Image-Processing-Based Subway Tunnel Crack Detection System.
With the increase in urban rail transit construction, instances of tunnel disease are on the rise, and cracks have become the focus of tunnel maintenance and management. Therefore, it is essential to carry out crack detection in a timely and efficient manner to not only prolong the service life of the tunnel but also reduce the incidence of accidents. In this paper, the design and structure of a tunnel crack detection system are analyzed. On this basis, this paper proposes a new method for crack identification and feature detection using image processing technology. This method fully considers the characteristics of tunnel images and the combination of these characteristics with deep learning, while a deep convolutional network (Single-Shot MultiBox Detector (SSD)) is proposed based on deep learning for object detection in complex images. The experimental results show that the test set accuracy and training set accuracy of the support vector machine (SVM) in the classification comparison test are up to 88% and 87.8%, respectively; while the test accuracy of Alexnet's deep convolutional neural network-based classification and identification is up to 96.7%, and the training set accuracy is up to 97.5%. It can be seen that this deep convolutional network recognition algorithm based on deep learning and image processing is better and more suitable for the detection of cracks in subway tunnels.


Editorial on "What is a potentially damaging vaccination delay in children younger than 2 years?"
Control of hepatitis B through routine infant immunization in more than 95% of countries has reduced the prevalence of chronic hepatitis carriers to less than 1%-2% in immunized cohorts of children even in high endemicity countries. In that context the authors of this editorial found the results of a paper by Gras et al in this issue concerning. They performed a Delphi survey of 37 French immunization experts and the results concluded that delayed hepatitis B immunization would cause "potential damage" only after 11 years. Large cohorts of French children and adolescents remain susceptible to hepatitis B infection. Given the high rates of immigration to France from areas of higher endemicity, the higher birth rate and degree of integration of these groups into the health system, plus the lower age of sexual debut and the use of injectable drugs in the general population, we cannot agree that a delay of 11 years is acceptable. Rates of adolescent immunization are quite low so relying on protection at this age will yield little in terms of population protection. Loss of confidence in Hepatitis B vaccine following disproved allegations that the vaccine caused Multiple Sclerosis persists in France, and we believe the results of this paper sends a damaging message to health workers and parents in France and beyond.


Impact of an education program on perinatal care practices.
Education of health professionals has an important role in improving health care. A media-based, self-instructional, perinatal education program was developed and field tested in rural and urban regions of Michigan. Cognitive tests, chart audits, and consultation/referral times were used to measure the impact on education and patient care. The program effectively increased knowledge and improved patient care practices by physicians and nurses. This study presents evidence that a targeted educational program in a media-based format can significantly improve perinatal care.


[Dilated cardiomyopathy: indication and role of endomyocardial biopsy].
In the last years, endomyocardial biopsy has become a useful diagnostic tool for the investigation and treatment of myocardial diseases. Mainly, its role is irreplaceable for the diagnosis of acute myocarditis. However, its routine use is criticized by some authors for the lack of therapeutic usefulness. Actual techniques enable to perform multiple drawings of tissue samples from both ventricles with low incidence of procedural complications. However, in consideration of the invasivity of the procedure, it is fundamental a correct selection of patients to undergo endomyocardial biopsy. In addition to some particular clinical contexts (after heart transplantation, specific myocardial disease), the more frequent indication to endomyocardial biopsy is suspected myocarditis in patients with "major" symptoms (recent-onset heart failure with relevant left ventricular dysfunction, sustained ventricular arrhythmias). In such cases, the correct analysis of tissue samples represents an important question to answer. The information derived from endomyocardial biopsy is increased by the number of methodologies that we apply. For this reason, it is necessary that traditional histology is followed by immunohistochemical, ultrastructural and molecular biological techniques, which can increase the accuracy of the analysis. About the therapeutic perspectives, endomyocardial biopsy has a role in few clinical cases, according to the persistent perplexity about the therapy of active myocarditis. However, we have to underline that in selected cases endomyocardial biopsy represents a useful method for a correct prognostic and therapeutic evaluation.


The interleukin-6 -147 g/c polymorphism is associated with increased risk of coronary artery disease in young South African Indian men.
Interleukin-6 (IL-6) is a proinflammatory cytokine involved in the pathogenesis of chronic inflammatory diseases such as coronary artery disease (CAD). The -174 IL-6 G/C promoter polymorphism influences mRNA levels and protein expression and is implicated in CAD. The Indian population in South Africa, unlike the black community, has a high prevalence of premature CAD. This polymorphism has not been fully explored in this population. The present study assessed the -174 IL-6 G/C polymorphism in young Indian patients with angiographically documented CAD and compared them with age- and gender-matched Indian and black control subjects. Polymorphic variants were assessed by polymerase chain reaction-restriction fragment length polymorphism, and IL-6 levels were measured using enzyme-linked immunosorbent assay (ELISA). The -174 IL-6 C allele was found with a higher frequency (23%) in the total Indian group compared to 2% in the black participants [P<0.0001, odds ratio (OR)=0.05, 95% confidence interval (CI) 0.018-0.14). The difference in frequency was more pronounced when Indian controls were compared to black controls (29% vs. 2%, respectively) (P<0.0001, OR=0.05, 95% CI 0.02-0.17). A significant association between the -174 IL-6 G allele and CAD was found in Indian patients compared to Indian controls (84% in cases vs. 71% in Indian controls; P=0.043, OR=0.47 95% CI 0.23-0.95). Levels of IL-6 in circulation were higher in black controls (6.62±0.63 pg/mL) compared to Indian controls (2.51±0.57 pg/mL) and CAD patients (1.46±0.36 pg/mL) (P<0.0001). Levels of IL-6 were higher in all groups with homozygous -174 IL-6 C alleles, but only significant in the healthy Indian control group (GG 3.73±0.94 pg/mL vs. GC/CC 0.89±0.5 pg/mL, P=0.0001). The presence of the IL-6 -174 G allele influences levels of IL-6 and increases the risk of CAD in South African Indians.


Mouse circulating extracellular vesicles contain virus-derived siRNAs active in antiviral immunity.
Induction and suppression of antiviral RNA interference (RNAi) has been observed in mammals during infection with at least seven distinct RNA viruses, including some that are pathogenic in humans. However, while the cell-autonomous immune response mediated by antiviral RNAi is gradually being recognized, little is known about systemic antiviral RNAi in mammals. Furthermore, extracellular vesicles (EVs) also function in viral signal spreading and host immunity. Here, we show that upon antiviral RNAi activation, virus-derived small-interfering RNAs (vsiRNAs) from Nodamura virus (NoV), Sindbis virus (SINV), and Zika virus (ZIKV) enter the murine bloodstream via EVs for systemic circulation. vsiRNAs in the EVs are biologically active, since they confer RNA-RNA homology-dependent antiviral activity in both cultured cells and infant mice. Moreover, we demonstrate that vaccination with a live-attenuated virus, rendered deficient in RNAi suppression, induces production of stably maintained vsiRNAs and confers protective immunity against virus infection in mice. This suggests that vaccination with live-attenuated VSR (viral suppressor of RNAi)-deficient mutant viruses could be a new strategy to induce immunity.


Computer-Aided Diagnosis Research of a Lung Tumor Based on a Deep Convolutional Neural Network and Global Features.
Based on the better generalization ability and the feature learning ability of the deep convolutional neural network, it is very significant to use the DCNN on the computer-aided diagnosis of a lung tumor. Firstly, a deep convolutional neural network was constructed according to the fuzzy characteristics and the complexity of lung CT images. Secondly, the relation between model parameters (iterations, different resolution) and recognition rate is discussed. Thirdly, the effects of different model structures for the identification of a lung tumor were analyzed by changing convolution kernel size, feature dimension, and depth of the network. Fourthly, the different optimization methods on how to influence the DCNN performance were discussed from three aspects containing pooling methods (maximum pooling and mean pooling), activation function (sigmoid and ReLU), and training algorithm (batch gradient descent and gradient descent with momentum). Finally, the experimental results verified the feasibility of DCNN used on computer-aided diagnosis of lung tumors, and it can achieve a good recognition rate when selecting the appropriate model parameters and model structure and using the method of gradient descent with momentum.


Sweeping-window arc therapy: an implementation of rotational IMRT with automatic beam-weight calculation.
Sweeping-window arc therapy (SWAT) is a variation of intensity-modulated radiation therapy (IMRT) with direct aperture optimization (DAO) that is initialized with a leaf sequence of sweeping windows that move back and forth periodically across the target as the gantry rotates. This initial sequence induces modulation in the dose and is assumed to be near enough to a minimum to allow successful optimization, done with simulated annealing, without requiring excessive leaf speeds. Optimal beam weights are calculated analytically, with easy extension to allow for variable beam weights. In this paper SWAT is tested on a phantom model and clinical prostate case. For the phantom, constant and variable beam weights are used. Although further work (in particular, improving the dose model) is required, the results show SWAT to be a feasible approach to generating deliverable dynamic arc treatments that are optimized.


Thoracic disc disease: experience with the transpedicular approach in twenty consecutive patients.
TWENTY CONSECUTIVE PATIENTS were treated for symptomatic thoracic disc herniation with the transpedicular approach. Most patients had severe, incapacitating local or radicular pain. Myelopathy was uncommon as magnetic resonance imaging allowed an early diagnosis. Computed tomography, after myelography, provided further information necessary for surgical planning. Three patients had disc disease at two levels. Nine central and 14 lateral disc herniations were found. Disc calcification or an associated osteophyte was identified in six instances. Although the size of the disc herniation correlated with the amount of cord compression, no radiological features were found to be correlated with neurological function. The transpedicular approach was used in all patients. New curettes, specifically designed for the procedure, allowed the removal of all discs, including central and calcified fragments. A modified arthroscope was used to confirm neural decompression in some instances. One year after surgery, all 20 patients were significantly improved and 8 patients were asymptomatic. Apart from the duration of the symptoms, no other factors were found to affect outcome. The findings suggest that the prognosis of thoracic disc herniation is excellent if the disease is recognized early. The transpedicular approach, using curettes specifically designed for the procedure, can be an effective and safe method of surgical decompression in carefully selected patients.


Modulation of corticospinal input to the legs by arm and leg cycling in people with incomplete spinal cord injury.
The spinal cervico-lumbar interaction during rhythmic movements in humans has recently been studied; however, the role of arm movements in modulating the corticospinal drive to the legs is not well understood. The goals of this study were to investigate the effect of active rhythmic arm movements on the corticospinal drive to the legs (study 1) and assess the effect of simultaneous arm and leg training on the corticospinal pathway after incomplete spinal cord injury (iSCI) (study 2). In study 1, neurologically intact (NI) participants or participants with iSCI performed combinations of stationary and rhythmic cycling of the arms and legs while motor evoked potentials (MEPs) were recorded from the vastus lateralis (VL) muscle. In the NI group, arm cycling alone could facilitate the VL MEP amplitude, suggesting that dynamic arm movements strongly modulate the corticospinal pathway to the legs. No significant difference in VL MEP between conditions was found in participants with iSCI. In study 2, participants with iSCI underwent 12 wk of electrical stimulation-assisted cycling training: one group performed simultaneous arm and leg (A&L) cycling and the other legs-only cycling. MEPs in the tibialis anterior (TA) muscle were compared before and after training. After training, only the A&L group had a significantly larger TA MEP, suggesting increased excitability in the corticospinal pathway. The findings demonstrate the importance of arm movements in modulating the corticospinal drive to the legs and suggest that active engagement of the arms in lower limb rehabilitation may produce better neural regulation and restoration of function.NEW & NOTEWORTHY This study aimed to demonstrate the importance of arm movements in modulating the corticospinal drive to the legs. It provides direct evidence in humans that active movement of the arms could facilitate corticospinal transmission to the legs and, for the first time, shows that facilitation is absent after spinal cord injury. Active engagement of the arms in lower limb rehabilitation increased the excitability of the corticospinal pathway and may produce more effective improvement in leg function.


Intrathecal midazolam for postoperative pain relief in caesarean section delivery.
Postoperative pain relief is a growing concern to an anaesthesiologist since no single analgesic is free from side-effects. Moreover, it becomes a challenge after caesarean section delivery to provide postoperative pain relief without much sedation, respiratory depression or problems like nausea, vomiting, so that early baby acceptance and care by mother is promoted. Antinociceptive effect of midazolam is well established by now and its safety is documented. This observation was made in a blind randomised study of 40 women of ASA I/II to evaluate postoperative pain relief using intrathecal midazolam in caesarean section delivery. Group A patients (n=20) received 1.5 ml of 5% lignocaine only and group B patients (n=20) received mixture of 1.5 ml 5% lignocaine with 2 mg midazolam (preservative free) through intrathecal route at L3.4 interspace; vital parameters were monitored intra-operatively and postoperatively and Apgar score of baby in 1st and 5th minute of deliverywas assessed. It was observed intrathecal midazolam produced highly significant (p<0.001) postoperative pain relief together with anti-emetic effect and tranquillity of patients of caesarean section delivery.


A review of biocompatible metal injection moulding process parameters for biomedical applications.
Biocompatible metals have been revolutionizing the biomedical field, predominantly in human implant applications, where these metals widely used as a substitute to or as function restoration of degenerated tissues or organs. Powder metallurgy techniques, in specific the metal injection moulding (MIM) process, have been employed for the fabrication of controlled porous structures used for dental and orthopaedic surgical implants. The porous metal implant allows bony tissue ingrowth on the implant surface, thereby enhancing fixation and recovery. This paper elaborates a systematic classification of various biocompatible metals from the aspect of MIM process as used in medical industries. In this study, three biocompatible metals are reviewed-stainless steels, cobalt alloys, and titanium alloys. The applications of MIM technology in biomedicine focusing primarily on the MIM process setting parameters discussed thoroughly. This paper should be of value to investigators who are interested in state of the art of metal powder metallurgy, particularly the MIM technology for biocompatible metal implant design and development.


Atypical protein kinase C regulates primary dendrite specification of cerebellar Purkinje cells by localizing Golgi apparatus.
Neurons have highly polarized structures that determine what parts of the soma elaborate the axon and dendrites. However, little is known about the mechanisms that establish neuronal polarity in vivo. Cerebellar Purkinje cells extend a single primary dendrite from the soma that ramifies into a highly branched dendritic arbor. We used the zebrafish cerebellum to investigate the mechanisms by which Purkinje cells acquire these characteristics. To examine dendritic morphogenesis in individual Purkinje cells, we marked the cell membrane using a Purkinje cell-specific promoter to drive membrane-targeted fluorescent proteins. We found that zebrafish Purkinje cells initially extend multiple neurites from the soma and subsequently retract all but one, which becomes the primary dendrite. In addition, the Golgi apparatus specifically locates to the root of the primary dendrite, and its localization is already established in immature Purkinje cells that have multiple neurites. Inhibiting secretory trafficking through the Golgi apparatus reduces dendritic growth, suggesting that the Golgi apparatus is involved in the dendritic morphogenesis. We also demonstrated that in a mutant of an atypical protein kinase C (aPKC), Prkci, Purkinje cells retain multiple primary dendrites and show disrupted localization of the Golgi apparatus. Furthermore, a mosaic inhibition of Prkci in Purkinje cells recapitulates the aPKC mutant phenotype. These results suggest that the aPKC cell autonomously controls the Golgi localization and thereby regulates the specification of the primary dendrite of Purkinje cells.


A novel homozygous missense mutation in FGF23 causes Familial Tumoral Calcinosis associated with disseminated visceral calcification.
Hyperphosphatemic Familial Tumoral Calcinosis (HFTC; MIM211900) is a rare autosomal recessive disorder characterized by the progressive deposition of calcified masses in cutaneous and subcutaneous tissues, associated with elevated circulating levels of phosphate. The disease was initially found to result from mutations in GALNT3 encoding a glycosyltransferase. However, more recently, the S71G missense mutation in FGF23, encoding a potent phosphaturic protein, was identified in two families. In the present report, we describe a second mutation in FGF23 underlying a severe case displaying calcifications of cutaneous and numerous extracutaneous tissues. The mutation (M96T) was found to affect a highly conserved methionine residue at position 96 of the protein. These observations illustrate the extent of genetic and phenotypic heterogeneity in HFTC.


Transcription factor and polymerase recruitment, modification, and movement on dhsp70 in vivo in the minutes following heat shock.
The uninduced Drosophila hsp70 gene is poised for rapid activation. Here we examine the rapid changes upon heat shock in levels and location of heat shock factor (HSF), RNA polymerase II (Pol II) and its phosphorylated forms, and the Pol II kinase P-TEFb on hsp70 in vivo by using both real-time PCR assays of chromatin immunoprecipitates and polytene chromosome immunofluorescence. These studies capture Pol II recruitment and progression along hsp70 and reveal distinct spatial and temporal patterns of serine 2 and serine 5 phosphorylation: in uninduced cells, the promoter-paused Pol II shows Ser5 but not Ser2 phosphorylation, and in induced cells the relative level of Ser2-P Pol II is lower at the promoter than at regions downstream. An early time point of heat shock activation captures unphosphorylated Pol II recruited to the promoter prior to P-TEFb, and during the first wave of transcription Pol II and the P-TEFb kinase can be seen tracking together across hsp70 with indistinguishable kinetics. Pol II distributions on several other genes with paused Pol II show a pattern of Ser5 and Ser2 phosphorylation similar to that of hsp70. These studies of factor choreography set important limits in modeling transcription regulatory mechanisms.


Variable-number tandem repeat typing of Mycobacterium tuberculosis isolates with low copy numbers of IS6110 by using mycobacterial interspersed repetitive units.
A study set of 180 Mycobacterium tuberculosis and Mycobacterium bovis isolates having low copy numbers of IS6110 were genotyped using the recently introduced method based on the variable-number tandem repeats of mycobacterial interspersed repetitive units (MIRU-VNTR). The results were compared with results of the more commonly used methods, IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping. The isolates were collected in Michigan from 1996 to 1999 as part of a project to genotype all isolates from new cases of tuberculosis in the state. Twelve MIRU loci were amplified, and the amplicons were analyzed by agarose gel electrophoresis to determine the copy number at each MIRU locus. MIRU-VNTR produced more distinct patterns (80 patterns) than did IS6110 RFLP (58 patterns), as would be expected in this study set. Spoligotyping identified 59 patterns. No single method defined all unique isolates, and the combination of all three typing methods generated 112 distinct patterns identifying 90 unique isolates and 90 isolates in 22 clusters. The results confirm the potential utility of MIRU-VNTR typing and show that typing with multiple methods is required to attain maximum specificity.


[Use of oxi-cardiotocography in twin delivery].
The intrapartum management of multiple gestation continues to challenge the obstetrics profession. Electronic fetal heart rate monitoring and fetal pulse oximetry are used in clinical research to assess fetal well-being during labor. The objective of this study was to establish values of fetal arterial oxygen saturation (FSpO2) and some other parameters by dint of computerised analysis of cardiotocogram during twin delivery. 5 cases of vaginal twin deliveries as the Clinic of Perinatology of the Institute of Gynecology and Obstetrics Lodz Mecical University over the years 1988-1999 were reviewed. In four cases vertex-vertex twins were delivered vaginally, while in one case an assisted breech extraction was conducted on vertex-non vertex twins. The foetus was monitored continually with a Oxford S 8000 systems and pulse oximeter Nellcor N-400. the medium parameters of computerised analysis of CTG and FSpO2 during progressive stages of labour were shown. Twin delivery needs modern fetal monitoring tools to provide the obstetrician information on well-being of the foetus. It is possible to monitor fetal oxygen saturation and CTG parameters in both of the twins during labour The ability to measure fetal oxygen saturation especially during the second stage of twin labor adds critical information about fetal status and refines the interpretation of abnormal FHR patterns. Only after examining a large number of twin fetuses we can assess whether pulse oximetry in useful in management of labor.


Identification of a macromolecule containing an anticarcinoembryonic antigen-reactive substance and immunoglobulin M in human pancreatic cancer.
Ascitic fluid from a patient with carcinoma of the pancreas was fractionated by ammonium sulfate precipitation. The fraction precipitated between 25 and 50% saturation of ammonium sulfate was sequentially chromatographed on Sephadex G-200 and Sepharose 6B. A macromolecular fraction (greater than 10(6) daltons) obtained was found to react with both antihuman IgM and antiserum to carcinoembryonic antigen (CEA). This fraction was further purified by adsorption with protein A-Sepharose CL-4B and chromatography on DEAE-Sephacel. The purified macromolecular fraction had a sedimentation value of 28S as determined by ultracentrifugation. Upon dissociation of the purified macromolecule at pH 2.3 and purification of the dissociated components on Sepharose CL-2B and BioGel A 1.5M, a 19S protein and a 5S protein were recovered. The 19S protein showed a complete line of identity with a reference human IgM when reacted with antihuman IgM in gel diffusion, whereas the 5S protein showed a partial immunologic identity with colon CEA against anti-CEA. These results indicated the existence of an IgM-containing macromolecular complex with an anti-CEA cross-reactive substance in the extracellular fluid of human pancreatic cancer.


Retinoic acid inhibits HIV-1-induced podocyte proliferation through the cAMP pathway.
HIV-associated nephropathy is characterized by renal podocyte proliferation and dedifferentiation. This study found that all-trans retinoic acid (atRA) reverses the effects of HIV-1 infection in podocytes. Treatment with atRA reduced cell proliferation rate by causing G1 arrest and restored the expression of the differentiation markers (synaptopodin, nephrin, podocin, and WT-1) in HIV-1-infected podocytes. It is interesting that both atRA and 9-cis RA increased intracellular cAMP levels in podocytes. Podocytes expressed most isoforms of retinoic acid receptors (RAR) and retinoid X receptors (RXR) with the exception of RXRgamma. RARalpha antagonists blocked atRA-induced cAMP production and its antiproliferative and prodifferentiation effects on podocytes, suggesting that RARalpha is required. For determination of the effect of increased intracellular cAMP on HIV-infected podocytes, cells were stimulated with either forskolin or 8-bromo-cAMP. Both compounds inhibited cell proliferation significantly and restored synaptopodin expression in HIV-infected podocytes. The effects of atRA were abolished by Rp-cAMP, an inhibitor of the cAMP/protein kinase A pathway and were enhanced by rolipram, an inhibitor of phosphodiesterase 4, suggesting that the antiproliferative and prodifferentiation effects of atRA on HIV-infected podocytes are cAMP dependent. Furthermore, both atRA and forskolin suppressed HIV-induced mitogen-activated protein kinase 1 and 2 and Stat3 phosphorylation. In vivo, atRA reduced proteinuria, cell proliferation, and glomerulosclerosis in HIV-1-transgenic mice. These findings suggest that atRA reverses the abnormal phenotype in HIV-1-infected podocytes by stimulating RARalpha-mediated intracellular cAMP production. These results demonstrate the mechanism by which atRA reverses the proliferation of podocytes that is induced by HIV-1.


Centennial record of anthropogenic impacts in Galveston Bay: Evidence from trace metals (Hg, Pb, Ni, Zn) and lignin oxidation products.
During the 20th century the impacts of industrialization and urbanization in Galveston Bay resulted in significant shifts in trace metals (Hg, Pb, Ni, Zn) and vascular plant biomarkers (lignin phenols) recorded within the surface sediments and sediment cores profile. A total of 22 sediment cores were collected in Galveston Bay in order to reconstruct the historical input of Hg, Pb, Ni, Zn and terrestrial organic matter. Total Hg (T-Hg) concentration ranged between 6 and 162 ng g-1 in surface sediments, and showed decreasing concentrations southward from the Houston Ship Channel (HSC) toward the open estuary. Core profiles of T-Hg and trace metals (Ni, Zn) showed substantial inputs starting in 1905, with peak concentrations between 1960 and 1970's, and decreasing thereafter with exception to Pb, which peaked around 1930-1940s. Stable carbon isotopes and lignin phenols showed an increasing input of terrestrial organic matter driven by urban development within the watershed in the early 1940s. Both the enrichment factor and the geoaccumulation index (Igeo) for T-Hg as a measure of the effectiveness of environmental management practices showed substantial improvements since the 1970s. The natural recovery rate in Galveston Bay since the peak input of T-Hg was non-linear and displayed a slow recovery during the twenty-first century.


Nitric oxide synthase histochemistry in insect nervous systems: Methanol/formalin fixation reveals the neuroarchitecture of formaldehyde-sensitive NADPH diaphorase in the cockroach Periplaneta americana.
Formaldehyde-insensitive NADPH diaphorase (NADPHd) activity is used widely as a histochemical marker for neuronal nitric oxide synthase (NOS). However, in several insects including the cockroach Periplaneta americana, NOS is apparently formaldehyde-sensitive; NADPHd fails to reveal neuron morphology and results in faint generalized staining. Here we have used a novel fixative, methanol/ formalin (MF), to reveal for the first time the neuroarchitecture of NADPHd in the cockroach, with intense selective staining occurring in neurons throughout the brain and thoracic ganglia. Immunocytochemical and histochemical analysis of cockroach and locust nervous systems indicated that neuronal NADPHd after MF fixation can be attributed to NOS. However, NADPHd in locust glial and perineurial cells was histochemically different from that in neurons and may thus be due to enzymes other than NOS. Histochemical implications of species-specific enzyme properties and of the transcriptional complexity of the NOS gene are discussed. The present findings suggest that MF fixation is a valuable new tool for the comparative analysis of the neuroarchitecture of NO signaling in insects. The Golgi-like definition of the staining enabled analysis of the NADPHd architecture in the cockroach and comparison with that in the locust. NADPHd in the tactile neuropils of the thoracic ganglia showed a similar organization in the two species. The olfactory glomeruli of the antennal lobes were in both species densely innervated by NADPHd-positive local interneurons that correlated in number with the number of glomeruli. Thus, the NADPHd architectures appear highly conserved in primary sensory neuropils. In the cockroach mushroom bodies, particularly dense staining in the gamma-layer of the lobes was apparently derived from Kenyon cells, whereas extrinsic arborizations were organized in domains across the lobes, an architecture that contrasts with the previously described tubular compartmentalization of locust mushroom bodies. These divergent architectures may result in different spatiotemporal dynamics of NO diffusion and suggest species differences in the role of NO in the mushroom bodies.


miR-145-5p Inhibits Neuroendocrine Differentiation and Tumor Growth by Regulating the SOX11/MYCN Axis in Prostate cancer.
Recent studies have shown that the downregulation of miR-145-5p in prostate cancer (PCa) is significantly associated with poor differentiation and prognosis. We aimed to investigate the biological role of miR-145-5p in the neuroendocrine differentiation (NED) of PCa. In this study, TheCancer Genome Atlas was used to identify the association of miR-145-5p with PCa. The functions of miR-145-5p were evaluated using the Cell Counting Kit-8 (CCK-8) assay and cell cycle analysis. We validated changes in cell cycle control by testing the expression of cyclin-related genes by western blot. The luciferase reporter assay was performed to test miR-145-5p-targeting genes and direct transcriptional targets of SOX11. The expression of miR-145-5p was found to be significantly downregulated in castration-resistant PCa, and this was correlated with higher Gleason score and prostate-specific antigen. We confirmed these results using PC3 and LNCaP cell lines depicted a gradual decline of miR-145-5p while the cells were cultured under androgen depletion conditions. Moreover, the knockdown of miR-145-5p significantly promoted NED and proliferation of LNCaP cells, whereas overexpression of miR-145-5p significantly inhibited NED and proliferation of LNCaP cells. Mechanistically, we found that SOX11 was a direct target of miR-145-5p, which regulates MYCN might mediate induction of NED and proliferation of LNCaP cells. Furthermore, knockdown of miR-145-5p promoted tumor growth in vivo. Our findings suggest that miR-145-5p can inhibit NED and tumor growth by targeting SOX11, which regulates the expression of MYCN, and that this could be a novel therapeutic strategy for preventing the progression of PCa.


Morphological and molecular characterisation of fungal populations possibly involved in the biological alteration of stones in historical buildings.
The deterioration process of historical building is progressive and irreversible, and the timing and mode of impact are different depending on the characteristics of building materials used, local microclimate, air pollution, presence of specific flora and fauna. The chemical and microbiological characterisation of building materials is mandatory in preventing and eventually recovering degradation effects. Ideally, the analysis of structural stones should be complete, efficient, rapid, and non destructive when dealing with a precious or unique construction. The investigation has been performed on a private historical building made using calcarenite stones and sited between the archaeological site of Lavello, a little town located in the Basilicata Region (South Italy), and the industrial area surrounding this town. To study in progress the degradation of stone materials, a new building sample (ca. 1 m3) was constructed by using the same stones (33 x 15cm), collected from a local quarry. The intact calcarenite stone was characterised by using different methods of surface analysis (XRD, XPS, SEM), and exposed to outdoor conditions. The analyses of the stone material were repeated after three and six months to early evaluate the progression of alterations and the forward modifications of calcarenite structure. After only three months of the new building sample exposure, the adopted analytical methods were able to provide a series of data, which allowed the assessment of the incipient modification of the stone surfaces. The degradation appeared worsened performing the same observations on sixth month replicates, suggesting that environmental conditions modified the structure and the compactness of stones and favoured the biological colonization of surfaces especially in the South-East direction of prevailing winds. For this reason the presence of fungi on the stones' surface was investigated and a morphological and molecular characterization of sampled fungi was performed. Several genera and species of fungi, possibly, involved in degradation were found. The most frequent colonies belonged to Alternaria (A. infectoria, A. citri and Alternaria sp.), Coprinopsis sp., Penicillium piceum, Fusatrium equiseti and Scytalidium termophilus.


Expression of D-amino acid oxidase in Rhodotorula gracilis under induction conditions: a biochemical and cytochemical study.
D-amino acid oxidase is expressed to a high level in the yeast Rhodotorula gracilis (0.3% of total cell protein) through induction by D-alanine in a defined growth medium. Monospecific polyclonal antibodies against pure enzyme were obtained. Western blot analysis showed that the enzyme is synthesized as the mature polypeptide. The localization of the enzyme was investigated by immunoelectron microscopy using the postembedding immunogold technique and by submicroscopic enzyme cytochemistry. D-Amino acid oxidase was detected in peroxisomes, and quantitation of immunoelectron microscopic data indicated that the enzyme is exclusively confined to these organelles. Immunoelectron microscopic observations are in complete agreement with biochemical data showing that the enzyme is not expressed in the absence of D-alanine. Morphometric analysis demonstrated that induction of D-amino acid oxidase synthesis is associated with a 241% increase of peroxisome volume density and with a 31% increase of peroxisome size as compared to cells grown on non-inducing medium.


Antibody-Drug Conjugates for the Treatment of Hematological Malignancies: A Comprehensive Review.
Antibody-drug conjugates (ADCs) are an emerging class of therapeutic agents that bring new opportunities for the treatment of hematological malignancies by meeting unmet medical needs. These drugs consist of a cytotoxic agent connected by a linker to a human, humanized, or chimeric antibody targeting a surface antigen specifically expressed by tumor cells. These ADCs are being developed to specifically deliver the cytotoxic agent into tumor cells. The cytotoxic payload is released from the ADC after internalization and cleavage of the linker, ultimately triggering the death of the cancer cell. Second- and even third-generation ADCs are currently being developed and have more stable linkers and more potent payloads, which should improve ADC efficacy even further. In this review, we analyze the results for the main ADCs currently developed and discuss the advantages and drawbacks of this therapeutic option.


[How much treatment does a person need? Addiction, spontaneous remission and "family" as biographical as leitmotiv].
Referring to theoretical and empirical examples taken from psychiatry, psychology and sociology, this paper first clarifies the dimensions and application of concepts such as 'natural recovery' and 'spontaneous remission'. In order to illustrate the general development of research in the area of therapy free drug remission from a purely quantitative approach to a qualitative, ethnographic one, two studies are referred to: the classic Vietnam veteran study done by Robins and a longitudinal analysis of a network of cocaine users. This is also supplemented by the latest results from Germany. Next, the empirical results of the ongoing spontaneous remitter follow-up study from the Swiss Institute for the Prevention of Alcohol and Drug Problems (SIPA) are presented. The analysis of the gender specific processes involved in the long-term stabilization of heroin autoremission, taking the support of social surroundings into consideration, shows the following: independent of gender, a high stability of autoremission was determined four years after the initial survey. In only three out of thirty cases has there been a relapse since the first interview. This result confirms the general significance of social support during the autoremission phase for both gender groups. Measured against dominant role expectations, it can be said that most of the men have successfully reintegrated into society. Women, on the other hand, seem to deviate more strongly from traditional professional and family roles. This might be due to particularly longlasting social sanctions during their former addiction. Finally, some practical consequences for the drug treatment system drawn from autoremission research are discussed as well as some drug and social policy conditions favorable to autoremission.


Diet and C-reactive protein.
C-reactive protein (CRP) is an independent predictor of cardiovascular events in healthy individuals and those with pre-existing disease. It also probably contributes to the disease process. CRP levels are higher in obese subjects and this link is almost certainly because of increased insulin resistance. Interventions that alter insulin resistance, such as weight loss, exercise, and conjugated linoleic acid, also alter CRP. Glycemic load is associated with CRP, but there have been no interventions with altered macronutrient composition. In the context of weight loss, macronutrient composition is probably not important. Alcohol lowers CRP, but the mechanism is unknown. The interaction between gender and obesity needs further work, but it appears that obesity has a greater effect on CRP levels in women.


Unravelling the Effects of A-Site Cations on Nonradiative Electron-Hole Recombination in Lead Bromide Perovskites: Time-Domain ab Initio Analysis.
Lead bromide perovskites APbBr3 (A = Cs, MA, FA) hold great promise in optoelectronics and photovoltaics. Because the band gaps of the three materials are similar, and also because the A-site cation does not contribute to band edges, one would expect a minor influence of A-site cation on the excited-state lifetime of the perovskites. Experiments defy that expectation. By performing ab initio nonadiabatic (NA) molecular dynamics combined with time-domain density functional simulations, we demonstrate that the nonradiative electron-hole recombination times are in the order FAPbBr3 > MAPbBr3 > CsPbBr3, which are determined by the NA electron-phonon coupling because decoherence times are similar. The simulations show that the larger A-site cation and the smaller NA coupling because larger A-site cation suppresses the Pb-Br cages' motion. The electron-hole recombination is slow, ranging from subnanosecond to nanoseconds, because the NA coupling is small, less than 3 meV, and because decoherence time is slow, less than 7 fs. Both the trend of recombination and the time scales show excellent agreement with experiments. The time-domain atomistic simulations rationalize the experimental observations and advance our understanding of the cations' influence on perovskite excited-state lifetimes.


[Combination therapy in primary hypertension].
Hypertension is an important risk factor for vascular disease. Primary goal of hypertension treatment is to prevent or delay the onset of blood pressure-related morbidity and mortality. It has been well demonstrated that the responses rate to any single class of antihypertensive agent, give as monotherapy is approximately 45% to 55%, and in half of hypertensive population a second will be required. The data from clinical trials clearly demonstrate that two-drug combination, usually with low-dose diuretics with any one of the other first-line agents increases the response rate to about 80% to 85% and reduces the likelihood of adverse events and alteration in lipid, carbohydrate and electrolyte metabolism. Of the various combinations being given that of an diuretic and ACE inhibitor, and ACE inhibitor and non-dihydropyridine calcium channel blockers seems particularly attractive. Some combinations are inappropriate, such as diuretic and calcium channel blockers, and beta-blocker with verapamil and diltiazem. Combination of ACE inhibitor and a non-dihydropyridine calcium channel blockers may provide benefit in regression left ventricular hypertrophy diabetic nephropathy, and post myocardial infarction.


Unraveling the web of viroinformatics: computational tools and databases in virus research.
The beginning of the second century of research in the field of virology (the first virus was discovered in 1898) was marked by its amalgamation with bioinformatics, resulting in the birth of a new domain--viroinformatics. The availability of more than 100 Web servers and databases embracing all or specific viruses (for example, dengue virus, influenza virus, hepatitis virus, human immunodeficiency virus [HIV], hemorrhagic fever virus [HFV], human papillomavirus [HPV], West Nile virus, etc.) as well as distinct applications (comparative/diversity analysis, viral recombination, small interfering RNA [siRNA]/short hairpin RNA [shRNA]/microRNA [miRNA] studies, RNA folding, protein-protein interaction, structural analysis, and phylotyping and genotyping) will definitely aid the development of effective drugs and vaccines. However, information about their access and utility is not available at any single source or on any single platform. Therefore, a compendium of various computational tools and resources dedicated specifically to virology is presented in this article.


Determination of the consensus binding sequence for the purified embryonic heat shock factor 2.
Heat shock transcription factors (HSFs) are characterized by their ability, upon activation, to bind to heat shock response elements (HSE) present in the promoter of their target genes. HSE are composed of inverted repeats of the pentamer nGAAm. In this study, we compare the embryonic HSF2 protein, purified from F9 embryonal carcinoma cells tumor, and the in vitro synthesized HSF2. We show that the context of HSF2 synthesis influences its thermosensitivity and DNA-binding properties. Therefore, we determined the consensus binding sequence for the purified embryonic HSF2 by the technique of systematic evolution of ligands by exponential enrichment (SELEX). We show that embryonic HSF2 prefers sites containing three or four nGAAm inverted pentamers and that its optimal binding sequence contains the 8-mer palindromic core 5'-TTCTAGAA-3'. The consensus binding sequence for the embryonic HSF2 will be very helpful to identify new targets for this factor, during developmental and differentiation processes.


Lesions of the posterior commissure disable the vertical neural integrator of the primate oculomotor system.
1. Spontaneous saccades, vestibuloocular responses (VOR), and optokinetic nystagmus were recorded in three squirrel monkeys before and after chemical or electrolytic lesion of the posterior commissure (PC). 2. PC lesions produced abnormal vertical eye movements, in particular, 1) Postsaccadic drifts, and 2) VOR gain reduction and phase advance more pronounced at lower frequencies of sinusoidal head rotation. Horizontal eye movements were much less affected (or normal). 3. We conclude that PC fibers are necessary for conveying the output of the vertical neural integrator to vertical oculomotor-neurons.


Preparation and characterization of 99mTc(CO)3-BPy-RGD complex as alphav beta3 integrin receptor-targeted imaging agent.
The aim of this study is to develop a novel arginine-glycine-aspartic acid (RGD) peptide-containing ligand for (99m)Tc labeling as alpha(v)beta(3) integrin receptor-targeted imaging agent. BPy-RGD conjugate was successfully synthesized by coupling of 5-carboxylate-2,2'-bipyridine and c(RGDyK) peptide through EDC/SNHS in aqueous solution and was characterized by MADLI-TOF-MS (m/z=802.72, C(38)H(48)N(11)O(9)). (99m)Tc(CO)(3)-BPy-RGD was prepared by exchange reaction between [(99m)Tc(H(2)O)(3)(CO)(3)](+) and BPy-RGD. Final product was purified by HPLC and tested for octanol/water partition coefficient. Cell-binding assays of BPy-RGD and unmodified c(RGDyK) were tested in MDA-MB-435 cells ((125)I-echistatin as radioligand). Preliminary biodistribution of the (99m)Tc(I)-labeled radiotracer in orthotopic MDA-MB-435 breast tumor xenograft model was also evaluated. The BPy-RGD conjugate had good integrin-binding affinity (50% inhibitory concentration (IC(50))=92.51+/-22.69 nM), slightly lower than unmodified c(RGDyK) (IC(50)=59.07+/-11.03 nM). The hydrophilic radiotracer also had receptor-mediated activity accumulation in MDA-MB-435 tumor (1.45+/-0.25 percentage of injected dose per gram (%ID/g) at 1.5h postinjection (p.i.)), which is known to be integrin positive. After blocking with c(RGDyK), the tumor uptake was reduced from 0.71+/-0.01%ID/g to 0.33+/-0.18%ID/g at 4h p.i. (99m)Tc(I) tricarbonyl complex of cyclic RGD peptide is a promising strategy for integrin targeting. Further modification of the bipyridine-conjugated RGD peptide by using more potent RGD peptides and fine tuning of the tether group between the RGD moiety and (99m)Tc(CO)(3)(+) core to improve the tumor targeting efficacy and in vivo kinetic profiles is currently in progress.


Solar powered oxygen systems in remote health centers in Papua New Guinea: a large scale implementation effectiveness trial.
Pneumonia is the largest cause of child deaths in Papua New Guinea (PNG), and hypoxaemia is the major complication causing death in childhood pneumonia, and hypoxaemia is a major factor in deaths from many other common conditions, including bronchiolitis, asthma, sepsis, malaria, trauma, perinatal problems, and obstetric emergencies. A reliable source of oxygen therapy can reduce mortality from pneumonia by up to 35%. However, in low and middle income countries throughout the world, improved oxygen systems have not been implemented at large scale in remote, difficult to access health care settings, and oxygen is often unavailable at smaller rural hospitals or district health centers which serve as the first point of referral for childhood illnesses. These hospitals are hampered by lack of reliable power, staff training and other basic services. We report the methodology of a large implementation effectiveness trial involving sustainable and renewable oxygen and power systems in 36 health facilities in remote rural areas of PNG. The methodology is a before-and after evaluation involving continuous quality improvement, and a health systems approach. We describe this model of implementation as the considerations and steps involved have wider implications in health systems in other countries. The implementation steps include: defining the criteria for where such an intervention is appropriate, assessment of power supplies and power requirements, the optimal design of a solar power system, specifications for oxygen concentrators and other oxygen equipment that will function in remote environments, installation logistics in remote settings, the role of oxygen analyzers in monitoring oxygen concentrator performance, the engineering capacity required to sustain a program at scale, clinical guidelines and training on oxygen equipment and the treatment of children with severe respiratory infection and other critical illnesses, program costs, and measurement of processes and outcomes to support continuous quality improvement. This study will evaluate the feasibility and sustainability issues in improving oxygen systems and providing reliable power on a large scale in remote rural settings in PNG, and the impact of this on child mortality from pneumonia over 3 years post-intervention. Taking a continuous quality improvement approach can be transformational for remote health services.


Genetic map of the Actinobacillus pleuropneumoniae RTX-toxin (Apx) operons: characterization of the ApxIII operons.
Actinobacillus pleuropneumoniae RTX-toxin III (ApxIII) is implicated as an important virulence factor of A. pleuropneumoniae, the causative agent of porcine pleuropneumonia. Recently, the genes coding for ApxIII (apxIIICA) of serotype 8 were cloned and characterized. The toxin appeared to be a member of the RTX-toxin family, as are the other two secreted toxins of A. pleuropneumoniae, i.e., ApxI and ApxII. In this report, we describe the cloning and sequencing of the remaining part of the ApxIII operon of serotype 8. This sequence coded for the RTX secretion proteins ApxIIIB and ApxIIID, which showed 86 and 63% similarity to ApxIB and ApxID, respectively, and 83 and 63% similarity to HlyB and HlyD of Escherichia coli, respectively. Potential functional domains, such as eight transmembrane regions and an ATP-binding cassette, were present in ApxIIIB. We examined the presence of apxIIICABD sequences in the 12 serotypes of A. pleuropneumoniae and found that these sequences were present only in serotypes 2, 3, 4, 6, and 8, the serotypes that secrete ApxIII. Comparison of the apxIIICABD gene sequences of the serotypes revealed very few serotype-specific differences. Only the C terminus of ApxIIIA of serotype 2 differed from ApxIIIA of the other serotypes. The differences were located between the glycine-rich repeats and the secretion signal. The analysis of the apxIIICABD genes completed our efforts to characterize the ApxI, ApxII, and ApxIII operons of the reference strains of the 12 serotypes of A. pleuropneumoniae. We present a complete map of the ApxI, ApxII, and ApxIII operons and discuss this in terms of gene expression and complementation and the role of the toxins in pathogenesis.


Antigenic heterogeneity of leukemia.
When reassessing the clonal origin of malignant tumors, it is argued that most malignant tumors are heterogeneous for a number of phenotypic characteristics including antigenicity. Taking the murine AkR leukemia as an example, how antigenic heterogeneity may affect immunologic approaches to diagnostics and treatment of leukemias is discussed. The consequences for use of monoclonal antibodies in relation to human leukemias are also discussed, especially in light of recent achievements in producing human monoclonal antibodies.


EPs® 7630 Stimulates Tissue Repair Mechanisms and Modifies Tight Junction Protein Expression in Human Airway Epithelial Cells.
Airway epithelium repair after infection consists of wound repair, re-synthesis of the extracellular matrix (ECM), and tight junction proteins. In humans, EPs® 7630 obtained from Pelargonium sidoides roots reduces the severity and duration of acute respiratory tract infections. The effect of EPs® 7630 on tissue repair of rhinovirus-16 (RV-16) infected and control human airway epithelial cells was assessed for: (i) epithelial cell proliferation by manual cell counts, (ii) epithelial wound repair by "scratch assay", (iii) ECM composition by Western-blotting and cell-based ELISA, and (iv) epithelial tight junction proteins by Western-blotting. EPs® 7630 stimulated cell proliferation through cAMP, CREB, and p38 MAPK. EPs® 7630 significantly improved wound repair. Pro-inflammatory collagen type-I expression was reduced by EPs® 7630, while fibronectin was increased. Virus-binding tight junction proteins desmoglein2, desmocollin2, ZO-1, claudin1, and claudin4 were downregulated by EPs® 7630. The RV16-induced shift of the ECM towards the pro-inflammatory type was prevented by EPs® 7630. Most of the effects of EPs® 7630 on tissue repair and regeneration were sensitive to inhibition of cAMP-induced signaling. The data suggest that EPs® 7630-dependent modification of epithelial cell metabolism and function might underlie the faster recovery time from viral infections, as reported by others in clinical studies.


Reliability in Standardization of Iron(III) and Titanium(III) Solutions in Volumetric Analysis.
Titanium(III) is a useful strong reductant and is usually standardized with iron(III) in volumetric analysis. Iron(III) is widely used as an oxidant and is usually standardized with thiosulfate ions through an iodine liberation reaction. The evaluation of the standardization procedure for iron(III) with thiosulfate ions is therefore essential to ensure the reliability of standardized titanium(III) solutions. To investigate the titration procedure for iron(III), two different titrations were performed: redox titration with thiosulfate ions through an iodine liberation reaction and chelatometric titration with disodium dihydrogen ethylenediaminetetraacetate. Subsequently, for the investigation of standardization of iron(III), titanium(III) was assayed through two titration paths: redox titration with standardized iron(III) and redox titration with standard potassium dichromate. The reliability of titrimetric procedures was evaluated by applying several different stoichiometric reactions to each chemical. All titrimetric procedures were consistent with each other within their expanded uncertainties and were capable of providing reliable volumetric standards with careful operations presented in this study.


Chronic bronchitis. A bronchologic evaluation.
214 patients with chronic bronchitis were subjected to bronchial studied at Päivärinne Chest Hospital during the period 1967-1973. Bronchoscopy alone gave a frequency of 30% for bronchiectases while the information from bronchography increased this figure to 44%. In addition, 24% of the patients showed bronchographic changes in the form of naked filling or mucus obstrcution. Sputum specimens were less reliable indicators of infection than bronichial aspirates in patients with chronic bronchitis. Tracheobroncho-malacia was found in 50 patients and greatly increased the gravity of the disease.


Molecular cloning and sequencing of a major Bacillus subtilis autolysin gene.
A major Bacillus subtilis 168S autolysin (N-acetylmuramoyl-L-alanine amidase [EC 184.108.40.206]) was purified and then cleaved with cyanogen bromide. The N-terminal amino acid sequence of one of the resultant peptides was determined in order to make synthetic oligonucleotides. A 2.5-kb EcoRI fragment was cloned into Escherichia coli JM109 and detected by colony hybridization by using the oligonucleotides as probes. Sequencing of the insert showed the presence of an open reading frame (designated cwlB), starting at a UUG codon, which encodes a polypeptide of 496 amino acids with a molecular mass of 52,623 Da. CWLB had a presumed signal peptide which is processed after Ala at position 24. Insertional inactivation of the cwlB gene of the B. subtilis chromosome led to an approximately 90% decrease in the total cell wall hydrolytic activity of stationary-phase cells and extraordinary resistance to cell lysis, even after 6 days of incubation at 37 degrees C. No apparent changes in cell morphology, motility, competence, sporulation, or germination were observed.


Risk Factors for Post-Acute Coronary Syndrome Depression: A Meta-analysis of Observational Studies.
The incidence of depression is very common among patients with post-acute coronary syndrome (ACS) and leads to adverse outcomes. The aim of this meta-analysis was to detect risk factors for depression among patients with ACS and to provide clinical evidence for its prevention. The authors followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guideline to search the PubMed, Web of Science, EMBASE, and EBSCO databases from January 1996 to March 2018. Data that met the inclusion criteria were extracted to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for the risk factors of post-ACS depression. A total of 30 articles met the inclusion criteria, and 25 risk factors were found to be associated with depression. The top 5 risk factors are as follows: antidepression treatment (OR, 4.25; 95% CI, 3.41-5.31), housewife status (OR, 4.17; 95% CI, 1.83-9.53), history of depressive disorders (OR, 3.52; 95% CI, 2.69-4.61), widow status (OR, 2.34; 95% CI, 1.05-5.21), and history of congestive heart failure (OR, 2.03; 95% CI, 1.04-3.97). The authors also found that a married status, high education level, and employment are protective factors. Clinical personnel should be alerted with regard to the high risk factors of depression, including female gender, low education level, unmarried status, living alone, unemployed status, unhealthy lifestyle, and complications such as cardiovascular, respiratory, and metabolic diseases. In particular, staff should pay attention to a history of previous depression, be concerned with the psychological condition of the patient, and monitor and perform early interventions to reduce the incidence of depression.


Progeria and the early aging in children: a case report.
The Hutchinson-Gilford syndrome or progeria is a rare autosomal dominant syndrome characterized by premature aging and involvement of internal systems, such as the circulatory and locomotor. The diagnosis is essentially clinical and the manifestations become more evident from the first year of life. Long term outcome data from Progeria Research Foundation clinical trials have demonstrated an increase in survival in recent years. Even though new trials are ongoing, the recognition of this syndrome is essential to prevent cardiovascular and cerebrovascular complications. A patient, initially asymptomatic, who developed characteristic signs of the syndrome at the age of 6 months is reported. She was referred for evaluation only when she was two years and eleven months old. The diagnosis of Hutchinson-Gilford syndrome was suspected owing to clinical characteristics. The diagnosis was confirmed by genetic testing. A mutation c.1824C> T in exon 11 of the LMNA gene was detected. She was registered in the Progeria Research Foundation and was invited to participate in the weighing and supplementation program. She was included in the lonafarnib protocol study. This medication is a farnesyl transferase inhibitor that prevents the production of progerina and slows cardiovascular and neurological complications of the syndrome. This case highlights the importance of diagnosing progeria patients because they may be referred to the Progeria Research Foundation, which offers genetic screening and inclusion in clinical and therapeutic follow-up protocols without any costs. Progeria trials and research may also contribute to new drug developments related to prevention of aging and atherosclerosis in the near future.


Gene expression and genetic control to cold tolerance during maize seed germination.
The study of cold tolerance in maize seeds and seedlings through physiological quality assessments, as well as the genetic control associated with this trait, allows an early characterization of genotypes. Here we studied the genetic control for cold tolerance during the germination process in maize seeds and genes influenced by this stress. Six maize lines were used, three classified as tolerant and three as susceptible to low germination temperature. A field was developed to produce the hybrid seeds, in a partial diallel scheme including the reciprocal crosses. For the expression analysis, seeds from two contrasting lines were used, as well as their hybrid combination and their reciprocal crosses, on dried and moistened seeds at 10 °C for 4 and 7 days. It was evaluated the catalase (CAT) and esterase (EST) enzymes, heat-resistant proteins and the genes Putative stearoyl-ACP desaturase (SAD), Ascorbate Peroxidase (APX), Superoxide Dismutase (SOD) and Mitogen Activated Protein Kinase (ZmMPK5). The estimated values for heterosis, general and specific combining abilities and reciprocal maternal and non-maternal effects were carried out and it showed that there is heterosis for germination at low temperatures, also the non-additive genes were more important and there was a reciprocal effect. There is a greater expression of the CAT and EST enzymes in moistened seeds at seven days and there is less expression of heat-resistant proteins and the SAD gene at seven days of moistening. Also, there are variations in the expression of the APX, SOD and ZmMPK5 genes in dried and moistened seeds, as well as among the genotypes studied.


The population bottleneck of the Iberian wolf impacted genetic diversity but not admixture with domestic dogs: A temporal genomic approach.
After decades of intense persecution, the Iberian wolf subspecies faced a severe bottleneck in the 1970s that considerably reduced its range and population size, nearly leading to its extinction in central and southern Iberian Peninsula. Such population decline could have impacted the genetic diversity of Iberian wolves through different processes, namely genetic drift and dynamics of hybridization with domestic dogs. By contrasting the genomes of 68 contemporary with 54 historical samples spanning the periods before and immediately after the 1970s bottleneck, we found evidence of its impact on genetic diversity and dynamics of wolf-dog hybridization. Our genome-wide assessment revealed that wolves and dogs form two well-differentiated genetic groups in Iberia and that hybridization rates did not increase during the bottleneck. However, an increased number of hybrid individuals was found over time during the population re-expansion, particularly at the edge of the wolf range. We estimated a low percentage of dog ancestry (~1.4%) in historical samples, suggesting that dog introgression was not a key driver for wolf extinction in central and southern Iberia. Our findings also unveil a significant decline in genetic diversity in contemporary samples, with the highest proportion of homozygous segments in the genome being recently inherited. Overall, our study provides unprecedented insight into the impact of a sharp decline on the Iberian wolf genome and refines our understanding of the ecological and evolutionary drivers of wolf-dog hybridization in the wild.


Interleukin-6 -174 G/C promoter gene polymorphism in nasal polyposis and asthma.
Interleukin-6 (IL-6) is an inflammatory mediator linked to nasal polyposis and asthma, with a single nucleotide poly- morphism -174 G/C that seems to promote an inflammatory status. We aimed to analyze the relationship between this poly-morhism and asthmatic nasal polyposis patients. Cross-sectional study to investigate IL-6 - 174 G/C genotypes of 45 nasal polyposis with asthma patients, 63 nasal polyposis-only patients, 45 asthma-only patients and 81 subjects without both diseases. Aspirin intolerance and atopy were main exclusion criteria. IL-6 genotyping was performed using the PCR method with specific primers followed by restriction enzyme analysis, classifying patients in GG, GC or CC genotype. The GG genotype was the most frequent in all inflammatory groups. Less than 40% of controls presented with the GG ge- notype. There were significant differences between inflammatory groups and control group. No significant differences were seen when comparing inflammatory groups to each other, other than between nasal polyposis-only group and asthma-only group. The IL-6 74 GG genotype was found more frequently in all inflammatory groups than in controls. This genotype could influence nasal polyposis and asthma, and seems to be more important in the latter.


Anharmonic densities of states: A general dynamics-based solution.
Density of states is a fundamental physical characteristic that lies at the foundation of statistical mechanics and theoretical constructs that derive from them (e.g., kinetic rate theories, phase diagrams, and others). Even though most real physical systems are anharmonic, the vibrational density of states is customarily treated within the harmonic approximation, or with some partial, often limited, account for anharmonicity. The reason for this is that the problem of anharmonic densities of states stubbornly resisted a general and exact, yet convenient and straightforward in applications, solution. Here we formulate such a solution within both classical and quantum mechanics. It is based on actual dynamical behavior of systems as a function of energy and as observed, or monitored, on a chosen time scale, short or long. As a consequence, the resulting anharmonic densities of states are fully dynamically informed and, in general, time-dependent. As such, they lay the ground for formulation of new statistical mechanical frameworks that incorporate time and are ergodic, by construction, with respect to actual dynamical behavior of systems.


Phytoplankton and Microzooplankton Community Structure and Assembly Mechanisms in Northwestern Pacific Ocean Estuaries with Environmental Heterogeneity and Geographic Segregation.
Phytoplankton and microzooplankton are crucial players in marine ecosystems and first responders to environmental changes, but their community structures and how they are shaped by environmental conditions have rarely been studied simultaneously. In this study, we conducted an eDNA metabarcoding sequencing combined with multiple statistical methods to simultaneously analyze the phytoplankton and microzooplankton in Liaohe (LH) and Yalujiang (YLJ) estuaries. The major objective was to examine how plankton community structure and assembly mechanism may differ between two estuaries with similar latitudinal position and climate but geographical segregation and differential level of urbanization (more in LH). Clear differences in diversity and composition of phytoplankton and microzooplankton communities between LH and YLJ estuaries were observed. Richness of phytoplankton was significantly higher in LH than YLJ, while richness of microzooplankton was higher in YLJ. The magnitude of intrahabitat variations in phytoplankton communities was significantly stronger than that of microzooplankton. Some phytoplankton and microzooplankton taxa also showed interhabitat differences in their relative abundances. Phytoplankton showed a stronger geographic distance-decay of similarity than microzooplankton, while significant environmental distance-decay of similarity in microzooplankton was found in the less urbanized YLJ estuary. Community assembly of phytoplankton was, based on the neutral community models, driven primarily by stochastic processes, while deterministic processes contributed more for microzooplankton. Furthermore, we detected wider habitat niche breadths and stronger dispersal abilities in phytoplankton than in microzooplankton. These results suggest that passive dispersal shapes the phytoplankton community whereas environmental selection shapes the microzooplankton community. IMPORTANCE Understanding the underlying mechanisms shaping a metacommunity is useful to management for improving the ecosystem function. The research presented in the manuscript mainly tried to address the effects of habitat geography and environmental conditions on the phytoplankton and microzooplankton communities, and the underlying mechanisms of community assembly in temperate estuaries. In order to achieve this purpose, we developed a metabarcoding sequencing method based on 18S rRNA gene. The phytoplankton and microzooplankton communities from two estuaries with similar latitude and climatic conditions but obvious geographical segregation and significant environmental heterogeneity were investigated. The results of our study could lay a solid foundation for ascertaining phytoplankton and microzooplankton communities in estuaries with obvious environmental heterogeneity and geographic segregation and mechanisms underlying community assembly.


Comparative study between skin prick tests and TOP-CAST allergen leukocyte stimulation in diagnosis of allergic status.
A recently developed CAST-ELISA method was applied to determine allergen-induced leukocyte stimulation. This method is based on the measurement of sulfidoleukotriene secretion by peripheral blood leukocytes previously stimulated with specific allergen in the presence of interleukin 3. 24 patients allergic to different aeroallergens and some food allergens were included in this study. Subjects received no medication 2 weeks before the testing. Leukocytes were isolated by dextran sedimentation. After incubation with allergen at two concentrations, the cells were centrifuged and the concentration of sulfidoleukotrienes in supernatant was determined. The results were expressed in pg/ml after subtraction of values of spontaneous sulfidoleukotriene production in portions incubated without allergen. We observed a wide range of sulfidoleukotriene secretion upon allergen stimulation. Concentrations of leukotrienes ranged from 0 to 5780 pg/ml at lower allergen concentration and from 210 to 5680 at higher allergen concentration. On the basis of observed results, we conclude that the better allergen concentration is the higher one because there was no appearance of negative cell stimulation. In the lower allergen concentration we observed negative results in two cases. Sixteen healthy control subjects were also included in this study. Eleven subjects had negative skin prick tests (SPT) as well as CAST results. In 5 healthy subjects with positive SPT, we also observed positive CAST results in 4 persons. In 1 healthy person with positive SPT results, CAST results were negative. We conclude that TOP-CAST allergen is a valuable mixture of different aero and food allergens for determining the allergic status in patients with suspicious allergic status, although it cannot differentiate between health and disease states.


[The psychiatrist and pain].
The psychiatrist might be called upon for states of pain on two occasions. The first is when the pain is mainly functioned, with or without a previous history of organic pain. It is most frequently met, with various diagnostic difficulties, in hypochondriac neurosis, complaining delirium, schizophrenia, multiple somatizations, isolated psychogenic pain and masked depression. The second is when the pain is organic, always progressive, with associated psychiatric disturbances which interact with it. We shall consider these various problems from both a clinical and a therapeutic point of view aiming at their practical rather than their theoretical aspects.


Multilayered LDH/Microcapsule Smart Epoxy Coating for Corrosion Protection.
A multilayered smart epoxy coating for corrosion prevention of carbon steel was developed and characterized. Toward this direction, as a first step, zinc-aluminum nitrate-layered double hydroxide (Zn/Al LDH) was synthesized using the hydrothermal crystallization technique and then loaded with dodecylamine (DOD), which was used as an inhibitor (pH-sensitive). Similarly, the synthesis of the urea-formaldehyde microcapsules (UFMCs) has been carried out using the in-situ polymerization method, and then the microcapsules (LAUFCs) were encapsulated with linalyl acetate (LA) as a self-healing agent. Finally, the loaded Zn/Al LDH (3 wt %) and modified LAUFCs (5 wt %) were reinforced into an epoxy matrix to develop a double-layer coating (DL-EP). For an exact comparison, pre-layer epoxy coatings comprising 3 wt % of the loaded Zn/Al LDH (referred to as LDH-EP), top-layer epoxy coatings comprising 5 wt % linalyl acetate urea-formaldehyde microcapsules (referred to as UFMLA COAT), and a blank epoxy coating (reference coating) were also developed. The developed epoxy coatings were characterized using various techniques such as XRD, XPS, BET, TGA, FTIR, EIS, etc. Electrochemical tests performed on the synthesized coatings indicate that the DL-EP demonstrates improved self-healing properties compared to LDH-EP and UFMLA COAT.


Tapentadol extended release for chronic pain patients.
Chronic pain reduces quality of life, utilizes healthcare resources, and increases healthcare costs. It is widespread, but generally inadequately treated or managed, partly due to several obstacles, including a limited number of mechanistic options for long-term pharmacologic agents. Opioids are generally the primary class of analgesic prescribed, but because of associated side effects during long-term treatment, many patients become noncompliant or discontinue treatment. A long-term use analgesic with a good benefit/risk ratio is advantageous. A literature search for randomized trials using tapentadol extended release (ER) for noncancer chronic pain patients was conducted. Databases searched included PubMed, MEDLINE, EMBASE, and Google Scholar, using key terms "tapentadol," "prolonged release," "extended release," and "chronic pain" individually or in combination. The results were synthesized and evaluated. A total of six randomized, controlled studies were identified. Chronic pain conditions analyzed included low back, osteoarthritis, and diabetic peripheral neuropathy. Treatment arms consisted most often of placebo, tapentadol ER (100-250 mg twice daily [b.i.d.]), and/or oxycodone CR (controlled release) (20-50 mg b.i.d.). Subjects treated with tapentadol ER had significant reduction in pain intensity compared to placebo controls and similar efficacy to oxycodone CR. Overall, the safety profile was superior to that of oxycodone CR in regards to reduction in side effects, reduced severity of side effects (particularly gastrointestinal related), and lower study discontinuation rates. The two mechanisms of analgesic action of tapentadol, combined with an ER, appears to provide equal efficacy to a strong controlled-release opioid while providing greater gastrointestinal tolerability. The reduction in incidence and severity of gastrointestinal side effects correlated with a higher compliance rate. These findings suggest that tapentadol ER might be a viable alternative to conventional strong opioids for pain management for chronic pain patients.


The morbidity and mortality of vermiculite miners and millers exposed to tremolite-actinolite: Part II. Mortality.
The vermiculite ore and concentrate of a mine and mill located near Libby, Montana was found to be contaminated with a fiber of the tremolite/acetinolite series. A study was conducted to estimate the exposure-response relationship for mortality for 575 men who had been hired prior to 1970 and employed at least 1 year at the Montana site. Individual cumulative fiber exposure (fiber-years) was calculated. Results indicated that mortality from nonmalignant respiratory disease (NMRD) and lung cancer was significantly increased compared to the U.S. white male population. For those workers more than 20 years since hire, the standard mortality rate (SMR) for lung cancer (ICDA 162-163) was 84.7, 225.1, 109.3, and 671.3 for less than 50, 50-99, 100-399, and more than 399 fiber-years respectively. Corresponding results for NMRD (ICDA 460-519) were 327.8, 283.5, 0, and 278.4. Based on a linear model for greater than 20 years since hire, the estimated percentage increase in lung cancer mortality risk was 0.6% for each fiber-year of exposure. At 5 fiber-years, the estimated percentage was 2.9% from an unrestricted (nonthreshold) linear model and 0.6% from a survival model.


Beliefs and Traditions Related to a Child's First Year of Life: A Study of the North-west of Portugal.
In this paper we propose an approach to investigate, in the North-west of Portugal, the parents' behaviour at birth and during the first year of life of their children. We compare the heritage, specifically the beliefs and traditions, with the changes that resulted from the recent and deep cultural transformations that have taken place in Portugal in the last few decades. In parallel, we tried to determine if the parents' behaviours, based on beliefs and traditions, can affect the children's health. We based our investigation on standardized interviews with 76 mothers of one-year-old children (born between January and December 2001) who lived in two parishes of Vizela city. This is a territory where a more traditional way of life prevails than in other territories of the centre and south of the country, where there is a strong attachment for religious and social values and where the influence of the ancestral traditions is still alive. The paper concludes that cultural heritage can have important impact on individual health. Health professionals, who work in primary care and in hospitals, must be aware of the responsibility they have to change this scenario.


Assessment of the feed additive consisting of Enterococcus lactis DSM 7134 and Lacticaseibacillus rhamnosus DSM 7133 (Provita LE) for calves for rearing for the renewal of its authorisation (Lactosan GmbH & Co.KG).
Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the assessment of the application for renewal of the authorisation of the additive Provita LE for calves for rearing, consisting of Enterococcus lactis DSM 7134 (formerly identified as Enterococcus faecium) and Lacticaseibacillus rhamnosus DSM 7133 (formerly identified as Lactobacillus rhamnosus) as a zootechnical feed additive. The applicant has provided evidence that the additive currently on the market complies with the existing terms of the authorisation. The FEEDAP Panel concluded that the use of the feed additive in animal nutrition remains safe for calves for rearing, consumers and the environment under the authorised conditions of use. The additive is not irritant to skin or eyes but should be considered a respiratory sensitiser. It was not possible to draw conclusions on the skin sensitisation potential of the additive under assessment. There was no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.


Vanilloid receptors mediate adrenergic nerve- and CGRP-containing nerve-dependent vasodilation induced by nicotine in rat mesenteric resistance arteries.
Previous studies showed that nicotine induces adrenergic nerve-dependent vasodilation that is mediated by endogenous calcitonin gene-related peptide (CGRP) released from CGRP-containing (CGRPergic) nerves. The mechanisms underlying the nicotine-induced vasodilation were further studied. Rat mesenteric vascular beds without endothelium were contracted by perfusion with Krebs solution containing methoxamine, and the perfusion pressure was measured with a pressure transducer. Perfusion of nicotine (1-100 microm) for 1 min caused concentration-dependent vasodilation. Capsazepine (vanilloid receptor-1 antagonist; 1-10 microm) and ruthenium red (inhibitor of vanilloid response; 1-30 microm) concentration-dependently inhibited the nicotine-induced vasodilation without affecting the vasodilator response to exogenous CGRP. Nicotine-induced vasodilation was not inhibited by treatment with 3,4-dihydroxyphenylalanine (DOPA) receptor antagonist (l-DOPA cyclohexyl ester; 0.001-10 microm), dopamine D1 receptor-selective antagonist (SCH23390; 1-10 microm), dopamine D2 receptor antagonist (haloperidol; 0.1-0.5 microm), ATP P2x receptor-desensitizing agonist (alpha,beta-methylene ATP; 1-10 microm), adenosine A2 receptor antagonist (8(p-sulfophenyl)theophylline; 10-50 microm) or neuropeptide Y (NPY)-Y1 receptor antagonist (BIBP3226; 0.1-0.5 microm). Immunohistochemical staining of the mesenteric artery showed dense innervation of CGRP- and vanilloid receptor-1-positive nerves, with both immunostainings appearing in the same neuron. The mesenteric artery was also densely innervated by NPY-positive nerves. Double immunostainings showed that both NPY and CGRP immunoreactivities appeared in the same neuron of the artery. These results suggest that nicotine acts on presynaptic nicotinic receptors to release adrenergic neurotransmitter(s) or related substance(s), which then stimulate vanilloid receptor-1 on CGRPergic nerves, resulting in CGRP release and vasodilation.


[Effect of anti-cancer drugs on the expression of BIC/miR-155 in human pancreatic cancer PANC-1 cells].
To investigate the effect of anti-cancer drugs on the expression of B-cell integration cluster (BIC) RNA/miRNA-155 in human pancreatic cancer PANC-1 cells. PANC-1 cells were treated with different concentrations of anti-cancer drugs. Total RNA of the treated cells were harvested and the expression levels of BIC RNA and mature miR-155 were quantified by using Taqman FAM/MGB probes on a real-time PCR system. Relative quantification was carried out using the DeltaDeltaCt method. A PI3K-related kinases inhibitor was used to determine whether these kinases were involved in the regulation of BIC RNA. The expression of BIC RNA was strongly induced by anti-cancer drugs. When PANC-1 cells were treated by gemcitabine with concentrations of 1.0, 2.5, 5.0, 10.0 mg/L for 48 h and 72 h, the level of BIC RNA (48 h: 37.1 +/- 4.1, 29.0 +/- 5.7, 21.0 +/- 7.6, 40.4 +/- 9.0, 72 h: 27.7 +/- 3.1, 43.1 +/- 1.2, 31.8 +/- 5.4, 23.1 +/- 1.4) were significantly higher than that of the control (48 h: 1.6 +/- 1.1, 72 h: 1.0 +/- 0.1, all P < 0.05). 5-FU (10 mg/L, 48 h) and bleomycin (100 mg/L, 48 h) also induced BIC RNA up-regulation (5.2 +/- 1.1 vs 1.7 +/- 0.7, 11.5 +/- 0.7 vs 1.7 +/- 0.7, both P < 0.05). When PANC-1 cells treated with 1.0, 2.5, 5.0, 10.0 mg/L gemcitabine for 72 h, the level of miR-155 (2.21 +/- 0.40, 1.86 +/- 0.03, 2.47 +/- 0.04, 3.24 +/- 0.05) also higher than that of the control (1.11 +/- 0.09, P < 0.05), while no change was observed when the cells only treated for 48 h. Further study showed gemcitabine-induced BIC RNA up-regulation was inhibited by wortmannin, a specific PI3K inhibitor, the expression levels of BIC RNA of 1 micromol/L wortmannin + 5 mg/L gemcitabine group and 10 micromol/L wortmannin + 5 mg/L gemcitabine group were 5.34 +/- 1.11 and 1.26 +/- 0.07, lower than that of 5 mg/L gemcitabine group (11.82 +/- 3.11, P < 0.05). BIC RNA is strongly induced by anti-cancer drugs in PANC-1 cells and the levels of miR-155 also slightly increase. PI3K pathway is involved in gemcitabine-induced BIC RNA up-regulation.


Delayed onset muscle soreness: Involvement of neurotrophic factors.
Delayed-onset muscle soreness (DOMS) is quite a common consequence of unaccustomed strenuous exercise, especially exercise containing eccentric contraction (lengthening contraction, LC). Its typical sign is mechanical hyperalgesia (tenderness and movement related pain). Its cause has been commonly believed to be micro-damage of the muscle and subsequent inflammation. Here we present a brief historical overview of the damage-inflammation theory followed by a discussion of our new findings. Different from previous observations, we have observed mechanical hyperalgesia in rats 1-3 days after LC without any apparent microscopic damage of the muscle or signs of inflammation. With our model we have found that two pathways are involved in inducing mechanical hyperalgesia after LC: activation of the B2 bradykinin receptor-nerve growth factor (NGF) pathway and activation of the COX-2-glial cell line-derived neurotrophic factor (GDNF) pathway. These neurotrophic factors were produced by muscle fibers and/or satellite cells. This means that muscle fiber damage is not essential, although it is sufficient, for induction of DOMS, instead, NGF and GDNF produced by muscle fibers/satellite cells play crucial roles in DOMS.


Soft tissue sarcomas of the hand.
The most common soft tissue sarcomas of the hand are epithelioid sarcoma, rhabdomyosarcoma, synovial sarcoma, fibrosarcoma, and clear cell sarcoma. The epidemiology and biology of these tumors are discussed and important unifying concepts, such as local recurrences and regional lymph node metastases, are stressed.


Extensive Bioinformatics Analyses Reveal a Phylogenetically Conserved Winged Helix (WH) Domain (Zτ) of Topoisomerase IIα, Elucidating Its Very High Affinity for Left-Handed Z-DNA and Suggesting Novel Putative Functions.
The dynamic processes operating on genomic DNA, such as gene expression and cellular division, lead inexorably to topological challenges in the form of entanglements, catenanes, knots, "bubbles", R-loops, and other outcomes of supercoiling and helical disruption. The resolution of toxic topological stress is the function attributed to DNA topoisomerases. A prominent example is the negative supercoiling (nsc) trailing processive enzymes such as DNA and RNA polymerases. The multiple equilibrium states that nscDNA can adopt by redistribution of helical twist and writhe include the left-handed double-helical conformation known as Z-DNA. Thirty years ago, one of our labs isolated a protein from Drosophila cells and embryos with a 100-fold greater affinity for Z-DNA than for B-DNA, and identified it as topoisomerase II (gene Top2, orthologous to the human UniProt proteins TOP2A and TOP2B). GTP increased the affinity and selectivity for Z-DNA even further and also led to inhibition of the isomerase enzymatic activity. An allosteric mechanism was proposed, in which topoII acts as a Z-DNA-binding protein (ZBP) to stabilize given states of topological (sub)domains and associated multiprotein complexes. We have now explored this possibility by comprehensive bioinformatic analyses of the available protein sequences of topoII representing organisms covering the whole tree of life. Multiple alignment of these sequences revealed an extremely high level of evolutionary conservation, including a winged-helix protein segment, here denoted as Zτ, constituting the putative structural homolog of Zα, the canonical Z-DNA/Z-RNA binding domain previously identified in the interferon-inducible RNA Adenosine-to-Inosine-editing deaminase, ADAR1p150. In contrast to Zα, which is separate from the protein segment responsible for catalysis, Zτ encompasses the active site tyrosine of topoII; a GTP-binding site and a GxxG sequence motif are in close proximity. Quantitative Zτ-Zα similarity comparisons and molecular docking with interaction scoring further supported the "B-Z-topoII hypothesis" and has led to an expanded mechanism for topoII function incorporating the recognition of Z-DNA segments ("Z-flipons") as an inherent and essential element. We further propose that the two Zτ domains of the topoII homodimer exhibit a single-turnover "conformase" activity on given G(ate) B-DNA segments ("Z-flipins"), inducing their transition to the left-handed Z-conformation. Inasmuch as the topoII-Z-DNA complexes are isomerase inactive, we infer that they fulfill important structural roles in key processes such as mitosis. Topoisomerases are preeminent targets of anti-cancer drug discovery, and we anticipate that detailed elucidation of their structural-functional interactions with Z-DNA and GTP will facilitate the design of novel, more potent and selective anti-cancer chemotherapeutic agents.


Protein-DNA footprinting by endcapped duplex oligodeoxyribonucleotides.
Oligodeoxyribonucleotides (5'-phosphorylated) of varying lengths were capped using a polyamide linker to form thermodynamically stable, endcapped DNA duplexes containing 8-14 bp. We have employed these endcapped DNA duplexes as tools to determine the DNA footprint of T4 DNA ligase. By high-performance liquid chromatography and PAGE analysis of the ligation mixtures of the endcapped DNA duplexes, we have found that by varying the lengths and the position of the nick, we can determine the minimal DNA-binding site as well as the mode of binding (symmetrical or asymmetrical binding) by the enzyme. The results of the study revealed that a 11 bp endcapped duplex was the shortest duplex effectively ligated. Dependence of ligation efficiency on nick position demonstrates that T4 DNA ligase bound asymmetrically to its DNA substrate. The use of a set of thermodynamically stable endcapped deoxyribonucleoside duplexes as a tool to elucidate the DNA footprint provides an efficient strategy for footprinting, which avoids ambiguities associated with chemical and biochemical footprinting methods.


Hyporesponsiveness of canine bronchoalveolar lymphocytes to mitogens: II. Analysis of macromolecular synthesis, T-cell antigen, and mitogen-binding.
Canine bronchoalveolar lymphocytes (BAL) and peripheral blood lymphocytes (PBL) were prepared by filtration over nylon wool columns and were studied for their ability to: (a) synthesize protein, RNA, and DNA after mitogen stimulation; (b) express T-cell antigen; and (c) bind radiolabeled mitogen. Bronchoalveolar lymphocytes were shown to be markedly hyporesponsive to mitogen stimulation as assessed by DNA synthesis after 72 hr of culture or by protein and RNA synthesis after 24 hr of culture. The hyporesponsiveness of BAL was not due to a decrease in the number of T-cells; results of an indirect immunofluorescent assay employing a rabbit antiserum indicated similar percentages of T-cells in BAL (78%) and PBL (79%). The hyporesponsiveness was not due to abnormalities in mitogen binding by BAL; results of a radiolabeled mitogen-binding assay indicated essentially no difference in mitogen receptor number or affinity between BAL and PBL. Thus, the initial interaction bronchoalveolar T lymphocytes and mitogen appears to be normal. This normal interaction with mitogen, however, fails to trigger an appropriate level of macromolecular synthesis by BAL. This suggests that the hyporesponsiveness of BAL is due to an abnormality in one of the early metabolic processes induced by the interaction of a mitogen with the cell membrane.


[Comparison of the effectiveness of omeprazole and pantoprazole treatment of gastroesophageal reflux disease in patients with asthma].
The combination of bronchial asthma with pathology of the digestive tract--one of the most frequent, clinically diverse and difficult, which complicates its course. The prevalence of gastroesophageal reflux disease in patients with asthma ranged from 33 to 90%. Aggravation of GERD is considered as one of the triggers of asthma attacks. The basis for treatment of GERD is acid suppression. Select proton pump inhibitors as the primary means of therapy related to their antisecretory action. Among the PPI pantoprazole can be identified that has high bioavailability, has minimal drug interactions with other drugs and a favorable safety profile. In patients taking pantoprazole pH-metry approached the norm, the proportion of time with pH < 4 decreased from 2 h 31 min to 52 min. On the background of pantoprazole indicators of lung function were better and significantly higher than in the groupwith omeprazole. Patients with combined pathology requiring compulsory treatment concomitant GERD, which will improve the course of asthma and help to achieve control.


Dose estimation to eye lens of industrial gamma radiography workers using the Monte Carlo method.
The ICRP Statement on Tissue Reactions (2011), based on epidemiological evidence, recommended a reduction for the eye lens equivalent dose limit from 150 to 20 mSv per year. This paper presents mainly the dose estimations received by industrial gamma radiography workers, during planned or accidental exposure to the eye lens, Hp(10) and effective dose. A Brazilian Visual Monte Carlo Dose Calculation program was used and two relevant scenarios were considered. For the planned exposure situation, twelve radiographic exposures per day for 250 days per year, which leads to a direct exposure of 10 h per year, were considered. The simulation was carried out using a 192Ir source with 1.0 TBq of activity; a source/operator distance between 5 and 10 m and placed at heights of 0.02 m, 1 m and 2 m, and an exposure time of 12 s. Using a standard height of 1 m, the eye lens doses were estimated as being between 16.3 and 60.3 mGy per year. For the accidental exposure situation, the same radionuclide and activity were used, but in this case the doses were calculated with and without a collimator. The heights above ground considered were 1.0 m, 1.5 m and 2.0 m; the source/operator distance was 40 cm, and the exposure time 74 s. The eye lens doses at 1.5 m were 12.3 and 0.28 mGy without and with a collimator, respectively. The conclusions were that: (1) the estimated doses show that the 20 mSv annual limit for eye lens equivalent dose can directly impact industrial gamma radiography activities, mainly in industries with high number of radiographic exposures per year; (2) the risk of lens opacity has a low probability for a single accident, but depending on the number of accidental exposures and the dose levels found in planned exposures, the threshold dose can easily be exceeded during the professional career of an industrial radiography operator, and; (3) in a first approximation, Hp(10) can be used to estimate the equivalent dose to the eye lens.


Neuropsychiatric Phenotypes Produced by GABA Reduction in Mouse Cortex and Hippocampus.
Whereas cortical GAD67 reduction and subsequent GABA level decrease are consistently observed in schizophrenia and depression, it remains unclear how these GABAergic abnormalities contribute to specific symptoms. We modeled cortical GAD67 reduction in mice, in which the Gad1 gene is genetically ablated from ~50% of cortical and hippocampal interneurons. Mutant mice showed a reduction of tissue GABA in the hippocampus and cortex including mPFC, and exhibited a cluster of effort-based behavior deficits including decreased home-cage wheel running and increased immobility in both tail suspension and forced swim tests. Since saccharine preference, progressive ratio responding to food, and learned helplessness task were normal, such avolition-like behavior could not be explained by anhedonia or behavioral despair. In line with the prevailing view that dopamine in anterior cingulate cortex (ACC) plays a role in evaluating effort cost for engaging in actions, we found that tail-suspension triggered dopamine release in ACC of controls, which was severely attenuated in the mutant mice. Conversely, ACC dopamine release by progressive ratio responding to reward, during which animals were allowed to effortlessly perform the nose-poking, was not affected in mutants. These results suggest that cortical GABA reduction preferentially impairs the effort-based behavior which requires much effort with little benefit, through a deficit of ACC dopamine release triggered by high-effort cost behavior, but not by reward-seeking behavior. Collectively, a subset of negative symptoms with a reduced willingness to expend costly effort, often observed in patients with schizophrenia and depression, may be attributed to cortical GABA level reduction.


Biomass composition: the "elephant in the room" of metabolic modelling.
Genome-scale stoichiometric models, constrained to optimise biomass production are often used to predict mutant phenotypes. However, for Saccharomyces cerevisiae, the representation of biomass in its metabolic model has hardly changed in over a decade, despite major advances in analytical technologies. Here, we use the stoichiometric model of the yeast metabolic network to show that its ability to predict mutant phenotypes is particularly poor for genes encoding enzymes involved in energy generation. We then identify apparently inefficient energy-generating pathways in the model and demonstrate that the network suffers from the high energy burden associated with the generation of biomass. This is tightly connected to the availability of phosphate since this macronutrient links energy generation and structural biomass components. Variations in yeast's biomass composition, within experimentally-determined bounds, demonstrated that flux distributions are very sensitive to such changes and to the identity of the growth-limiting nutrient. The predictive accuracy of the yeast metabolic model is, therefore, compromised by its failure to represent biomass composition in an accurate and context-dependent manner.