening tests will in all probability become the gold standard in the diagnosis of HNPCC. As a result, these developments will undoubtedly have profound implications for early detection and subsequent management of affected individuals.


Quantitative proteomic and transcriptomic analyses of molecular mechanisms associated with low silk production in silkworm Bombyx mori.
To investigate the molecular mechanisms underlying the low fibroin production of the ZB silkworm strain, we used both SDS-PAGE-based and gel-free-based proteomic techniques and transcriptomic sequencing technique. Combining the data from two different proteomic techniques was preferable in the characterization of the differences between the ZB silkworm strain and the original Lan10 silkworm strain. The correlation analysis showed that the individual protein and transcript were not corresponded well, however, the differentially changed proteins and transcripts showed similar regulated direction in function at the pathway level. In the ZB strain, numerous ribosomal proteins and transcripts were down-regulated, along with the transcripts of translational related elongation factors and genes of important components of fibroin. The proteasome pathway was significantly enhanced in the ZB strain, indicating that protein degradation began on the third day of fifth instar when fibroin would have been produced in the Lan10 strain normally and plentifully. From proteome and transcriptome levels of the ZB strain, the energy-metabolism-related pathways, oxidative phosphorylation, glycolysis/gluconeogenesis, and citrate cycle were enhanced, suggesting that the energy metabolism was vigorous in the ZB strain, while the silk production was low. This may due to the inefficient energy employment in fibroin synthesis in the ZB strain. These results suggest that the reason for the decreasing of the silk production might be related to the decreased ability of fibroin synthesis, the degradation of proteins, and the inefficiency of the energy exploiting.


Quality of reporting of studies evaluating time to diagnosis: a systematic review in paediatrics.
An ever-increasing number of studies analyses the distribution, determinants and consequences of time to diagnosis and delays. Weaknesses in their reporting can impede the assessment of the risks of bias and variation and thus create a risk of invalid conclusions and counterproductive clinical and public health efforts. This study sought to assess systematically the quality of reporting of articles about time to diagnosis in paediatrics. Two authors identified and analysed the quality of reporting of 50 consecutive articles assessing these intervals published from 2005 through October 2011, according to a checklist we developed of 35 items potentially associated with risks of bias and variation. Frequency of articles reporting each item. Symptoms that should trigger a diagnostic procedure were reported in 28% of the articles; only two articles reported whether all patients with these symptoms underwent that procedure. Only 44% of the articles defined the beginning of the illness, 46% the date of diagnosis and 60% the distribution of time to diagnosis. Two studies met the criteria for all 11 items considered essential for assessing the risks of bias and variation in this type of study. This study identified many weaknesses in the quality of reporting of studies of time to diagnosis in paediatrics, especially for items potentially related to risks of bias and variation. This finding underlines the need for the development of new (or the refinement of existing) guidelines for reporting this type of study.


PF20 gene product contains WD repeats and localizes to the intermicrotubule bridges in Chlamydomonas flagella.
The central pair of microtubules and their associated structures play a significant role in regulating flagellar motility. To begin a molecular analysis of these components, we generated central apparatus-defective mutants in Chlamydomonas reinhardtii using insertional mutagenesis. One paralyzed mutant recovered in our screen contains an allele of a previously identified mutation, pf20. Mutant cells have paralyzed flagella, and the entire central apparatus is missing in isolated axonemes. We have cloned the wild-type PF20 gene and confirmed its identity by rescuing the pf20 mutant phenotype upon transformation. Rescued transformants were wild type in motility and in axonemal ultrastructure. A cDNA clone containing a single, long open reading frame was obtained and sequenced. Database searches using the predicted 606-amino acid sequence of PF20 indicate that the protein contains five contiguous WD repeats. These repeats are found in a number of proteins with diverse cellular functions including beta-transducin and dynein intermediate chains. An antibody was raised against a fusion protein expressed from the cloned cDNA. Immunogold labeling of wild-type axonemes indicates that the PF20 protein is localized along the length of the C2 microtubule on the intermicrotubule bridges connecting the two central microtubules. We suggest that the PF20 gene product is a new member of the family of WD repeat proteins and is required for central microtubule assembly and/or stability and flagellar motility.


Recent insights on risk factors of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a disease prevalent in many populations worldwide. It initiates many economic and health problems in management modalities and leads to increasing mortality rates. Worldwide, trials have attempted to discover specific early markers for detection and prediction of the disease, hoping to set a more precise strategy for liver cancer prevention. Unfortunately, many economic, cultural and disciplinary levels contribute to confounding preventive strategies. Many risk factors contribute to predisposition to HCC, which can present individually or simultaneously. Previous articles discussed many risk factors for hepatocellular carcinogenesis; however, most of them didn't consider collectively the most recent data relating to causes. In this article, the pathogenesis and risk factors of HCC are discussed. Most of the intermediary steps of HCC involve molecular and transcriptional events leading to hepatocyte malignant transformation. These steps are mainly triggered by hepatitis B, C or transfusion-transmitted virus, either alone, or with other factors. Diabetes seems to be a major contributing risk factor. Schistosomiasis, a blood infestation, mostly affects Nile basin inhabitants leading to bladder, renal and hepatic cancers. Alcoholism, food and water pollutants and some drugs can also lead to HCC. Additionally, some hereditary diseases, as hemochromatosis, α-1-antitrypsin deficiency and tyrosinaemia are known to lead to the development of HCC, if not well managed.


Opinion formation in evaluating sanity at the time of the offense: an examination of 5175 pre-trial evaluations.
Sanity evaluations are high-stake undertakings that explicitly examine the defendant's culpability for a crime and implicitly explore clinical information that might inform a plea agreement. Despite the gravity of such evaluations, relatively little research has investigated the process by which evaluators form their psycholegal opinions. In the current study, we explore this process by examining 5175 sanity evaluations conducted by a cohort of forensic evaluators in Virginia over a ten-year period. Our analyses focus on (i) the clinical, criminal, and demographic attributes of the defendant correlated with opinions indicative of insanity; (ii) the clinical content of the evaluations and the legal criteria referenced as the basis for the psycholegal opinion; (iii) the process and outcome differences in the sanity evaluations conducted by psychologists versus psychiatrists; and (iv) the consistency in these opinions over a ten year period. Analyses predicting an opinion of insanity indicate a positive relationship with psychotic, organic, and affective diagnoses and previous psychiatric treatment. Analyses also indicate a negative relationship with prior criminal history, drug charges, personality disorder diagnosis, and intoxication at the time of the offense. Modest racial disparities were observed with evaluators offering opinions that the defendant was insane more often for white than for minority defendants despite comparable psychiatric and criminal characteristics.


On the Relation between Chemical Oscillations and Self-Replication.
One proposed scenario for the emergence of biochemical oscillations is that they may have provided the basic mechanism behind cellular self-replication by growth and division. However, alternative scenarios not requiring any chemical oscillation have also been proposed. Each of the various protocell models proposed to support one or another scenario comes with its own set of specific assumptions, which makes it difficult to ascertain whether chemical oscillations are required or not for cellular self-replication. This article compares these two cases within a single whole-cell model framework. This model relies upon a membrane embedding a chemical reaction network (CRN) synthesizing all the cellular constituents, including the membrane, by feeding from an external nutrient. Assuming the osmolarity is kept constant, the system dynamics are governed by a set of nonlinear differential equations coupling the chemical concentrations and the surface-area-to-volume ratio. The resulting asymptotic trajectories are used to determine the cellular shape by minimizing the membrane bending energy (within an approximate predefined family of shapes). While the stationary case can be handled quite generally, the oscillatory one is investigated using a simple oscillating CRN example, which is used to identify features that are expected to hold for any network. It is found that cellular self-replication can be reached with or without chemical oscillations, and that a requirement common to both stationary and oscillatory cases is that a minimum spontaneous curvature of the membrane is required for the cell to divide once its area and volume are both doubled. The oscillatory case can result in a greater variety of cellular shape trajectories but raises additional constraints for cellular division and self-replication: (i) the ratio of doubling time to oscillation period should be an integer, and (ii) if the oscillation amplitude is sufficiently high, then the spontaneous curvature must be below a maximum value to avoid early division before the end of the cycle. Because of these additional stringent constraints, it is likely that early protocells did not rely upon chemical oscillations. Biochemical oscillations typical of modern evolved cells may have emerged later through evolution for other reasons (e.g., metabolic advantage) and must have required additional feedback mechanisms for such a self-replicating system to be robust against even slight environmental variations (e.g., temperature fluctuations).


Two cases of acneiform eruption induced by inhibitor of epidermal growth factor receptor.
Cetuximab is a member of a new family of antineoplastic agents that inhibit the epidermal growth factor receptor (EGF-R), and which are increasingly being used in the treatment of solid tumors. We have observed new secondary side effects. We present here 2 patients with acneiform eruption secondary to the administration of cetuximab (IMC-C225, Erbitux). The diagnoses of these patients were adenocarcinoma. Histologically, a superficial purulent folliculitis and disordered differentiation with focal parakeratosis were observed. The follicular eruption responded favorably to treatment with 5% benzoyl peroxide and 4% erythromycin gel. These lesions healed within a few days after treatment. The cutaneous adverse effects of cetuximab are similar to other EGF-R-targeted agents and result from direct interference with the functions of EGF-R signaling in the skin.


CT characteristics of a transplantable canine glioma model: preliminary kinetic analysis.
Experimental brain tumors can be produced in dogs through the intracerebral injection of 3 X 10(6) live tumor cells in either neonates or adult animals. Tumors are visible by computed tomography on day 8 postinjection. Most tumors appear as ring lesions with central lucencies and shaggy borders. By postinjection day 12, tumor volumes increase more than 10 times; the cell cycling time is about 1-3 days. The initial doubling time is about 1-2 days and corresponds to the in vitro doubling time of about 24 hr. The use of computed tomography to perform noninvasive kinetic analysis deserves further study. The transplantable canine glioma model would appear to be ideal for this purpose.


A model based on linguistic 2-tuples for dealing with multigranular hierarchical linguistic contexts in multi-expert decision-making.
In those problems that deal with multiple sources of linguistic information we can find problems defined in contexts where the linguistic assessments are assessed in linguistic term sets with different granularity of uncertainty and/or semantics (multigranular linguistic contexts). Different approaches have been developed to manage this type of contexts, that unify the multigranular linguistic information in an unique linguistic term set for an easy management of the information. This normalization process can produce a loss of information and hence a lack of precision in the final results. In this paper, we shall present a type of multigranular linguistic contexts we shall call linguistic hierarchies term sets, such that, when we deal with multigranular linguistic information assessed in these structures we can unify the information assessed in them without loss of information. To do so, we shall use the 2-tuple linguistic representation model. Afterwards we shall develop a linguistic decision model dealing with multigranular linguistic contexts and apply it to a multi-expert decision-making problem.


[Relation between cognitive activity of excitable psychopathic personalities and the pattern of reinforcement].
Excitable psychopathic personalities manifest qualitative and quantitative disorders of cognitive activity directly unrelated to emotional reactions. If the result of the activity is estimated by some other person with the aid of positive and negative reinforcement stimuli, deficit of the training is marked to a much greater degree than in cases where the excitable personality itself draws, on the basis of information available in the feedback stimulus, the conclusion whether its decision was right or erroneous.


Observation of a nonlinear microfiber resonator.
Measurements of the intensity transfer function of a silica microfiber resonator are shown to follow a wide variety of hysteresis cycles, depending on the cavity detuning and the scanning frequency of the range of input powers. We attribute these observations to a nonlinear phase shift of thermal origin and provide a simple model that reproduces well our measurements. The response time is found to be around 0.6 ms.


In vivo and in situ rhizosphere respiration in Acer saccharum and Betula alleghaniensis seedlings grown in contrasting light regimes.
A perfusive method combined with an open-system carbon dioxide measurement system was used to assess rhizosphere respiration of Acer saccharum Marsh. (sugar maple) and Betula alleghaniensis Britton (yellow birch) seedlings grown in 8-l pots filled with coarse sand. We compared in vivo and in situ rhizosphere respiration between species, among light regimes (40, 17 and 6% of full daylight) and at different times during the day. To compute specific rhizosphere respiration, temperature corrections were made with either species-specific coefficients (Q10) based on the observed change in respiration rate between 15 and 21 degrees C or an arbitrarily assigned Q10 of 2. Estimated, species-specific Q10 values were 3.0 and 3.4 for A. saccharum and B. alleghaniensis, respectively, and did not vary with light regime. Using either method of temperature correction, specific rhizosphere respiration did not differ either between A. saccharum and B. alleghaniensis, or among light regimes except in A. saccharum at 6% of full daylight. At this irradiance, seedlings were smaller than in the other light treatments, with a larger fine root fraction of total root dry mass, resulting in higher respiration rates. Specific rhizosphere respiration was significantly higher during the afternoon than at other times of day when temperature-corrected on the basis of an arbitrary Q10 of 2, suggesting the possibility of diurnal variation in a temperature-independent component of rhizosphere respiration.


Tissue adenosine content in active soleus and gracilis muscles of cats.
Vascularly isolated cat soleus and gracilis muscles were stimulated to contract isometrically and were then frozen in situ. Adenosine, inosine, and hypoxanthine (nucleosides), and lactate were measured in neutralized, perchloric acid extracts of muscle. During contraction, nucleoside content increased in soleus muscle but changed little in gracilis muscle. However, adenosine content did not correlate with vascular conductance or oxygen consumption in either soleus or gracilis muscle. Adenosine content did correlate with lactate content in soleus but not gracilis muscle. The activity of AMP deaminase was highest in cat gracilis muscle and lowest in dog cardiac muscle. The activity of 5'-nucleotidase was lowest in cat gracilis muscle and highest in dog cardiac muscle. Cat soleus and dog gracilis muscles had intermediate activities of both enzymes. The findings of the present study do not support a role for adenosine in mediating prolonged active hyperemia in fast-twitch gracilis muscle of cats and cast doubt on such a role in slow-twitch soleus muscle of cats. Differences in the activities of AMP deaminase and 5'-nucleotidase provide a qualitative, biochemical explanation for apparent differences in net adenosine production among muscles composed of different fiber types and between skeletal and cardiac muscle.


Observation of vortex dipoles in an oblate Bose-Einstein condensate.
We report experimental observations and numerical simulations of the formation, dynamics, and lifetimes of single and multiply charged quantized vortex dipoles in highly oblate dilute-gas Bose-Einstein condensates (BECs). We nucleate pairs of vortices of opposite charge (vortex dipoles) by forcing superfluid flow around a repulsive Gaussian obstacle within the BEC. By controlling the flow velocity we determine the critical velocity for the nucleation of a single vortex dipole, with excellent agreement between experimental and numerical results. We present measurements of vortex dipole dynamics, finding that the vortex cores of opposite charge can exist for many seconds and that annihilation is inhibited in our trap geometry. For sufficiently rapid flow velocities, clusters of like-charge vortices aggregate into long-lived multiply charged dipolar flow structures.


Bowman Layer Onlay Transplantation to Manage Herpes Corneal Scar.
To introduce the concept of using a Bowman layer (BL) onlay graft to manage superficial herpetic corneal scarring and to describe the clinical outcomes of the first 2 cases undergoing this procedure. Two patients with a quiescent superficial corneal scar after herpes (varicella zoster virus [n = 1] and herpes simplex virus [n = 1]) keratitis underwent BL onlay transplantation. After the removal of the host epithelium and limited superficial keratectomy, an isolated BL graft was placed onto the host corneal surface. The cornea was then covered with an amniotic membrane and a bandage contact lens. Best spectacle-corrected visual acuity (VA) and/or best contact lens-corrected VA (BCLVA), biomicroscopy, corneal tomography, and anterior segment optical coherence tomography were recorded at 1 week, 1 month, and 3, 6, 9, 12, and 18 months postoperatively. In both cases, the surgical and postoperative courses were uneventful. An improvement of the corneal clarity was observed at biomicroscopy, and no varicella zoster virus/herpes simplex virus reactivation occurred throughout the follow-up period. Biomicroscopy, Scheimpflug imaging, and anterior segment optical coherence tomography showed a completely epithelialized and well-integrated graft postoperatively. In case 1, BCLVA with a scleral lens improved from 20/100 (0.1) preoperatively to 20/32 (0.6) postoperatively. For case 2, no preoperative BCLVA was available, but a BCLVA of 20/36 (0.55) was achieved after the procedure. A BL onlay graft may be a feasible surgical procedure, which may have the potential to reduce superficial corneal scarring and/or anterior corneal irregularities without resorting to deeper keratoplasty in these complex cases.


Habitat disturbance and the diversity and abundance of ants (Formicidae) in the Southeastern Fall-Line Sandhills.
We examined habitat disturbance, species richness, equitability, and abundance of ants in the Fall-Line Sandhills, at Fort Benning, Georgia. We collected ants with pitfall traps, sweep nets, and by searching tree trunks. Disturbed areas were used for military training; tracked and wheeled vehicles damaged vegetation and soils. Highly disturbed sites had fewer trees, diminished ground cover, warmer soils in the summer, and more compacted soils with a shallower A-horizon. We collected 48 species of ants, in 23 genera (141,468 individuals), over four years of sampling. Highly disturbed areas had fewer species, and greater numbers of ants than did moderately or lightly disturbed areas. The ant communities in disturbed areas were also less equitable, and were dominated by Dorymyrmex smithi.


alpha1-Antitrypsin deficiency and liver disease in children.
This report describes the clinical, biochemical, and hepatic morphologic findings in ten children with severe serum alpha1-antitrypsin deficiency. Genetic protease inhibitor (Pi) phenotyping, using acid-starch gel and crossed antigen-antibody electrophoresis, demonstrated Pi phenotype ZZ in all our cases. In eight patients, manifestations of liver disease appeared during the first year of life. The case reports show that alpha1-antitrypsin deficiency should be suspected in any child with neonatal hepatitis, unexplained hepatomegaly or splenomegaly, or cirrhosis. In our report, one infant is normal at age 6 months, and one infant had progressive hepatic damage that culminated in liver failure and death at age 6 months. The variable clinical course and prognosis for infants with severe alpha1-antitrypsin deficiency is well illustrated by these two infants.


GTP cyclohydrolase I and tyrosine hydroxylase gene mutations in familial and sporadic dopa-responsive dystonia patients.
Dopa-responsive dystonia (DRD) is a rare inherited dystonia that responds very well to levodopa treatment. Genetic mutations of GTP cyclohydrolase I (GCH1) or tyrosine hydroxylase (TH) are disease-causing mutations in DRD. To evaluate the genotype-phenotype correlations and diagnostic values of GCH1 and TH mutation screening in DRD patients, we carried out a combined study of familial and sporadic cases in Chinese Han subjects. We collected 23 subjects, 8 patients with DRD, 5 unaffected family members, and 10 sporadic cases. We used PCR to sequence all exons and splicing sites of the GCH1 and TH genes. Three novel heterozygous GCH1 mutations (Tyr75Cys, Ala98Val, and Ile135Thr) were identified in three DRD pedigrees. We failed to identify any GCH1 or TH mutation in two affected sisters. Three symptom-free male GCH1 mutation carriers were found in two DRD pedigrees. For those DRD siblings that shared the same GCH1 mutation, symptoms and age of onset varied. In 10 sporadic cases, only two heterozygous TH mutations (Ser19Cys and Gly397Arg) were found in two subjects with unknown pathogenicity. No GCH1 and TH mutation was found in 40 unrelated normal Han Chinese controls. GCH1 mutation is the main etiology of familial DRD. Three novel GCH1 mutations were identified in this study. Genetic heterogeneity and incomplete penetrance were quite common in DRD patients, especially in sporadic cases. Genetic screening may help establish the diagnosis of DRD; however, a negative GCH1 and TH mutation test would not exclude the diagnosis.


The efficacy of inflammatory markers in diagnosing infected diabetic foot ulcers and diabetic foot osteomyelitis: Systematic review and meta-analysis.
Diabetes foot ulcer (DFU) is a complication of diabetes mellitus. Accurate diagnosis of DFU severity through inflammatory markers will assist in reducing impact on quality of life. We aimed to ascertain the diagnostic test accuracy of commonly used inflammatory markers such as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), procalcitonin (PCT), and white cell count (WCC) for the diagnosis and differentiation between DFU grades based on the International Working Group on the Diabetic Foot classification system. This systematic review explored studies that investigated one or more of the above-listed index tests aiding in diagnosing infected DFU. This review was registered on PROSPERO database (ID = CRD42021255618) and searched 5 databases including an assessment of the references of included studies. Records were manually screened as per Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. A total of 16 studies were included which were assessed for quality using QUADAS-2 tool and meta-analysed using Meta-Disc v1.4. CRP had the greatest area under the curve (AUC) of 0.893 for diagnosing grade 2 DFU. This returned a pooled sensitivity and specificity of 77.4% (95% CI: 72% to 82%) and 84.3% (95% CI: 79% to 89%) respectively. In terms of diagnosing grade 3 DFU, procalcitonin had the highest AUC value of 0.844 when compared with other markers. The pooled sensitivity of PCT was calculated as 85.5% (95% CI: 79% to 90%) and specificity as 68.9% (95% CI: 63% to 75%). CRP and PCT are the best markers for diagnosing grade 2 and grade 3 DFU respectively. Other markers are also valuable when used in conjunction with clinical judgement. The findings accentuate the necessity of further research to establish standardised cut-off values for these inflammatory markers in diagnosing diabetic foot ulcers.


Bcl-xL mediates RIPK3-dependent necrosis in M. tuberculosis-infected macrophages.
Virulent Mycobacterium tuberculosis (Mtb) triggers necrosis in host Mϕ, which is essential for successful pathogenesis in tuberculosis. Here we demonstrate that necrosis of Mtb-infected Mϕ is dependent on the action of the cytosolic Receptor Interacting Protein Kinase 3 (RIPK3) and the mitochondrial Bcl-2 family member protein B-cell lymphoma-extra large (Bcl-xL). RIPK3-deficient Mϕ are able to better control bacterial growth in vitro and in vivo. Mechanistically, cytosolic RIPK3 translocates to the mitochondria where it promotes necrosis and blocks caspase 8-activation and apoptosis via Bcl-xL. Furthermore, necrosis is associated with stabilization of hexokinase II on the mitochondria as well as cyclophilin D-dependent mitochondrial permeability transition. Collectively, these events upregulate the level of reactive oxygen species to induce necrosis. Thus, in Mtb-infected Mϕ, mitochondria are an essential platform for induction of necrosis by activating RIPK3 function and preventing caspase 8-activation.


Structural and functional characterization of the human NBC3 sodium/bicarbonate co-transporter carboxyl-terminal cytoplasmic domain.
The sodium bicarbonate co-transporter, NBC3, is expressed in a range of tissues including heart, skeletal muscle and kidney, where it modulates intracellular pH and bicarbonate levels. NBC3 has a three-domain structure: 67 kDa N-terminal cytoplasmic domain, 57 kDa membrane domain and an 11 kDa C-terminal cytoplasmic domain (NBC3Ct). The role of C-terminal domains as important regulatory regions is an emerging theme in bicarbonate transporter physiology. This study determined the functional role of human NBC3Ct and characterized its structure using biochemical techniques. The NBC3 C-terminal domain deletion mutant (NBC3DeltaCt) had only 12 +/- 5% of wild-type transport activity. This low activity is attributable to low steady-state levels of NBC3DeltaCt and almost complete retention inside the cell, as assessed by immunoblots and confocal microscopy, suggesting a role of NBC3Ct in cell surface processing. To characterize the structure of NBC3Ct, amino acids 1127-1214 of NBC3 were expressed as a GST fusion protein (GST.NBC3Ct). GST.NBC3Ct was cleaved with PreScission Protease and native NBC3Ct could be purified to 94% homogeneity. Gel permeation chromatography and sedimentation velocity ultracentrifugation of NBC3Ct indicated a Stokes radius of 26 and 30 angstroms, respectively. Shape modelling revealed NBC3Ct as a prolate shape with long and short axes of 19 and 2 nm, respectively. The circular dichroism spectra of NBC3Ct did not change over the pH 6.2-7.8 range, which rules out a large change of secondary structure as a component of pH sensor function. Proteolysis with trypsin and chymotrypsin identified two proteolytically sensitive regions, R1129 and K1183-K1186, which could form protein interaction sites.


Health promotion for mild frailty based on behaviour change: Perceptions of older people and service providers.
Mild frailty is common among older people, but it is potentially reversible with health promotion interventions. Behaviour change may be a key to preventing progression of frailty; however, we know little about what interventions work best and how a behaviour change approach would be perceived by this group. The aim of this study was to explore how mildly frail older people perceive health promotion based on behaviour change and what factors affect engagement with this approach. We conducted semi-structured interviews with 16 older people with mild frailty who received a pilot home-based behaviour change health promotion service, including a dyad of older person/family carer, and two service providers delivering the service in two diverse areas of South England. Interviews were audio-recorded, transcribed and thematically analysed. The concept of goal setting was acceptable to most participants, though the process of goal setting needed time and consideration. Goals on maintaining independence, monitoring of progress and receiving feedback were reported to increase motivation. Physical/mental capability and knowledge/perception of own needs were main determinants of the type of goals chosen by participants as well as the approach used by the project workers. Older people with complex needs benefited from care coordination, with a combination of goal setting and elements of social, practical and emotional support in varying proportions. Mildly frail older people responded well to a behaviour change approach to promote health and well-being. Further consideration is needed of the most effective strategies based on complexity of needs, and how to overcome barriers among people with cognitive impairment.


Resonance Raman study of the J-type aggregation process of a water soluble perylene bisimide.
Perylene bisimides (PBIs) are dyes known for combining high absorption and emission in the visible region with thermal and photochemical stability. H-bond-directed aggregation driven by free imide groups has been reported to promote the uncommon J-type aggregate formation of PBIs. J-aggregates are highly desired thanks to their bathochromically shifted narrow absorption and fluorescence due to excitonic coupling, together with hyperchromicity and superradiance compared to the monomer. Herein we present the water soluble MEG-PBI showing interesting aggregation in water and in the solid state. Unlike its hydrophobic counterparts, MEG-PBI aggregates in water with increasing temperature, indicating entropy-driven self-assembly. Temperature-dependent Resonance Raman (RR) spectroscopy was employed for the structural characterization of MEG-PBI in aqueous solution versus toluene and in aggregated thin films, employing excitation at different wavelengths to probe the contribution of various chromophores to the supramolecular structure of the aggregate. We find that the perylene core distorts upon aggregation, where the bonds along the perylene long N-N axis lengthen and the ones perpendicular to that shorten, suggesting a head-to-tail arrangement due to H-bonding between neighboring units.


Results of late surgical intervention in children with early-onset bilateral cataracts.
Cataracts are a major cause of childhood blindness globally. Although surgically treatable, it is unclear whether children would benefit from such interventions beyond the first few years of life, which are believed to constitute 'critical' periods for visual development. To study visual acuity outcomes after late treatment of early-onset cataracts and also to determine whether there are longitudinal changes in postoperative acuity. We identified 53 children with dense cataracts with an onset within the first half-year after birth through a survey of over 20,000 rural children in India. All had accompanying nystagmus and were older than 8 years of age at the time of treatment. They underwent bilateral cataract surgery and intraocular lens implantation. We then assessed their best-corrected visual acuity 6 weeks and 6 months after surgery. 48 children from the pool of 53 showed improvement in their visual acuity after surgery. Our longitudinal assessments demonstrated further improvements in visual acuity for the majority of these children proceeding from the 6-week to 6-month assessment. Interestingly, older children in our subject pool did not differ significantly from the younger ones in the extent of improvement they exhibit. Our results demonstrate that not only can significant vision be acquired until late in childhood, but that neural processes underlying even basic aspects of vision like resolution acuity remain malleable until at least adolescence. These data argue for the provision of cataract treatment to all children, irrespective of their age.


100 YEARS OF VITAMIN D: Dose-response for change in 25-hydroxyvitamin D after UV exposure: outcome of a systematic review.
A systematic review of publications addressing change in vitamin D status (25-hydroxyvitamin D (25OHD)) after exposure to UV radiation identified 2001 independent peer-reviewed publications. Of these, 21 used artificial sources of UV radiation, met all inclusion criteria and were quality assured; 13 publications used solar radiation and met sufficient inclusion criteria to be retained as supporting evidence; 1 further included publication used both solar and artificial sources. The review consistently identified that low dose, sub-erythemal doses are more effective for vitamin D synthesis than doses close to a minimum erythema dose; increasing skin area exposed increases the amount of vitamin D synthesised although not necessarily in a linear manner; constant dosing leads to a dose-dependent plateau in 25OHD, and dose-response is greatest at the start of a dosing regime; there is a large interpersonal variation in response to UV exposure. Fourteen of the studies using artificial sources of radiation were used to determine a dose-response relationship for change in 25OHD on whole-body exposure to repeated sub-erythemal doses of UV radiation, taking the form Δ25OHD (nmol/L) = A ln(standard vitamin D dose) + B. This helps quantify our understanding of UV as a source of vitamin D and enables exposure regimes for safe synthesis of vitamin D to be assessed. Specific studies of people with pigmented skin (Fitzpatrick skin types 5 and 6) were rare, and this dose-response relationship is only applicable to white-skinned individuals as skin type is a determinant of response to UV radiation. Findings provide information for vitamin D guidance updates.


Crystallization and preliminary diffraction analysis of the HincII restriction endonuclease-DNA complex.
Crystals of the 60 kDa dimeric HincII restriction enzyme bound to a 12 base-pair dyad-symmetric duplex DNA carrying the specific 5'-GTCGAC recognition site have been obtained. Crystals grew by hanging-drop vapor diffusion from solutions containing polyethylene glycol 4000 as precipitating agent. The rod-shaped crystals belong to space group I222 (or I2(1)2(1)2(1)), with unit-cell dimensions a = 66.9, b = 176.7, c = 256.0 A. There are most likely to be two dimeric complexes in the asymmetric unit. A complete native data set has been collected from a high-energy synchrotron source to a resolution of 2.5 A at 100 K, with an R(merge) of 4.8%.


Is the effect of aerobic exercise on cognition a placebo effect?
A number of studies and meta-analyses conclude that aerobic fitness (walking) interventions improve cognition. Such interventions typically compare improvements from these interventions to an active control group in which participants engage in non-aerobic activities (typically stretching and toning) for an equivalent amount of time. However, in the absence of a double-blind design, the presence of an active control group does not necessarily control for placebo effects; participants might expect different amounts of improvement for the treatment and control interventions. We conducted a large survey to explore whether people expect greater cognitive benefits from an aerobic exercise intervention compared to a control intervention. If participants expect greater improvement following aerobic exercise, then the benefits of such interventions might be due in part to a placebo effect. In general, expectations did not differ between aerobic and non-aerobic interventions. If anything, some of the results suggest the opposite (e.g., respondents expected the control, non-aerobic intervention to yield bigger memory gains). These results provide the first evidence that cognitive improvements following aerobic fitness training are not due to differential expectations.


White matter abnormalities and structural hippocampal disconnections in amnestic mild cognitive impairment and Alzheimer's disease.
The purpose of this project was to evaluate white matter degeneration and its impact on hippocampal structural connectivity in patients with amnestic mild cognitive impairment, non-amnestic mild cognitive impairment and Alzheimer's disease. We estimated white matter fractional anisotropy, mean diffusivity and hippocampal structural connectivity in two independent cohorts. The ADNI cohort included 108 subjects [25 cognitively normal, 21 amnestic mild cognitive impairment, 47 non-amnestic mild cognitive impairment and 15 Alzheimer's disease]. A second cohort included 34 subjects [15 cognitively normal and 19 amnestic mild cognitive impairment] recruited in Montreal. All subjects underwent clinical and neuropsychological assessment in addition to diffusion and T1 MRI. Individual fractional anisotropy and mean diffusivity maps were generated using FSL-DTIfit. In addition, hippocampal structural connectivity maps expressing the probability of connectivity between the hippocampus and cortex were generated using a pipeline based on FSL-probtrackX. Voxel-based group comparison statistics of fractional anisotropy, mean diffusivity and hippocampal structural connectivity were estimated using Tract-Based Spatial Statistics. The proportion of abnormal to total white matter volume was estimated using the total volume of the white matter skeleton. We found that in both cohorts, amnestic mild cognitive impairment patients had 27-29% white matter volume showing higher mean diffusivity but no significant fractional anisotropy abnormalities. No fractional anisotropy or mean diffusivity differences were observed between non-amnestic mild cognitive impairment patients and cognitively normal subjects. Alzheimer's disease patients had 66.3% of normalized white matter volume with increased mean diffusivity and 54.3% of the white matter had reduced fractional anisotropy. Reduced structural connectivity was found in the hippocampal connections to temporal, inferior parietal, posterior cingulate and frontal regions only in the Alzheimer's group. The severity of white matter degeneration appears to be higher in advanced clinical stages, supporting the construct that these abnormalities are part of the pathophysiological processes of Alzheimer's disease.


Methylation of ribonucleic acid by the carcinogens dimethyl sulphate, N-methyl-N-nitrosourea and N-methyl-N'-nitro-N-nitrosoguanidine. Comparisons of chemical analyses at the nucleoside and base levels.
1. The following methods for hydrolysis of methyl-(14)C-labelled RNA, and for chromatographic isolation and determination of the products, were investigated: enzymic digestion to nucleosides at pH6 or 8; alkaline hydrolysis and conversion into nucleosides; hydrolysis by acid to pyrimidine nucleotides and purine bases, or completely to bases; chromatography on Dowex 50 (NH(4) (+) form) at pH6 or 8.9, or on Dowex 50 (H(+) form), or on Sephadex G-10. 2. The suitability of the various methods for determination of methylation products was assessed. The principal product, 7-methylguanosine, was unstable under the conditions used for determinations of nucleosides. 3- and 7-Methyladenine and 3- and 7-methylguanine are best determined as bases; 1-methyladenine and 3-methylcytosine can be isolated as either nucleosides or bases; O(6)-methylguanine is unstable under the acid hydrolysis conditions used and can be determined as the nucleoside; 3-methyluracil was detected, but may be derived from methylation of the ionized form of uracil. 3. Differences between the patterns of methylation of RNA and homopolyribonucleotides by the N-methyl-N-nitroso compounds and dimethyl sulphate were found: the nitroso compounds were able to methylate O-6 of guanine, were relatively more reactive at N-7 of adenine and probably at N-3 of guanine, but less reactive at N-1 of adenine, N-3 of cytosine and probably at N-3 of uridine. They probably reacted more with the ribose-phosphate chain, but no products from this were identified. 4. The possible influences of these differences on biological action of the methylating agents is discussed. Nitroso compounds may differ principally in their ability to induce miscoding in the Watson-Crick sense by reaction at O-6 of guanine. Both types of agent may induce miscoding to a lesser extent through methylation at N-3 of guanine; both can methylate N atoms, presumably preventing Watson-Crick hydrogen-bonding. N-Methyl-N-nitrosourea can degrade RNA, possibly through phosphotriester formation, but this mechanism is not proven.


Helical and β-Turn Conformations in the Peptide Recognition Regions of the VIM1 PHD Finger Abrogate H3K4 Peptide Recognition.
The PHD finger-containing VARIANT IN METHYLATION/ORTHRUS (VIM/ORTH) family of proteins in Arabidopsis consists of functional homologues of mammalian UHRF1 and is required for the maintenance of DNA methylation. Comparison of the sequence with those of other PHD fingers implied that VIM1 and VIM3 PHD could recognize lysine 4 of histone H3 (H3K4) through interactions mediated by a conserved aspartic acid. However, our calorimetric and modified histone peptide array binding studies suggested that neither H3K4 nor other histone marks are recognized by VIM1 and VIM3 PHD fingers. Here, we report a 2.6 Å resolution crystal structure of the VIM1 PHD finger and demonstrate significant structural changes in the putative H3 recognition segments in contrast to canonical H3K4 binding PHD fingers. These changes include (i) the H3A1 binding region, (ii) strand β1 that forms an intermolecular β-sheet with the H3 peptide, and (iii) an aspartate-containing motif involved in salt bridge interaction with H3K4, which together appear to abrogate recognition of H3K4 by the VIM1 PHD finger. To understand the significance of the altered structural features in the VIM1 PHD that might prevent histone H3 recognition, we modeled a chimeric VIM1 PHD (chmVIM1 PHD) by grafting the peptide binding structural features of the BHC80 PHD onto the VIM1 PHD. Molecular dynamics simulation and metadynamics analyses revealed that the chmVIM1 PHD-H3 complex is stable and also showed a network of intermolecular interactions similar to those of the BHC80 PHD-H3 complex. Collectively, this study reveals that subtle structural changes in the peptide binding region of the VIM1 PHD abrogate histone H3 recognition.


Learning sensory maps with real-world stimuli in real time using a biophysically realistic learning rule.
We present a real-time model of learning in the auditory cortex that is trained using real-world stimuli. The system consists of a peripheral and a central cortical network of spiking neurons. The synapses formed by peripheral neurons on the central ones are subject to synaptic plasticity. We implemented a biophysically realistic learning rule that depends on the precise temporal relation of pre- and postsynaptic action potentials. We demonstrate that this biologically realistic real-time neuronal system forms stable receptive fields that accurately reflect the spectral content of the input signals and that the size of these representations can be biased by global signals acting on the local learning mechanism. In addition, we show that this learning mechanism shows fast acquisition and is robust in the presence of large imbalances in the probability of occurrence of individual stimuli and noise.


Increases in Physical Activity Result in Diminishing Increments in Daily Energy Expenditure in Mice.
Exercise is a common component of weight loss strategies, yet exercise programs are associated with surprisingly small changes in body weight [1-4]. This may be due in part to compensatory adaptations, in which calories expended during exercise are counteracted by decreases in other aspects of energy expenditure [1, 5-10]. Here we examined the relationship between a rodent model of voluntary exercise- wheel running- and total daily energy expenditure. Use of a running wheel for 3 to 7 days increased daily energy expenditure, resulting in a caloric deficit of ∼1 kcal/day; however, total daily energy expenditure remained stable after the first week of wheel access, despite further increases in wheel use. We hypothesized that compensatory mechanisms accounted for the lack of increase in daily energy expenditure after the first week. Supporting this idea, we observed a decrease in off-wheel ambulation when mice were using the wheels, indicating behavioral compensation. Finally, we asked whether individual variation in wheel use within a group of mice would be associated with different levels of daily energy expenditure. Despite a large variation in wheel running, we did not observe a significant relationship between the amount of daily wheel running and total daily energy expenditure or energy intake across mice. Together, our experiments support a model in which the transition from sedentary to light activity is associated with an increase in daily energy expenditure, but further increases in physical activity produce diminishingly small increments in daily energy expenditure.


Effect of N-acyl-phosphatidylethanolamine on the membrane fusion between Sendai virus and liposome.
We have compared the properties of two N-acyl derivatives of dilauryl phosphatidylethanolamine on lipid polymorphism, vesicle leakage and Sendai virus fusion. The derivatives contained either an N-lauroyl group (NLPE) or an N-acetyl group (NAcPE). Only the NAcPE markedly affected the bilayer to hexagonal transition temperature of dielaidoyl phosphatidylethanolamine, shifting it to higher values. In contrast the NLPE slightly lowered this phase transition temperature. The two lipids also have opposite effects on leakage from small unilamellar vesicles of egg phosphatidylcholine. The NLPE inhibits leakage, while the NAcPE promotes it. This vesicle stabilizing effect of NLPE against leakage is not manifested in alterations of rates or extents of Sendai virus fusion to liposomes of egg phosphatidylethanolamine plus 2% ganglioside GD1a. The NLPE has no effect, while the NAcPE reduces the observed fusion, at least in part as a consequence of a reduction in the final extent of fusion. These results demonstrate that the bilayer stabilizing effects of NLPE do not result in a lower rate of viral fusion. Furthermore, these bilayer stabilizing effects against leakage are not solely a function of the lipid headgroup but also require a structure with three long acyl chains. The reduced leakage is not related to a loss in monolayer curvature strain.


Genetic salivary protein polymorphism in Mexican population.
Genetic polymorphism is the major contributor that affects human salivary composition. In order to determine the molecular phenotypes in saliva, it is important to know the distribution of proteins with specific functions which allows the clinical diagnosis of specific diseases. Unstimulated human whole saliva samples from 120 subjects were subjected to sodium dodecyl sulfate polyacrylamide slab gel electrophoresis (SDS-PAGE). The phenotype distribution of several molecules including MG1, MG2, alpha-Amylase, PRP-I and cystatins were similar. Qualitative and quantitative characteristics were specific in each subject.


Optimal Encoding in Stochastic Latent-Variable Models.
In this work we explore encoding strategies learned by statistical models of sensory coding in noisy spiking networks. Early stages of sensory communication in neural systems can be viewed as encoding channels in the information-theoretic sense. However, neural populations face constraints not commonly considered in communications theory. Using restricted Boltzmann machines as a model of sensory encoding, we find that networks with sufficient capacity learn to balance precision and noise-robustness in order to adaptively communicate stimuli with varying information content. Mirroring variability suppression observed in sensory systems, informative stimuli are encoded with high precision, at the cost of more variable responses to frequent, hence less informative stimuli. Curiously, we also find that statistical criticality in the neural population code emerges at model sizes where the input statistics are well captured. These phenomena have well-defined thermodynamic interpretations, and we discuss their connection to prevailing theories of coding and statistical criticality in neural populations.


Cellular interaction and in vitro antitumor activity of mitomycin C-dextran conjugate.
Cellular interaction and in vitro antitumor activity of a polymeric prodrug of mitomycin C (MMC), mitomycin C-dextran conjugate (MMC-D), were studied in relation to its physicochemical characteristics. MMC-D with cationic and anionic charges were examined. The cationic MMC-D was synthesized using a spacer, epsilon-aminocaproic acid and dextrans with molecular weights of 10,000, 70,000, or 500,000 [MMC(C6)Dcat]. The anionic MMC-D was synthesized using 6-bromohexanoic acid as a spacer and dextran with a molecular weight of 70,000 [MMC(C6)Dan]. Cellular adsorption was determined by measuring the concentration of the drug in the medium after incubation with three tumor cell lines, Ehrlich ascites carcinoma, L1210 leukemia, and AH66 ascites hepatoma cells. MMC(C6)Dcat was adsorbed more readily than MMC or MMC(C6)Dan on the tumor cell surface by an electrostatic force. The percentage of adsorption remained almost constant during the course of incubation and no significant difference was observed between the incubation at 4 degrees C and that at 37 degrees C. A corresponding increase in the amounts of MMC(C6)Dcat adsorbed on with higher molecular weights was noted, which conformed to Langmuir's adsorption isotherm. In vitro antitumor activity was evaluated using L1210 and EAC cell culture systems and human tumor colony forming assay. MMC(C6)Dcat showed growth inhibition essentially equal to that of MMC in continuous drug exposure experiments. In a 1-h drug exposure experiment, MMC(C6)Dcat with a molecular weight of 70,000 or 500,000 was more active than MMC, and a good correlation was observed between the effects of MMC(C6)Dcat and the extent of cellular interaction. These results show that cellular interaction played an important role in the manifestation of the antitumor effect of MMC-D and that these phenomena are governed by the physicochemical properties of macromolecular prodrugs, such as electric charge and molecular weight.


Differential Expression of miRNAs and Their Predicted Target Genes Indicates That Gene Expression in Atlantic Salmon Gill Is Post-Transcriptionally Regulated by miRNAs in the Parr-Smolt Transformation and Adaptation to Sea Water.
Smoltification (parr-smolt transformation) is a complex developmental process consisting of developmental changes that lead to remodeling of the Atlantic salmon gill. Here, the expression changes of miRNAs and mRNAs were studied by small-RNA sequencing and microarray analysis, respectively, to identify miRNAs and their predicted targets associated with smoltification and subsequent sea water adaptation (SWA). In total, 18 guide miRNAs were identified as differentially expressed (gDE miRNAs). Hierarchical clustering analysis of expression changes divided these into one cluster of 13 gDE miRNAs with decreasing expression during smoltification and SWA that included the miRNA-146, miRNA-30 and miRNA-7132 families. Another smaller cluster that showed increasing expression consisted of miR-101a-3p, miR-193b-5p, miR-499a-5p, miR-727a-3p and miR-8159-5p. The gDE miRNAs were predicted to target 747 of the genes (DE mRNAs), showing expression changes in the microarray analysis. The predicted targets included genes encoding NKA-subunits, aquaporin-subunits, cystic fibrosis transmembrane conductance regulator and the solute carrier family. Furthermore, the predicted target genes were enriched in biological processes associated with smoltification and SWA (e.g., immune system, reactive oxygen species, stress response and extracellular matrix organization). Collectively, the results indicate that remodeling of the gill involves the post-transcriptional regulation of gene expression by the characterized gDE miRNAs.


Biological evaluation of 131I- and CF750-labeled Dmab(scFv)-Fc antibodies for xenograft imaging of CD25-positive tumors.
A Dmab(scFv)-Fc antibody containing the single chain variable fragment of a humanized daclizumab antibody and the Fc fragment of a human IgG1 antibody was produced via recombinant expression in Pichia pastoris. The Dmab(scFv)-Fc antibody forms a dimer in solution, and it specifically binds CD25-positive tumor cells and tumor tissues. For tumor imaging, the Dmab(scFv)-Fc antibody was labeled with the 131I isotope and CF750 fluorescent dye, respectively. After intravenous injection of mice bearing CD25-positive tumor xenografts, tumor uptake of the (131)I-Dmab(scFv)-Fc antibody was visible at 1 h, and clear images were obtained at 5 h using SPECT/CT. After systemic administration of the CF750-Dmab(scFv)-Fc antibody, tumor uptake was present as early as 1 h, and tumor xenografts could be kinetically imaged within 9 h after injection. These results indicate that the Dmab(scFv)-Fc antibody rapidly and specifically targets CD25-positive tumor cells, suggesting the potential of this antibody as an imaging agent for the diagnosis of lymphomatous-type ATLL.


Does melatonin protect or treat brain damage from traumatic oxidative stress?
A variety of experimental studies have demonstrated the neuroprotective effects of melatonin, based on its antioxidant activity. In a prospective randomized study, the effects of melatonin were investigated in experimental head trauma-induced oxidative stress in rabbits. The experimental study was performed on 30 rabbits. The animals were divided into three groups. Group I (sham procedure): a right parietal craniotomy was performed on each animal, and the dura mater was left intact. Group II: experimental brain trauma (EBT) was performed on each animal using a 1 cm inner diameter x 10 cm long glass tube, through which a 20 g weight (0.5 cm diameter) was dropped onto the brain at the craniotomy site, causing a contusional head trauma. Group III: the same EBT model was performed, but 2.5 mg/kg melatonin was injected intraperitoneally four times (total dose 10 mg/kg); these injections were performed 20 min before the operation, during the trauma, 1 h later and 2 h later. The rabbits were sacrificed after the EBT at 24 h after the brain trauma. The activities of the three principal antioxidant enzymes-catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px)-were determined, and the levels of malondialdehyde (MDA), a product of lipid peroxidation, and glutathione (GSH) were measured in brain homogenates. MDA levels were found to be higher in the EBT group than in the EBT+melatonin group or the sham procedure group. The SOD activity was found to be higher in the EBT group than in the sham procedure group. Enzymatic parameters (except for SOD) were significantly higher in melatonin-treated animals than in EBT animals. GSH levels in melatonin-treated animals were decreased compared with EBT animals. In conclusion, the data indicate that melatonin protects against free radical-mediated oxidative changes in brain tissue by boosting antioxidant enzymes, and in particular lowering lipid peroxidation in rabbits with EBT.


Abnormalities of ejaculation.
The normal physiologic processes of emission and ejaculation require coordination of neurophysiologic, anatomic, and, in certain cases, psychological phenomena. Disruption of any component, from the embryologic development of the müllerian duct through the medications used for nonrelated systemic disease, can alter the efficient function of ejaculation. Evaluation of the urologic patient who has any of a number of abnormal ejaculatory states requires an understanding of the many possible mechanisms of failure. The majority of these men need evaluation because of a possible male-factor infertile marriage. The potential for improvement is significant, given the development of improved techniques to stimulate ejaculation and the promise shown by extracorporeal fertilization techniques such as in vitro fertilization and gamete intrafallopian tube transfer. These patients deserve complete assessment and optimization of any factors that will enable them to achieve their goal of procreation.


Cloning and sequencing of protochlorophyllide reductase.
Putative protochlorophyllide reductase cDNA clones (252 and 113) were isolated from an etiolated-oat (Avena sativa) cDNA library. These were used to indirectly characterize a further clone, p127, isolated from a lambda-phage gt11 cDNA library. The latter (1.15 kb in length) was sequenced, and the derived amino acid sequence was shown to be remarkably similar to that derived from chemical analysis of a CNBr-cleavage fragment of the purified reductase, p127 codes for more than 95% of the reductase protein.


Inhibition of human acetyl- and butyrylcholinesterase by novel carbamates of (-)- and (+)-tetrahydrofurobenzofuran and methanobenzodioxepine.
A new enantiomeric synthesis utilizing classical resolution provided two novel series of optically active inhibitors of cholinesterase: (-)- and (+)-O-carbamoyl phenols of tetrahydrofurobenzofuran and methanobenzodioxepine. An additional two series of (-)- and (+)-O-carbamoyl phenols of pyrroloindole and furoindole were obtained by known procedures, and their anticholinesterase actions were similarly quantified against freshly prepared human acetyl- (AChE) and butyrylcholinesterase (BChE). Both enantiomeric forms of each series demonstrated potent cholinesterase inhibitory activity (with IC(50) values as low as 10 nM for AChE and 3 nM for BChE), with the exception of the (+)-O-carbamoyl phenols of pyrroloindole, which lacked activity (IC(50) values >1 microM). Based on the biological data of these four series, a structure-activity relationship (SAR) analysis was provided by molecular volume calculations. In addition, a probable transition-state model was established according to the known X-ray structure of a transition-state complex of Torpedo californica AChE-m-(N,N,N-trimethylammonio)-2,2,2-trifluoroacetophenone (TcAChE-TMTFA). This model proved valuable in explaining the enantioselectivity and enzyme subtype selectivity of each series. These carbamates are more potent than, or similarly potent to, anticholinesterases in current clinical use, providing not only inhibitors of potential clinical relevance but also pharmacological tools to define drug-enzyme binding interactions within an enzyme crucial in the maintenance of cognition and numerous systemic physiological functions in health, aging, and disease.


Absolute intensities of NH-stretching transitions in dimethylamine and pyrrole.
Vibrational spectra of vapor-phase dimethylamine (DMA) and pyrrole have been recorded in the 1000 to 13000 cm(-1) region using long path conventional spectroscopy techniques. We have focused on the absolute intensities of the NH-stretching fundamental and overtone transitions; Δν(NH) = 1-4 regions for DMA and the Δν(NH) = 1-3 regions for pyrrole. In the Δν(NH) = 1-3 regions for DMA, evidence of tunneling splitting associated with the NH-wagging mode is observed. For DMA, the fundamental NH-stretching transition intensity is weaker than the first NH-stretching overtone. Also, the fundamental NH-stretching transition in DMA is much weaker than the fundamental transition in pyrrole. We have used an anharmonic oscillator local mode model with ab initio calculated local mode parameters and dipole moment functions at the CCSD(T)/aug-cc-pVTZ level to calculate the NH-stretching intensities and explain this intensity anomaly in DMA.


[Laser surgery in the conservative management of vaginal intraepithelial neoplasms].
The increase finding of vaginal intraepithelial neoplasia (VaIN) in young women has led to the need to develop personalized conservative treatment. The authors studied the use of CO2 laser in the treatment of VaIN, evaluating the efficacy, repeatability, complications and the conservation of sexual function. A total of 37 consecutive patients with histologically demonstrated VaIN (10 VaIN 1, 14 VaIN 2 and 13 VaIN 3): mean age 45.3 years, 37.8% aged < 35 years, 20/37 in childbearing age; VaIN was isolated in two patients, synchronous in 22 and in 13 cases it was metachronous with other intraepithelial or invasive genital neoplasia. 30/37 patients were treated with laser surgery as outpatients for a total of 41 operations (37 vaporisations and four excisions) using colposcopic guided CO2 laser with a constant emission power of 35 watts or 30 watts in the pulsed mode, under local anaesthesia, following the application of 5% acetic acid. Twenty-one (87.5%) of the 24 patients evaluated at 12 months were negative after one (16 patients) or more treatments (from 2 to 5 in 5 patients). Following the second recidivation, one elderly patient preferred traditional demolitive surgery; 2 patients are still awaiting further laser surgery. Five patients with multifocal lesions associated topical treatment with 5-fluorouracil. None of the sexually active patients complained of dyspareunias after treatment. Given the lack of other equally effective treatment and with the aim of preserving genital integrity, the authors regard laser surgery as the elective treatment for VaIN 2 and 3.


Central venous catheter associated upper extremity deep vein thrombosis in cancer patients: Diagnosis and therapeutic management.
Catheter-related thrombosis (CRT) is a relatively frequent and potentially fatal complication arising in patients with cancer who require a central catheter placement for intravenous treatment. In everyday practice, CRT remains a challenge for management; despite its frequency and its negative clinical impact, few data are available concerning diagnosis and treatment of CRT. In particular, no diagnostic studies or clinical trials have been published that included exclusively patients with cancer and a central venous catheter (CVC). For this reason, many questions regarding optimal management of CRT remain unanswered. Due to the paucity of high-grade evidence regarding CRT in cancer patients, guidelines are derived from upper extremity DVT studies for diagnosis, and from those for lower limb DVT for treatment. This article addresses the issues of diagnosis and management of CRT through a review of the available literature and makes a number of proposals based on the available evidence. In symptomatic patients, venous ultrasound is the most appropriate choice for first-line diagnostic imaging of CRT because it is noninvasive, and its diagnostic performance is high (which is not the case in asymptomatic patients). In the absence of direct comparative clinical trials, we suggest treating patients with CRT with a therapeutic dose of either a LMWH or a direct oral factor Xa inhibitor, with or without a loading dose. These anticoagulants should be given for a total of at least three months, including at least one month after catheter removal following initiation of therapy.


Characterization of methods for mechanistic inference of the gut microbiome in disease.
Knowledge graphs have found broad biomedical applications, providing useful representations of complex knowledge. Although plentiful evidence exists linking the gut microbiome to disease, mechanistic understanding of those relationships remains generally elusive. A structured analysis of existing resources is necessary to characterize the resources and methodologies needed to facilitate mechanistic inference. Here we demonstrate the potential of knowledge graphs to hypothesize plausible mechanistic accounts of host-microbe interactions in disease and define the need for semantic constraint in doing so. We constructed a knowledge graph of linked microbes, genes and metabolites called MGMLink, and one of microbial traits, environments, and human pheno-types called KG-microbe-phenio. Using a shortest path search and a pattern based semantically constrained path search through the graphs, we highlight the need for a microbiome-disease resource and semantically informed search methods to enable mechanistic inference. The software to create MGMLink is openly available at https://github.com/bsantan/MGMLink , and KG-microbe is available at https://github.com/Knowledge-Graph-Hub/kg-microbe and KG-phenio is available at https://github.com/Knowledge-Graph-Hub/kg-phenio . email@example.com.


Prognostic significance of exercise-induced left bundle-branch block.
Approximately 0.5% of all patients who undergo exercise testing develop a transient left bundle-branch block (LBBB) during exercise, but its prognostic significance is unclear. To determine whether exercise-induced LBBB is an independent predictor of mortality and cardiac morbidity. Matched control cohort study. Between September 1990 and February 10, 1994, 17277 exercise stress tests were performed on patients. Tertiary care, academic medical center. From the cohort, 70 cases of exercise-induced LBBB were identified. The controls comprised 70 individuals without LBBB at rest or during exercise that matched the 70 cases based on age, test date, sex, prior history of coronary artery disease, hypertension, diabetes, smoking, and beta-blocker use. All-cause mortality, percutaneous coronary intervention, open heart surgery, nonfatal myocardial infarction, documented symptomatic or sustained ventricular tachydysrhythmia, or implantation of a permanent pacemaker or an implantable cardiac defibrillator. A total of 37 events (28 events from the exercise-induced LBBB cases and 9 from the control cohort) occurred in 25 patients (17 exercise-induced LBBB patients and 8 control patients) during a mean follow-up period of 3.7 (0.9 years) (median, 3.8 years [range, 0.9-5.2 years]). There were 7 deaths, of which 5 occurred among patients with exercise-induced LBBB. Four-year Kaplan-Meier event rates were 19% among exercise-induced LBBB patients and 10% among controls (log-rank chi2, 5.2; P=.02). After further adjusting for small differences in age, exercise-induced LBBB remained associated with a higher risk of primary events (adjusted relative risk, 2.78; 95% confidence interval, 1.16-6.65; P=.02). Exercise-induced LBBB independently predicts a higher risk of death and major cardiac events.


Massively parallel sequencing for chromosomal abnormality testing in trophectoderm cells of human blastocysts.
Preimplantation genetic diagnosis and screening are widely accepted for chromosomal abnormality identification to avoid transferring embryos with genetic defects. Massively parallel sequencing (MPS) is a rapidly developing approach for genome analysis with increasing application in clinical practice. The purpose of this study was to use MPS for identification of aneuploidies and unbalanced chromosomal rearrangements after blastocyst biopsy. Trophectoderm (TE) samples of 38 blastocysts from 16 in vitro fertilization cycles were subjected to analysis. Low-coverage whole genome sequencing was performed using the Illumina HiSeq2000 platform with a novel algorithm purposely created for chromosomal analysis. The efficiency of this MPS approach was estimated by comparing results obtained by an Affymetrix single-nucleotide polymorphism (SNP) array. Whole genome amplification (WGA) products of TE cells were detected by MPS, with an average of 0.07× depth and 5.5% coverage of the human genome. Twenty-six embryos (68.4%) were detected as euploid, while six embryos (15.8%) contained uniform aneuploidies. Four of these (10.5%) were with solely unbalanced chromosomal rearrangements, whereas the remaining two embryos (5.3%) showed both aneuploidies and unbalanced rearrangements. Almost all these results were confirmed by the SNP array, with the exception of one sample, where different sizes of unbalanced rearrangements were detected, possibly due to chromosomal GC bias in array analysis. Our study demonstrated MPS could be applied to accurately detect embryonic chromosomal abnormality with a flexible and cost-effective strategy and higher potential accuracy.


The Effects of a Single Versus Three Consecutive Sessions of Football Training on Postprandial Lipemia: a Randomized, Controlled Trial in Healthy, Recreationally Active Males.
Exercise frequency is important for maintaining health; however, its effects on postprandial responses remain largely unknown. Better understanding this during popular sports activities such as football may influence exercise habits. Therefore, the aim of the present study was to examine the effects of playing one single versus three consecutive days of 60-min small-sided football matches on postprandial lipemia. Fifteen males performed either one (1FOOT; n = 7) or three 60-min football (3FOOT; n = 8) sessions across an 8-day trial period. On day 1, a blood sample was collected at fasted (0 min) and 0.75, 2, 4, 6 h after a high-fat meal. Participants were then randomly allocated to the 1FOOT (day 7) or 3FOOT (days 5, 6, 7) condition. On day 8, they repeated the high-fat meal and blood sampling for 6 h following the meal. Postprandial total and incremental area under the curve (AUC, iAUC, respectively) were calculated. The postprandial triglyceride iAUC was 41% lower from pre- to post-measures for the 1FOOT (p < 0.05; ES = 1.02) and 15.7% lower for the 3FOOT (ns; ES = 0.41). Total triglyceride AUC was lower (26%) post-football matches in the 3FOOT group only (p < 0.01; ES = 1.23). In 3FOOT, insulin concentration was lower for post- compared to pre-measures at 0.75 and 2 h, respectively (p < 0.001). One single 60-min small-sided football match lowered postprandial TG incremental area under the curve while performing three consecutive days of football matches did not result in a greater attenuation. ISRCTN17934193 , registered 06 April 2019.


[Surgical treatment of bronchiectases in elderly patients].
The authors present experiences with surgical treatment of 29 patients (aged 50-64 years) with bronchiectases. Early and late results were analyzed. It was shown that complex approach to the estimation of the findings of radiography, spiral computed tomography, investigation of the external respiration function, fibrobronchoscopy and bronchoscopy, if necessary, allowed operating the patients older than 50 years with local forms of bronchiectases which gave good results.


Social Media Use for Health Purposes: Systematic Review.
Social media has been widely used for health-related purposes, especially during the COVID-19 pandemic. Previous reviews have summarized social media uses for a specific health purpose such as health interventions, health campaigns, medical education, and disease outbreak surveillance. The most recent comprehensive review of social media uses for health purposes, however, was conducted in 2013. A systematic review that covers various health purposes is needed to reveal the new usages and research gaps that emerge in recent years. This study aimed to provide a systematic review of social media uses for health purposes that have been identified in previous studies. The researchers searched for peer-reviewed journal articles published between 2006 and 2020 in 12 databases covering medicine, public health, and social science. After coding the articles in terms of publication year, journal area, country, method, social media platform, and social media use for health purposes, the researchers provided a review of social media use for health purposes identified in these articles. This study summarized 10 social media uses for various health purposes by health institutions, health researchers and practitioners, and the public. Social media can be used for various health purposes. Several new usages have emerged since 2013 including advancing health research and practice, social mobilization, and facilitating offline health-related services and events. Research gaps exist regarding advancing strategic use of social media based on audience segmentation, evaluating the impact of social media in health interventions, understanding the impact of health identity development, and addressing privacy concerns.


Au16 Cd16 (SC6 H11 )20 : A Glance at Structure-Property Relationship.
Doping Cd atom(s) into gold clusters is very promising in both theoretical study and practical applications. However, it has long been a challenge to synthesize heavily Cd-doped AuCd bimetallic clusters and thereby reveal their structure-property correlations. Herein a novel AuCd bimetallic cluster: Au16 Cd16 (SC6 H11 )20 (SC6 H11 denotes deprotonated cyclohexanethiol) with a Cd to Au atomic ratio of 1:1 is reported. The precise structure of the cluster determined by single crystal X-ray diffraction demonstrates that it has a unique hexatetrahedron Au14 core and a distinctive shell. Intriguingly, due to the special protecting motifs, the cluster exhibits high stability in various conditions studied, indicating that the geometric structure is crucial in determining the stability of the cluster. Most importantly, the photothermal property of the cluster has been investigated in comparison with those of M13 -kernel (M denotes metal atoms) clusters, and the results imply that the compactness and the Cd atom doping of the core play important roles in dictating the photothermal effect of the cluster. The authors believe that this work will provide some ideas for the rational design of clusters with high stability and excellent photothermal property.


Integrated Proteomics-Based Physical and Functional Mapping of AXL Kinase Signaling Pathways and Inhibitors Define Its Role in Cell Migration.
To better understand the signaling complexity of AXL, a member of the tumor-associated macrophage (TAM) receptor tyrosine kinase family, we created a physical and functional map of AXL signaling interactions, phosphorylation events, and target-engagement of three AXL tyrosine kinase inhibitors (TKI). We assessed AXL protein complexes using proximity-dependent biotinylation (BioID), effects of AXL TKI on global phosphoproteins using mass spectrometry, and target engagement of AXL TKI using activity-based protein profiling. BioID identifies AXL-interacting proteins that are mostly involved in cell adhesion/migration. Global phosphoproteomics show that AXL inhibition decreases phosphorylation of peptides involved in phosphatidylinositol-mediated signaling and cell adhesion/migration. Comparison of three AXL inhibitors reveals that TKI RXDX-106 inhibits pAXL, pAKT, and migration/invasion of these cells without reducing their viability, while bemcentinib exerts AXL-independent phenotypic effects on viability. Proteomic characterization of these TKIs demonstrates that they inhibit diverse targets in addition to AXL, with bemcentinib having the most off-targets. AXL and EGFR TKI cotreatment did not reverse resistance in cell line models of erlotinib resistance. However, a unique vulnerability was identified in one resistant clone, wherein combination of bemcentinib and erlotinib inhibited cell viability and signaling. We also show that AXL is overexpressed in approximately 30% to 40% of nonsmall but rarely in small cell lung cancer. Cell lines have a wide range of AXL expression, with basal activation detected rarely. Our study defines mechanisms of action of AXL in lung cancers which can be used to establish assays to measure drug targetable active AXL complexes in patient tissues and inform the strategy for targeting it's signaling as an anticancer therapy.


Automated Wormscan.
There has been a recent surge of interest in computer-aided rapid data acquisition to increase the potential throughput and reduce the labour costs of large scale Caenorhabditis elegans studies. We present Automated WormScan, a low-cost, high-throughput automated system using commercial photo scanners, which is extremely easy to implement and use, capable of scoring tens of thousands of organisms per hour with minimal operator input, and is scalable. The method does not rely on software training for image recognition, but uses the generation of difference images from sequential scans to identify moving objects. This approach results in robust identification of worms with little computational demand. We demonstrate the utility of the system by conducting toxicity, growth and fecundity assays, which demonstrate the consistency of our automated system, the quality of the data relative to manual scoring methods and congruity with previously published results.


Variability of four limb blood pressure in normal neonates.
To assess the variation in blood pressure (BP) between limbs in normal neonates. To assess whether comparison of arm and leg BP in neonates is reproducible enough to allow the difference to raise suspicion of coarctation of the aorta. Infants recruited from the postnatal wards and the postnatal murmur clinic underwent echocardiography and BP measurement in each limb using a Dinamap Compact T 482210. The method of BP measurement was guided by a telephone survey of 40 UK neonatal units. Forty healthy neonates underwent echocardiography and all had a normal aortic arch. BP was measured in 39. In three, BP in the arms was 20 mm Hg higher than in the legs. This gave a specificity of comparison of the upper and lower limb BPs of 92 (36/39) or a false positive rate of 8% (3/39). The standard deviation in BPs was 15.7 mm Hg between arms, 14.5 mm Hg between legs, and 11 mm Hg when the nearest arm and leg were compared. With current measurement techniques, normal neonates may have a wide variation in BP between limbs. A difference of 20 mm Hg in isolation is more likely to be due to random variability in measurement than to coarctation of the aorta. If coarctation of the aorta is suspected, it can only be excluded or confirmed by echocardiography.


Contrasting resting-state fMRI abnormalities from sickle and non-sickle anemia.
Sickle cell disease (SCD) is a chronic blood disorder that is often associated with acute and chronic cerebrovascular complications, including strokes and impaired cognition. Using functional resting state magnetic resonance images, we performed whole-brain analysis of the amplitude of low frequency fluctuations (ALFF), to detect areas of spontaneous blood oxygenation level dependent signal across brain regions. We compared the ALFF of 20 SCD patients to that observed in 19 healthy, age and ethnicity-matched, control subjects. Significant differences were found in several brain regions, including the insula, precuneus, anterior cingulate cortex and medial superior frontal gyrus. To identify the ALFF differences resulting from anemia alone, we also compared the ALFF of SCD patients to that observed in 12 patients having comparable hemoglobin levels but lacking sickle hemoglobin. Increased ALFF in the orbitofrontal cortex and the anterior and posterior cingulate cortex and decreased ALFF in the frontal pole, cerebellum and medial superior frontal gyrus persisted after accounting for the effect of anemia. The presence of white matter hyperintensities was associated with depressed frontal and medial superior frontal gyri activity in the SCD subjects. Decreased ALFF in the frontal lobe was correlated with decreased verbal fluency and cognitive flexibility. These findings may lead to a better understanding of the pathophysiology of SCD.


Functional Role of Internal and External Visual Imagery: Preliminary Evidences from Pilates.
The present study investigates whether a functional difference between the visualization of a sequence of movements in the perspective of the first- (internal VMI-I) or third- (external VMI-E) person exists, which might be relevant to promote learning. By using a mental chronometry experimental paradigm, we have compared the time or execution, imagination in the VMI-I perspective, and imagination in the VMI-E perspective of two kinds of Pilates exercises. The analysis was carried out in individuals with different levels of competence (expert, novice, and no-practice individuals). Our results showed that in the Expert group, in the VMI-I perspective, the imagination time was similar to the execution time, while in the VMI-E perspective, the imagination time was significantly lower than the execution time. An opposite pattern was found in the Novice group, in which the time of imagination was similar to that of execution only in the VMI-E perspective, while in the VMI-I perspective, the time of imagination was significantly lower than the time of execution. In the control group, the times of both modalities of imagination were significantly lower than the execution time for each exercise. The present data suggest that, while the VMI-I serves to train an already internalised gesture, the VMI-E perspective could be useful to learn, and then improve, the recently acquired sequence of movements. Moreover, visual imagery is not useful for individuals that lack a specific motor experience. The present data offer new insights in the application of mental training techniques, especially in field of sports. However, further investigations are needed to better understand the functional role of internal and external visual imagery.


Oxytocin system alleviates intestinal inflammation by regulating macrophages polarization in experimental colitis.
Inflammatory bowel disease (IBD) is a chronic intestinal inflammation, but the accurate etiology remains to be elucidated. Increasing evidence has shown that macrophages polarize to different phenotypes depending on the intestinal microenvironment and are associated with the progression of IBD. In the present study, we investigated the effect of oxytocin, a neuroendocrinal, and pro-health peptide, on the modulation of macrophages polarization and the progression of experimental colitis. Our data demonstrated that oxytocin decreased the sensitivity of macrophages to lipopolysaccharide stimulation with lower expression of inflammatory cytokines, like IL-1β, IL-6, and TNF-α, but increased the sensitivity to IL-4 stimulation with enhanced expression of M2-type genes, arginase I (Arg1), CD206, and chitinase-like 3 (Chil3). This bidirectional modulation was partly due to the up-regulation of β-arrestin2 and resulted in the inhibition of NF-κB signaling and reinforcement of Signal transducer and activator of transcription (STAT) 6 phosphorylation. Moreover, oxytocin receptor (OXTR) myeloid deficiency mice were more susceptible to dextran sulfate sodium (DSS) intervention compared with the wild mice. For the first time, we reveal that oxytocin-oxytocin receptor system participates in modulating the polarization of macrophages to an anti-inflammatory phenotype and alleviates experimental colitis. These findings provide new potential insights into the pathogenesis and therapy of IBD.


Identification of a gene that shortens clonal life span of Paramecium tetraurelia.
We have isolated a Paramecium tetraurelia mutant that divides slowly in daily reisolation cultures and repeats short clonal life spans after successive autogamies. Here we show, using breeding analysis, that a recessive mutation is responsible for the low fission rate and that this low rate is closely related to the short clonal life span. We conclude that a single pleiotropic gene controls these traits and have named it jumyo. In an attempt to further characterize the jumyo mutant, we have revealed that it has a culture life span similar to that of the wild-type cells and that, when mass cultured, it can divide as rapidly as wild-type cells. There was strong evidence that the mutant cells excreted into culture medium some substance that promotes their cell division. These findings may not only present supporting evidence for the hypothesis that the cellular life span is genetically programmed but also give a material basis for the study of the controlling mechanism of cell division in relation to the clonal life span.


RhoJ interacts with the GIT-PIX complex and regulates focal adhesion disassembly.
RhoJ is a Rho GTPase expressed in endothelial cells and tumour cells, which regulates cell motility, invasion, endothelial tube formation and focal adhesion numbers. This study aimed to further delineate the molecular function of RhoJ. Using timelapse microscopy RhoJ was found to regulate focal adhesion disassembly; small interfering RNA (siRNA)-mediated knockdown of RhoJ increased focal adhesion disassembly time, whereas expression of an active mutant (daRhoJ) decreased it. Furthermore, daRhoJ co-precipitated with the GIT-PIX complex, a regulator of focal adhesion disassembly. An interaction between daRhoJ and GIT1 was confirmed using yeast two-hybrid experiments, and this depended on the Spa homology domain of GIT1. GIT1, GIT2, β-PIX (also known as ARHGEF7) and RhoJ all colocalised in focal adhesions and depended on each other for their recruitment to focal adhesions. Functionally, the GIT-PIX complex regulated endothelial tube formation, with knockdown of both GIT1 and GIT2, or β-PIX phenocopying RhoJ knockdown. RhoJ-knockout mice showed reduced tumour growth and diminished tumour vessel density, identifying a role for RhoJ in mediating tumour angiogenesis. These studies give new insight into the molecular function of RhoJ in regulating cell motility and tumour vessel formation.


CRF and urocortin 3 protect the heart from hypoxia/reoxygenation-induced apoptosis in zebrafish.
Fish routinely experience environmental hypoxia and have evolved various strategies to tolerate this challenge. Given the key role of the CRF system in coordinating the response to stressors and its cardioprotective actions against ischemia in mammals, we sought to characterize the cardiac CRF system in zebrafish and its role in hypoxia tolerance. We established that all genes of the CRF system, the ligands CRFa, CRFb, urotensin 1 (UTS1), and urocortin 3 (UCN3); the two receptor subtypes (CRFR1 and CRFR2); and the binding protein (CRFBP) are expressed in the heart of zebrafish: crfr1 > crfr2 = crfbp > crfa > ucn3 > crfb > uts1 In vivo, exposure to 5% O2 saturation for 15 min and 90 min of recovery resulted in four- to five-fold increases in whole heart crfb and ucn3 mRNA levels but did not affect the gene expression of other CRF system components. In vitro, as assessed by monitoring caspase 3 activity and the number of terminal deoxynucleotidyl transferase dUTP nick-end labeling-positive cells, pretreatment of excised whole hearts with CRF or UCN3 for 30 min prevented the increase in apoptosis associated with exposure to 1% O2 saturation for 30 min with a 24-h recovery. Lastly, the addition of the nonselective CRF receptor antagonist αh-CRF(9-41) prevented the cytoprotective effects of CRF. We show that the CRF system is expressed in fish heart, is upregulated by hypoxia, and is cytoprotective. These findings identify a novel role for the CRF system in fish and a new strategy to tolerate hypoxia.


Dynamic (live) 3D roadmap as navigational tool in multiplug brain arteriovenous malformation embolization: technical note.
Treatment of brain arteriovenous malformation (bAVM) includes microsurgical excision, stereotactic radiosurgery, endovascular embolization, or combination. With bAVM embolization, complete angiographic obliteration ranges from 12.5 to 51%, and higher total occlusion rate is seen in SM grades I to III, ranging from 96 to 100%. In this paper, we illustrate the use of 3D rotational angiography and dynamic (live) 3D roadmap functions in endovascular treatment of bAVM. A single dynamic 3D roadmap or two dynamic 3D roadmaps obtained help tremendously in navigation of microcatheters and wires along the parent artery and bAVM feeders. This method eliminates the need for repeated 2D angiograms and roadmaps for new working projections every time the C-arm position is changed for cannulation of different feeders, thereby reducing radiation dose. No instances of misalignment error, vascular perforation, or thromboembolic phenomena were observed in the 21 embolization cases performed within the previous 2 years while utilizing this feature. The dynamic 3D roadmap is an extremely useful tool for multiple-feeder cannulation, by reducing the use of multiple 2D angiograms, providing intraprocedural live and adjustable 3D roadmap for better mental orientation to angioarchitecture of the bAVM, which further aids in the overall complete angiographic obliteration rate of bAVM in a single session especially in multiplug embolization technique.