The historical development of academic journals in occupational medicine, 1901-2009.
Academic journals in a specialist field provide an interesting historical record of its development and progression over time. This article describes the evolution of some major international journals of occupational medicine, including some historical background on their editorial board. As North America, the United Kingdom, and Northern Europe are known to have the highest contribution to scientific production, it was considered appropriate to investigate the main occupational medicine periodicals in these regions. Given the remarkable improvements in Japanese occupational health following the Second World War, it was also considered worthwhile to investigate the two English-language journals of occupational medicine from this country.


Vascular Calcification and Stone Disease: A New Look towards the Mechanism.
Calcium phosphate (CaP) crystals are formed in pathological calcification as well as during stone formation. Although there are several theories as to how these crystals can develop through the combined interactions of biochemical and biophysical factors, the exact mechanism of such mineralization is largely unknown. Based on the published scientific literature, we found that common factors can link the initial stages of stone formation and calcification in anatomically distal tissues and organs. For example, changes to the spatiotemporal conditions of the fluid flow in tubular structures may provide initial condition(s) for CaP crystal generation needed for stone formation. Additionally, recent evidence has provided a meaningful association between the active participation of proteins and transcription factors found in the bone forming (ossification) mechanism that are also involved in the early stages of kidney stone formation and arterial calcification. Our review will focus on three topics of discussion (physiological influences-calcium and phosphate concentration-and similarities to ossification, or bone formation) that may elucidate some commonality in the mechanisms of stone formation and calcification, and pave the way towards opening new avenues for further research.


Variations of the iliolumbar and ascending lumbar veins.
A cadaveric retrospective study. To verify the variation of tributaries draining into the iliac system at the lumbosacral junction. Special attention to vascular anatomy at the great venous bifurcation during anterior lumbar surgery is needed as more tributaries may increase the rate of operative bleeding. Although important, there have been very few detailed anatomic studies about variations of the ascending and iliolumbar veins. The latter tributary forms a horizontal system draining blood into the iliac vein. A total of 116 formaldehyde-preserved specimens (61 male and 55 female) were studied. All specimens were dissected and simply classified into patterns and subpatterns according to the similar or different tributaries and their entry points. There were 2 patterns of tributaries: symmetric and asymmetric. More variants were found in the asymmetric group. Each pattern comprising 58 specimens was further classified into 3 subpatterns according to their main tributary trunks. One to 3 tributaries may enter into any trunk of the iliac vein. The most common trunk was the common iliac vein seen in 75.9% of specimens. Draining into the external and internal veins was found in 21.6% and 2.6% of specimens, respectively. One common drainage of the ascending and iliolumbar veins on 1 or both sides was found in 0.9% and > 50% of specimens, respectively (right, 51.7%; left, 56%), whereas 2 or more tributaries were seen in < 50% of specimens (right, 47.4%; left, 43.1%). There was no significant statistical difference among sexes and body sides. Draining into the iliac system has a high frequency of variations through a number of tributaries and their entry points. The iliolumbar vein will mainly drain into the common iliac vein with frequently 2 or 3 tributaries, whereas drainage into the external iliac or the internal vein is less often observed.


[Blood platelets metabolic and activation processes and options for their inhibition].
A range of activation processes and confirmatory and metabolic changes take place in blood platelets following their activation. The paper discusses some of the blood platelet activation steps and describes the specific processes that take place on this level. Furthermore, the points at which it is possible to actively interfere with the blood platelet metabolism or activation processes are described. Antiplatelet treatment focuses on these points and there are efforts to identify active substances that either inhibit access to certain receptors or targets or, alternatively, inhibit the enzymes participating in these processes.


Chiari I Malformation and Spinal Cord Ischemia in a Cadaver.
With advanced imaging, the Chiari I malformation (CIM) is more frequently diagnosed than in the past when this entity was identified most commonly at autopsy. Herein, we report the rare case of an adult cadaver found not only to have CIM but also adjacent spinal cord ischemia. This case is discussed in the context of chronic compression of the spinal cord by a CIM and the need for close monitoring of these patients.


Identification of a novel laccase gene EuLAC1 and its potential resistance against Botrytis cinerea.
In this study, a novel laccase gene, EuLAC1, was cloned from Eucommia ulmoides Oliver (E. ulmoides). An overexpression vector harboring the EuLAC1 was constructed and introduced into the tobacco (Nicotiana tabacum cv. Xanthi). The laccase activity, resistance to Botrytis cinerea (B. cinerea) and lignin level in wild-type and transgenic plants were thereafter investigated. Interestingly, the transgenic tobacco displayed a significantly higher laccase activity and resistance to gray mold as compared to the wild-type tobacco. Additionally, the lignin contents in the leaves and stems of the transgenic tobacco were significantly higher in comparison to the wild-type tobacco. Scanning electron microscopy was used to observe the cross sections of wild-type and transgenic tobacco stems and it was noted that the cell wall near the xylem catheter of the transgenic tobacco was substantially thicker and the outline clearer than that of the wild-type. Thus, the EuLAC1 gene can significantly increase laccase activity and lignin content in tobacco, leading to an increase in the physical defenses, thereby increasing tobacco resistance to gray mold.


Inhibition of hcv replication in HCV replicon by shRNAs.
We show that the vector-derived long dsRNA specifically inhibits the replication of HCV RNA in HCV replicon. We designed a long dsRNA targeted to the full-length HCV IRES/core elements (1-to 377-nt). Our results revealed that the replication of HCV RNA was reduced to near background levels in a sequence-specific manner by the long dsRNAs in the HCV replicon. We also designed four shRNAs against several regions (120- to 139-nt, 260- to 279-nt, 330- to 349-nt, and 340- to 359-nt) of the HCV IRES/Core elements. The two HCV IRES/core-specific shRNAs, 330- to 349-nt and 340- to 359-nt, containing the AUG initiation codon sequence showed stronger HCV inhibitory effects than the other two shRNAs, 120- to 139-nt and 260- to 279-nt.


Essential role of Swp73p in the function of yeast Swi/Snf complex.
Swi/Snf protein was purified previously from the yeast Saccharomyces cerevisiae as an 11-polypeptide complex, including five novel Swp polypeptides. Here we present evidence concerning the role of Swp73p in the function of the complex. Deletion mutants in the SWP73 gene display phenotypes similar to those of swi and snf mutants, and in addition are temperature-sensitive. Swp73p is required for transcriptional activation by full-length glucocorticoid receptor (GR), but not by all GR derivatives. Swp73p is also required for activation with an enhancer element that binds the transcription factors Swi5p and Pho2p, which may underlie the defects in HO expression observed with swi and snf mutants. A single amino acid change in the protein confers phenotypes that are similar to those observed in the swp73 delta strain, but in some cases the two strains behave differently. The extent to which Swp73p is required for assisting transcriptional activation depends on the activator and promoter tested. Homologs of SWP73 are present in S. cerevisiae, Ashbya gossypii, Caenorhabditis elegans, and mice, indicating that SWP73 may belong to a family of related genes encoding proteins with analogous functions.


A new species of deep-water Lethrinops (Cichlidae) from Lake Malawi.
A new species of cichlid fish, Lethrinops atrilabris is described from specimens collected by trawling at a depth of around 90 m off Monkey Bay, southern Lake Malawi. It is assigned to the genus Lethrinops on the basis of its vertical flank barring, lack of enlarged cephalic lateral line canal pores and the form of the lower jaw dental arcade. It can be distinguished from congeneric species by its male breeding dress of contrasting flank barring and dark ventral surface, most strikingly on the lips, throat and chest, its relatively small known maximum size [<75 mm standard length (SL)], large eyes (38%-41% head length), laterally compressed body (depth 2.5-2.7 times max head width) and lower gill raker count (13-14).


Neurogenic stress cardiomyopathy following aneurysmal subarachnoid hemorrhage in a very elderly patient--case report.
A 90-year-old woman presented with aneurysmal subarachnoid hemorrhage (SAH) corresponding to Hunt and Hess grade II. Acute congestive heart failure and pulmonary edema developed following uneventful surgical clipping. Serial electrocardiography and echocardiography led to a diagnosis of neurogenic stress cardiomyopathy (NSC), also known as tako-tsubo cardiomyopathy. The outcome was favorable after supportive therapy with respiratory management and diuretic administration. Neurosurgeons treating SAH must take into account the various general treatment strategy options on a case by case basis after prompt recognition of NSC.


[Laparoscopic management of an infected oesophageal cyst].
Oesophageal cysts are rare congenital anomalies. Most commonly, they are located in the posterior mediastinum, approximately at the level of the distal third of the oesophagus. In most cases, they are diagnosed during childhood as an accidental finding of a symptomatic mediastinal lesion. A CT or MRI, oesophagoscopy and endosonography of the oesophagus are the essential diagnostic methods. In the adulthood, most of the cysts cause problems pushing against the surrounding organs and, therefore, surgical removal is recommended. In this report, a case of a potential complication is described: infection of the oesophageal cyst of the distal part of the oesophagus following a transoesophageal punction. The case resulted in sepsis, which was managed laparoscopically. Entering through the diaphragmatic hiatus,a cyst in the posterior mediastinum was revealed. The cyst was partially resected and its content evacuated. No complications followed. The laparoscopic method is feasible and safe even if the oesophageal cysts are infected. A complete resection of the cyst would be complicated due to a possibility of injuring the oesophagus. A significance of the peroperative oesophagoscopy to verify the oesophagus position in an unclear anatomic situation due to inflammatory infiltration, is highlighted. The above decreases the risk of the oesophageal wall lesion.


Effects of raloxifene on the renin-angiotensin-aldosterone system and blood pressure in hypertensive and normotensive osteoporotic postmenopausal women.
An increase in blood pressure after menopause has been documented. The renin-angiotensin-aldosterone system (RAAS) plays a central role in the regulation of blood pressure and in the pathophysiology of hypertension. This study investigated the effects of raloxifene, a selective estrogen receptor modulator, on components of the RAAS and blood pressure in hypertensive and normotensive osteoporotic postmenopausal women. A total of 41 hypertensive or normotensive postmenopausal women with osteoporosis or osteopenia were divided into four groups. Eleven hypertensive and eight normotensive women received raloxifene hydrochloride (60 mg/day) p.o. for 6 months, and 12 hypertensive and 10 normotensive women did not receive raloxifene hydrochloride for 6 months. In all of the hypertensive women, blood pressure had been controlled prior to the start of the study using a variety of antihypertensive drugs other than angiotensin-converting enzyme (ACE) inhibitors, angiotensin (Ang)II type 1 receptor antagonists or diuretics. Plasma renin activity (PRA), serum ACE activity, plasma AngI, AngII and aldosterone concentrations, and blood pressure were measured before and 6 months after the start of the study. No significant changes in PRA, ACE activity, or the AngI, AngII or aldosterone levels were observed in any of the groups. In all the groups, blood pressure remained unchanged. Raloxifene may have no significant effect on the RAAS or blood pressure in hypertensive and normotensive osteoporotic postmenopausal women.


[Gastric sarcoidosis -- a case report].
Gastrointestinal sarcoidosis is very rare manifestation of the illness. This article presents patient who had been diagnosed for several months because of abdominal pain, vomiting and significant weight loss. Numerous gastric endoscopy examinations showed the difficult healing ulcers in the stomach, progressive thickening of the mucosal folds with narrowing of the lumen, lack of peristalsis and the rigidity of the walls. Histological examination of the specimen of the ventricular mucosa revealed chronic inflammation, as well as suspicion of tumor infiltration. Finally gastric sarcoidosis has been diagnosed on the basis of CT with double contrast, and histopathological examination of biopsis of stomach collected during laparotomy.


Electron multiplier-scintillator detector for pulse counting positive or negative ions.
A general-purpose ion detector has been developed for fast pulse counting in mass spectrometry. It consists of a six-stage electron multiplier joined to part of a scintillator detector. This novel arrangement combines the virtually noiseless pulse-amplification response of an electron multiplier with the bipolar voltage flexibility of a scintillator detector. The resulting hybrid detector is superior in performance to either an electron multiplier or a conventional scintillator detector. It is designed to detect a beam of positive ions or negative ions focused to a line image or to a spot at the focal plane of a mass analyzer. Background counting noise is less than 1 count/min when the detector is used in either the positive ion or negative ion detecting mode. Mass-resolved ions are post-accelerated through an electric field to give them an additional kinetic energy of 10 keV before they strike the ion-to-electron converter in the electron multiplier. This allows low-energy ions to be detected with an efficiency close to unity, over a range of ion masses from 1 amu up to several hundred amu. Randomly spaced pulses can be counted at rates from 0 Hz to 10 MHz. The design of the detector and its operating characteristics are discussed.


Glyphosate Use, Toxicity and Occurrence in Food.
Glyphosate is a systemic, broad-spectrum and post-emergent herbicide. The use of glyphosate has grown in the last decades, and it is currently the most used herbicide worldwide. The rise of glyphosate consumption over the years also brought an increased concern about its possible toxicity and consequences for human health. However, a scientific community consensus does not exist at the present time, and glyphosate's safety and health consequences are controversial. Since glyphosate is mainly applied in fields and can persist several months in the soil, concerns have been raised about the impact that its presence in food can cause in humans. Therefore, this work aims to review the glyphosate use, toxicity and occurrence in diverse food samples, which, in certain cases, occurs at violative levels. The incidence of glyphosate at levels above those legally allowed and the suspected toxic effects of this compound raise awareness regarding public health.


Selecting the Antigen.
The classical method for generating polyclonal or monoclonal antibodies relies on the in vivo humoral response of animals. Here we describe the factors that antigens can have that might influence the strength and quality of an antibody response. This introduction is divided into three sections: (1) an overview of immunogenicity, (2) choosing the best form for the immunogen, and (3) methods for modifying antigens to make them more immunogenic.


SH479, a Betulinic Acid Derivative, Ameliorates Experimental Autoimmune Encephalomyelitis by Regulating the T Helper 17/Regulatory T Cell Balance.
CD4+ T helper cells, especially T helper 17 (TH17) cells, combined with immune regulatory network dysfunction, play key roles in autoimmune diseases including multiple sclerosis (MS). Betulinic acid (BA), a natural pentacyclic triterpenoid, has been reported to be involved in anti-inflammation, in particular having an inhibitory effect on proinflammatory cytokine interleukin 17 (IL-17) and interferon-γ (IFN-γ) production. In this study, we screened BA derivatives and found a BA derivative, SH479, that had a greater inhibitory effect on TH17 differentiation. Our further analysis showed that SH479 had a greater inhibitory effect on TH17 and TH1, and a more stimulatory effect on regulatory T (Treg) cells. To evaluate the effects of SH479 on autoimmune diseases in vivo, we employed the extensively used MS mouse model experimental autoimmune encephalomyelitis (EAE). Our results showed that SH479 ameliorated clinical and histologic signs of EAE in both prevention and therapeutic protocols by regulating the TH17/Treg balance. SH479 dose-dependently reduced splenic lymphocyte proinflammatory factors and increased anti-inflammatory factors. Moreover, SH479 specifically inhibited splenic lymphocyte viability from EAE mice but not normal splenic lymphocyte viability. At the molecular level, SH479 inhibited TH17 differentiation by regulating signal transducer and activator of transcription-3 (STAT3) phosphorylation, DNA binding activity, and recruitment to the Il-17a promoter in CD4+ T cells. Furthermore, SH479 promoted the STAT5 signaling pathway and inhibited the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway. Together, our data demonstrated that SH479 ameliorated EAE by regulating the TH17/Treg balance through inhibiting the STAT3 and NF-κB pathways while activating the STAT5 pathway, suggesting that SH479 is a potential novel drug candidate for autoimmune diseases including MS.


Enzymuria in aminoglycoside-induced kidney damage. Comparative study of gentamicin, amikacin, sisomicin and netilmicin.
Forty-one patients with urinary tract infections were randomly assigned to receive for six days gentamicin, amikacin, sisomicin or netilmicin. The dose for each patient was calculated according to creatinine clearance and lean body mass in order to avoid overdosages. Urinary enzymes (alpha-glucosidase, gamma-glutamyltranspeptidase and muramidase), serum creatinine and creatinine clearance, proteinuria and urinary sediment were evaluated for nephrotoxicity. None of the patients developed nephrotoxicity, but urinary enzymes rose significantly in all. The statistical analysis of enzymuria during the treatment permitted the definition of a rank order of the nephrotoxic potential of the aminoglycosides studied.


Translational control of gene expression noise and its relationship to ageing in yeast.
Gene expression noise influences organism evolution and fitness but is poorly understood. There is increasing evidence that the functional roles of components of the translation machinery influence noise intensity. In addition, modulation of the activities of at least some of these same components affects the replicative lifespan of a broad spectrum of organisms. In a novel comparative approach, we modulate the activities of the translation initiation factors eIFG1 and eIF4G2, both of which are involved in the process of recruiting ribosomal 43S preinitiation complexes to the 5' end of eukaryotic mRNAs. We show that tagging of the cell wall using a fluorescent dye allows us to follow gene expression noise as different yeast strains progress through successive cycles of replicative ageing. This procedure reveals a relationship between global protein synthesis rate and gene expression noise (cell-to-cell heterogeneity), which is accompanied by a parallel correlation between gene expression noise and the replicative age of mother cells. An alternative approach, based on microfluidics, confirms the interdependence between protein synthesis rate, gene expression noise and ageing. We additionally show that it is important to characterize the influence of the design of the microfluidic device on the nutritional state of the cells during such experiments. Analysis of the noise data derived from flow cytometry and fluorescence microscopy measurements indicates that both the intrinsic and the extrinsic noise components increase as a function of ageing.


Modification of enteral diets in inflammatory bowel disease.
The provision of food is thought to promote the maintenance of gut integrity. Nutrients are able to elicit and affect both systemic and mucosal immune responses. Enteral diet therapy has long been known to be efficacious in inflammatory bowel disease (IBD), particularly in childhood Crohn's disease. However, the mechanisms of action of these diets are not clear. Nutritional repletion, direct effects on the gut mucosa or decreased intestinal permeability have all been postulated as being important in nutritional therapy. There is some evidence that the enteral diet has a direct effect on the gut mucosa by reducing cytokine production and the accompanying inflammation, thus leading to decreased intestinal permeability. Modifications of enteral diet composition have been evaluated in many studies. Such modifications include fat and/or protein content and the addition of bioactive peptides. The fatty acid composition of the enteral diet seems to have a much greater impact on its efficacy than modification of the N source. As specific fatty acids are precursors of inflammatory mediators derived from arachidonic acid, the reduction in these components may be beneficial in nutritional therapy for IBD. Addition of bioactive peptides to enteral diet formulas may also have a role; such peptides may have specific growth factor or anti-inflammatory actions. There is still much work to be done to define disease-specific enteral diet formulas that are effective as therapies for both Crohn's disease and ulcerative colitis.


Voltage-dependent conformational changes in human Ca(2+)- and voltage-activated K(+) channel, revealed by voltage-clamp fluorometry.
Large conductance voltage- and Ca(2+)-activated K(+) (BK(Ca)) channels regulate important physiological processes such as neurotransmitter release and vascular tone. BK(Ca) channels possess a voltage sensor mainly represented by the S4 transmembrane domain. Changes in membrane potential displace the voltage sensor, producing a conformational change that leads to channel opening. By site-directed fluorescent labeling of residues in the S3-S4 region and by using voltage clamp fluorometry, we have resolved the conformational changes the channel undergoes during activation. The voltage dependence of these conformational changes (detected as changes in fluorescence emission, fluorescence vs. voltage curves) always preceded the channel activation curves, as expected for protein rearrangements associated to the movement of the voltage sensor. Extremely slow conformational changes were revealed by fluorescent labeling of position 202, elicited by a mutual interaction of the fluorophore with the adjacent tryptophan 203.


Increased waiting time for liver transplantation results in higher mortality.
Waiting time to liver transplantation (LTx) has dramatically lengthened, but the proportion of candidates who die awaiting transplantation has not increased. We evaluated whether longer waiting time for LTx candidates increases mortality. A cohort of candidates listed for LTx between 1990 and 1993 by three large transplantation programs was followed for 2 years. The exposure measure was ABO blood type, which is not inherently related to outcome, but is a major determinant of waiting time. The main outcome measure was 2-year mortality, as evaluated by logistic regression analysis that controlled for differences in clinical status at the time of evaluation for LTx. The 308 candidates with type O blood waited longer for LTx (median 109 days) than the 399 candidates with other blood types (median 58 days) (P=0.001). Candidates listed for LTx with type O blood had better clinical status at evaluation, but then had higher pretransplantation mortality (13.3%) than other candidates (7.0%) (P=0.005). Blood group O candidates had higher 2-year mortality (26.6%) than other candidates (22.1%), which on multivariate analysis resulted in a mortality odds ratio at 2 years of 1.52 (95% confidence interval=1.04-2.23). With the difference in median waiting time between blood groups increasing from 44 days in the first year to 108 days in the third year, the 2-year mortality odds ratio also rose from 0.94 to 1.97. When compared with LTx candidates with other blood types, blood type O candidates have longer waiting times and higher pretransplantation mortality, which results in higher 2-year mortality.


HIV seroconversion illness.
HIV seroconversion is associated with characteristic clinical findings which reflect the first interaction of HIV with the host immune system. Recognition of the characteristic clinical illness associated with HIV seroconversion helps to establish an early diagnosis and obviates the need for further investigation or empiric therapy. It may also afford a unique opportunity to institute counselling to prevent the further spread of HIV in the community.


Thermoresponsive fluconazole gels for topical delivery: rheological and mechanical properties, in vitro drug release and anti-fungal efficacy.
The aim of this study was to develop thermosensitive gels using poloxamers for topical delivery of fluconazole (FLZ). Eight different formulations containing 1% FLZ in poloxamer and a particular co-solvent (propylene glycol (PG) or Transcutol-P) of various concentrations were prepared. The gels were characterized for transition temperatures, rheological and mechanical properties. FLZ permeability and antifungal effect of the gels were also evaluated. Except for one formulation, all gels exhibited thermosensitive property, i.e. transformed from Newtonian (liquid-like) behavior at 20 °C to non-Newtonian (gel-like) behavior at 37 °C. Transcutol-P increased the transition temperature of the formulations, while the opposite effect was observed for PG. At 37 °C, formulations with high poloxamer concentrations (17%) resulted in high viscosity, compressibility and hardness. Formulations containing 17% poloxamer and 20% Transcutol-P and 10% PG, respectively, exhibited high adhesiveness. No significant differences in the in vitro antifungal activity of FLZ were observed among the formulations suggesting that the gel vehicles did not influence the biological effect of FLZ. FLZ permeability decreased with increasing poloxamer concentration. Formulations containing 17% poloxamer and 20% Transcutol-P and 10% PG seemed to be promising in situ gelling systems for the topical delivery of FLZ.


[Evaluation by proton magnetic resonance spectroscopy of carotid-cavernous fistula with cortical venous drainage].
Cases with carotid-cavernous fistula (CCF) associated with cortical venous drainage through the sylvian veins are rather rare. However, such cases involve risk for subarachnoid hemorrhage, subcortical hemorrhage and venous infarction due to venous hypertension in the brain. Even without these symptoms, CCF under these conditions provokes disturbance in cerebral metabolism. We report two cases of CCF associated with cortical venous drainage evaluated by proton magnetic resonance spectroscopy (1H-MRS). Case 1: A 56 year-old female suffered from a right CCF associated with cortical venous drainage through the sylvian veins after trauma. Before embolization with a detachable balloon catheter, the ratios of N-acetyl-aspartate (NAA)/Choline (Cho) and NAA/Creatine (Cr) in 1H-MRS on the right temporo-basal ganglia were lower than those in normal volunteers. After curative balloon embolization of the CCF, serial 1H-MRS still demonstrated laterality (NAA/Cho and NAA/Cr ratios on the right side were lower than those on the left). Six months after embolization, these ratios on the right became closer to those on the left. Case 2: A 48 year-old female suffered from spontaneous CCF associated with cortical venous drainage. Before embolization of the CCF, ratios of NAA/Cho and NAA/Cr on the temporo-basal ganglia of the drainage side (left side) were lower than those on the contralateral side. After partial embolization of the CCF, which caused the angiographical disappearance of the cortical venous drainage, NAA/Cho and NAA/Cr ratios on the affected side increased to almost the same levels as those on the contralateral side. We consider that 1H-MRS is noninvasive and is a useful method to generate data evaluation of affected brain metabolism by venous reflux in cases of CCF associated with cortical venous drainage.


Is the transvenous extraction of cardioverter-defibrillator leads more hazardous than that of pacemaker leads?
Leads used for low-voltage and high-voltage therapy delivered by implantable cardioverter-defibrillators (ICD) differ from low-voltage pacemaker (PM) leads in their diameter and complexity of structure. Although there are reports showing that the extraction of ICD leads may be hazardous, due to firm adhesions to the vascular and chamber walls of high-voltage therapy coils, clinical evidence suggests that such procedures are safe. To compare the efficacy and safety of transvenous extraction of ICD and PM leads in patients enrolled in a single tertiary centre. We compared the results of lead extraction procedures in 345 patients with PM and in 79 patients submitted for the lead removal including at least one ICD lead. We analysed ingrown leads i.e. over 12 month-old PM leads and over 6 month-old ICD leads, which were removed using Cook's device. Patients in the two groups differed significantly in age and gender. The ICD systems were significantly younger, less complex (fewer leads per patient) and presented higher efficacy of extraction and fewer technical difficulties. The number of major complications was similar to the encountered during extraction of PM leads. However, minor complications were significantly more frequent in the ICD group. 1. Extraction of ICD and PM leads is associated with a similar risk for developing major complications, however minor complications are more often during extraction of ICD leads. 2. A larger number of double coil leads may be the cause of complications despite a shorter time period elapsing from ICD implantation. 3. A probable cause of complications during ICD lead extraction is the pronounced growth of the connective tissue around the coils. However, further studies are required to clarify this phenomenon. 4. The success rate of ICD leads extraction using our own surgical technique is similar to that reported by other investigators using laser systems.


The possible route of introduction of bluetongue virus serotype 3 into Sicily by windborne transportation of infected Culicoides spp.
In October 2017, the first outbreak of bluetongue virus serotype 3 (BTV-3) began in Italy, specifically in western Sicily. The route of entrance remains unclear, although since 2016 the same strain had been circulating only 150 km away, on the Tunisian peninsula of Cape Bon. The present analysis assessed the feasibility that wind could have carried BTV-3-infected Culicoides spp. from Tunisia to Sicily. An advection-deposition-survival (ADS) model was used to estimate when and where Culicoides spp. were likely to be introduced prior to the first BTV-3 report in Italy. Additionally, the Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) model was used to support ADS outputs. The modelling suggests that during September 2017, strong wind currents and suitable climatic conditions could have allowed the transportation of Culicoides spp. from BTV-3-infected areas in Tunisia into Sicily. ADS simulations suggest that particles could have reached the province of Trapani in western Sicily on 2 and 12 September. These simulations suggest the feasibility of aerial transportation of infected Culicoides spp. from Tunisia into Sicily. They demonstrate the suitability of the ADS model for retrospective studies of long-range transportation of insects across large water bodies, which may enhance the early detection of vectorial disease introduction in a region.


Detection by rapd of genetic alterations in vitro: amplification and conservation conditions of DNA extracts.
The technique of random amplified polymorphic DNA (RAPD) permits the study of genetic ecotoxicology without having to face the problem posed by the high number of chromosomes generally presented by the fish species used in traditional methods. Additionally, the use of in vitro systems allows the study of a large number of samples in environmental-risk assessment. Good standardization of the parameters involved in the RAPD reaction, such as primer concentration and the DNA template used and its integrity, is fundamental for obtaining reliable and repeatable results. This is especially important when the differences in DNA fingerprints between control cells and cells exposed to genotoxic agents are interpreted as toxic-dependent alterations. The use of more than one primer increases sensitivity in the detection of such differences, provided the amplification is carried out under optimum conditions. This article demonstrates that the conditions established for certain primers in previous studies can be acceptable for others, independent of the complexity of band patterns generated. Furthermore, the integrity of the DNA is shown to be stable for several months in the genomic extracts stored at 4 degrees C, which to a large extent facilitates the application of this methodology.


The Battle is On: Factors that Motivate People to Combat Anti-Vaccine Misinformation.
This study proposes a theory-driven model to concurrently examine the cognitive and emotional factors that motivate vaccine supporters to combat erroneous online anti-vaccination information. The model was tested using data from a web survey of 599 vaccination supporters in the United States. The vaccine supporters reported greater support for government regulation of misinformation when they perceived greater susceptibility among the general public to the influence of misinformation. Surprisingly, the perceived severity of the influence was inversely related to respondents' intention to correct misinformation. In addition, perceived susceptibility to the influence of anti-vaccine misinformation and perceived severity of its influence on others induced negative emotions that included anticipated guilt and anger. The negative emotions in turn motivate vaccine supporters to attitudinally support government's media restriction or behaviorally correct the online misinformation.


Immune regulatory cytokines in the milk of lactating women from farming and urban environments.
Children living on farms have fewer allergies. It is unclear whether breastfeeding in different environments contributes to preventing allergies by exposing offspring to different cytokines that can modulate immune responses. The aim of this study was to quantify and compare levels of Transforming Growth Factor-beta1 (TGF-beta1) and Interleukin-10 (IL-10) in the colostrum and mature milk of mothers living in towns at sea level (references) and mothers on farms. Milk samples were collected within 3 days postpartum (colostrum) and at the first month of the baby's life (mature milk). Sixty-nine reference mothers and 45 farm mothers participated in the study. TGF-beta1 concentrations were significantly higher both in the colostrum (p < 0.05) and in mature milk (p < 0.05) of farm mothers. In the reference mothers, a significant decrease in TGF-beta1 concentrations was observed between colostrum (650, range 0-8000 pg/ml) and mature milk (250, range 0-8000 pg/ml) (p < 0.05). In farm mothers, TGF-beta1 concentrations were 1102 pg/ml (range 0-14,500) in colostrum and remained high in mature milk (821 pg/ml, range 0-14,650). IL-10 concentrations were higher in the mature milk of farm mothers (p < 0.05). No significant differences in IL-10 were observed between colostrum and mature milk in the control group (15 pg/ml, range 0-1800, and 0 pg/ml, range 0-230) or in farm mothers (9.5 pg/ml, range 0-1775, and 14.2 pg/ml, range 0-930), respectively. Exposure to a farm environment is associated with higher concentrations of TGF-beta1 and IL-10 in breast milk when compared to exposure to an urban environment. Higher cytokine concentrations in breast milk may influence early modulation of the development of an immune response, leading to a reduced prevalence of allergy-related diseases in farm children.


[Influence of age, growth potential and functionality in the management of bone tumours in children : example of the rotationplasty].
Malignant bone tumours in children are rare diseases whose survival rate has progressively improved in recent years thanks to advances in pharmacology, surgery and radiotherapy. We present the particular case of a very young child with Ewing's sarcoma of the thigh. We discuss the influence of this young age on short and long term care. Limb skeletal immaturity and the residual growth potential need to be integrated to define the safest oncological and functional strategy. For this purpose, we summarize the different possible reconstructive options. We describe the Van Nes rotationplasty that was proposed in our patient's case and we will detail the issues in terms of functional well-being and self-image.


Enhanced H Transport Capacity and ATP Hydrolysis Activity of the Tonoplast H-ATPase after NaCl Adaptation.
Tonoplast enriched membrane vesicle fractions were isolated from unadapted and NaCl (428 millimolar) adapted tobacco cells (Nicotiana tabacum L. var Wisconsin 38). Polypeptides from the tonoplast enriched vesicle fractions were separated by SDS-PAGE and analyzed by Western blots using polyclonal antibodies to the 70 kilodalton subunit of the red beet tonoplast H(+)-ATPase. These antibodies cross-reacted exclusively to a tobacco polypeptide of an apparent molecular weight of 69 kilodaltons. The antibodies inhibited ATP-dependent, NO(3) (-) sensitive H(+) transport into vesicles in tonoplast enriched membrane fractions from both unadapted and NaCl adapted cells. The relative H(+) transport capacity per unit of 69 kilodalton subunit of the tonoplast ATPase of vesicles from NaCl adapted cells was fourfold greater than that observed for vesicles from unadapted cells. The increase in specific H(+) transport capacity after adaptation was also observed for ATP hydrolysis.


HDAC7 promotes NSCLC proliferation and metastasis via stabilization by deubiquitinase USP10 and activation of β-catenin-FGF18 pathway.
Histone deacetylases (HDACs) play crucial roles in cancers, but the role and mechanism of HDAC7 in NSCLC have not been fully understood. A total of 319 patients with non-small cell lung cancer (NSCLC) who underwent surgery were enrolled in this study. Immunohistochemistry and Kaplan-Meier survival analysis were performed to investigate the relationship between HDAC7, fibroblast growth factor 18 (FGF18) expression, and clinicopathologic characteristics. Cell functional experiments were implemented both in vivo and in vitro to investigate the effects on NSCLC cell proliferation and metastasis. Recombinant lentivirus-meditated in vivo gene overexpression or knockdown, real-time polymerase chain reaction (PCR), western blotting, and coimmunoprecipitation assays were applied to clarify the underlying molecular mechanism of HDAC7 in promoting NSCLC progression. The elevated expression of HDAC7 or FGF18 was positively correlated with poor prognosis, tumor-node-metastasis (TNM) stage, and tumor differentiation of NSCLC patients. NSCLC patients with co-expressed HDAC7 and FGF18 suffered the worst prognosis. HDAC7 overexpression promoted NSCLC proliferation and metastasis by upregulating FGF18. Conversely, overexpression of FGF18 reversed the attenuated ability in tumor growth and metastasis mediated by downregulating HDAC7. In terms of mechanism, our results suggested that the interaction of HDAC7 with β-catenin caused decreased β-catenin acetylation level at Lys49 and decreased phosphorylation level at Ser45. As a consequence, the HDAC7-mediated posttranslational modification of β-catenin facilitated nuclear transfer and activated FGF18 expression via binding to TCF4. Furthermore, deubiquitinase USP10 interacted with and stabilized HDAC7. The suppression of USP10 significantly accelerated the degradation of HDAC7 and weakened NSCLC growth and migration. Our findings reveal that HDAC7 promotes NSCLC progression through being stabilized by USP10 and activating the β-catenin-FGF18 pathway. Targeting this novel pathway may be a promising strategy for further developments in NSCLC therapy.


The epigenetic alterations of human sperm cells caused by heroin use disorder.
The molecular mechanisms of drug use on sexual health are largely unknown. We investigated, the relationship between heroin use disorder and epigenetic factors influencing histone acetylation in sperm cells. The volunteers included twenty-four 20- to 50-year-old men with a normal spermogram who did not consume any drugs and twenty-four age- to BMI-matched men who consume only the drug heroin for more than last four months. HDAC1 and HDAC11 mRNA expression levels in spermatozoa and miR-34c-5p and miR-125b-5p expression levels in seminal plasma were measured. The heroin-user group showed significantly increased white blood cell counts and decreased sperm motility and survival rates (8.61 ± 1.73, 21.50 ± 3.11, 69.90 ± 4.69 respectively) as compared to the control group (1.49 ± 0.32, 38.82 ± 3.05, 87.50 ± 0.99 respectively) (p ≤ .001). An increase in DNA fragmentation index (DFI) (heroin-user group: 41.93 ± 6.59% and control group: 10.14 ± 1.43%, p = .003), a change in frequency of HDAC1 (heroin-user group: 1.69 ± 0.55 and control group: 0.45 ± 0.14, p = .045) and HDAC11 (heroin-user group: 0.29 ± 0.13 and control group: 2.36 ± 0.76, p = .019) in spermatozoa and a significant decrease in seminal miR-125b-5p abundance (heroin-user group: 0.37 ± 0.11 and control group: 1.59 ± 0.47, p = .028) were reported in heroin consumers. Heroin use can lead to male infertility by causing leukocytospermia, asthenozoospermia, DFI elevation in sperm cells and alterations in seminal RNA profile.


Incidence and risk factors of dementia in a defined elderly Japanese population: the Hisayama study.
Vascular dementia (VaD) is more common than Alzheimer's disease (AD) in Japan, while AD is much more frequent in Western countries. The Hisayama study, an epidemiological study on dementia in a Japanese elderly population aged 65 years or older, has shown that the prevalence of VaD decreased for men, while AD remained unchanged in both sexes during a 7-year follow-up period (1985-1992). Decreased prevalence of VaD seems to be due to decreased incidence of stroke in recent years, resulting from the management of hypertension. The age-adjusted incidence of total dementia was approximately 20 per 1000 person-years in either sex; VaD was more frequent for men and AD for women. Risk factors for VaD were age, hypertension, previous stroke, and alcohol consumption, while age was only a significant risk factor for AD. Although the Hisayama study failed to demonstrate the relationship of vascular factors to AD, the previously reported studies suggest that either hypertension per se or blood pressure changes appear to partially participate in the pathogenesis of AD.


The IgA nephropathy Biobank. An important starting point for the genetic dissection of a complex trait.
IgA nephropathy (IgAN) or Berger's disease, is the most common glomerulonephritis in the world diagnosed in renal biopsied patients. The involvement of genetic factors in the pathogenesis of the IgAN is evidenced by ethnic and geographic variations in prevalence, familial clustering in isolated populations, familial aggregation and by the identification of a genetic linkage to locus IGAN1 mapped on 6q22-23. This study seems to imply a single major locus, but the hypothesis of multiple interacting loci or genetic heterogeneity cannot be ruled out. The organization of a multi-centre Biobank for the collection of biological samples and clinical data from IgAN patients and relatives is an important starting point for the identification of the disease susceptibility genes. The IgAN Consortium organized a Biobank, recruiting IgAN patients and relatives following a common protocol. A website was constructed to allow scientific information to be shared between partners and to divulge obtained data (URL: http://www.igan.net). The electronic database, the core of the website includes data concerning the subjects enrolled. A search page gives open access to the database and allows groups of patients to be selected according to their clinical characteristics. DNA samples of IgAN patients and relatives belonging to 72 multiplex extended pedigrees were collected. Moreover, 159 trios (sons/daughters affected and healthy parents), 1068 patients with biopsy-proven IgAN and 1040 healthy subjects were included in the IgAN Consortium Biobank. Some valuable and statistically productive genetic studies have been launched within the 5th Framework Programme 1998-2002 of the European project No. QLG1-2000-00464 and preliminary data have been published in "Technology Marketplace" website: http://www.cordis.lu/marketplace. The first world IgAN Biobank with a readily accessible database has been constituted. The knowledge gained from the study of Mendelian diseases has shown that the genetic dissection of a complex trait is more powerful when combined linkage-based, association-based, and sequence-based approaches are performed. This Biobank continuously expanded contains a sample size of adequately matched IgAN patients and healthy subjects, extended multiplex pedigrees, parent-child trios, thus permitting the combined genetic approaches with collaborative studies.


Cholestatic features in focal nodular hyperplasia of the liver.
Twenty specimens of focal nodular hyperplasia were studied with special attention to the histological features of chronic cholestasis (grouped under the headings of cholestasis, cholate-stasis and signs of ductular reabsorption). In all specimens, evidence was found for one or more features of cholestasis and cholate-stasis. Signs of ductular reabsorption were less constant, and apparently varied according to the developmental stage of the lesion. The cholestatic features emphasize the bile secretory capacity of the lesional parenchyma, and are apparently due to the lack of real bile ducts in the portal tract equivalents of the lesional tissue. Evidence is presented that the "ductular component" in FNH is not due to proliferation of pre-existing ductules, but rather derives from ductular metaplasia of liver cell plates in zone 1 equivalents. This metaplastic development of a ductular network may serve the function of reabsorbing the biliary constituents produced by the lesional parenchyma, leading to periductular inflammation and progressive fibrosis, thus producing an equivalent of biliary fibrosis and biliary cirrhosis.


Race, genes and preterm delivery.
High rates of preterm delivery (PTD) among African Americans are the leading cause of excess infant mortality among African Americans. Failure to fully explain racial disparity in PTD has led to speculation that genetic factors might contribute to this disparity. Current evidence suggests that genetic factors contribute to PTD, but this does not imply that genetic factors contribute to racial disparity in PTD. Environmental factors clearly contribute to PTD. Many of these factors acting over a women's life prior to pregnancy disproportionately affect African Americans and contribute significantly to racial disparity in PTD. Thus, inferring genetic contribution to racial disparity in PTD by attempting to control for environmental factors measured at a single point in time is flawed. There is emerging evidence of gene-environment interactions for PTD, some of which disproportionately affect African Americans. There is also evidence of racial differences in the prevalence of polymorphisms potentially related to PTD. However, to date there is no direct evidence that these differences contribute significantly to racial disparity in PTD. Given the complexity of polygenic conditions such as PTD, the possibility of any single gene contributing substantially to racial disparity in PTD seems remote.


[Acupuncture, animal magnetism, and French medical orthodoxy (1780-1830)].
This article analyses why the French phenomenon of acupuncture was confined to the 1810s-1820s. It argues that the French medical orthodoxy played a decisive role. First, we recount the history of the French reception of Japanese acupuncture from the late 17th century to the 1820s. Second, we go back to the animal magnetism trial to find some explanatory tools for the decline of French acupuncture. Third, we show that the oppositions to both therapies were not mere juxtapositions, but due to the growing strength of medical orthodoxy. Finally, we suggest a model of analysis of the French medical orthodoxy of the early 19th century through a set of multidimensional oppositions: anthropological (imagination/reason), epistemological (to heal/to explain), therapeutic (drug/fluid), nosological (organic disease/functional disease), and lastly, economic, moral and political oppositions (doctor/charlatan).


Isolated lymphocytic infiltration of pituitary stalk preceding the diagnosis of germinoma in 2 prepubertal children treated with growth hormone.
We report the clinical course of 2 patients with central diabetes insipidus and evolving to panyhypopituitarism which prompted the diagnosis of an isolated pituitary stalk thickening (PST). In both patients, all etiological investigations were normal and the first biopsy revealed an isolated lymphocytic infiltrate with no sign of malignancy. Close clinical follow-up accompanied by serial brain MRIs was proposed to determine a precise diagnosis and for early detection and treatment of neoplastic disease. In our first case, the diagnosis of germinoma was made 9 months after the PST diagnosis owing to tumor progression. In the second case, the time course was even longer with the diagnosis of germinoma 6 years following initial presentation. In these cases, it is speculated that the lymphocytic infiltrates represent the first sign of a host reaction to an occult germinoma. To our knowledge, this is the third case reported of lymphocytic infiltrates preceding a germinoma in a prepubertal girl, and the only case reported in a prepubertal boy. These cases underline the difficulties in establishing the diagnosis of germinoma in a patient with isolated PST.


Diet-induced obesity in the selenocysteine lyase knockout mouse.
Selenocysteine lyase (Scly) mediates selenocysteine decomposition. It was previously demonstrated that, upon adequate caloric intake (12% kcal fat) and selenium deficiency, disruption of Scly in mice leads to development of metabolic syndrome. In this study, we investigate the effect of a high-fat (45% kcal) selenium-adequate diet in Scly knockout (KO) mice on development of metabolic syndrome. Involvement of selenoproteins in energy metabolism after Scly disruption was also examined in vitro in the murine hepatoma cell line, Hepa1-6, following palmitate treatment. Scly KO mice were more susceptible to diet-induced obesity than their wild-type counterparts after feeding a high-fat selenium-adequate diet. Scly KO mice had aggravated hyperinsulinemia, hypercholesterolemia, glucose, and insulin intolerance, but unchanged inflammatory cytokines and expression of most selenoproteins, except increased serum selenoprotein P (Sepp1). Scly KO mice also exhibited enhanced hepatic levels of pyruvate and enzymes involved in the regulation of pyruvate cycling, such as pyruvate carboxylase (Pcx) and pyruvate dehydrogenase (Pdh). However, in vitro silencing of Scly in Hepa1-6 cells led to diminished Sepp1 expression, and concomitant palmitate treatment decreased Pdh expression. The role of selenium in lipid metabolism is recognized, but specific selenium-dependent mechanisms leading to obesity are unclear. This study uncovers that Scly has a remarkable effect on obesity and metabolic syndrome development triggered by high-fat exposure, independent of the expression of most selenoproteins. Diet-induced obesity in Scly KO mice is aggravated, with effects on pyruvate levels and consequent activation of energy metabolism independent of selenoprotein levels.


A randomised trial of an eight-week, once weekly primaquine regimen to prevent relapse of plasmodium vivax in Northwest Frontier Province, Pakistan.
Vivax malaria remains a major cause of morbidity in the subtropics. To undermine the stability of the disease, drugs are required that prevent relapse and provide reservoir reduction. A 14-day course of primaquine (PQ) is effective but cannot safely be used in routine practice because of its interaction with glucose-6-phosphate dehydrogenase (G6PD) deficiency for which testing is seldom available. Safe and effective use of PQ without the need for G6PD testing would be ideal. The efficacy and safety of an 8-week, once weekly PQ regimen was compared with current standard treatment (chloroquine alone) and a 14-day PQ regimen. 200 microscopically confirmed Plasmodium vivax patients were randomly assigned to either once weekly 8-week PQ (0.75 mg/kg/week), once weekly 8-week placebo, or 14-day PQ (0.5mg/kg/day) in North West Frontier Province, Pakistan. All patients were treated with a standard chloroquine dose and tested for G6PD deficiency. Deficient patients were assigned to the 8-week PQ group. Failure was defined as any subsequent episode of vivax malaria over 11 months of observation. There were 22/71 (31.0%) failures in the placebo group and 1/55 (1.8%) and 4/75 (5.1%) failures in the 14-day and 8-week PQ groups, respectively. Adjusted odds ratios were: for 8-week PQ vs. placebo-0.05 (95%CI: 0.01-0.2, p<0.001) and for 14-day PQ vs. placebo-0.01 (95%CI: 0.002-0.1, p<0.001). Restricted analysis allowing for a post-treatment prophylactic effect confirmed that the 8-week regimen was superior to current treatment. Only one G6PD deficient patient presented. There were no serious adverse events. A practical radical treatment for vivax malaria is essential for control and elimination of the disease. The 8-week PQ course is more effective at preventing relapse than current treatment with chloroquine alone. Widespread use of the 8-week regimen could make an important contribution to reservoir reduction or regional elimination where G6PD testing is not available. ClinicalTrials.gov NCT00158587.


Epithelial-Mesenchymal Transition and Metabolic Switching in Cancer: Lessons From Somatic Cell Reprogramming.
Epithelial-mesenchymal transition (EMT) and its critical roles during cancer progression have long been recognized and extensively reviewed. Recent studies on the generation of induced pluripotent stem cells (iPSCs) have established the connections among EMT, energy metabolism, DNA methylation, and histone modification. Since energy metabolism, DNA methylation, and histone modification are important for cancer development and there are common characteristics between cancer cells and stem cells, it is reasonable to identify mechanisms that have been established during both reprogramming and cancer progression. In the current review, we start from a brief review on EMT and related processes during cancer progression, and then switch to the EMT during somatic cell reprogramming. We summarize the connection between EMT and metabolic switch during reprogramming, and further review the involvements of DNA methylation and cell proliferation. The connections between EMT and mesenchymal-epithelial transition (MET) and cellular aspects including DNA methylation, histone modification and energy metabolism may provide potential new targets for cancer diagnosis and treatment.


Effect of Gabaculine on the Synthesis of Heme and Cytochrome f in Etiolated Wheat Seedlings.
The effect of gabaculine (3-amino 2,3-dihydrobenzoic acid), an inhibitor of tetrapyrrole synthesis, on the accumulation of heme and cytochrome f in etiolated wheat (Triticum aestivum var Mardler) seedlings has been examined. Gabaculine treatment resulted in decreased amounts of heme and of holocytochrome f detected spectroscopically and by peroxidase activity after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amount of the cytochrome f polypeptide detected immunochemically on Western blots was much less affected by gabaculine treatment, indicating that apocytochrome f synthesis was not tightly coupled to heme availability. Gabaculine treatment did not affect the size of the cytochrome f polypeptide, indicating that heme addition is not required for proteolytic removal of the presequence.


ABA Uptake in Source and Sink Tissues of Sugar Beet.
The mode of abscisic acid (ABA) uptake was studied in excised leaf and root tissue discs of sugar beet (Beta vulgaris L.). Discs were incubated in buffered medium that contained 1 mm CaCl(2) and [(14)C]ABA. The sensitivity of ABA uptake to metabolic inhibitors and temperature indicated that the ABA transport system had an energy-dependent component. Energy-dependent uptake was greater in leaf than in root tissue (70% and 50%, respectively). Energy-dependent uptake by both tissues and passive uptake by root tissues were highly pH dependent. Maximal uptake was observed at pH 5.5. Leaf tissue incubated in the dark showed a 50% reduction of uptake as compared with tissue under light. The decrease was due to reduced passive uptake.The results suggest that ABA moves across membranes as the undissociated lipophilic species. As a weak acid, ABA would dissociate and accumulate in the more alkaline compartment. Therefore, the distribution of ABA within the tissue is regulated by the pH differential between any two compartments. Although diffusion may be the predominant form of transport, the uptake of ABA is dependent on metabolic energy for the establishment of a pH gradient across the membrane.


Mutations in MBOAT7, Encoding Lysophosphatidylinositol Acyltransferase I, Lead to Intellectual Disability Accompanied by Epilepsy and Autistic Features.
The risk of epilepsy among individuals with intellectual disability (ID) is approximately ten times that of the general population. From a cohort of >5,000 families affected by neurodevelopmental disorders, we identified six consanguineous families harboring homozygous inactivating variants in MBOAT7, encoding lysophosphatidylinositol acyltransferase (LPIAT1). Subjects presented with ID frequently accompanied by epilepsy and autistic features. LPIAT1 is a membrane-bound phospholipid-remodeling enzyme that transfers arachidonic acid (AA) to lysophosphatidylinositol to produce AA-containing phosphatidylinositol. This study suggests a role for AA-containing phosphatidylinositols in the development of ID accompanied by epilepsy and autistic features.


Visual performance and optical quality with soft lenses in keratoconus patients.
To assess visual performance and ocular aberrations in keratoconic patients using toric soft contact lenses (SCL), rigid-gas-permeable (RGP) contact lenses and spectacle lens correction. Twenty-two keratoconus patients (16 RGP lens wearers and six spectacle wearers) were fitted with toric SCL. Ocular aberrations were measured with and without the patient's habitual RGP lenses and with the SCL in place. In the spectacle wearers, aberrations were measured with and without the SCL. Visual performance (high- and low-contrast visual acuity) was evaluated with the patient's habitual correction and with the SCL. In the RGP lens wearers both the habitual lenses and the toric SCL significantly reduced coma, trefoil, 3rd-order, 4th-order cylinder and higher-order root-mean-square (RMS) aberrations (p ≤ 0.015). In the spectacle wearers the toric SCL significantly reduced coma, 3rd-order and higher-order RMS aberrations (p ≤ 0.01). The patients' habitual RGP lenses gave better low-contrast acuity (p ≤ 0.006) compared to the toric SCL; however, no significant difference was found between lens types for high-contrast acuity (p = 0.10). In the spectacle wearers no significant differences in visual performance measurements were found between the patients' spectacles and the toric SCL (p ≥ 0.06). The results show that RGP lenses provided superior visual performances and greater reduction of 3rd-order aberrations compared to toric SCL in this group of keratoconic patients. In the spectacle-wearing group, visual performance with the toric SCL was found to be comparable to that measured with spectacles. Nevertheless, with the exception of spherical aberration, the toric SCL were successful in significantly reducing uncorrected higher-order aberrations.


Phosphorylation of cellular proteins in response to treatment with Clostridium difficile toxin B and Clostridium sordellii toxin L.
Toxin B from Clostridium difficile induces typical morphological changes of cultured cells consisting of rounding up and arborization, which are associated with a dramatic disruption of microfilaments. In this study, we show that toxin L, a cytotoxin produced by bacterial strain Clostridium sordellii, has similar effects on cultured cells including the redistribution of F-actin and of the adhesion plaque protein vinculin. It has been assumed that the mechanisms involved in cytopathic effects of toxin B are related to the function of an unidentified component that regulates the organization of the actin cytoskeleton. We demonstrate that the treatment of cultured astrocytes with toxin B or toxin L alters the incorporation of inorganic phosphate into several proteins. Immunoblot analysis revealed that one of these proteins is tropomyosin. Since tropomyosin stabilizes microfilaments and inhibits the severing activity of gelsolin, the toxin-induced phosphorylation may counteract this inhibition resulting in severing of microfilaments and capping of short filaments. A decrease in the radioactivity associated with intermediate filament protein vimentin was also detected using a monoclonal antibody which specifically recognizes a phosphorylated epitope of vimentin. Since vimentin is an in vivo substrate for various protein kinases, these data are in favor of broad effects of these toxins. Direct measurement of protein kinase C in cells exposed to toxin B or to toxin L did not reveal a significant change in protein kinase C activity. Furthermore, treatments with toxins do not increase cAMP levels, suggesting that toxins do not activate protein kinase A. Although further studies are required to determine the primary target site for the clostridial cytotoxin B and L, our results show that they provoke the alteration in the phosphorylation of cellular proteins.


Correlated expression of gfp and Bt cry1Ac gene facilitates quantification of transgenic hybridization between Brassicas.
Gene flow from transgenic oilseed rape (BRASSICA NAPUS) might not be avoidable, thus, it is important to detect and quantify hybridization events with its relatives in real time. Data are presented showing the correlation between genetically linked green fluorescent protein (GFP) with BACILLUS THURINGIENSIS (Bt) CRY1AC gene expression in hybrids formed between transgenic B. NAPUS "Westar" and a wild Chinese accession of wild mustard (B. JUNCEA) and hybridization between transgenic B. NAPUS and a conspecific Chinese landrace oilseed rape. Hybrids were obtained either by spontaneous hybridization in the field or by hand-crossing in a greenhouse. In all cases, transgenic hybrids were selected by GFP fluorescence among seedlings originating from seeds harvested from B. JUNCEA and the Chinese oilseed rape plants. Transgenicity was confirmed by PCR detection of transgenes. GFP fluorescence was easily and rapidly detected in the hybrids under greenhouse and field conditions. Results showed that both GFP fluorescence and Bt protein synthesis decreased as either plant or leaf aged, and GFP fluorescence intensity was closely correlated with Bt protein concentration during the entire vegetative lifetime in hybrids. These findings allow the use of GFP fluorescence as an accurate tool to detect gene-flow in time in the field and to conveniently estimate BT CRY1AC expression in hybrids on-the-plant.


Amphiphilic Graft Copolymers as a Versatile Binder for Various Electrodes of High-Performance Lithium-Ion Batteries.
It is known that grafting one polymer onto another polymer backbone is a powerful strategy capable of combining dual benefits from each parent polymer. Thus amphiphilic graft copolymer precursors (poly(vinylidene difluoride)-graft-poly(tert-butylacrylate) (PVDF-g-PtBA)) have been developed via atom transfer radical polymerization, and demonstrated its outstanding properties as a promising binder for high-performance lithium-ion battery (LIB) by using in situ pyrolytic transformation of PtBA to poly(acrylic acid) segments. In addition to its superior mechanical properties and accommodation capability of volume expansion, the Si anode with PVDF-g-PtBA exhibits the excellent charge and discharge capacities of 2672 and 2958 mAh g(-1) with the capacity retention of 84% after 50 cycles. More meaningfully, the graft copolymer binder shows good operating characteristics in both LiN0.5 M1.5 O4 cathode and neural graphite anode, respectively. By containing such diverse features, a graft copolymer-loaded LiN0.5 M1.5 O4 /Si-NG full cell has been successfully achieved, which delivers energy density as high as 546 Wh kg(-1) with cycle retention of ≈70% after 50 cycles (1 C). For the first time, this work sheds new light on the unique nature of the graft copolymer binders in LIB application, which will provide a practical solution for volume expansion and low efficiency problems, leading to a high-energy-density lithium-ion chemistry.


Optimising long-term athletic development: An investigation of practitioners' knowledge, adherence, practices and challenges.
Long-term athletic development practices have been recommended for the past two decades. However, limited research exists exploring the knowledge and skills required by practitioners to optimise long-term athletic development. Therefore, this study aimed to evaluate the knowledge, adherence, practices, and challenges of practitioners responsible for delivering long-term athletic development. A mixed methods survey was completed by 236 practitioners (e.g., sport coaches, physical education teachers) consisting of four parts; 1) demographics, 2) knowledge, 3) adherence, and 4) practices and challenges. Quantitative and qualitative data were analysed by Friedman's analysis of variance and thematic analyses, respectively. Quantitative findings showed practitioners 1) recognised their responsibility for delivering long-term athletic development, 2) have a familiarity with existing developmental models, and 3) had high adherence, focused upon health and wellbeing, to delivering long-term athletic development. However, practices associated with growth and maturity, monitoring and assessment, and the systematic progression and individualisation of training had lower adherence. Qualitative analysis indicated that practitioner's perceived definitions of athleticism and long-term athletic development were inconsistent, especially according to the psychological components (i.e., confidence). Practitioners' descriptions of their long-term athletic development practices identified two higher order themes; 1) goals, in which long-term athletic development "is for life" and the importance of "an individual centered journey" highlighted as sub-themes; and 2) realities of delivering long-term athletic development, whereby variety in programme delivery, monitoring development and practical challenges were noted as key priorities. Eight practical challenges were identified including governance and priorities, resources, education, early specialization, high training volumes, staff communication, parents and youth motivation. This mixed method survey highlighted a multitude of knowledge, adherence, practices and challenges towards long-term athletic development. These novel findings can help inform policy to optimise long-term athletic development and to support the complex problem of developing a healthier, fitter and more physically active youth population.


Indications for Salivary Gland Radiotherapy.
There is an established role for post-operative radiotherapy in the treatment of benign and malignant salivary gland tumours. For benign disease, the addition of radiotherapy improves local tumour control in cases with incomplete excision, involved surgical margins or multi-focal disease recurrence. After capsule rupture or spillage alone, surveillance should usually be advised. For malignant disease, post-operative radiotherapy is recommended for an advanced tumour stage, high-grade tumour, perineural or lympho-vascular invasion, close or positive resection margins, extra-parotid extension or lymph node involvement. The main benefit is increased loco-regional tumour control, although this may translate into a modest improvement in survival. The possible late side effects of parotid bed irradiation include skin changes, chronic otitis externa, sensorineural hearing loss, osteoradionecrosis and secondary malignancy. Severe complications are rare, but patients should be counselled carefully about the risks. Primary radiotherapy is unlikely to be curative and is reserved to cases in which resection would cause unacceptable functional or cosmetic morbidity or would likely result in subtotal resection (R2) or to patients with distant metastases to gain local tumour control. There are provisional data on the use of charged particle radiotherapy in this setting. Some patients may benefit from synchronous chemotherapy with radiotherapy, but this group is not defined, and data from comparative prospective studies are required before routine clinical use of this treatment.


Bacterial helicases.
Helicases are proteins that use the energy of ATP hydrolysis to open double-stranded DNA, RNA, or RNA-DNA hybrids into two single strands. Based upon sequence analysis, at least 12 helicases exist in Escherichia coli. We know that these proteins play important roles in DNA replication, recombination, repair, and transcription, as well as in RNA processing. Recent crystallographic studies have revealed a highly conserved catalytic core in the helicases, shared with the RecA protein and the F1-ATPase. However, evidence suggests that the functional divergence may be large.


Nurse-physician communication during labor and birth: implications for patient safety.
To describe communication between nurses and physicians during labor within the context of the nurse-managed labor model in community hospitals and its relationship to teamwork and patient safety. Multicenter qualitative study involving focus groups and in-depth interviews. Labor and birth units in 4 Midwestern community hospitals. 54 labor nurses and 38 obstetricians. Focus groups and in-depth interviews were conducted using open-ended questions. Data were analyzed using inductive coding methods to gain understanding from the perspective of those directly involved. Description of interdisciplinary interactions during labor. Nurses and physicians shared the common goal of a healthy mother and baby but did not always agree on methods to achieve that goal. Two clinical situations critical to patient safety (fetal assessment and oxytocin administration) were frequent areas of disagreement and sources of mutual frustration, often leading to less than optimal teamwork. Minimal communication occurred when the mother and fetus are doing well, and this seemed to be purposeful and considered normal. Physicians and nurses had distinct opinions concerning desirable traits of members of the other discipline. Interdisciplinary communication and teamwork could be improved to promote a safer care environment during labor and birth.


Stent placement in hemodialysis access: historical lessons, the state of the art and future directions.
Vascular access stenosis in patients undergoing chronic hemodialysis is a major issue that is associated with increased morbidity, mortality, and cost of medical care. Recent data have emphasized that endovascular stents could be used in the treatment of central as well as peripheral stenotic lesions. In general, a peripheral or central vein lesion that is elastic or recurs within a three-month period after an initially successful balloon angioplasty or a stenosis where surgical revision is not possible are some indications for intravascular stent placement. Recent reports have expanded the role of stents in the management of pseudoaneurysms associated with dialysis access. In this context, the utilization of these devices must take into account a fair comparison with the traditional (surgical) approaches regarding effectiveness as well as costs. This report describes the role of stents in arteriovenous dialysis access. In addition, some of the recent advances in the structure and complicating issues such as stent fracture, migration, and infection, as well cannulation through the stent, are discussed.


Continual representation learning for evolving biomedical bipartite networks.
Many real-world biomedical interactions such as 'gene-disease', 'disease-symptom' and 'drug-target' are modeled as a bipartite network structure. Learning meaningful representations for such networks is a fundamental problem in the research area of Network Representation Learning (NRL). NRL approaches aim to translate the network structure into low-dimensional vector representations that are useful to a variety of biomedical applications. Despite significant advances, the existing approaches still have certain limitations. First, a majority of these approaches do not model the unique topological properties of bipartite networks. Consequently, their straightforward application to the bipartite graphs yields unsatisfactory results. Second, the existing approaches typically learn representations from static networks. This is limiting for the biomedical bipartite networks that evolve at a rapid pace, and thus necessitate the development of approaches that can update the representations in an online fashion. In this research, we propose a novel representation learning approach that accurately preserves the intricate bipartite structure, and efficiently updates the node representations. Specifically, we design a customized autoencoder that captures the proximity relationship between nodes participating in the bipartite bicliques (2 × 2 sub-graph), while preserving both the global and local structures. Moreover, the proposed structure-preserving technique is carefully interleaved with the central tenets of continual machine learning to design an incremental learning strategy that updates the node representations in an online manner. Taken together, the proposed approach produces meaningful representations with high fidelity and computational efficiency. Extensive experiments conducted on several biomedical bipartite networks validate the effectiveness and rationality of the proposed approach.


Evidence for the association between FTO gene variants and vitamin B12 concentrations in an Asian Indian population.
Low vitamin B12 concentrations have been associated with major clinical outcomes, including adiposity, in Indian populations. The Fat mass and obesity-associated gene (FTO) is an established obesity-susceptibility locus; however, it remains unknown whether it influences vitamin B12 status. Hence, we investigated the association of two previously studied FTO polymorphisms with vitamin B12 concentrations and metabolic disease-related outcomes and examined whether these associations were modified by dietary factors and physical activity. A total of 176 individuals with type 2 diabetes, 152 with pre-diabetes, and 220 normal glucose-tolerant individuals were randomly selected from the Chennai Urban Rural Epidemiology Study. Anthropometric, clinical, and biochemical investigations, which included body mass index (BMI), waist circumference, vitamin B12, homocysteine, and folic acid were measured. A validated food frequency questionnaire was used for dietary assessment and self-reported physical activity measures were collected. An unweighted genetic risk score (GRS) was calculated for two FTO single-nucleotide polymorphisms (rs8050136 and rs2388405) by summation of the number of risk alleles for obesity. Interaction analyses were performed by including the interaction terms in the regression model. The GRS was significantly associated with increased BMI (P = 0.009) and risk of obesity (P = 0.023). Individuals carrying more than one risk allele for the GRS had 13.13% lower vitamin B12 concentrations, compared to individuals carrying zero risk alleles (P = 0.018). No associations between the GRS and folic acid and homocysteine concentrations were observed. Furthermore, no statistically significant GRS-diet or GRS-physical activity interactions with vitamin B12, folic acid, homocysteine or metabolic-disease outcomes were observed. The study shows for the first time that a genetic risk score using two FTO SNPs is associated with lower vitamin B12 concentrations; however, we did not identify any evidence for the influence of lifestyle factors on this association. Further replication studies in larger cohorts are warranted to investigate the association between the GRS and vitamin B12 concentrations.


Dental abnormalities in children after chemotherapy treatment for acute lymphoid leukemia.
The frequency of dental abnormalities, such as delayed dental development, microdontia, hypoplasia, agenesis, V-shaped root and shortened root was evaluated in 76 acute lymphoblastic leukemia (ALL) pediatric patients who had been off chemotherapy for 6 months. These children had been subjected to one of the three Brazilian Protocols or the BFM86 Protocol. The patients were divided into three groups: Group I (GI; high risk) treated with one of the three Brazilian Protocols who received high-dose chemotherapy, intensive maintenance and cranial radiotherapy; Group II (GII; low risk) who were also treated with one of the three Brazilian Protocols using low-intensive chemotherapy with no radiotherapy; and Group III (GIII) based on the BFM86 Protocol. Of 76 children, 13 showed no dental abnormalities (8 were at the age of tooth formation). The remaining 63 children (82.9%) showed at least one dental anomaly. The abnormalities were probably caused by the type, intensity, frequency of the treatment and age of the patients at ALL diagnosis and this might have important consequences for the children's dental development.


Distortion of terahertz signals due to imperfect synchronization with chirped probe pulses.
Terahertz (THz) signals measured by means of the spectral-encoding technique with different temporal discrepancies between probe pulses and THz signals are investigated. It is found that imperfect synchronization between the chirped probe and THz pulses induce a distortion and this distortion affects significantly the retrieved THz spectrum if the temporal discrepancy is large. The distortion becomes more prominent if the probe pulse length is less than the optimal chirped probe pulse duration. A simple approach is proposed to realize the synchronization and minimize the distortion. THz signals from a high-voltage-biased air plasma filament are measured with this approach and distortion similar to the simulation results is observed.


Clinical Potential of miR-451 and miR-506 as a Prognostic Biomarker in Patients with Breast Cancer.
The incidence and mortality of breast cancer in the world remain high. The function and important role of miR-451 and miR-506 in a series of cancers have been proved. The purpose of this research was to explore the clinical diagnosis and prognostic significance of miR-451 and miR-506 expression in breast cancer. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect miR-451 and miR-506 expression in serum and tissues. The relationship of miR-451 and miR-506 with clinical parameters was determined by the chi-square test. Receiver operating characteristics (ROC) analysis was conducted to evaluate the diagnostic accuracy of miR-451 and miR-506 in breast cancer. In addition, we determined the prognostic performance of miR-451 and miR-506 using Kaplan-Meier survival assay. The expression of miR-451 and miR-506 in breast cancer patients was significantly lower than that in healthy people. miR-451 and miR-506 expression decreased in breast cancer tissues compared with paracancerous tissue. High expression of miR-451 and miR-506 was associated with positive lymph node metastasis and late tumor node metastasis stage. Breast cancer patients with high miR-451 and miR-506 expression had lower five-year survival rate. The level of miR-451 and miR-506 expression showed high diagnostic accuracy for distinguishing breast cancer patients and healthy people. miR-451 and miR-506 could be used as biomarker for the diagnosis and prognosis of breast cancer.


Decentralized Adaptive Neural Inverse Optimal Control of Nonlinear Interconnected Systems.
Existing methods on decentralized optimal control of continuous-time nonlinear interconnected systems require a complicated and time-consuming iteration on finding the solution of Hamilton-Jacobi-Bellman (HJB) equations. In order to overcome this limitation, in this article, a decentralized adaptive neural inverse approach is proposed, which ensures the optimized performance but avoids solving HJB equations. Specifically, a new criterion of inverse optimal practical stabilization is proposed, based on which a new direct adaptive neural strategy and a modified tuning functions method are proposed to design a decentralized inverse optimal controller. It is proven that all the closed-loop signals are bounded and the goal of inverse optimality with respect to the cost functional is achieved. Illustrative examples validate the performance of the methods presented.


Changes in clinical trials mandated by the advent of meta-analysis.
Service on the Data Monitoring Committee of the CPEP (Calcium for Pre-eclampsia Prevention) has led us to four conclusions about clinical trials which we should like to present to this gathering of biostatisticians for their reactions: (i) meta-analyses of the pertinent published trials of the same therapy should always be undertaken before the start of a new trial, and the results examined to help determine the design of a new trial or determine if a trial should be undertaken at all; (ii) assuming that a decision is made to go ahead, the results of the past trials should be used in sizing the new one; (iii) in the course of the new one, regardless of the size estimates, stopping early should be considered if the trends conform to the results of the meta-analysis; and (iv) heterogeneity of patients entering clinical trials is desirable and should be specifically studied, and it should never be concluded that an average outcome is applicable to all future patients.


Early embryonic losses in mice induced by diethylstilbestrol.
Estrogens cause embryonic lethality and the disturbance of early placental development in mice. Diethylstilbestrol (DES) at 1, 10, or 100 microg/kg was orally administered to Institute of Cancer Research mice on gestational days (GD) 4 through 8, and the uterus and placenta were examined histopathologically on GD 9. Decidua of DES-treated mice showed insufficient development, and the uterine lumen at the implantation site did not effectively minimize. The trophoblast giant cell layer was not separated from the uterine lumen by the decidua capsularis, and hemorrhage from the denuded trophoblast giant cell layer into the uterine lumen was noted at the peripheral part of the decidua basalis. The results of the present study suggest that decidual hypoplasia and subsequent placental hemorrhage causes fetal death due to the administration of DES during the early stage of pregnancy.


Analysis of CYP450 gene expression and function in white-rot fungus, Lenzites gibbosa, treated with Congo red.
We investigated the function of cytochrome P450 (CYP450) genes in degradation of the diazo dye, Congo red, by white-rot fungus Lenzites gibbosa. Hyphae treated with Congo red at different times were sequenced to obtain transcription data. CYP450 genes in transcriptomes were identified using a gene-encoding protein functional search and analyzed in Cluster of Orthologous Genes, Gene Ontology, Kyoto Encyclopedia of Genes and Genomes Databases. Differentially expressed genes (DEGs) in different groups were analyzed using EdgeR. We present the relation between transcription factors (TFs) and CYP450 regulation by analysis of the co-expression network. One hundred sixty CYP450 genes obtained by a functional annotation search were related to the oxido-reduction reaction of secondary metabolism, defense mechanism and aromatic compound degradation. The fastest decolorization and the greatest expression of CYP450 genes, which were related to the decolorization effect, occurred at 0-3 h. Seven CYP450 genes (7522, 6568, 4482, 9118, 10935, 7521 and 10926) were identified. The key TFs that regulate these genes belong to the zinc finger family. CYP450 genes and their products in L. gibbosa participated in degradation of Congo red and stress resistance. We provide a reference value for degradation and decolorization of Congo red.


Expression of survivin, YB-1, and KI-67 in sporadic adenomas and dysplasia-associated lesions or masses in ulcerative colitis.
Sporadic adenomas are said to exhibit an orderly growth pattern with a reversal of proliferative and apoptotic cell distribution as compared with normal colonic crypts. Dysplastic polyps of patients with ulcerative colitis (UC) may represent dysplasia-associated lesions or masses (DALM) with a high associated cancer risk, or, alternatively, may represent sporadic adenomas. Histologic criteria to differentiate between sporadic adenomas and DALM have not focused on the balance between cell renewal and cell loss. The expression of the novel anti-apoptosis gene product, survivin, and the proliferation markers, Ki-67 and Y-box binding protein (YB-1), were investigated by immunohistochemical localization in sporadic adenomas and DALM lesions of patients with UC. In adenomas, KI-67 was expressed preponderantly at the luminal aspect of the polyp, whereas its expression was diffuse in DALM. Survivin was detected diffusely in both adenomas and DALM. YB-1 showed positive staining in the deep aspect of adenomatous glands but only to a minor degree at the surface, whereas both deep and diffuse expression patterns of YB-1 were seen in DALM. The authors conclude that DALM and sporadic adenomas exhibit different patterns of cellular proliferation and that molecular markers of cell proliferation, Ki-67 and YB-1, may be useful to distinguish sporadic adenomas from DALM. However, the similar expression of survivin suggests that the underlying mechanisms that regulate apoptotic cell death are uniform in these lesions.


Immunogenicity and protection efficacy of enhanced fitness recombinant Salmonella Typhi monovalent and bivalent vaccine strains against acute toxoplasmosis.
The proficiency of Salmonella Typhi to induce cell-mediated immunity has allowed its exploitation as a live vector against the obligate intracellular protozoan Toxoplasma gondii. T. gondii vaccine research is of great medical value due to the lack of a suitable toxoplasmosis vaccine. In the present work, we integrated T. gondii antigen into a growth-dependent chromosome locus guaBA of S. Typhi CVD910 strain to form recombinant S. Typhi monovalent CVD910-SAG1 expressed T. gondii SAG1 antigen and monovalent CVD910-GRA2 expressed T. gondii GRA2 antigen. Furthermore, a low-copy stabilized recombinant plasmid encoding SAG1 antigen was transformed into CVD910-GRA2 to form bivalent CVD910-GS strain. An osmolarity-regulated promoter was also incorporated to control the gene transcription, whereas clyA export protein was included to translocate the antigen out of the cytoplasm. Both CVD910-GRA2 and CVD910-GS displayed healthy growth fitness and readily expressed the encoded T. gondii antigens. When administered in vivo, CVD910-GS successfully induced both humoral and cellular immunity in the immunized BALB/c mice, and extended mice survival against virulent T. gondii. In particular, the mice immunized with bivalent CVD910-GS presented the highest titers of IgG, percentages of CD4+ T, CD8+ T, B cells and memory T cells, and total IgG+ memory B cells as compared to the CVD910-GRA2 and control strains. The CVD910-GS group also generated mixed Th1/Th2 cytokine profile with secretions of IFN-ɣ, IL-2 and IL-10. This study demonstrated the importance of enhancing live vector fitness to sustain heterologous antigen expression for eliciting robust immune responses and providing effective protection against pathogen.


Participation of the molecular chaperone DnaK in intracellular growth of Brucella suis within U937-derived phagocytes.
In the intracellular bacterium Brucella suis, the molecular chaperone DnaK was induced under heat-shock conditions and at low pH. Insertional inactivation of dnaK and dnaJ within the dnaK/J locus led to the conclusion that DnaK, but not DnaJ, was required for growth at 37 degrees C in vitro. Viability of the dnaK null mutant was also greatly affected at low pH. Under conditions allowing intracellular multiplication, the infection of U937-derived phagocytes resulted in long-lasting DnaK induction in the wild-type bacteria. In infection experiments performed with both mutants at the reduced temperature of 30 degrees C, the dnaK mutant of B. suis survived but failed to multiply within U937 cells, whereas the wild-type strain and the dnaJ mutant multiplied normally. Complementation of the dnaK mutant with the cloned dnaK gene restored growth at 37 degrees C, increased resistance to acid pH, and increased intracellular multiplication. This is the first report of the effects of dnaK inactivation in a pathogenic species, and of the temperature-independent contribution of DnaK to intracellular multiplication of the pathogen B. suis.


Integrating deep learning with microfluidics for biophysical classification of sickle red blood cells adhered to laminin.
Sickle cell disease, a genetic disorder affecting a sizeable global demographic, manifests in sickle red blood cells (sRBCs) with altered shape and biomechanics. sRBCs show heightened adhesive interactions with inflamed endothelium, triggering painful vascular occlusion events. Numerous studies employ microfluidic-assay-based monitoring tools to quantify characteristics of adhered sRBCs from high resolution channel images. The current image analysis workflow relies on detailed morphological characterization and cell counting by a specially trained worker. This is time and labor intensive, and prone to user bias artifacts. Here we establish a morphology based classification scheme to identify two naturally arising sRBC subpopulations-deformable and non-deformable sRBCs-utilizing novel visual markers that link to underlying cell biomechanical properties and hold promise for clinically relevant insights. We then set up a standardized, reproducible, and fully automated image analysis workflow designed to carry out this classification. This relies on a two part deep neural network architecture that works in tandem for segmentation of channel images and classification of adhered cells into subtypes. Network training utilized an extensive data set of images generated by the SCD BioChip, a microfluidic assay which injects clinical whole blood samples into protein-functionalized microchannels, mimicking physiological conditions in the microvasculature. Here we carried out the assay with the sub-endothelial protein laminin. The machine learning approach segmented the resulting channel images with 99.1±0.3% mean IoU on the validation set across 5 k-folds, classified detected sRBCs with 96.0±0.3% mean accuracy on the validation set across 5 k-folds, and matched trained personnel in overall characterization of whole channel images with R2 = 0.992, 0.987 and 0.834 for total, deformable and non-deformable sRBC counts respectively. Average analysis time per channel image was also improved by two orders of magnitude (∼ 2 minutes vs ∼ 2-3 hours) over manual characterization. Finally, the network results show an order of magnitude less variance in counts on repeat trials than humans. This kind of standardization is a prerequisite for the viability of any diagnostic technology, making our system suitable for affordable and high throughput disease monitoring.


Dynamics of the stochastic chemostat with Monod-Haldane response function.
The stochastic chemostat model with Monod-Haldane response function is perturbed by environmental white noise. This model has a global positive solution. We demonstrate that there is a stationary distribution of the stochastic model and the system is ergodic under appropriate conditions, on the basis of Khasminskii's theory on ergodicity. Sufficient criteria for extinction of the microbial population in the stochastic system are established. These conditions depend strongly on the Brownian motion. We find that even small scale white noise can promote the survival of microorganism populations, while large scale noise can lead to extinction. Numerical simulations are carried out to illustrate our theoretical results.


One Cell, Two Gears: Extensive Somatic Genome Plasticity Accompanies High Germline Genome Stability in Paramecium.
Mutation accumulation (MA) experiments are conventionally employed to study spontaneous germline mutations. However, MA experiments can also shed light on somatic genome plasticity in a habitual and genetic drift-maximizing environment. Here, we revisit an MA experiment that uncovered extraordinary germline genome stability in Paramecium tetraurelia, a single-celled eukaryote with nuclear dimorphism. Our re-examination of isogenic P. tetraurelia MA lines propagated in nutrient-rich medium for >40 sexual cycles reveals that their polyploid somatic genome accrued hundreds of intervening DNA segments (IESs), which are normally eliminated during germline-soma differentiation. These IESs frequently occupy a fraction of the somatic DNA copies of a given locus, producing IES excision/retention polymorphisms, and preferentially fall into a class of epigenetically controlled sequences. Relative to control lines, retained IESs are flanked by stronger cis-acting signals and interrupt an excess of highly expressed coding exons. These findings suggest that P. tetraurelia's elevated germline DNA replication fidelity is associated with pervasive somatic genome plasticity. They show that MA regimes are powerful tools for investigating the role that developmental plasticity, somatic mutations, and epimutations have in ecology and evolution.


ADAR1-mediated RNA editing links ganglioside catabolism to glioblastoma stem cell maintenance.
Glioblastoma (GBM) is the most common and lethal primary malignant brain tumor, containing GBM stem cells (GSCs) that contribute to therapeutic resistance and relapse. Exposing potential GSC vulnerabilities may provide therapeutic strategies against GBM. Here, we interrogated the role of adenosine-to-inosine (A-to-I) RNA editing mediated by adenosine deaminase acting on RNA 1 (ADAR1) in GSCs and found that both ADAR1 and global RNA editomes were elevated in GSCs compared with normal neural stem cells. ADAR1 inactivation or blocking of the upstream JAK/STAT pathway through TYK2 inhibition impaired GSC self-renewal and stemness. Downstream of ADAR1, RNA editing of the 3'-UTR of GM2A, a key ganglioside catabolism activator, proved to be critical, as interference with ganglioside catabolism and disruption of ADAR1 showed a similar functional impact on GSCs. These findings reveal that RNA editing links ganglioside catabolism to GSC self-renewal and stemness, exposing a potential vulnerability of GBM for therapeutic intervention.


ASI1, a gene encoding a novel leucine zipper protein, is induced during development of the macronucleus in Tetrahymena.
Sexual reproduction in the ciliate Tetrahymena follows a complex developmental program involving the sequential regulation of dozens of genes. Genes that are up-regulated during post-zygotic development in Tetrahymena were isolated by subtractive hybridization. Anlagen stage induced gene 1 (ASI1) encodes a 2.8 kb transcript that contains a single intron and is induced during macronuclear development. ASI1 is a single copy gene in both the micronucleus and the macronucleus. It encodes a 95 kDa conceptual protein with a leucine zipper near the amino terminus.