Human INHBB Gene Variant (c.1079T>C:p.Met360Thr) Alters Testis Germ Cell Content, but Does Not Impact Fertility in Mice.
Testicular-derived inhibin B (α/β B dimers) acts in an endocrine manner to suppress pituitary production of follicle-stimulating hormone (FSH), by blocking the actions of activins (β A/B/β A/B dimers). Previously, we identified a homozygous genetic variant (c.1079T>C:p.Met360Thr) arising from uniparental disomy of chromosome 2 in the INHBB gene (β B-subunit of inhibin B and activin B) in a man suffering from infertility (azoospermia). In this study, we aimed to test the causality of the p.Met360Thr variant in INHBB and testis function. Here, we used CRISPR/Cas9 technology to generate InhbbM364T/M364T mice, where mouse INHBB p.Met364 corresponds with human p.Met360. Surprisingly, we found that the testes of male InhbbM364T/M364T mutant mice were significantly larger compared with those of aged-matched wildtype littermates at 12 and 24 weeks of age. This was attributed to a significant increase in Sertoli cell and round spermatid number and, consequently, seminiferous tubule area in InhbbM364T/M364T males compared to wildtype males. Despite this testis phenotype, male InhbbM364T/M364T mutant mice retained normal fertility. Serum hormone analyses, however, indicated that the InhbbM364T variant resulted in reduced circulating levels of activin B but did not affect FSH production. We also examined the effect of this p.Met360Thr and an additional INHBB variant (c.314C>T: p.Thr105Met) found in another infertile man on inhibin B and activin B in vitro biosynthesis. We found that both INHBB variants resulted in a significant disruption to activin B in vitro biosynthesis. Together, this analysis supports that INHBB variants that limit activin B production have consequences for testis composition in males.


The synthesis and structure of [Zn(TEMPO)2]2 and [Zn(μ-H)(μ(2)-η(1):η(1)-TEMPO)]6.
The commercially available radical TEMPO (2,2,6,6-tetramethylpiperidin-1-yloxy) reacts with [ZnCp*2] (1) to yield the homoleptic compound [Zn(TEMPO)2]2 (2) through coupling of two Cp* radicals. Compound 1 reacts with H2 to afford the hydride complex [Zn(μ-H)(μ(2)-η(1)-η(1)-TEMPO)]6 (3) featuring a planar Zn6H6 ring in the solid state. Preliminary data suggest that the formation of 3 proceeds via a radical mechanism.


Ab initio investigation of quantum size effects on the adsorption of CO2, CO, H2O, and H2 on transition-metal particles.
Adsorption is a crucial preliminary step for the conversion of CO2 into higher-value chemicals, nonetheless, the atomistic understanding of how substrate particle size affects this step is still incomplete. In this study, we employed density functional theory to investigate the effects of particle size on the adsorption of model molecules involved in the CO2 transformations (CO2, CO, H2O and H2) on Con, Nin and Cun particles with different sizes (n = 13, 55, 147) and on the respective close-packed surfaces. We found significant size-dependence of the adsorption properties for physisorbed (linear) and chemisorbed (bent) CO2 on the substrates and distinct (symmetric or asymmetric) stretching of the C-O bonds, which can play a crucial role to understand the CO2 dissociation pathways. For CO and H2, some properties showed small oscillations, due to size effects that induced alternation of the adsorption site preference for different particle sizes; for H2O, the adsorption properties were almost independent of particle size. The presence of low-coordinated adsorption sites resulted in a trend for stronger adsorption and greater charge transfer for smaller clusters. Fixing the size-independent factors (e.g., type of metal), our results show that CO2 adsorption on transition-metal clusters is significantly affected by particle size, suggesting that substrate particle size could be a key factor to understand and control the catalytic transformations of CO2.


Surveying wearable human assistive technology for life and safety critical applications: standards, challenges and opportunities.
In this survey a new application paradigm life and safety for critical operations and missions using wearable Wireless Body Area Networks (WBANs) technology is introduced. This paradigm has a vast scope of applications, including disaster management, worker safety in harsh environments such as roadside and building workers, mobile health monitoring, ambient assisted living and many more. It is often the case that during the critical operations and the target conditions, the existing infrastructure is either absent, damaged or overcrowded. In this context, it is envisioned that WBANs will enable the quick deployment of ad-hoc/on-the-fly communication networks to help save many lives and ensuring people's safety. However, to understand the applications more deeply and their specific characteristics and requirements, this survey presents a comprehensive study on the applications scenarios, their context and specific requirements. It explores details of the key enabling standards, existing state-of-the-art research studies, and projects to understand their limitations before realizing aforementioned applications. Application-specific challenges and issues are discussed comprehensively from various perspectives and future research and development directions are highlighted as an inspiration for new innovative solutions. To conclude, this survey opens up a good opportunity for companies and research centers to investigate old but still new problems, in the realm of wearable technologies, which are increasingly evolving and getting more and more attention recently.


Hard to diagnose and potentially fatal: slow aortic erosion post spinal fusion.
Delayed aortic injuries are a rare, but well-recognized complication of spinal surgery. They are a result of slow erosion of osteosynthesis material into the aorta. Although this is a life-threatening complication, patients might present years later with nonspecific symptoms. A complex case of slow aortic injury after thoracic spinal surgery is presented, which highlights the challenges involved in diagnosis and treatment. A 62-year-old man had a T6 vertebrectomy and T5-7 anterior spinal fusion for multiple myeloma 5 years earlier. Two years postoperatively, the patient developed intermittent hemoptysis that triggered several presentations to the emergency department and consecutive hospital admissions during a 3-year period. All investigations, including endoscopy, bronchoscopy, and repeated chest computed tomography (CT) scans, were unremarkable. Eventually, the patient presented with frank hemoptysis associated with severe left-sided chest pain. Urgent CT angiography revealed a pseudoaneurysm measuring 34 × 20 mm at the level of the vertebrectomy. The patient underwent emergency surgery and an endoluminal stent graft was successfully placed. The patient remains well after 6 months. The close proximity of the aorta and spine entertains the risk of aortic injury associated with vertebral osteosynthesis. Long-term complications of slow aortic erosion are extremely difficult to diagnose. The presented patient suffered from an undetected bronchio-aortic fistula with consecutive pseudoaneurysm formation and rupture. Awareness of slow aortic erosion is important for correct diagnostic pathways and subsequent early diagnosis to ensure a positive outcome for the patient.


Collective dynamics of microtubule-based 3D active fluids from single microtubules.
Self-organization of kinesin-driven, microtubule-based 3D active fluids relies on the collective dynamics of single microtubules. However, the connection between macroscopic fluid flows and microscopic motion of microtubules remains unclear. In this work, the motion of single microtubules was characterized by means of 2D gliding assays and compared with the flows of 3D active fluids. While the scales of the two systems differ by ∼1000×, both were driven by processive, non-processive or an equal mixture of both molecular motor proteins. To search for the dynamic correlation between both systems, the motor activities were tuned by varying temperature and ATP concentration, and the changes in both systems were compared. Motor processivity played an important role in active fluid flows but only when the fluids were nearly motionless; otherwise, flows were dominated by hydrodynamic resistance controlled by sample size. Furthermore, while the motors' thermal reaction led active fluids to flow faster with increasing temperature, such temperature dependence could be reversed by introducing temperature-varying depletants, emphasizing the potential role of the depletant in designing an active fluid's temperature response. The temperature response of active fluids was nearly immediate (⪅10 s). Such a characteristic enables active fluids to be controlled with a temperature switch. Overall, this work not only clarifies the role of temperature in active fluid activity, but also sheds light on the underlying principles of the relationship between the collective dynamics of active fluids and the dynamics of their constituent single microtubules.


Comparison of physiological and behavioral responses to chronic restraint stress between C57BL/6J and BALB/c mice.
Rodent models of chronic restraint stress (CRS) are often used as simple models of depressive disorder. However, these models of stress have been mainly developed in rats, and the behavioral phenotypes of CRS models are still controversial. In this study, we compared the physiological and behavioral responses of C57BL/6J (B6) and BALB/c mice, which are commonly used in genetic and behavioral studies, to CRS. In addition to measuring physiological parameters and the levels of corticosterone (a stress hormone) in response to stress, we also examined changes in the levels of testosterone (an anti-stress hormone), which have rarely been studied in stressed mice. The mice were exposed to CRS for 6 h a day for 21 days. In both B6 and BALB/c mice, CRS elicited several physiological stress responses, including decreased body weight gain and changes in the tissue weights of stress-related organs. Accumulated corticosterone in the hair was measured, and BALB/c mice had significantly greater levels than control mice and B6 mice after CRS. On the other hand, in the case of accumulated testosterone in the hair, both B6 mice and BALB/c mice showed significantly higher concentrations than control mice, but the degree of change was not different between the two strains. In the sucrose preference test, BALB/c mice, but not B6 mice, showed anhedonia-like behavior after CRS. However, neither strain showed depressive-like behavior in the forced swim or tail suspension test. Our results show that the physiological and behavioral stress responses of BALB/c mice are greater than those of B6 mice, although anti-stress responses to CRS are similar in both strains. This suggests that BALB/c mice are likely to be advantageous for use as a CRS-induced depression model.


[Effectiveness of cefpirome on Klebsiella pneumoniae producing beta-lactamases with an extended spectrum of activity].
In the submitted paper the author investigated the effectiveness of cefpirome on Klebsiella pneumoniae strains producing beta-lactamases with an extended spectrum (ESBL). From a total of 1348 examined strains of Klebsiella pneumoniae ESBL production was confirmed in 59 strains (4.4%). Resistance of ESBL-positive strains of Klebsiella pneumoniae to ceftazidime was 83.1%, to cefotaxime 93.2%, while resistance to cefpirome was recorded only in 45.8%.


Chronic monarticular synovitis. Diagnostic and prognostic features.
Data have been analysed from a retrospective review of 151 patients with monarthritis of more than 3 months' duration, usually involving the knee joint. The largest group, 49 patients (32%), had synovitis of unknown cause, 44 (29%) had synovitis probably associated with osteoarthrosis and 13 (9%) were diagnosable at presentation as having rheumatoid arthritis according to American Rheumatism Association (1959) criteria, which include serological and histological findings. There was only one case of tuberculous synovitis. Twelve of the thirteen patients diagnosed as rheumatoid arthritis developed involvement in other joints. In most other conditions, however, including synovitis of uknown cause, the prognosis was favourable, with either improvement or complete remission.


Physiologic variations in dural venous sinus flow on phase-contrast MR imaging.
Our study quantifies normal physiologic variations of dural sinus flow using phase-contrast MR imaging. Fifteen volunteers were imaged using nontriggered and triggered phase-contrast MR venography of the superior sagittal and transverse sinuses. Triggered scans were obtained during regular breathing; nontriggered scans were obtained during regular breathing, breath-holding, deep inspiratory breath-holding, and deep expiratory breath-holding. Analysis of variance, Bonferroni method, and Dunn post hoc analysis were used to determine any significant differences in the mean flow and velocity between the different breathing maneuvers. A paired t test was used to compare flow between sinuses during regular breathing. Deep inspiratory breath-holding and deep expiratory breath-holding resulted in a significant decrease in blood flow and velocity in all dural sinuses compared with regular breathing. During deep inspiratory breath-holding, blood flow decreased 30.8% in the superior sagittal sinus, 19.7% in the left transverse sinus, and 19.1% in the right transverse sinus. Similarly, during deep expiratory breath-holding, blood flow decreased 30.2% in the superior sagittal sinus, 20.8% in the left transverse sinus, and 20.3% in the right transverse sinus. The sum of the flow in the transverse sinuses was significantly greater than in the sagittal sinus. Normal pulsatility of dural sinus blood velocity was also characterized for all measured sinuses. Characterization of variations in dural sinus velocity and flow as a function of the cardiac cycle and breathing maneuvers, using phase-contrast MR imaging, may help separate physiologic from pathologic changes of flow resulting from conditions that influence the cerebrovascular circulation.


Surgical Considerations in Total Anomalous Pulmonary Venous Connection.
Total anomalous pulmonary venous connection (TAPVC) is predominantly a neonatal congenital cardiac diagnosis in which the pulmonary veins fail to return to the left atrium. The presence or absence of obstruction defines the clinical presentation, and in the setting of obstructed pulmonary veins, this diagnosis represents a true surgical emergency. Improvements in perioperative care and refinement of surgical techniques continue to allow for improved survival and decreased morbidity. A description of the surgical anatomy and key concepts in the perioperative and surgical management of TAPVC are presented in this review.


Evaluation of skin cancer screening in Germany - Spatiotemporal associations between skin cancer screening and skin cancer mortality based on ambulatory claims data.
There are regional differences in skin cancer screening uptake in Germany. So far, it is unclear whether a high uptake of screening services leads to a reduction in mortality. This article presents study results on the investigation of spatiotemporal associations between skin cancer screening and mortality. The methods used are discussed regarding their suitability. The basis is ambulatory claims data on the utilization of early skin cancer detection as well as data on skin cancer mortality from the cause-of-death statistics of the years 2011-2015 at county level in Germany. In addition to a descriptive evaluation, spatiotemporal cluster analyses and regression models were used to investigate the relationship between the uptake of early detection and mortality. In addition to age, adjustments were also made for other selected socio-economic and socio-graphical variables. The descriptive results show striking spatial patterns of skin cancer screening and mortality. Cluster analyses identified regions with significantly higher and lower cases of early detection and skin cancer mortality. The spatiotemporal regression analyses show no clear association. Only early detection by a dermatologist, adjusted for age, shows an association with mortality. No clear association between early skin cancer detection and mortality can be derived from the results. However, the study design used with a spatiotemporal cluster and regression analysis has shown that these methods allow in-depth statements about the relationship between early skin cancer detection and mortality.


A role for P2X7 in microglial proliferation.
Microglia, glial cells with an immunocompetent role in the CNS, react to stimuli from the surrounding environment with alterations of their phenotypic response. Amongst other activating signals, the endotoxin lipopolysaccharide (LPS) is widely used as a tool to mimic bacterial infection in the CNS. LPS-activated microglia undergo dramatic changes in cell morphology/activity; in particular, they stop proliferating and differentiate from resting to effector cells. Activated microglia also show modifications of purinoreceptor signalling with a significant decrease in P2X(7) expression. In this study, we demonstrate that the down-regulation of the P2X(7) receptor in activated microglia may play an important role in the antiproliferative effect of LPS. Indeed, chronic blockade of the P2X(7) receptor by antagonists (oxidized ATP, KN62 and Brilliant Blue G), or treatment with the ATP-hydrolase apyrase, severely decreases microglial proliferation, down-regulation of P2X(7) receptor expression by small RNA interference (siRNA) decreases cell proliferation, and the proliferation of P2X(7)-deficient N9 clones and primary microglia, in which P2X(7) expression is down-regulated by siRNA, is unaffected by either LPS or P2X(7) antagonists. Furthermore, flow cytometric analysis indicates that exposure to oxidized ATP or treatment with LPS reversibly decreases cell cycle progression, without increasing the percentage of apoptotic cells. Overall, our data show that the P2X(7) receptor plays an important role in controlling microglial proliferation by supporting cell cycle progression.


Carbachol-induced changes in excitability and [Ca2+]i signalling in projection cells of medial entorhinal cortex layers II and III.
The entorhinal cortex (EC) is a major gateway for sensory information into the hippocampus and receives a cholinergic input from the forebrain. Therefore, we studied muscarinic effects on excitability and intracellular Ca2+ signalling in layer II stellate and layer III pyramidal projection neurons of the EC. In both classes of neurons, local pressure-pulse application of carbachol (1 mM) caused small, atropine-sensitive membrane depolarizations that were not accompanied by any detectable changes in [Ca2+]i. At a higher concentration (10 mM), carbachol induced a larger membrane depolarization associated with synaptic oscillations and epileptiform activity in both classes of neurons. In contrast to the intrinsic theta rhythm in stellate cells with one dominant peak frequency at approximately 7 Hz, the synaptically mediated oscillation induced by carbachol showed three characteristic peaks in the theta and gamma frequency range at approximately 11, 23 and 40 Hz. Although carbachol-induced epileptiform activity was associated with increases in intracellular free Ca2+ in both layer II and III cells, the observed [Ca2+]i accumulation was significantly larger in layer III than in layer II cells. Responses to intracellular current injections showed differences in Ca2+ accumulation in layer II and III cells at the same membrane potentials, suggesting a dominant expression of low- and high-voltage-activated Ca2+ channels in these layer II and III cells, respectively. In conclusion, we present evidence for significant differences in the [Ca2+]i regulation between layer II stellate and layer III pyramidal cells of the medial EC.


Association Between Nonselective Beta-Blocker Use and Hepatocellular Carcinoma in Patients With Chronic Hepatitis B Without Cirrhosis and Decompensation.
Background: Nonselective beta-blockers (NSBBs) can reduce the incidence or mortality of certain types of cancers, and NSBBs exert a protective effect on hepatocellular carcinoma (HCC) in patients with cirrhosis. However, the potential preventive effect of NSBBs has not yet been investigated in patients with chronic hepatitis B (CHB) who have a high HCC risk regardless of the presence of underlying cirrhosis. Aim: This study evaluated the association between NSBB use and HCC incidence in patients with CHB without cirrhosis and decompensation. Methods: From the 2000 Longitudinal Generation Tracking Database, we enrolled patients who were newly diagnosed as having CHB from January 2001 to December 2011 and then followed them up for at least 5 years. To estimate the causal effect of NSBBs on the time-to-event outcomes of HCC, a marginal Cox proportional hazards model was used to calculate hazard ratios (HRs) and 95% confidence intervals (CIs). Results: After adjustment, no significant benefit of HCC risk reduction was observed between the NSBB users and nonusers (adjusted HR, 0.82; 95% CI, 0.52-1.31). The cumulative defined daily dose (cDDD) analysis revealed no significant dose correlation among the three groups [adjusted HR (95% CI): 1.08, (0.56-2.05), 0.54 (0.17-1.77), and 0.76 (0.40-1.42) in the <90 cDDD, 90 to <180 cDDD, and ≥180 cDDD groups, respectively]. Duration-dependent associations were not observed. Multivariable stratified analysis results demonstrated that HCC risk markedly decreased in the patients aged >55 years (adjusted HR, 0.49; 95% CI, 0.25-0.96; p = 0.04). Conclusion: NSBB did not significantly prevent HCC in the patients with CHB infection without cirrhosis and decompensation. This study provided one of valuable results that it is not clinically required to use NSBBs as recommended chemoprevention for HCC in high-risk patients who have CHB.


Hypothyroidism modulates beta adrenergic receptor adenylate cyclase interactions in rat reticulocytes.
We have investigated alterations in beta adrenergic receptor binding sites of rat reticulocytes occurring in animals rendered hypothyroid by thyroidectomy. Beta adrenergic receptor interactions were assessed by measuring the number of (-)[3H]-dihydroalprenolol binding sites and the ability of an agonist to compete for occupancy of the receptors. The number of receptors was significantly reduced in cells from the hypothyroid animals. In addition, there were significant agonist-specific alterations in binding. Using computer assisted curve fitting techniques, it was found that the ability of (-)isoproterenol to stabilize a high affinity guanine nucleotide sensitive "coupled" form of the receptor was impaired. Reticulocytes from hypothyroid animals have, in addition, a reduction in the concentration of the nucleotide regulatory protein as assessed by the number of 42,000 Mr substrates for cholera toxin catalyzed ADP ribosylation. These alterations are associated with reductions in catecholamine and NaF stimulated adenylate cyclase activity. Diminished coupling of beta adrenergic receptors with other regulatory components of the adenylate cyclase system represents a mechanism by which altered thyroid states modulate beta adrenergic receptor function and beta adrenergic responsiveness of tissues.


Histories of childhood trauma and the effects on risky HIV behaviors in a sample of women drug users.
A cross-sectional study was conducted to examine the association between childhood trauma and current human immunodeficiency virus (HIV) risk behaviors of 181 active illicit-drug-using women in San Antonio, Texas. We found very few statistically significant associations, which could he explained by (a) childhood trauma subtypes not being mutually exclusive, (b) clustering of mild-to-severe forms of abuse. and (c) childhood trauma having an indirect, rather than direct, effect on HIV risk behavior. Public health implications from this study are that prevention programs need to consider past and current individual and environmental factors that influence HIV sexual risk behaviors in women drug users.


Control of follicular development during and after lactation in sows.
Follicular development during early lactation in the sow is characterized by a large population of small-sized follicles and a small population of medium-sized follicles. As lactation progresses there is a gradual shift in number of follicles into medium- or large-sized categories and the percentage of follicles classified as atretic declines. Weaning at birth often leads to aberrant follicular development, apparently because the positive feedback response of LH to oestradiol does not occur during the first week post partum. Secretion of LH during lactation is primarily controlled by suckling intensity of the litter while FSH is controlled by a nonsteroidal ovarian factor, presumably inhibin. Suckling apparently limits secretion of GnRH and weaning leads to an increase of GnRH within the hypothalamus coincident with an increase of LH in the anterior pituitary and plasma and increased follicular growth. Follicular development during both lactation and post-weaning anoestrus can be stimulated by exogenous gonadotrophins and pulsatile administration of GnRH. Factors such as nutrition, season, boar exposure, litter size and split weaning affect follicular development during lactation and after weaning, probably because they affect secretion of LH.


Exploiting the "Hot-Spots" of Hsp70-Bim Protein-Protein Interaction to Optimize the 1-Oxo-1H-phenalene-2,3-dicarbonitrile Analogues as Specific Hsp70-Bim Inhibitors.
Selectively targeting the cancer-specific protein-protein interaction (PPI) between Hsp70 and Bim has been discovered as a promising strategy for treating chronic myeloid leukemia (CML). The first Hsp70-Bim PPI inhibitor, S1g-2, has been identified to overcome the on-target toxicity of known Hsp70 inhibitors when it induces apoptosis of CML cells. Herein, we carried out a hit-to-lead optimization of S1g-2, yielding S1g-10, which exhibited a 10-fold increase in Hsp70/Bim suppressing potency. Furthermore, S1g-10 not only exhibited a 5- to 10-fold stronger antitumor activity in the sub-μM range against CML cells than S1g-2 in vitro, but it also overcame BCR-ABL-independent tyrosine kinase inhibitor resistance in CML in vivo depending on the Hsp70-Bim signaling pathway. Moreover, through structure-activity relationship analysis, TROSY-HSQC NMR, molecular dynamics simulation, and point mutation validation, two hydrophobic pockets composed of eight key residues were demonstrated to produce predominant interactions with either Bim or S1g-10, regarded as the "hot-spots" in the Hsp70-Bim PPI interface.


Complement mediates attenuation of endothelium-dependent relaxations in canine coronary arteries after porcine serum exposure: a mechanism for vascular thrombosis in xenograft hyperacute rejection.
Evidence for complement activation in xenograft hyperacute rejection includes prolongation of graft survival after complement inactivation as well as component deposition and consumption during hyperacute rejection. The current investigations examined the endothelial production of vasoactive substances during heterologous serum exposure. Segments of canine coronary artery were exposed to either autologous canine serum for 90 minutes or heterologous porcine serum for 30, 60, or 90 minutes. After replacement of the serum with buffered saline, segments were contracted with phenylephrine (10(-6) mol/L) in the presence of indomethacin (10(-5) mol/L). Compared with responses of vessels exposed to autologous canine serum, receptor-dependent relaxation to acetylcholine was impaired in arteries after 60 or 90 minutes of exposure to porcine serum. Receptor-independent relaxation to calcium ionophore A23187 was not significantly impaired at any length of porcine serum exposure. Endothelial-independent relaxation to sodium nitroprusside was not impaired with either canine or porcine serum exposure. Oxyhemoglobin (10(-6) mol/L) abolished acetylcholine-mediated relaxation, indicating that nitric oxide was the predominant mediator of the impaired pathway. Basal release of nitric oxide after a 60-minute porcine serum exposure was reduced by half compared with coronary arteries exposed to autologous canine serum. Serum pretreated by either heat inactivation of complement or immunoadsorption with anti-C3 antibodies failed to depress endothelial-dependent relaxation on 60 minutes of exposure to canine coronary arteries. Scanning electron microscopy revealed an intact endothelial layer in coronary arteries exposed to either porcine or canine serum for 60 minutes. Hyperacute xenograft rejection impairs receptor-dependent relaxation of canine coronary arteries at 60 and 90 minutes. These data strongly suggest that impairment of endothelial production of nitric oxide during acute xenograft rejection is mediated by complement activation.


The expression of transforming growth factor-beta by cultured chick growth plate chondrocytes: differential regulation by 1,25-dihydroxyvitamin D3.
1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) and transforming growth factor-beta (TGF-beta) are both important regulators of chondrocyte growth and differentiation. We report here that 1,25(OH)2D3 differentially regulates the expression of the genes for TGF-beta 1 to -beta 3 and the secretion of the corresponding proteins in cultured chick chondrocytes. Confluent growth plate chondrocytes were serum-deprived and cultured in varying concentrations of 1,25(OH)2D3. Cells were assayed for TGF-beta mRNA and conditioned medium was assayed for TGF-beta activity and isoform composition. Active TGF-beta was only detected in 10(-8) M 1,25(OH)2D3-treated cultures (8.37 ng active TGF-beta/mg protein). There was a significant decrease in total (latent-active) TGF-beta activity in conditioned medium of 10(-12) M (23.4%; P < 0.05) and 10(-10) M (20.7%; P < 0.05) 1,25(OH)2D3-treated cultures but 10(-8) M 1,25(OH)2D3 significantly increased (30.9%; P < 0.01) TGF-beta activity. The amounts of TGF-beta 1, -beta 2 and -beta 3 isoforms produced were similar in control, 10(-10) or 10(-12) M 1,25(OH)2D3-treated cultures but the conditioned medium of 10(-8) M 1,25(OH)2D3-treated cultures contained significantly higher amounts of all three isoforms. Quantification of TGF-beta mRNA demonstrated differential control of TGF-beta gene expression with TGF-beta 1 and -beta 3 mRNA levels reduced by all concentrations of 1,25(OH)2D3 examined (10(-8), 10(-10) and 10(-12) M) whilst TGF-beta 2 mRNA concentrations were elevated. Our results indicated that 1,25(OH)2D3 regulates chick growth plate chondrocyte TGF-beta secretion and mRNA expression in a concentration-dependent and isoform-specific manner. This interaction may be important in the regulation of chondrocyte metabolism and endochondral bone growth.


Jeune's asphyxiating thoracic dystrophy of the newborn.
A 13-month-old patient with Jeune's thoracic asphyxiating dystrophy, was surgically treated using a methyl-methacrylate (acrylic) prosthesis. Although postoperative recovery was fast, the patient died of respiratory distress two months following surgery. Limitations of currently available surgical techniques and the need for long-term results will be discussed.


Meta-analysis of depressive symptoms in dual-diagnosis schizophrenia.
Substance abuse is highly prevalent in schizophrenia and associated with numerous negative consequences. While studies have regularly reported more severe depressive symptoms in addicted schizophrenia patients relative to non-abusing patients, some studies have not corroborated this finding. The current meta-analysis was performed to quantify the relative severity of depressive symptoms in dual-diagnosis schizophrenia. A search of the literature using computerized engines was undertaken. Studies were retained in the analysis if (i) they assessed depressive symptoms using validated scales specific to depression (e.g. Hamilton Depression Rating Scale); and (ii) groups of schizophrenia patients were divided according to substance use disorders (alcohol, amphetamines, cannabis, cocaine, hallucinogens, heroin and/or phencyclidine). According to the inclusion criteria, 20 studies were available for mathematical analysis. A small, positive and significant effect size estimate (n =3283; 1680 dual diagnosis; 1603 single diagnosis; adjusted Hedges's g =0.292; p =0.003) was obtained, within a random-effect model, suggesting that some dual-diagnosis patients experience more severe depressive symptoms than single-diagnosis patients. This significant difference was found only for studies using the Hamilton Depression Rating Scale but not for other depression scales. The results of the present meta-analysis suggest that addicted schizophrenia patients experience more severe depressive symptoms compared to non-abusing patients, but that the difference is smaller than commonly assumed. The meta-analysis also shows that the significance of results is related to the scale used to measure depressive symptoms. These results have methodological implications for future studies of depressive symptoms in dual-diagnosis patients, and potential implications for the prevention and treatment of depressive symptoms in schizophrenia.


Visual observations of the sea floor subduction line in the middle-america trench.
Four dives were made to the floor of the Middle-America Trench with the U.S. Navy's deep research submersible DSV Turtle. The area investigated is located between Costa Rica and the Cocos Ridge where the depth of the trench floor does not exceed the 2000-meter capability of the submersible. At the axis of the trench floor a series of steep northeast-facing scarps 10 to 20 meters high lie parallel to the trench axis. Here oceanic crust appears to have been carried down by near-vertical normal vaults of small displacement. Between these small scarps and the landward wall of the trench a narrow line of recent deformation interrupts a smooth apron. Unconsolidated sediments are thrust in sharply serrated piles and cut by sharp-edged chasms. This line of deformation is interpreted as the present sea floor trace of crustal subduction.


Contemporary unconventional clinical use of co-trimoxazole.
In the late 1960s, the combination of trimethoprim and sulphamethoxazole (co-trimoxazole) was introduced into clinical practice and used to treat many infectious diseases, such as urinary tract infections, respiratory infections, sexually transmitted diseases, Gram-negative sepsis, enteric infections and typhoid fever. Subsequently, co-trimoxazole was reported to be effective against numerous bacterial, fungal and protozoal pathogens, including Nocardia, Listeria monocytogenes, Brucella, Stenotrophomonas maltophilia, Burkholderia, Coxiella burnetii, Tropheryma whipplei, atypical mycobacteria, and Pneumocystis jirovecii. Among protozoal infections, in addition to toxoplasmosis, co-trimoxazole has been used to treat susceptible Plasmodium falciparum, Cyclospora and Isospora infections. Several retrospective and prospective studies have demonstrated good clinical outcome with co-trimoxazole in treating invasive methicillin-resistant Staphylococcus aureus infections. We summarize herein the accumulated evidence in the literature on the new, 'unconventional' clinical use of co-trimoxazole during the last three decades. In the era of widespread antibiotic resistance and shortage of new antibiotic options, large-scale, well-designed studies are needed to explore the tremendous potential concealed in this well-established drug.


Impact of psychiatric disorders on the HIV epidemic.
Mental illness continues to fuel the HIV epidemic. There is a high prevalence of mental disorders in the HIV-infected population and a high prevalence of HIV infection in the mentally ill. Without effective treatment of mental disorders, HIV treatment outcome is poor, and transmission of disease continues. High frequencies of mental illness are found in corrections facilities, among the homeless, among injection drug users, and in patients attending sexually transmitted disease clinics. Such populations must be targeted for effective mental health treatment if overall outcomes of HIV treatment are to be improved and transmission of HIV is to be reduced. This article summarizes a presentation on mental illness and the HIV epidemic made by Andrew F. Angelino, MD, at the 10th Annual Ryan White HIV/AIDS Program Clinical Update in June 2007.


Touchscreen-based cognitive tasks reveal age-related impairment in a primate aging model, the grey mouse lemur (Microcebus murinus).
Mouse lemurs are suggested to represent promising novel non-human primate models for aging research. However, standardized and cross-taxa cognitive testing methods are still lacking. Touchscreen-based testing procedures have proven high stimulus control and reliability in humans and rodents. The aim of this study was to adapt these procedures to mouse lemurs, thereby exploring the effect of age. We measured appetitive learning and cognitive flexibility of two age groups by applying pairwise visual discrimination (PD) and reversal learning (PDR) tasks. On average, mouse lemurs needed 24 days of training before starting with the PD task. Individual performances in PD and PDR tasks correlate significantly, suggesting that individual learning performance is unrelated to the respective task. Compared to the young, aged mouse lemurs showed impairments in both PD and PDR tasks. They needed significantly more trials to reach the task criteria. A much higher inter-individual variation in old than in young adults was revealed. Furthermore, in the PDR task, we found a significantly higher perseverance in aged compared to young adults, indicating an age-related deficit in cognitive flexibility. This study presents the first touchscreen-based data on the cognitive skills and age-related dysfunction in mouse lemurs and provides a unique basis to study mechanisms of inter-individual variation. It furthermore opens exciting perspectives for comparative approaches in aging, personality, and evolutionary research.


A giraffe neck sign of the medial meniscus: A characteristic finding of the medial meniscus posterior root tear on magnetic resonance imaging.
The posterior root ligament of the medial meniscus (MM) has a critical role in regulating the MM movement. An accurate diagnosis of the MM posterior root tear (MMPRT) using magnetic resonance imaging (MRI) is important for preventing sequential osteoarthritis following the MMPRT. However, diagnosis of the MMPRT is relatively difficult even after using several characteristic MRI findings. The aim of this study was to identify a useful meniscal body sign of the MMPRT for improving diagnostic MRI reading. Eighty-five patients who underwent surgical treatments for the MMPRT (39 knees) and other types of MM tears (49 knees) were included. The presence of characteristic MRI findings such as cleft sign, ghost sign, radial tear sign, medial extrusion sign, and new meniscal body shape-oriented "giraffe neck sign" was evaluated in 120 MRI examinations. Giraffe neck signs were observed in 81.7% of the MMPRTs and in 3.3% of other MM tears. Cleft, ghost, and radial tear signs were highly positive in the MMPRTs compared with other MM tears. Medial extrusion signs were frequently observed in both groups. Coexistence rates of any 2 MRI signs, except for medial extrusion sign, were 91.7% in the MMPRT group and 5% in other MM tears. This study demonstrated that a new characteristic MRI finding "giraffe neck sign" was observed in 81.7% of the MMPRT. Our results suggest that the combination of giraffe neck, cleft, ghost, and radial tear signs may be important for an accurate diagnostic MRI reading of the MMPRT.


Essential Oils and Their Constituents: An Alternative Source for Novel Antidepressants.
Depression is a disease that has affected a high proportion of the world's population and people of different ages, incapacitating them from good performance at work and in social relationships, and causing emotional disorders to millions of families. Therefore, the search for new therapeutic agents is considered a priority for the discovery of more effective forms of treatment. In this review, studies of essential oils and their constituents in experimental models related to depression are discussed. The mechanisms of action of the oils and the presence of psychoactive constituents in their chemical compositions are discussed. The data in the review show the therapeutic potential of essential oils and their chemical constituents for use in depressive disorders. Advanced studies using humans are needed to confirm the antidepressant properties described in animals.


The effect of lipophilicity and dose on the frequency of statin-associated muscle symptoms: A systematic review and meta-analysis.
Addressing the factors which lead to the development of statin-associated muscle symptoms (SAMS) is vital for maintaining patient compliance with these pharmaceuticals, and thus improving patient outcomes. This study aimed to clarify the relationship between statin lipophilicity, or dose, and the frequency of adverse muscle symptoms using a systematic review of randomised controlled trials (RCTs). RCTs, including statin monotherapy and placebo groups, which reported data on muscle adverse events were identified through the PubMed and Scopus databases. Risk ratios (RRs) and 95% confidence intervals (CI) were pooled using a random-effects meta-analysis. A total of 135 RCTs were included in this review. Statin therapy was associated with a significant, but modest, increase in the risk of adverse muscle symptoms compared to placebo (RR=1.050; 95% CI=1.014-1.089; P=0.007; I2=3.291%). This significant association was primarily due to the inclusion of RCTs recruiting participants with a history of statin intolerance. Lipophilic statins had no appreciable impact on the development of SAMS compared to hydrophilic formulations. A univariate meta-regression of dose (standardised to atorvastatin dose equivalents) and the risk of musculoskeletal complaints also showed no significant association. The results obtained from this meta-analysis indicate that there is a slight increase in the risk of SAMS, especially in individuals with a history of statin intolerance. There is limited evidence to suggest that the risk of SAMS would differ between the use of lipophilic and hydrophilic statins, or high- and low-dose therapy.


Dixdc1 is a critical regulator of DISC1 and embryonic cortical development.
The psychiatric illness risk gene Disrupted in Schizophrenia-1 (DISC1) plays an important role in brain development; however, it is unclear how DISC1 is regulated during cortical development. Here, we report that DISC1 is regulated during embryonic neural progenitor proliferation and neuronal migration through an interaction with DIX domain containing-1 (Dixdc1), the third mammalian gene discovered to contain a Disheveled-Axin (DIX) domain. We determined that Dixdc1 functionally interacts with DISC1 to regulate neural progenitor proliferation by co-modulating Wnt-GSK3beta/beta-catenin signaling. However, DISC1 and Dixdc1 do not regulate migration via this pathway. During neuronal migration, we discovered that phosphorylation of Dixdc1 by cyclin-dependent kinase 5 (Cdk5) facilitates its interaction with the DISC1-binding partner Ndel1. Furthermore, Dixdc1 phosphorylation and its interaction with DISC1/Ndel1 in vivo is required for neuronal migration. Together, these data reveal that Dixdc1 integrates DISC1 into Wnt-GSK3beta/beta-catenin-dependent and -independent signaling pathways during cortical development and further delineate how DISC1 contributes to neuropsychiatric disorders.


The diurnal transcriptome reveals reprogramming of lung adenocarcinoma cells under a time-restricted feeding mimicking regimen.
The processes of tumor growth and circadian rhythm are intimately intertwined; thus, rewiring circadian metabolism by time-restricted feeding (TRF) may contribute to delaying carcinogenesis. However, research on the effect of a TRF cellular regimen on cancer is lacking. Investigate the circadian signatures of TRF in lung cancer in vitro. We first developed a cellular paradigm mimicking in vivo TRF and collected cells for transcriptome analysis. We further confirmed the effect on tumor cells upon 6 h TRF-mimicking (6 h TRFM) by real-time PCR, Lumicycle experiments, CCK-8 and flow cytometry assays. We found that A549 lung adenocarcinoma cells treated with 6 h TRFM conditions displayed robust diurnal rhythms of transcriptomes, as well as modulation of the core clock genes relative to other different cellular regimens used in this study, including the fasting-mimicking conditions (i.e., short-term starvation, STS) and the serum-free regime. Notably, pathway analysis of oscillating genes exclusively in 6 h TRFM showed that some circadian genes were enriched in tumor-related pathways, such as the oxytocin signaling pathway, HIF-1 signaling pathway and pentose and glucuronate interconversions. Moreover, in line with the circadian pathway enrichment results, 6 h TRFM robustly inhibited cell proliferation and induced cell apoptosis and cell cycle arrest in lung adenocarcinoma A549 cells, lung adenocarcinoma H460 cells, esophageal carcinoma Eca-109 cells and breast adenocarcinoma MCF-7 cells. Our findings provide the first in vitro mimicking medium for TRF intervention and indicate that 6 h TRFM is sufficient to reprogram the circadian signatures of lung adenocarcinoma cells and inhibit the progression of multiple tumors.


Transseptal puncture for radiofrequency catheter ablations of left-sided arrhythmias in a paediatric population.
Puncturing the atrial septum is frequently used in adults. In children, the transseptal puncture is less common, technically more demanding, and the rate of complications is not well described. We studied the feasibility and safety of this procedure in a retrospective analysis of 157 consecutive children undergoing transseptal puncture for radiofrequency catheter ablation of left atrial targets in two tertiary-care centres between 2005 and 2013. The median age of the patients at intervention was 12.5 years (1.1-18 years), with median weight of 42 kg (range 9.0-97.0 kg). Pre-excitation was found in 102 procedures, accessory pathway with exclusively retrograde conduction in 41, focal atrial tachycardia in nine, left-sided permanent junctional/reciprocating tachycardia-like accessory pathways in three, and atypical atrioventricular nodal re-entry tachycardia in two. All the procedures were guided by fluoroscopy. Additional imaging by transoesophageal echocardiography was used in three patients. Successful transseptal puncture was possible in 99.4% of the cases, ablation in 97.4%. The median time, including mapping and radiofrequency ablation, was 120 minutes (range 60-450), the median fluoroscopy time 10.8 minutes (range 1.8-75), and the median radiation dose 3 Gy cm2 (range 0.3-35). In total, five patients (3.2%) had a recurrent arrhythmia during the observation period of a median of 40 months (range 1-103). No complications associated with the transseptal puncture were observed. Transseptal puncture is a feasible and safe procedure in children. This access allows successful and efficient radiofrequency ablation of arrhythmia of the left atrium in the vast majority of the patients and might be considered as the first-line approach in this population.


Vaccination during pregnancy: Canadian maternity care providers' opinions and practices.
A number of countries have implemented vaccination in pregnancy as a strategy to reduce the burden of influenza and pertussis. The aim of this study was to assess the involvement of Canadian maternity care providers in administration of vaccines to their pregnant patients. A cross-sectional web-based survey was sent to family physicians, obstetricians-gynecologists, midwives, pharmacists, and nurses. A multivariable logistic regression model was used to determine variables independently associated with offering vaccination services in pregnancy in providers' practice. A total of 1,135 participants participated. Overall, 64% (n = 724) of the participants reported offering vaccines in their practice and 56% (n = 632) reported offering vaccines to pregnant patients. The main reasons reported for not offering vaccination services in pregnancy were the belief that vaccination was outside of the scope of practice; logistical issues around access to vaccines; or lack of staff to administer vaccines. In multivariable analysis, the main factors associated with vaccination of pregnant patients in practices where vaccination services were offered were: providers' confidence in counseling pregnant patients about vaccines, seeing fewer than 11 pregnant patients on average each week, and being a nurse or a family physician. Although the majority of participants expressed strong support for vaccination during pregnancy, half were not offering vaccination services in their practice. Many were not equipped to offer vaccines in their practice or felt that it was not their role to do so. To enhance vaccine acceptance and uptake in pregnancy, it will be important to address the logistical barriers identified in this study.


Lung cancer associated with cystic airspaces: Characteristic morphological features on CT in a series of 11 cases.
To familiarize the reader with the entity 'lung cancer associated with cystic airspaces' (LC-CAS) and create an awareness about the potential for slow progressive development of cancer within these nonaggressive appearing cystic airspaces (CAS) encountered in routine radiology practice. Morphological appearances of (n = 11) LC-CAS detected during routine radiological reporting of chest CT scans were studied. Patient demographics, clinical history, characteristics of LC-CAS including location, size, wall thickening, diffuse nodularity, eccentric nodule, ground glass change, emphysema and pathology results were collected from the hospital's internal database. Patients with LC-CAS (9F/2M) were between 49 and 77 years (mean 63.18 years). All patients (n = 11) had a history of smoking. LC-CAS had a characteristic multicystic bubbly appearance. Average size of CAS at initial detection of LC was 2.52 cm (range 1.3-4 cm). Lesions were located in the RLL (n = 4), RML (n = 2), RUL (n = 1), LLL (n = 1) and LUL (n = 3) with no lobar predilection and were more commonly peripheral (n = 7) than central (n = 4). Ground glass change (n = 2), extrinsic nodules (n = 4), diffuse wall nodularity (n = 3) and intrinsic nodules (n = 2) were observed and prompted biopsy. Lesions ranged between T1a to T4. Most cancers were T1a N0 (n = 5). Adenocarcinomas formed the majority of cases (n = 9). LC-CAS present as new development of diffuse nodularity, eccentric nodules or ground glass change associated with CAS. These are more commonly adenocarcinomas on histology. Recognition of CAS and appropriate malignancy workup when suspicious features are observed is essential to enable early detection of lung cancer.


Self-reported changes in the professional singing voice after surgical intervention treatment for breast cancer: a survey pilot study of female professional singers.
The effects of breast cancer surgical treatment on the professional singing voice are unknown. The purpose of this study was to discover whether there are self-perceived changes in the quality and/or process of singing experienced by professional female singers who have undergone surgical intervention for the treatment of diagnosed breast cancer-including any changes perceived from the use of radiation, chemotherapy, and other drug treatments related to those surgeries. A voluntary subject pool comprised female professional singers who have undergone surgery for breast cancer was recruited from professional singing networks. Participants underwent evaluation through an anonymous online survey, psychometrically vetted for content and instrument reliability/validity before administration. Valid participants (N=56) responded to 45 questions regarding surgical procedures, related therapies, and self-perceived vocal effects. Analysis of results produced a preliminary description of types of voice change, duration of changes, and qualitative self-perceptions. This initial report reveals that there are self-perceived singing voice changes experienced by professional singers treated for breast cancer. However, additional research is needed to determine the degree of vocal impact perceived to be attributable to individual surgical interventions and related therapies.


Structural and luminescent properties of Eu2+ and Nd3+-doped mixed alkaline earth aluminates prepared by the sol-gel method.
Alkaline earth aluminates with the overall nominal compositions Mg0.5Sr0.5Al2O4 (MSA), Ca0.5Mg0.5Al2O4 (CMA) and Ca0.5Sr0.5Al2O4 (CSA) doped with 0.5 mol% of Eu2+ and 0.25 mol% of Nd3+ ions were obtained by a modified aqueous sol-gel method and annealed in a reducing atmosphere at 900, 1000, 1100 and 1300 °C. The sample structures were investigated by XRD. Solid solubility was only confirmed for the CSA samples. UV-excited luminescence was observed in the blue region (λ = 440 nm) in the samples of CMA containing the monoclinic CaAl2O4 phase and in the green region (λ = 512 nm) in the samples of MSA containing hexagonal or monoclinic phases of SrAl2O4. The CSA samples, besides the blue region, exhibited an extended shoulder in the green region, which proved the existence of some pure strontium phases. Co-doped Nd3+ ions did not affect the wavelength of the emitted light, but the persistent luminescence at room temperature was greatly extended with respect to the aluminates doped with Eu2+ ions only.


Western Spruce Budworm Effects on Forest Resilience.
Western spruce budworm (Choristoneura freemani Razowski) is the most destructive defoliator of forests in the western US. Forests in northern New Mexico experienced high levels of WSBW-caused defoliation and subsequent mortality between the 1980s and 2010s. The effects of severe western spruce budworm outbreaks on stand dynamics in the US Southwest are still relatively unknown, but understanding the impacts is important to the management and resilience of these forests. To begin addressing this knowledge gap, we conducted a study along two gradients: an elevational gradient from mixed-conifer to spruce-fir forests and a gradient of WSBW-caused defoliation intensity. We recorded overstory and understory stand conditions (size structure, species composition, damaging agents). Western spruce budworm was the primary damaging agent of host trees in all stands andcaused host tree mortality across all size classes, particularly in spruce-fir stands. Results indicate an unsustainable level of mortality in spruce-fir stands and a transition towards non-host species in mixed-conifer stands. Low levels of regeneration coupled with high overstory mortality rates indicate a potential lack of resilience in spruce-fir stands, whereas resilience to future western spruce budworm defoliation events may have increased in mixed-conifer stands affected by these outbreaks.


Splitting of the circadian rhythm of activity is abolished by unilateral lesions of the suprachiasmatic nuclei.
The circadian rhythm of activity in vertebrates often splits into two components after continuous exposure to constant light. This observation suggests that at least two circadian pacemakers underlie the activity rhythm. After unilateral ablation of the hypothalamic suprachiasmatic nuclei in hamsters, the splitting phenomenon was eliminated and a single rhythm of activity was established. The period of the new circadian activity rhythm different from the periods of the split rhythm and that preceding the split. These results suggest an interaction between the bilaterally paired suprachiasmatic nuclei in the generation of the circadian rhythm of activity.


Countermovement Jump Training Is More Effective Than Drop Jump Training in Enhancing Jump Height in Non-professional Female Volleyball Players.
The aim of the present study was to compare the effects of countermovement jump (CMJ) and drop jump (DJ) training on the volleyball-specific jumping ability of non-professional female volleyball players. For that purpose, 26 female volleyball players (15-32 years) were assigned to either a CMJ (20.4 ± 3.1 years, 171.0 ± 3.0 cm) or a DJ training group (22.0 ± 4.4 years, 168.2 ± 5.0 cm), which performed a six-week jump training (two sessions per week, 60 jumps per session). Each group performed 20% of the jumps in the jump type of the other group in order to minimize the influence of enhanced motor coordination on the differences between groups regarding the improvements of jump performance. Before and after the training, jump height was assessed in four jump types, including the trained and volleyball-specific jump types. Although both training forms substantially improved jump height, the CMJ training was significantly more effective in all jump types (17 vs. 7% on average; p < 0.001). This suggests that, at least for non-professional female volleyball players and a training duration of six weeks, training with a high percentage of CMJs is more effective than one with a high percentage of DJs. We hypothesize that this might be related to the slower stretch-shortening cycle during CMJs, which seems to be more specific for these players and tasks. These findings should support volleyball coaches in designing optimal jump trainings.


The sequential expression of CD40 and Icam2 defines progressive steps in the formation of blood precursors from the mesoderm germ layer.
During embryogenesis, the hematopoietic program is specified from the mesodermal germ layer through the formation of hemangioblast. This precursor gives rise to a hemogenic endothelium that later on matures to generate primitive and definitive hematopoietic precursors. A lack of specific cell surface markers to identify cells with discrete developmental potential is a major hurdle in the quest to further understand the cellular and molecular program governing blood formation. In the present study, we identify CD40 and Icam2, two markers typically associated with the adult immunological compartment, as expressed at the earliest stages of blood specification both in vitro and in vivo. Using in vitro serum-free culture conditions that support the efficient and directed differentiation of embryonic stem cells, we show that the sequential expression of CD40 and Icam2 delineate a transition in the acquisition of the blood potential from hemangioblast to hemogenic endothelium leading to the formation of primitive and definitive hematopoietic progenitors. CD40 is transiently expressed at the onset of blood development and marks first the hemangioblast then the hemogenic endothelium but is no longer expressed on fully committed hematopoietic precursors within the fetal liver. In contrast, Icam2 is first expressed on the hemogenic endothelium and its expression persists on fetal liver hematopoietic progenitors. Taken together, our data identify novel cell surface markers allowing us to further refine our understanding of the events marking progressive hematopoietic commitment from the mesoderm germ layer.


Provoked Narcissistic Aggression: Examining the Role of De-Escalated and Escalated Provocations.
Theories have postulated why provocation is particularly likely to incite aggression for narcissists, but key propositions from such theories dealing with psychological process or boundary conditions have seldom been tested. Here, we investigated narcissists' cognitive, emotional, and motivational experience and aggression following escalated and de-escalated provocation. Participants (N = 680) completed measures of grandiose narcissism (normal and pathological expressions) and vulnerable narcissism. Next, participants simulated provocation via imagining everyday scenarios in which a provocateur either de-escalates (apologizes and expresses concern) or escalates (makes a spiteful remark) the provocation and then rated anger and humiliation, perceived "narcissistic injury," goals, and aggression. Expressions of grandiose narcissism, but vulnerable narcissism, more strongly related to aggression following escalated (vs. de-escalated) provocation. Path modeling revealed that perceived narcissistic injury and narcissistic-identity goals explained this relation for pathological grandiose narcissists, and revenge goals and narcissistic-identity goals explained this relation for normal grandiose narcissists. Conversely, vulnerable narcissism related more strongly to anger, perceived narcissistic injury, and narcissistic-identity goals following de-escalated (vs. escalated) provocation. Path modeling revealed that vulnerable narcissists' enhanced anger-induced aggression is bounded to de-escalated provocation. We discuss the findings in relation to various theories of provoked narcissistic aggression.


Review of the COVID-19 Risk in Multiple Sclerosis.
The ongoing pandemic of the novel coronavirus of 2019 (COVID-19) has resulted in over 1 million deaths, primarily affecting older patients with chronic ailments. Multiple sclerosis (MS) patients have been deemed particularly vulnerable given their high rates of disability and increased susceptibility to infections. There have also been concerns regarding disease-modifying therapy (DMT) during the pandemic as many DMTs may increase the risk of infection due to some of their immunosuppressive properties. Furthermore, due to MS-related chronic inflammatory damage within the central nervous system, there have been concerns for worsening neurological injury by COVID-19. This has resulted in an alarmingly high level of anxiety and stress among the MS community leading to a lack of compliance with medications and routine check-ups, and even failure to obtain treatment for relapse. However, there is currently substantial evidence that MS and most DMT usage is not associated with increased COVID-19 severity. MS patients who suffer worse outcomes were more likely to be older and suffer from significant disabilities and comorbid conditions, which would also be expected from those in the general population. Likewise, there is little if any evidence demonstrating an increased susceptibility of MS patients to COVID-19-related neurological complications. Therefore, we aim to summarize the most recent findings related to COVID-19 and MS demonstrating that MS and most DMTs do not appear as risk factors for severe COVID-19.


Acute promyelocytic leukaemia presenting as necrotising fasciitis of the perineum (Fournier gangrene).
We present a case of an unusual presentation of acute promyelocytic leukaemia (APML), which presented with Fournier gangrene (FG). A 38-year-old man presented with malaise, groin swelling, anal bleeding, fever and was found to have FG. Initial workup revealed pancytopaenia, borderline low fibrinogen, prolonged international normalized ratio (INR), which raised the suspicion for leukaemia. The peripheral blood differential revealed leucopaenia with absolute neutropaenia and a 5% abnormal promyelocytes but no blasts, suspicious for APML. Bone marrow biopsy was performed and fluorescence in situ hydridization (FISH), karyotype and PCR confirmed a t(15;17) translocation, establishing a diagnosis of APML. After 1 month of therapy for intermediate risk APML with All-trans retinoic acid (ATRA) and arsenic trioxide (ATO), repeat chromosomal analysis and repeat bone marrow biopsy revealed no evidence of residual APML. After the consolidation phase was started with ATRA and ATO regimen, the wound healed after 2 months and the patient achieved complete remission.


Role of the DksA-like protein in the pathogenesis and diverse metabolic activity of Campylobacter jejuni.
DksA is well known for its regulatory role in the transcription of rRNA and genes involved in amino acid synthesis in many bacteria. DksA has also been reported to control expression of virulence genes in pathogenic bacteria. Here, we elucidated the roles of a DksA-like protein (CJJ81176_0160, Cj0125c) in the pathogenesis of Campylobacter jejuni. As in other bacteria, transcription of stable RNA was repressed by the DksA-like protein under stress conditions in C. jejuni. Transcriptomic and proteomic analyses of C. jejuni 81-176 and an isogenic mutant lacking the DksA-like protein showed differential expression of many genes involved in amino acid metabolism, iron-related metabolism, and other metabolic reactions. Also, the C. jejuni DksA-like protein mutant exhibited a decreased ability to invade intestinal cells and induce release of interleukin-8 from intestinal cells. These results suggest that the DksA-like protein plays an important regulatory role in diverse metabolic events and the virulence of C. jejuni.


Follow-up of treated osteosarcoma patient.
To evaluate the value of post-treatment follow-up in osteosarcoma patients. Data were collected through a clinical record, with socio-demographic and clinical data, and information relating to the medical appointment. Descriptive analysis of the data was carried out. The Chi-squared test was used to associate the independent variables with attendance at scheduled follow-up appointments. We found a recurrence in 59.6% of cases, of which 58% were lung related; 44% presented clinical complaints and arrived on the scheduled date of the appointment. There was no statistically significant association between the demographic characteristics and early attendance of follow-up visits. 81.3% of the cases who came for the appointment earlier than originally scheduled presented complaints compared to those who did not (p=0.005). Of the cases who presented recurrence, 12.9% attended an appointment late and those who did not present recurrence, 47.6% were late for the appointment (p=0.006). It is seen that the patients who came for an earlier appointment presented more complaints and were associated with the positive result of the exams carried out. The patients who had recurrence and came for an earlier appointment did not present a statistically significant difference in recurrence-free survival. It was observed that distance was not a predominant factor in late attendance at appointments. Level of Evidence II, Retrospective Study.


Roles of Dicer-Like Proteins 2 and 4 in Intra- and Intercellular Antiviral Silencing.
RNA silencing is an innate antiviral mechanism conserved in organisms across kingdoms. Such a cellular defense involves DICER or DICER-LIKEs (DCLs) that process plant virus RNAs into viral small interfering RNAs (vsiRNAs). Plants encode four DCLs that play diverse roles in cell-autonomous intracellular virus-induced RNA silencing (known as VIGS) against viral invasion. VIGS can spread between cells. However, the genetic basis and involvement of vsiRNAs in non-cell-autonomous intercellular VIGS remains poorly understood. Using GFP as a reporter gene together with a suite of DCL RNAi transgenic lines, here we show that despite the well-established activities of DCLs in intracellular VIGS and vsiRNA biogenesis, DCL4 acts to inhibit intercellular VIGS whereas DCL2 is required (likely along with DCL2-processed/dependent vsiRNAs and their precursor RNAs) for efficient intercellular VIGS trafficking from epidermal to adjacent cells. DCL4 imposed an epistatic effect on DCL2 to impede cell-to-cell spread of VIGS. Our results reveal previously unknown functions for DCL2 and DCL4 that may form a dual defensive frontline for intra- and intercellular silencing to double-protect cells from virus infection in Nicotiana benthamiana.


Distribution and sequence of gonadotropin-inhibitory hormone and its potential role as a molecular link between feeding and reproductive systems in the Pekin duck (Anas platyrhynchos domestica).
The reproductive status of adult Pekin drakes is very sensitive to nutritional status. Thus, the purpose of this study was to increase our understanding of the neurobiology underlying the depressive effect of fasting on the secretion of reproductive hormones. It was hypothesized that this effect was mediated by gonadotropin-inhibitory hormone (GnIH). Networks of GnIH fibers were present throughout the diencephalon, and cell bodies were present primarily, in the hypothalamic paraventricular nucleus (PVN). The duck GnIH gene was cloned and sequenced and found to encode GnIH and two GnIH-related peptides (GnIH-RP1, GnIH-RP2) which have a similar identity to those found in other avian species. Intracerebroventricular injection of GnIH, but not of GnIH-RP1, depressed plasma LH and stimulated feeding. Fasting for 48h depressed plasma LH and induced fos expression in about half the population of GnIH-ir neurons. These data suggest that GnIH neurons are mediators between feeding and reproductive systems in Pekin drakes.


IL-21 and IL-12 inhibit differentiation of Treg and TH17 cells and enhance cytotoxicity of peripheral blood mononuclear cells in patients with cervical cancer.
Interleukin 21 (IL-21) and IL-12 have been known to be effective antitumor agents. In this study, we evaluated whether IL-21 in combination with IL-12 could enhance the cytotoxicity of peripheral blood mononuclear cells (PBMCs) in patients with cervical intraepithelial neoplasia III and cervical cancer. Peripheral blood mononuclear cells were isolated from peripheral blood of cervical intraepithelial neoplasia III patients (n = 17) and cervical cancer patients (n = 18). Peripheral blood mononuclear cells were cultured with IL-2 in low concentration as control group. Interleukin 2-stimulated PBMCs were cocultured with anti-human IL-21 neutralizing antibody, IL-21 alone, IL-12 alone, and IL-21 plus IL-12, respectively, as test groups. The cytotoxicity of PBMCs against SiHa tumor cells was examined by lactate dehydrogenase released assay. CD4CD25FOXP3 T regulatory (Treg) cells and CD4IL-17A T helper 17 (TH17) cells were analyzed by flow cytometry. Proliferation and apoptosis were detected by CCK-8 (cell counting kit 8) assay and flow cytometry, respectively. Compared with controls, IL-21 and IL-12 significantly elevated PBMC cytotoxicity against SiHa cells. Moreover, IL-21 plus IL-12 significantly elevated PBMC cytotoxicity in comparison to IL-21 alone and IL-12 alone. We also found that IL-21 plus IL-12 significantly decreased Treg and TH17 cell proportion in comparison to controls. Notably, IL-21 plus IL-12 significantly decreased TH17 cell proportion in comparison to IL-21 alone. Both IL-21 and IL-12 significantly decreased the apoptosis rate of PBMCs, whereas neither IL-21 nor IL-12 had significant effect on PBMC proliferation. The combination of IL-21 and IL-12 could efficiently stimulate PBMCs with cytotoxicity against SiHa cells, and the possible mechanisms may be due to down-regulated Treg and TH17 cell differentiation.


Can metal ions be used as gas-phase disulfide bond cleavage reagents? A survey of coinage metal complexes of model peptides containing an intermolecular disulfide bond.
The role that a metal ion can have in promoting disulfide bond cleavage has been assessed by surveying the tandem mass spectra of the following metal complexes of model peptides containing an intermolecular disulfide bond: [M--H+Cu(II)](+); [M--H+Cu(II)(bipy)](+); [M+Ag(I)](+); and [M+Au(I)(PMe(3))](+). In comparison to previously studied protonated peptides, these binary and ternary metal complexes generally yield more abundant S--S and/or C--S bond cleavage. In general, [M--H+Cu(II)](+) ions cleave the adjacent C--S bond more readily, while the [M+Au(I)(PMe(3))](+) ion cleaves the S--S bond more readily. The ternary metal complex [M--H+Cu(II)(bipy)](+), on the other hand, fragments by exclusive loss of the bipyridyl ligand for the larger model peptides studied. Of all coinage metal systems studied, Me(3)PAu(+) is superior in promoting disulfide bond cleavage.


Transfection of avian LMH-2A hepatoma cells with cationic lipids.
LMH-2A is an estrogen-responsive avian hepatoma cell line whose susceptibility to cationic-lipid-mediated transfection is poorly described. 3 beta[N-N',N'-dimethylaminoethane)-carbamoyl] cholesterol (DCC) requires a one-step synthesis, and can be used to formulate transfection-grade liposomes when combined with dioleoylphosphatidyl-ethanolamine (DOPE) 1/1 (wt/wt). Luciferase activities in LMH-2A cells were 8.5-fold and 87.5-fold greater than those in HepG2 and FTO2B cells, respectively, following DCC-liposome-mediated transfection with a reporter consisting of the human cytomegalovirus immediate-early promoter (CMV), joined to Photinus pyralis luciferase (L) cDNA, designated pCMVL. Using pCMVL, N-(2-bromoethyl)-N,N-dimethyl-2,3-bis(9-octadecenyloxy)-propana minimun bromide) (BMOP)/DOPE 1/1 (wt/wt), at a 7.5:1 ratio with DNA, produced luciferase activities that were 2.9-fold higher than those of DCC-liposomes, at an optimal 10:1 lipid:DNA ratio. At optimal lipid:DNA ratios, commercially available liposomes, Transfectam, Lipofectamine, and Lipofectin, produced luciferase activities that were 1.39, 1.03, and 0.47-fold those of DCC-liposomes. The effect of 0, 10, 100, or 500 nM/L 17 beta-estradiol on the expression of pCMVL and a second luciferase reporter containing the -593/+48 promoter region of the estrogen-responsive avian apo VLDL-II gene, designated pApoL, was tested in cells cultured in the presence or absence of 10% chicken serum. The CMV promoter supported a high level of expression in LMH-2A cells that was unaffected by serum alone, but was weakly responsive to estrogen. Estrogen responses of both reporters reached a plateau at 10 nM/L. Estrogen increased the expression of pApoL 24-fold and 79-fold in the absence and presence of serum, respectively. The -593/+48 region of the apo VLDL-II promoter may not contain previously reported negative insulin response elements, but chicken serum contains factors that enhance estrogen responsiveness of this region.


Combining SDAE Network with Improved DTW Algorithm for Similarity Measure of Ultra-Weak FBG Vibration Responses in Underground Structures.
Quantifying structural status and locating structural anomalies are critical to tracking and safeguarding the safety of long-distance underground structures. Given the dynamic and distributed monitoring capabilities of an ultra-weak fiber Bragg grating (FBG) array, this paper proposes a method combining the stacked denoising autoencoder (SDAE) network and the improved dynamic time wrapping (DTW) algorithm to quantify the similarity of vibration responses. To obtain the dimensionality reduction features that were conducive to distance measurement, the silhouette coefficient was adopted to evaluate the training efficacy of the SDAE network under different hyperparameter settings. To measure the distance based on the improved DTW algorithm, the one nearest neighbor (1-NN) classifier was utilized to search the best constraint bandwidth. Moreover, the study proposed that the performance of different distance metrics used to quantify similarity can be evaluated through the 1-NN classifier. Based on two one-dimensional time-series datasets from the University of California, Riverside (UCR) archives, the detailed implementation process for similarity measure was illustrated. In terms of feature extraction and distance measure of UCR datasets, the proposed integrated approach of similarity measure showed improved performance over other existing algorithms. Finally, the field-vibration responses of the track bed in the subway detected by the ultra-weak FBG array were collected to determine the similarity characteristics of structural vibration among different monitoring zones. The quantitative results indicated that the proposed method can effectively quantify and distinguish the vibration similarity related to the physical location of structures.


Prenatal Exposure to Progesterone Affects Sexual Orientation in Humans.
Prenatal sex hormone levels affect physical and behavioral sexual differentiation in animals and humans. Although prenatal hormones are theorized to influence sexual orientation in humans, evidence is sparse. Sexual orientation variables for 34 prenatally progesterone-exposed subjects (17 males and 17 females) were compared to matched controls (M age = 23.2 years). A case-control double-blind design was used drawing on existing data from the US/Denmark Prenatal Development Project. Index cases were exposed to lutocyclin (bioidentical progesterone = C21H30O2; M W : 314.46) and no other hormonal preparation. Controls were matched on 14 physical, medical, and socioeconomic variables. A structured interview conducted by a psychologist and self-administered questionnaires were used to collect data on sexual orientation, self-identification, attraction to the same and other sex, and history of sexual behavior with each sex. Compared to the unexposed, fewer exposed males and females identified as heterosexual and more of them reported histories of same-sex sexual behavior, attraction to the same or both sexes, and scored higher on attraction to males. Measures of heterosexual behavior and scores on attraction to females did not differ significantly by exposure. We conclude that, regardless of sex, exposure appeared to be associated with higher rates of bisexuality. Prenatal progesterone may be an underappreciated epigenetic factor in human sexual and psychosexual development and, in light of the current prevalence of progesterone treatment during pregnancy for a variety of pregnancy complications, warrants further investigation. These data on the effects of prenatal exposure to exogenous progesterone also suggest a potential role for natural early perturbations in progesterone levels in the development of sexual orientation.


[Left ventricular myocardial function in ischemic heart disease in combination with hypertension].
The work discusses the echocardiographic data on the function of the left-ventricular myocardium in patients with ischemic heart disease and hypertensive disease. Computer treatment of the echocardiograms allowed the authors to study not only the systole but the diastole also. It is shown that in patients in whom hypertensive disease is combined with ischemic heart disease, the process of myocardial relaxation is disturbed. The function of the left atrium is disturbed to a greater measure and earlier, and these disturbances preceed the disorders of th function of the left ventricle.


[Potential of prevention of metastasizing with the aid of tumor-specific transfer-factor].
The report discusses our experimental data in support of biotherapy which uses chemotherapy and antitumor immune treatment with in vivo xenogenic transfer-factor polypeptides (TFP) isolated from lymphocytes sensitized to antigens of given tumor. After excision of primary tumor--lung carcinoma of Lewis--mice C57BL/6 were injected intraperitoneally with xenogenic TFP (200 pg/body, twice) and a cytostaic dose of cyclophosphamide. Such adjuvant chemotherapy was found to prevent metastases from spreading to the lung in 100%. The marked anti-metastatic effect of the treatment correlated with recovery of splenic cell mass and its cellular structure, higher levels of large granular lymphocytes in peripheral blood and enhanced functional activity of cytotoxic cells in vitro. Our results point to a possibility of raising efficacy of treating solid malignancies with adjuvant chemotherapy in combination with adoptive immune therapy.


The transformation of continuing medical education (CME) in the United States.
This article describes five major themes that inform and highlight the transformation of continuing medical education in the USA. Over the past decade, the Institute of Medicine (IOM) and other national entities have voiced concern over the cost of health care, prevalence of medical errors, fragmentation of care, commercial influence, and competence of health professionals. The recommendations from these entities, as well as the work of other regulatory, professional, academic, and government organizations, have fostered discussion and development of strategies to address these challenges. The five themes in this paper reflect the changing expectations of multiple stakeholders engaged in health care. Each theme is grounded in educational, politico-economic priorities for health care in the USA. The themes include (1) a shift in expectation from simple attendance or a time-based metric (credit) to a measurement that infers competence in performance for successful continuing professional development (CPD); (2) an increased focus on interprofessional education to augment profession-specific continuing education; (3) the integration of CPD with quality improvement; (4) the expansion of CPD to address population and public health issues; and (5) identification and standardization of continuing education (CE) professional competencies. The CE profession plays an essential role in the transformation of the US CPD system for health professionals. Coordination of the five themes described in this paper will foster an improved, effective, and efficient health system that truly meets the needs of patients.


Prevention of falls in hospital.
Falls among inpatients are the most frequently reported safety incident in NHS hospitals. 30-50% of falls result in some physical injury and fractures occur in 1-3%. No fall is harmless, with psychological sequelae leading to lost confidence, delays in functional recovery and prolonged hospitalisation. Yet falls are not true accidents and there is evidence that a coordinated multidisciplinary clinical team approach can reduce their incidence. Identification of multiple underlying risk factors coupled with clear interventions to ameliorate the impact of each has been shown to reduce the incidence of inpatient falls by 20-30%. The implementation of complex multiprofessional interventions is challenging and successful schemes seek to nurture a culture of vigilant safety consciousness in all staff at the clinical interface. Strong leadership and organisational oversight help to combine this cultural evolution with relevant evidence and rigorous measurement of performance in order to improve patient safety. The results of national audit suggest that NHS acute hospitals could do more to reduce the incidence of falls among inpatients.


[An advanced case of myopathy and dementia with a new mutation in the valosin-containing protein gene].
We report a 51-year-old man with myopathy and dementia probably caused by a novel mutation of the valosin-containing protein (VCP) gene, in the form of a p.Ala439Pro substitution. At 43 years old, he presented at least 2-year history of weakness of right ankle dorsiflexion. Findings from muscle biopsy suggested distal myopathy with rimmed vacuoles. However, no mutation in the GNE gene was identified. He complained of giving way of the knee, and muscle imaging study showed adipose tissue infiltration in the quadriceps. Ten years later, he was confined to a wheelchair and became reticent and antisocial with slightly impaired memory. A muscle CT revealed atrophy or replacement by adipose tissue in the muscles of neck, trunks and extremities muscles with laterality and variation of the degree. The magnetic resonance imaging of the brain showed bilateral frontal and temporal lobe atrophy with left dominance. Findings were compatible with inclusion body myopathy with Paget's disease of bone and frontotemporal dementia.


Elaborate visual and acoustic signals evolve independently in a large, phenotypically diverse radiation of songbirds.
The concept of a macroevolutionary trade-off among sexual signals has a storied history in evolutionary biology. Theory predicts that if multiple sexual signals are costly for males to produce or maintain and females prefer a single, sexually selected trait, then an inverse correlation between sexual signal elaborations is expected among species. However, empirical evidence for what has been termed the 'transfer hypothesis' is mixed, which may reflect different selective pressures among lineages, evolutionary covariates or methodological differences among studies. Here, we examine interspecific correlations between song and plumage elaboration in a phenotypically diverse, widespread radiation of songbirds, the tanagers. The tanagers (Thraupidae) are the largest family of songbirds, representing nearly 10% of all songbirds. We assess variation in song and plumage elaboration across 301 species, representing the largest scale comparative study of multimodal sexual signalling to date. We consider whether evolutionary covariates, including habitat, structural and carotenoid-based coloration, and subfamily groupings influence the relationship between song and plumage elaboration. We find that song and plumage elaboration are uncorrelated when considering all tanagers, although the relationship between song and plumage complexity varies among subfamilies. Taken together, we find that elaborate visual and vocal sexual signals evolve independently among tanagers.


Patterns of temporal lobe atrophy in semantic dementia and Alzheimer's disease.
Volumetric magnetic resonance imaging analyses of 30 subjects were undertaken to quantify the global and temporal lobe atrophy in semantic dementia and Alzheimer's disease. Three groups of 10 subjects were studied: semantic dementia patients, Alzheimer's disease patients, and control subjects. The temporal lobe structures measured were the amygdala, hippocampus, entorhinal cortex, parahippocampal gyrus, fusiform gyrus, and superior, middle, and inferior temporal gyri. Semantic dementia and Alzheimer's disease groups did not differ significantly on global atrophy measures. In semantic dementia, there was asymmetrical temporal lobe atrophy, with greater left-sided damage. There was an anteroposterior gradient in the distribution of temporal lobe atrophy, with more marked atrophy anteriorly. All left anterior temporal lobe structures were affected in semantic dementia, with the entorhinal cortex, amygdala, middle and inferior temporal gyri, and fusiform gyrus the most severely damaged. Asymmetrical, predominantly anterior hippocampal atrophy was also present. In Alzheimer's disease, there was symmetrical atrophy of the entorhinal cortex, hippocampus, and amygdala, with no evidence of an anteroposterior gradient in the distribution of temporal lobe or hippocampal atrophy. These data demonstrate that there is a marked difference in the distribution of temporal lobe atrophy in semantic dementia and Alzheimer's disease. In addition, the pattern of atrophy in semantic dementia suggests that semantic memory is subserved by anterior temporal lobe structures, within which the middle and inferior temporal gyri may play a key role.


Defining the cytosolic pathway of glutathione degradation in Arabidopsis thaliana: role of the ChaC/GCG family of γ-glutamyl cyclotransferases as glutathione-degrading enzymes and AtLAP1 as the Cys-Gly peptidase.
Glutathione homoeostasis is critical to plant life and its adaptation to stress. The γ-glutamyl cycle of glutathione biosynthesis and degradation plays a pre-eminent role in glutathione homoeostasis. The genes encoding two enzymatic steps of glutathione degradation, the γ-glutamyl cyclotransferase (GGCT; acting on γ-glutamyl amino acids) and the Cys-Gly dipeptidase, have, however, lacked identification. We have investigated the family of GGCTs in Arabidopsis thaliana. We show through in vivo functional assays in yeast that all three members of the ChaC/GCG subfamily show significant activity towards glutathione but no detectable activity towards γ-glutamyl methionine. Biochemical characterization of the purified recombinant enzymes GGCT2;2 and GGCT2;3 further confirmed that they act specifically to degrade glutathione to yield 5-oxoproline and Cys-Gly peptide and show no significant activity towards γ-glutamyl cysteine. The Km for glutathione was 1.7 and 4.96 mM for GGCT2;2 and GGCT2;3 respectively and was physiologically relevant. Evaluation of representative members of other subfamilies indicates the absence of GGCTs from plants showing significant activity towards γ-glutamyl-amino acids as envisaged in the classical γ-glutamyl cycle. To identify the Cys-Gly peptidase, we evaluated leucine aminopeptidases (LAPs) as candidate enzymes. The cytosolic AtLAP1 (A. thaliana leucine aminopeptidase 1) and the putative chloroplastic AtLAP3 displayed activity towards Cys-Gly peptide through in vivo functional assays in yeast. Biochemical characterization of the in vitro purified hexameric AtLAP1 enzyme revealed a Km for Cys-Gly of 1.3 mM that was physiologically relevant and indicated that AtLAP1 represents a cytosolic Cys-Gly peptidase activity of A. thaliana. The studies provide new insights into the functioning of the γ-glutamyl cycle in plants.


Biological activity of FGF-23 fragments.
The phosphaturic activity of intact, full-length, fibroblast growth factor-23 (FGF-23) is well documented. FGF-23 circulates as the intact protein and as fragments generated as the result of proteolysis of the full-length protein. To assess whether short fragments of FGF-23 are phosphaturic, we compared the effect of acute, equimolar infusions of full-length FGF-23 and various FGF-23 fragments carboxyl-terminal to amino acid 176. In rats, intravenous infusions of full-length FGF-23 and FGF-23 176-251 significantly and equivalently increased fractional phosphate excretion (FE Pi) from 14 +/- 3 to 32 +/- 5% and 15 +/- 2 to 33 +/- 2% (p < 0.001), respectively. Chronic administration of FGF-23 176-251 reduced serum Pi and serum concentrations of 1alpha,25-dihydroxyvitamin D. Shorter forms of FGF-23 (FGF-23 180-251 and FGF-23 184-251) retained phosphaturic activity. Further shortening of the FGF-23 carboxyl-terminal domain, however, abolished phosphaturic activity, as infusion of FGF-23 206-251 did not increase urinary phosphate excretion. Infusion of a short fragment of the FGF-23 molecule, FGF-23 180-205, significantly increased FE Pi in rats and reduced serum Pi in hyperphosphatemic Fgf-23 ( -/- ) knockout mice. The activity of FGF-23 180-251 was confirmed in opossum kidney cells in which the peptide reduced Na(+)-dependent Pi uptake and enhanced internalization of the Na(+)-Pi IIa co-transporter. We conclude that carboxyl terminal fragments of FGF-23 are phosphaturic and that a short, 26-amino acid fragment of FGF-23 retains significant phosphaturic activity.


Multi-session electrical neuromodulation effects on craving, relapse and cognitive functions in cocaine use disorder: A randomized, sham-controlled tDCS study.
The use of transcranial Direct Current Stimulation (tDCS) has previously shown promising results for reducing craving in cocaine use disorder. In this study we further explored the potential of tDCS as add-on intervention in the treatment of cocaine use disorder. In a randomized, placebo-controlled, between subject study, we applied tDCS bilaterally with the anodal electrode targeting the right dorsolateral prefrontal cortex (DLPFC; https://clinicaltrials.gov/ct2/show/NCT03025321). Patients with cocaine use disorder were allocated to ten sessions of either active tDCS (n = 29) or sham (n = 30) on five consecutive days. Inhibitory control and risky decision-making were measured via a Go-NoGo task and a two-choice gambling task, respectively, each at baseline, one day after all tDCS sessions and after three months. Relapse at follow-up and craving were also assessed. There was no significant effect of active tDCS on the number of cocaine use days and craving. Relapse was frequent among patients who had received either active or sham tDCS (48.0 % and 69.2 %, respectively), despite an overall decrease in craving during the first two weeks of treatment. No effects were found on cognitive functions. An exploratory analysis for crack cocaine use only revealed that relapse rates were significantly reduced after active tDCS (n = 17) as compared to sham (n = 19). No beneficial effects of tDCS on number of cocaine use days, craving and cognitive functions were found in the present study, but somewhat promising results were obtained regarding relapse rates among crack-cocaine users specifically. Further research is required to determine the efficacy of tDCS as a complementary treatment in cocaine use disorder.


Resolution of cassava-infecting alphaflexiviruses: Molecular and biological characterization of a novel group of potexviruses lacking the TGB3 gene.
Several potexviruses (Family Alphaflexiviridae) have been reported infecting cassava (Manihot esculenta Crantz) in the Americas. They were isolated from severely diseased plants during the last 30-40 years and include: Cassava common mosaic virus (CsCMV), Cassava Caribbean mosaic virus (CsCaMV), Cassava Colombian symptomless virus (CsCSV) and Cassava virus X (CsVX). However, their definitive classification as distinct species remains unresolved for several reasons, including the lack of sequence data and unavailability of samples from original isolates. This complicates disease diagnostics, cassava germplasm exchange certification, evaluation of virus cleaning protocols and epidemiological studies. Furthermore, a recently detected novel alphaflexivirus, indicates that cassava-infecting potexviruses may be more diverse. To solve the identity of these viruses, we started indexing samples from different parts of Colombia using different sets of PCR primers, antisera available and inoculation to indicator plants. Results show that there are three major phylogenetic groups of potexviruses infecting cassava, and they correspond to CsCMV, CsVX and the newly identified Cassava new alphaflexivirus (CsNAV). Bioassays and sequence analysis established that isolates of CsNAV and CsVX cause latent infections in different cassava landraces, they are not efficiently transmitted to the indicator plant Nicotiana benthamiana and they lack the gene 3 of the conserved potexviral 'triple gene block' (TGB). In contrast, all isolates of CsCMV (which have a characteristic potexvirus genome arrangement) caused Cassava Common Mosaic Disease (CCMD) in single infections and were efficiently transmitted to N. benthamiana. Although phylogenetic analysis of the replicase sequence placed CsNAV and CsVX as members of the Potexvirus genus, their distinct genome arrangement and biological characteristics suggest they can be considered as members of a separate taxonomic group.


Resistance of Fasciola hepatica against triclabendazole in cattle and sheep in The netherlands.
In the winter of 1998/1999, sheep on a farm in the province of North Holland, The Netherlands, died from subacute and chronic liver fluke disease despite four previous treatments with triclabendazole (TCBZ). Faecal examinations of sheep and cattle on the farm showed high number of liver fluke eggs. In a randomised clinical trial, the fluke egg output was monitored weekly for 3 weeks in sheep which were treated with TCBZ or with closantel; in dairy cows treated with TCBZ or with clorsulon; and in heifers treated with TCBZ or clorsulon. The results showed a significant reduction of 99.7, 98.1 and 99.2%, respectively, in fluke egg output at 21 days in all non-TCBZ treated animals. TCBZ treatment produced percentage decreases of 15.3, 4.3 and 36.6%, respectively. These results are highly indicative of the presence of TCBZ-resistant Fasciola hepatica in sheep and cattle on this farm.


Discretization provides a conceptually simple tool to build expression networks.
Biomarker identification, using network methods, depends on finding regular co-expression patterns; the overall connectivity is of greater importance than any single relationship. A second requirement is a simple algorithm for ranking patients on how relevant a gene-set is. For both of these requirements discretized data helps to first identify gene cliques, and then to stratify patients.We explore a biologically intuitive discretization technique which codes genes as up- or down-regulated, with values close to the mean set as unchanged; this allows a richer description of relationships between genes than can be achieved by positive and negative correlation. We find a close agreement between our results and the template gene-interactions used to build synthetic microarray-like data by SynTReN, which synthesizes "microarray" data using known relationships which are successfully identified by our method.We are able to split positive co-regulation into up-together and down-together and negative co-regulation is considered as directed up-down relationships. In some cases these exist in only one direction, with real data, but not with the synthetic data. We illustrate our approach using two studies on white blood cells and derived immortalized cell lines and compare the approach with standard correlation-based computations. No attempt is made to distinguish possible causal links as the search for biomarkers would be crippled by losing highly significant co-expression relationships. This contrasts with approaches like ARACNE and IRIS.The method is illustrated with an analysis of gene-expression for energy metabolism pathways. For each discovered relationship we are able to identify the samples on which this is based in the discretized sample-gene matrix, along with a simplified view of the patterns of gene expression; this helps to dissect the gene-sample relevant to a research topic--identifying sets of co-regulated and anti-regulated genes and the samples or patients in which this relationship occurs.


Adipokines as a novel link between obesity and atherosclerosis.
The traditional perception of adipose tissue as a storage organ of fatty acids has been replaced by the notion that adipose tissue is an active endocrine organ, releasing various adipokines that are involved in the pathogenesis of obesity-related metabolic disturbances. Obesity is a well-known risk factor for atherosclerosis, and accelerates atherosclerosis by many mechanisms such as increase in blood pressure and glucose level, abnormal lipid profiles, and systemic inflammation. Furthermore, growing evidence suggests that some adipokines directly mediate the process of atherosclerosis by influencing the function of endothelial cells, arterial smooth muscle cells, and macrophages in vessel walls. In obese patients, the secretion and coordination of such adipokines is abnormal, and the secretion of specific adipokines increases or decreases. Accordingly, the discovery of new adipokines and elucidation of their functions might lead to a new treatment strategy for metabolic disorders related to obesity, including cardiovascular diseases.