The burden of acute respiratory infections in crisis-affected populations: a systematic review.
Crises due to armed conflict, forced displacement and natural disasters result in excess morbidity and mortality due to infectious diseases. Historically, acute respiratory infections (ARIs) have received relatively little attention in the humanitarian sector. We performed a systematic review to generate evidence on the burden of ARI in crises, and inform prioritisation of relief interventions. We identified 36 studies published since 1980 reporting data on the burden (incidence, prevalence, proportional morbidity or mortality, case-fatality, attributable mortality rate) of ARI, as defined by the International Classification of Diseases, version 10 and as diagnosed by a clinician, in populations who at the time of the study were affected by natural disasters, armed conflict, forced displacement, and nutritional emergencies. We described studies and stratified data by age group, but did not do pooled analyses due to heterogeneity in case definitions. The published evidence, mainly from refugee camps and surveillance or patient record review studies, suggests very high excess morbidity and mortality (20-35% proportional mortality) and case-fatality (up to 30-35%) due to ARI. However, ARI disease burden comparisons with non-crisis settings are difficult because of non-comparability of data. Better epidemiological studies with clearer case definitions are needed to provide the evidence base for priority setting and programme impact assessments. Humanitarian agencies should include ARI prevention and control among infants, children and adults as priority activities in crises. Improved data collection, case management and vaccine strategies will help to reduce disease burden.


Assessment of cardiac troponin I (cTnI) and tissue velocity imaging (TVI) in 14 dogs with malignant lymphoma undergoing chemotherapy treatment with doxorubicin.
Doxorubicin has been shown to be cardiotoxic at high doses but is an efficacious chemotherapeutic agent in the treatment of canine lymphoma. Echocardiographic measurements and serum ultrasensitive cardiac troponin I (cTnI) levels were obtained before and after doxorubicin administration in 14 dogs diagnosed with lymphoma. The aim of this prospective study was to evaluate changes in cTnI concentrations and tissue velocity imaging (TVI) values in dogs with lymphoma undergoing chemotherapy with doxorubicin. A total of 182 cTnI and 1017 TVI measurements were performed. Standard echocardiographic parameters, tissue Doppler indices and cTnI concentrations did not differ at any time point within a 12-week cyclic combination protocol. In conclusion, the use of doxorubicin at standard doses in the treatment of canine lymphoma may not be associated with significant myocardial damage.


Calcium signal transmission in chick sensory neurones is diffusion based.
In many cells, the cytosol is an excitable medium through which calcium waves propagate by calcium induced calcium release (CICR). Many labs. have reported CICR in neurones subsequent to calcium influx through voltage gated channels. However, these have used long depolarizations. We have imaged calcium within chick sensory neurones following 50 ms depolarizations. Calcium signals travelled rapidly throughout the cell, such that changes at the cell centre were delayed by 24 ms compared to regions 3 microm from the plasma membrane. The nuclear envelope imposed a delay of 9 ms. A simple diffusion model with few unknowns gave good fits to the measured data, indicating that passive diffusion is responsible for signal transmission in these neurones. Simulations run without indicator dye did not reveal markedly different spatiotemporal dynamics, although concentration changes were larger. Simulations of calcium changes during action potentials revealed that large calcium transients occurring in the cytosol close to the nucleus are significantly attenuated by the nuclear envelope. Our results indicate that for the brief depolarisations that neurones will experience during normal signal processing calcium signals are transmitted by passive diffusion only. Diffusion is perfectly capable of transmitting the calcium signal into the interior of nerve cell bodies, and into the nucleoplasm.


A novel neutrophil chemoattractant generated during an inflammatory reaction in the rabbit peritoneal cavity in vivo. Purification, partial amino acid sequence and structural relationship to interleukin 8.
An inflammatory reaction was induced in vivo by injection of zymosan into the peritoneal cavity of the rabbit. The inflammatory exudate was found to contain oedema-inducing and neutrophil chemoattractant activity when assayed in rabbit skin in vivo, using 125I-albumin and 111In-neutrophils. This activity was additional to that of complement fragment C5a, which was removed by an affinity gel. Two chemoattractants were isolated by cation-exchange, gel-filtration and reversed-phase h.p.l.c. One of these, which ran as a single band of 6-8 kDa on SDS/PAGE, was subjected to N-terminal sequence analysis without reduction and alkylation of cysteine residues. Positive identification of 28 of the first 31 amino acids revealed a rabbit homologue of interleukin-8 (75% sequence identity with human interleukin-8). The demonstration of interleukin-8 as a major neutrophil chemoattractant in an inflammatory reaction in vivo provides the basis for further investigations into the role of this cytokine in the inflammatory process.


Role of potassium levels in pkBADH heterogeneity of NAD+ binding site.
Betaine aldehyde dehydrogenase (BADH) catalyzes the oxidation of betaine aldehyde to glycine betaine using NAD+ as a coenzyme. Studies in porcine kidney BADH (pkBADH) suggested that the enzyme exhibits heterogeneity of active sites and undergoes potassium-induced conformational changes. This study aimed to analyze if potassium concentration plays a role in the heterogeneity of pkBADH active sites through changes in NAD+ affinity constants, in its secondary structure content and stability. The enzyme was titrated with NAD+ 1 mM at fixed-variable KCl concentration, and the interaction measured by Isothermal Titration Calorimetry (ITC) and Circular Dichroism (CD). ITC data showed that K+ increased the first active site affinity in a manner dependent on its concentration; KD values to the first site were 14.4, 13.1, and 10.4 μM, at 25, 50, and 75 mM KCl. ΔG values showed that the coenzyme binding is a spontaneous reaction without changes between active sites or depending on KCl concentration. ΔH and TΔSb values showed that NAD+ binding to the active site is an endothermic process and is carried out at the expense of changes in entropy. α-Helix content increased as KCl increased, enzyme (Tm)app values were 2.6 °C and 3.3 °C higher at 20 mM and 200 mM K+. PkBADH molecular model showed three different interaction K+ sites. Results suggested K+ can interact with pkBADH and cause changes in the secondary structure, it provokes changes in the enzyme affinity by the coenzyme, and in the thermostability.


Intersections of Adolescent Well-Being: School, Work, and Weight Status in Brazil.
Socioeconomic and health disadvantages can emerge early in the life course, making adolescence a key period to examine the association between socioeconomic status and health. Past research on obesity in adolescence has focused on family measures of socioeconomic status, overlooking the role of individual-level nascent indicators of socioeconomic disadvantage. Using measured height and weight from nationally representative data from Brazil, we estimate sibling fixed effects models to examine the independent effects of nascent socioeconomic characteristics-school enrollment and work status-on adolescent overweight and obesity, accounting for unobserved genetic and environmental factors shared by siblings. Results show that school enrollment is associated with lower odds of overweight and obesity. Working is not significantly associated with overweight/obesity risk. However, adolescents not enrolled but working face the highest risk of overweight/obesity. Findings suggest that adolescents face added layers of disadvantage from being out of school, with important implications for the accumulation of health disadvantages.


Intranigral microinjection of neurotensin suppresses feeding in food deprived rats.
This study was conducted to determine whether the substantia nigra is involved in the anorectic effect of centrally administered neurotensin. Microinjection of neurotensin (2.5 - 10.0 micrograms) into the substantia nigra produced a significant suppression of feeding in 18 hour food deprived rats. To determine if the anorectic effect resulted from a general impairment of sensory-motor functioning, eight behaviors were measured in a separate study. Significant effects were found in only two of the eight behavioral categories (sniffing and eating), and only at the highest neurotensin dose (10.0 micrograms). These results suggest that the substantia nigra may be involved in the anorectic effect of neurotensin.


Clinical validation of IFNγ/IL-10 and IFNγ/IL-2 FluoroSpot assays for the detection of Tr1 T cells and influenza vaccine monitoring in humans.
The type of T cell polarization and simultaneous production of multiple cytokines have been correlated with vaccine efficacy. ELISpot is a T cell detection technique optimized for the measurement of a secreted cytokine at the single cell level. The FluoroSpot assay differs from ELISpot by the use of multiple fluorescent-labeled anticytokine detection antibodies, allowing optimal measurement of multiple cytokines. In the present study, we show that an IFNγ/IL-10 FluoroSpot assay is more sensitive than flow cytometry to detect Tr1 regulatory T cells, an immunosuppressive T cell population characterized by the production of IL-10 and IFNγ. As many tolerogenic vaccines are designed to induce these Tr1 cells, this FluoroSpot test could represent a standard method for the detection of these cells in the future. The use of an IFNγ/IL-2 FluoroSpot assay during influenza vaccine monitoring showed that the influenza-specific IL-2-producing T-cell response was the dominant response both before and after vaccine administration. This study therefore questions the rationale of using the single-color IFNγ ELISpot as the standard technique to monitor vaccine-specific T-cell response. Using this same test, a trend was also observed between baseline levels of IFNγ T cell response and T cell vaccine response. In addition, a lower IFNγ+IL-2+ T-cell response after vaccine was observed in the group of patients treated with TNFα inhibitors (P=0.08). This study therefore supports the use of the FluoroSpot assay due to its robustness, versatility and the complementary information that it provides compared with ELISpot or flow cytometry to monitor vaccine-specific T-cell responses.


A Rare Cause of Placental Abruption: Uterine Torsion.
Uterine torsion is defined as a rotation on its long axis and it is a dangerous, unexpected obstetric emergency. We report a case of uterine torsion at 32 weeks of gestation in a singleton pregnancy. A 37-year-old woman with multiple prior cesarean deliveries referred to emergency unit of our hospital at 32 weeks of gestation with severe abdominal pain and mild vaginal bleeding. Ultrasonography showed a single fetus in vertex position, with a normal amniotic fluid. Fetal biometer was appropriate for 32 weeks of gestation. Placental location was anterior with a subchorionic hypoechogenic small area which was suspected to be a sign of placental abruption. An emergency cesarean section was performed under general anesthesia. The 180° uterine torsion was diagnosed and it was not possible to perform detorsion of the gravid uterus by exteriorization by pfannenstiel incision. Posterior hysterotomy was performed and a male baby of 1830 grams weight was delivered. The newborn was transported to Neonatal Intensive Care Unit (NICU) of another hospital and discharged within two weeks. Patient recovered well and was discharged on second postoperation day. Uterine torsion is a very rare and life threatening situation. In unexpected cases posterior low transuerse hysterotomy is generally performed and it is suggested as a safe choice when detorsion was not accomplished. It is not easy to keep in mind the possibility of uterine torsion in cases of abdominal pain during pregnancy. Because it generally causes abruption, management of abruption is vitally important to prevent fetal mortality.


Clearance by the rat of inhaled fly ash from fluidized-bed coal combustion.
Fly ash from a fluidized-bed coal combustor was neutron-activated and administered to male Fischer 344 rats by a single nose-only inhalation exposure. The activated fly ash contained 46Sc and smaller amounts of other gamma-emitting radionuclides. Whole-body clearance of radioactivity to 127 d was described by an exponential equation, with the long-term component having a biological half-life of 78 d. High-resolution gamma spectra of the lungs were obtained with a Ge(Li) detector and the relative activities of several fly ash constituents were compared. The activities of 152Eu, 134Cs, 54Mn, and 60Co significantly decreased with time relative to those of 46Sc and 59Fe. These results indicate that the clearance of fly ash is similar to that of other relatively insoluble particles and that some elements may have been preferentially dissolved from the fly ash particles in vivo.


Diagnosis of Tooth Agenesis in Childhood and Risk for Neoplasms in Adulthood.
Tooth agenesis, the congenital absence of one or more teeth, can be diagnosed in children in the first decade of life. Tooth agenesis is a phenotypic feature of conditions such as ectodermal dysplasia, cleft lip, cleft palate, Down syndrome, and Van der Woude syndrome. Tooth agenesis can also be nonsyndromic. Studies have shown an association between the genetic determinants of nonsyndromic tooth agenesis and neoplasms in adulthood. This review of the implications of tooth agenesis as a risk indicator for neoplasms in adulthood is based on a search of PubMed to identify published case series, case reports, and review articles. The reference articles were manually searched. The search was limited to articles published in the English language. Neoplasms reported in patients with tooth agenesis include colorectal neoplasms and epithelial ovarian cancer, as well as family histories of breast cancer, prostate cancer, and cancers of the brain and nervous system. Although odontogenesis and tumorigenesis may seem to be unrelated processes, the clinical association between the two highlights the overlap of genetic determinants and molecular pathways. Tooth agenesis can be diagnosed during childhood and should be considered a marker for risk of neoplasms in adulthood. Healthcare providers should identify tooth agenesis and provide appropriate anticipatory guidance.


The synergy of the refugee crisis and the financial crisis in Greece: Impact on mental health.
The current global financial crisis that started in 2008 resulted in a significant decline in global trade, slowing/reversing economic growth worldwide, and a dramatic increase in public sector debt. At the same time, the global migrant/refugee crisis has reached extreme rates, with millions of people being forced to abandon their homes and communities because of war, political violence or related threats. There is a broad consensus about the deleterious consequences of these crises on psychological well-being, depression, anxiety disorders, insomnia, alcohol abuse and suicidal behavior. Although the separate consequences of economic recession and immigration are extensively discussed in previous research, we know very little about the processes through which the intersection of economic crisis and migrant crisis contributes to the vulnerabilities of natives and migrants during these crises. Of particular concern is the status of children, adolescents and their families, who constitute one of the most vulnerable groups in society. To discuss the contexts that economic and migrant crises shape and suggest possible effects of this intersection on mental health risks, especially among children, adolescents and their families, through reflecting on the recent experience in Greece. Review of the literature and critical analysis of the effects of the confluent crises. The interactive effects of these two crises need further exploration. Novel and diverse models of psychological understanding need to be developed in order to manage the effects of the confluent crises. The role of mental health professionals is crucial in this respect, offering culturally flexible, accommodating and empathetic approaches, allowing healing and acceptance in the face of adversity.


Effect of muscle relaxation on the oxygenation of human skeletal muscle: a prospective in-vivo experiment using an isolated forearm technique.
Total oxygen consumption has been found to be reduced under deep neuromuscular blockade due to a lower rate of metabolism of skeletal muscles. However, the magnitude of this effect in individual muscles has not been investigated. Thus the aim of this study was to compare the oxygenation of paralyzed versus non-paralyzed forearm muscle under tourniquet-provoked ischemia. After ethics approval and written informed consent, 30 patients scheduled for elective hand and wrist surgery were included. Ischemia was provoked by inflation of bilateral upper arm tourniquets and muscle relaxation was achieved via intravenous administration of rocuronium 0.9 mg/kg. Bilateral tourniquets were applied to both upper arms before induction of anesthesia and near infrared spectrometry (NIRS) electrodes applied on both forearms. Muscular ischemia in an isolated (= non-paralyzed, NP) as well as a paralyzed forearm (P) was created by sequential inflation of both tourniquets before and after intravenous administration of rocuronium. Muscle oxygen saturations (SmO2) of NIRS in both forearms and their changes were determined and compared. Data of 30 patients (15 male, 15 female; 41.8 ± 14.7 years) were analyzed. The speed of SmO2 decrease (50% decrease of SmO2 from baseline (median [percentiles]: NP 210 s [180/480s] vs. P 180 [180/300]) as well as the maximum decrease in SmO2 (minimum SmO2 in % (median [percentiles]: NP 20 [19/24] vs. P 21 [19/28]) were not significantly affected by neuromuscular paralysis. No significant effect of muscle relaxation on NIRS-assessed muscle oxygenation under tourniquet-induced ischemia was found in human forearm muscles.


Effect of excipients on the particle size of precipitated pioglitazone in the gastrointestinal tract: impact on bioequivalence.
This study sought to understand the reasons for the bioinequivalence of a newly developed generic product of pioglitazone hydrochloride and to improve its formulation so that it is equivalent to that of the reference listed drug (RLD). In this clinical study, despite a similar in vitro dissolution profile, the new oral product exhibited a lower plasma concentration of pioglitazone compared to the RLD. The strong pH-dependency of pioglitazone solubility as a weak base indicates that pioglitazone would precipitate in the small intestine after being dissolved in the stomach. Thus, in vitro experiments were performed to investigate the effect of excipients on the particle size distribution of precipitated pioglitazone. Then, the impact of particle size on in vivo absorption was discussed. The precipitated pioglitazone from the RLD showed a peak for small particles (less than 1 μm), which was not observed in the precipitate from the new product. As an excipient, hydroxypropyl cellulose (HPC) influenced the particle size of precipitated pioglitazone, and the amount of HPC in the formulation was increased to the same level as that in the RLD. The precipitate from this improved product showed approximately the same particle size distribution as that of the RLD and successfully demonstrated bioequivalence in the clinical study. In conclusion, for drugs with low solubility, this type of analysis of the particle size distribution of precipitated drugs, in addition to the dissolution test, may help to obtain a better in vitro-in vivo correlation for oral absorption and to develop a bioequivalent product.


GABA immunoreactivity in hypothalamic neurons and growth cones in early development in vitro before synapse formation.
GABA (gamma-aminobutyrate) is the most prevalent inhibitory transmitter in the mature hypothalamus. In contrast, in the developing hypothalamus, GABA may exert depolarizing actions leading to neuronal excitation. To determine whether GABA is present in hypothalamic neurons early in development, and whether there is a preferential expression in axonal growth cones, immunogold and peroxidase studies were used with light and whole mount transmission electron microscopy. At embryonic day 15, a stage of development at the beginning of hypothalamic neurogenesis, histological sections showed GABA immunoreactivity in fibers and weakly stained perikarya. Hypothalamic neurons (13%) cultured at embryonic day 15 were immunoreactive after 1 day in vitro. The percentage of neurons stained, and the intensity of staining increased during the next few days to 39% at 4 days in vitro. Neuritic growth cones, including lamellipodia and long filopodia, showed strong immunoreactivity before synaptogenesis. By using neuronal whole mounts studied with transmission electron microscopy and GABA silver-enhanced immunogold staining, a quantitative comparison of growth cones after a day and a half in culture revealed that the growth cone of the longest process, the putative axon, had a greater level of immunogold labeling than that of the shorter processes, the putative dendrites. This finding is one of the earliest biochemical differences between putative axons and dendrites. Astrocytes in the same cultures showed no immunolabeling. These results indicate that GABA is present very early in the development of hypothalamic neurons and is in a position to be released.


Effects of manual acupuncture with sparrow pecking on muscle blood flow of normal and denervated hindlimb in rats.
In clinical practice, it has been thought that acupuncture might serve to wash out pain-generating metabolic end-products by improving blood circulation in muscles. We investigated the effects of manual acupuncture (MA) on muscle blood flow (MBF) of normal and denervated hindlimbs in rats. Sprague-Dawley rats (n=100) anaesthetised with urethane (1.2g/kg ip) were used. Manual acupuncture with sparrow pecking (SP) at different doses (1, 10 or 30 pecks) was given to the right ventral hindlimb muscles (tibial anterior and extensor digitorum longus muscles) or the right dorsal hindlimb muscles (gastrocnemius, plantaris and soleus muscles). MBF with or without MA was measured using the radiolabelled microsphere technique. The blood pressure was recorded through the right common carotid artery until MBF measurement started. Denervation of hindlimb was conducted by cutting the sciatic and femoral nerves. In normal rats, significantly increased MBF after MA were observed only in muscles which were penetrated by an acupuncture needle. The size of the increase depended on the number of times of pecking and seemed to be sustained at least until 60 minutes after MA. However, the increase was observed after both acute and chronic denervation. On the other hand, the mean arterial blood pressure (MAP) did not change significantly before, during or after MA. These results suggest that MA could increase muscle blood flow locally in a dose-dependent manner and that this increase may be caused by local vasodilators, as well as the axon reflex. A further study is needed to elucidate the mechanism.


Caspase-cleaved glial fibrillary acidic protein within cerebellar white matter of the Alzheimer's disease brain.
Although the cerebellum is generally thought of as an area spared of Alzheimer's disease (AD) pathology, recent evidence suggests that balance and mobility dysfunction may be magnified in affected individuals. In the present study, we sought to determine the degree of pathological changes within the cerebellum utilizing an antibody that specifically detects caspase-cleaved GFAP within degenerating astrocytes. Compared to control subjects, application of this antibody, termed the GFAP caspase-cleavage product (GFAPccp) antibody, revealed widespread labeling in cerebellar white matter with little staining observed in grey matter. Staining was observed within damaged astrocytes, was often localized near blood vessels and co-localized with other markers of apoptosis including TUNEL and caspase-cleaved tau. Of interest was the association of beta-amyloid deposition in white matter together with GFAPccp in cerebellar AD sections. In contrast, utilizing the tangle marker, PHF-1, neuritic pathology was completely absent in AD cerebellar sections. It is suggested that the observed pathological changes found in the white matter of the cerebellum may contribute to the declined motor performance in AD.


A case of non-resolving cough and weight loss.
Sarcoidosis is a chronic granulomatous multi-system disease with a clinical picture often mimicking tuberculosis. We present a case of a patient who presented with a clinical picture akin to both of these granulomatous disorders and was started on anti-tuberculous regimen despite the lack of any solid evidence pointing towards tuberculosis. As a result her clinical condition continued to deteriorate for months until finally a bronchoscopic biopsy established her disease process as sarcoidosis. She was then started on systemic corticosteroid therapy for sarcoidosis and during the ensuing period has shown marked improvement in her clinical picture with near normalization of the biochemical and radiographic parameters of her pathology. This case illustrates the need for vigilant interpretation of the clinical scenario in patients such as these where a misdiagnosis may lead to significant patient distress as well as weighing down on the economic and health resources.


A neuromorphic VLSI device for implementing 2-D selective attention systems.
Selective attention is a mechanism used to sequentially select and process salient subregions of the input space, while suppressing inputs arriving from nonsalient regions. By processing small amounts of sensory information in a serial fashion, rather than attempting to process all the sensory data in parallel, this mechanism overcomes the problem of flooding limited processing capacity systems with sensory inputs. It is found in many biological systems and can be a useful engineering tool for developing artificial systems that need to process in real-time sensory data. In this paper we present a neuromorphic hardware model of a selective attention mechanism implemented on a very large scale integration (VLSI) chip, using analog circuits. The chip makes use of a spike-based representation for receiving input signals, transmitting output signals and for shifting the selection of the attended input stimulus over time. It can be interfaced to neuromorphic sensors and actuators, for implementing multichip selective attention systems. We describe the characteristics of the circuits used in the architecture and present experimental data measured from the system.


Clonidine and the primate locus coeruleus: evidence suggesting anxiolytic and anti-withdrawal effects.
One may conclude from this series of studies and the data of others reviewed at this symposium that NE systems are involved in fear or anxiety and in mediating the actions of opioids and endorphins. It is rational pharmacotherapy, therefore, to utilize anatomically and neurophysiologically characterized receptors to produce those desired specific effects on NE systems that are expected to reduce anxiety. These same receptor interactions could also be used to antagonize side effects or to reverse withdrawal syndromes via alternative receptor sites. This strategy of using different receptors to produce synergistic or antagonistic effects on a common neuronal pathway has benefited the treatment of pathological anxiety, pain, drug withdrawal symptoms, and other classical psychosomatic conditions. It should be emphasized, of course, that the LC-noradrenergic system has a much broader function than the mediation of fear or anxiety, and that the endorphins, opiates, and clonidine influence many other systems, as do their abstinence syndromes. Anatomical projections of the LC and similar central NE systems also produce the observed effects. Detailed studies are needed to discover precisely how the abnormalities observed in numerous biochemical systems [Eidelberg, 1976] are related to the alterations in NE function we have studied and discussed in this chapter. Finally, careful clinical studies are needed to determine, define, and delimit the potential therapeutic uses of clonidine in the areas suggested by the preliminary studies reviewed here.


Structure of an aryl esterase from Pseudomonas fluorescens.
The structure of PFE, an aryl esterase from Pseudomonas fluorescens, has been solved to a resolution of 1.8 A by X-ray diffraction and shows a characteristic alpha/beta-hydrolase fold. In addition to catalyzing the hydrolysis of esters in vitro, PFE also shows low bromoperoxidase activity. PFE shows highest structural similarity, including the active-site environment, to a family of non-heme bacterial haloperoxidases, with an r.m.s. deviation in 271 C(alpha) atoms between PFE and its five closest structural neighbors averaging 0.8 A. PFE has far less similarity (r.m.s. deviation in 218 C(alpha) atoms of 5.0 A) to P. fluorescens carboxyl esterase. PFE favors activated esters with small acyl groups, such as phenyl acetate. The X-ray structure of PFE reveals a significantly occluded active site. In addition, several residues, including Trp28 and Met95, limit the size of the acyl-binding pocket, explaining its preference for small acyl groups.


Aryl diazonium salts: a new class of coupling agents for bonding polymers, biomacromolecules and nanoparticles to surfaces.
This critical review summarizes existing knowledge on the use of diazonium salts as a new generation of surface modifiers and coupling agents for binding synthetic polymers, biomacromolecules, and nanoparticles to surfaces. Polymer grafts can be directly grown at surfaces through the so-called grafting from approaches based on several polymerization methods but can also be pre-formed in solution and then grafted to surfaces through grafting onto strategies including "click" reactions. Several routes are also described for binding biomacromolecules through aryl layers in view of developing biosensors and protein arrays, while the use of aryl diazonium coupling agents is extended to the attachment of nanoparticles. Patents and industrial applications of the surface chemistry of diazonium compounds are covered. This review stresses the paramount role of aryl diazonium coupling agents in adhesion, surface and materials sciences (114 references).


Comprehensive Characterization of the Human Endogenous Retrovirus HERV-K(HML-6) Group: Overview of Structure, Phylogeny, and Contribution to the Human Genome.
Eight percent of the human genome is composed of human endogenous retroviruses (HERVs), remnants of ancestral germ line infections by exogenous retroviruses, which have been vertically transmitted as Mendelian characters. The HML-6 group, a member of the class II betaretrovirus-like viruses, includes several proviral loci with an increased transcriptional activity in cancer and at least two elements that are known for retaining an intact open reading frame and for encoding small proteins such as ERVK3-1, which is expressed in various healthy tissues, and HERV-K-MEL, a small Env peptide expressed in samples of cutaneous and ocular melanoma but not in normal tissues.IMPORTANCE We reported the distribution and genetic composition of 66 HML-6 elements. We analyzed the phylogeny of the HML-6 sequences and identified two main clusters. We provided the first description of a Rec domain within the env sequence of 23 HML-6 elements. A Rec domain was also predicted within the ERVK3-1 transcript sequence, revealing its expression in various healthy tissues. Evidence about the context of insertion and colocalization of 19 HML-6 elements with functional human genes are also reported, including the sequence 16p11.2, whose 5' long terminal repeat overlapped the exon of one transcript variant of a cellular zinc finger upregulated and involved in hepatocellular carcinoma. The present work provides the first complete overview of the HML-6 elements in GRCh37(hg19), describing the structure, phylogeny, and genomic context of insertion of each locus. This information allows a better understanding of the genetics of one of the most expressed HERV groups in the human genome.


The impact of liquefaction disaster on farming systems at agriculture land based on technical and psychosocial perspectives.
This research aims to determine the attitudes of the farmers whose lands are affected by liquefaction in Jono Oge, Central Sulawesi Province, The Republic of Indonesia. The methods used here were integrated survey and experimental design. The survey approach was intended to figure out the attitudes of the farmers viewpoints: (1) to return to their activities on the agricultural lands affected by liquefaction; (2) to consume their own agricultural products; and (3) of their willingness to be relocated. The experimental design approach was used to figure out the effectiveness of organic material input combined with the SP-36 fertilizer. The obtained results were analyzed using the Likert Scale, diversity test, correlational test, and regression test. The results showed that the farmers persevered farming on the lands affected by liquefaction (Index = 88.82%) yet refused to consume their own agricultural products with the reason that corpses remained buried beneath their lands (Index = 27.82%); and they also refused to be relocated (Index = 28.80%). The continued production suitability of the affected land was also investigated. Terrain profile identification results in Jono Oge showed the disaster impact was dominantly landslide as it still showed a clear characteristic horizon between the topsoil and the sub soil. This contrasts to terrain at Petobo, Central Sulawesi Province, where the high mix of the topsoil with the sub soil of agricultural land affected by liquefaction, prevented demarcation of the horizon. The land treatment of organic material and SP-36 fertilizer showed that the combined dose (M) of 40-kg ha-1 with P 300-kg ha-1 had the highest effect by changing the field pH from 5.7 to 6.41, increased the availability of P and increased the corncob indicator plant weight. Based on these indications, the lands affected by the liquefaction in Jono Oge can still be used as agricultural lands through restoration, from both social and technical aspects.


Neuronal and glial cell number in the hippocampus after experimental traumatic brain injury: analysis by stereological estimation.
Fluid percussion (FP) brain injury causes spatial memory dysfunction in rats regardless of injury location (midline vs. lateral). Standard histological analysis of the injured brains shows hippocampal neuronal loss after lateral, but not midline FP injury. We have used the optical volume fractionator (OVF) stereological procedure to quantify neuronal loss and glial proliferation within specific subregions of the hippocampus after midline or lateral FP injury. The OVF method is a design-based cell counting procedure, which combines cellular numerical density estimates (from the optical disector) with volume estimates (generated by point counting and the fractionator stereology method) to produce an estimate of the absolute cell number. Fifteen adult male Sprague-Dawley rats were randomly divided into 3 groups (n = 5/group): midline injury, lateral injury and naive. A single fluid percussion pulse was delivered to anesthetized rats in the injured groups. At 14 days post-injury, strict morphological criteria enabled the estimation of neurons, astrocytes, oligodendrocytes, and microglia in defined hippocampal subregions. The results confirm that hippocampal neurons are selectively vulnerable to brain injury, particularly observed as a significant loss in the hilus following both types of injury and in area CA3 after lateral injury. In contrast, the number of astrocytes and oligodendrocytes remains unaffected by brain injury, regardless of subregion. However, the significant increase in microglia number (bilaterally after midline and ipsilateral following lateral injury) suggests that underlying cellular processes continue weeks following injury. The implications of the observed cell population changes are discussed in relation to the reported cognitive deficits associated with both lateral and midline FP brain injury.


Benzotriazole UV stabilizers (BUVs) as an emerging contaminant of concern: a review.
Benzotriazole UV stabilizers (BUVs) are a group of industrial chemicals used in various consumer products and industrial applications. Due to its large-scale production and use, BUVs have been detected in all environmental matrices. Humans are exposed to BUVs from environmental media, food, personal care products (PCPs), and consumer products. As a result, BUVs are detected in human breast milk, attracting researchers and regulatory bodies worldwide. BUVs such as UV-328 exhibit the characteristics of persistent organic pollutants (POPs); hence, it has been recently listed under Stockholm Convention POP list. The current review focuses on the occurrence of BUVs in the environment with emphasis on persistency, bioaccumulation, and toxicity (PBT). Scarcity of scientific data on BUVs' properties, environmental occurrence, exposure levels, and effects on organisms poses significant challenges to the policymakers and regulatory bodies in adopting management strategies. The need for a science-based integrated framework for risk assessment and management of BUVs is recommended. Considering the potential threat of BUVs to human health and the environment, it is recommended that BUVs should be taken as a subject of priority research. Studies on the degradation and transformation route of BUVs need to be explored for the sound management of BUVs.


Nurses' Awareness and Perceptions towards Disaster Situations.
Man-made or natural disasters are occurring more frequently in India and around the world. It is important that all nurses be prepared to care for people affected by disasters. An exploratory survey was conducted to assess the nurses' awareness regarding disaster nursing and their perceived abilities to react when disaster strikes. In this study, conducted in Aurangabad (MS), a total of 60 nurses of various cadres were selected by simple random sampling method. Semi-structured questionnaire was prepared with 15 questions to assess the awareness regarding disaster nursing and 15 questions related to perceived abilities of the nurses. Overall awareness level of Nurses was good, still majority of the nurses had gaps in the awareness. The overall ability of the all cadres of nurses was not satisfactory. Awareness about disaster nursing is independent of cadre of nurses. To test this we applied standard F test for collective comparison of means using ANOVA table. Since the computed F ratio was less than the table value the mean awareness of four cadres did not differ significantly accepting null hypothesis. Looking at the means for four cadres it was found that mean ability of teachers, staff nurses, D7 year students and IV year students did not differ significantly; however their ability score was substantially low as compared to the awareness score.


Surgical coronary angioplasty of both coronary ostia after chest radiotherapy. Is it good alternative to conventional coronary bypass surgery?
Isolated coronary ostial stenosis of both ostia is a rare, potentially life-threatening condition, occurring in 0.1%-0.2% of patients undergoing coronary angiography. We present a case of a 69-year-old woman with a past medical history of breast cancer, who had been treated with radiotherapy, which most likely caused significant stenosis of both coronary ostia and likely accelerated aortic stenosis. Surgical angioplasty with autopericardium patch reconstruction of the left main coronary artery and right coronary arteries due to proximal stenotic disease was performed instead of venous or arterial bypasses with concomitant aortic valve replacement. The postoperative course was uneventful. There were no cardiovascular events 5 years after operation, and the patient remained free of any symptoms. Surgical coronary angioplasty offers an alternative to conventional coronary artery bypass grafting in isolated coronary ostial lesions and is advantageous in restoring more physiological myocardial perfusion, especially in those cases when conduits are suspected to be fibrotic, scarred or stenosed after radiation therapy or if there is the need to preserve conduits for future myocardial revascularisation in young patients.


Tryptophan at position 181 of the D2 protein of photosystem II confers quenching of variable fluorescence of chlorophyll: implications for the mechanism of energy-dependent quenching.
The lumenal CD loop region of the D2 protein of photosystem II contains residues that interact with a reaction center chlorophyll and the redox-active Tyr(D). Using combinatorial mutagenesis, photoautotrophic mutants of Synechocystis sp. PCC 6803 have been generated with multiple amino acid changes in this region. The CD loop mutations were transferred into a photosystem I-less Synechocystis strain to facilitate characterization of photosystem II properties in the mutants. Most of the combinatorial photosystem I-less mutants obtained had a high yield of variable fluorescence, F(V). However, in three mutants, which shared a replacement of Phe181 by Trp, the F(V) yield was dramatically reduced although a high rate of oxygen evolution was maintained. A site-directed F181W D2 mutant shared similar properties. Picosecond time-resolved fluorescence measurements revealed that in the combinatorial F181W mutants the fluorescence lifetimes in closed and open photosystem II centers were essentially identical and were similar to the fluorescence lifetime in open centers of the control strain. These results are explained by quenching of variable fluorescence in the mutants by charge separation between Trp181 and excited reaction center chlorophyll. This reaction competes efficiently with fluorescence and nonradiative decay in closed photosystem II centers, where the lifetime of the excitation in the chlorophyll antenna is long. Thermodynamic considerations favor the formation of oxidized tryptophan and reduced chlorophyll in the quenching reaction, presumably followed by charge recombination. A possible role of tryptophan-chlorophyll charge separation in the mechanism of energy-dependent quenching of excitations in photosynthesis is discussed.


Homodimeric reverse transcriptases from rous sarcoma virus mutated within the polymerase or RNase H active site of one subunit are active.
Heterodimeric reverse transcriptase (RT) alphabeta from Rous sarcoma virus (RSV) possesses an asymmetric subunit organization with the polymerase and RNase H active sites localized in the alpha subunit. To determine whether homodimeric RSV RTs alpha (63 kDa) or beta (95 kDa) assume alpha subunit organization similar to that of the heterodimer, an essential aspartic acid residue was mutated in the active site of either the polymerase (Asp181 > Asn) or the RNase H (Asp505 > Asn). Homodimeric alpha or beta RT consisting of one wild-type and one mutated subunit exhibit polymerase or RNase H activity, respectively, whereas the corresponding doubly mutated enzymes are inactive, indicating that the catalytic sites of the polymerase and RNase H domains are formed by only one subunit of the homodimer.


Repeated low-dose exposures to sarin disrupted the homeostasis of phospholipid and sphingolipid metabolism in guinea pig hippocampus.
Repeated low-level exposure to sarin results to hippocampus dysfunction. Metabonomics involves a holistic analysis of a set of metabolites in an organism in the search for a relationship between these metabolites and physiological or pathological changes. The objective of the present study was to evaluate the effects of repeated exposure to low-level sarin on the metabonomics in hippocampus of a guinea pig model. Guinea pigs were divided randomly into control and sarin treated groups (n = 14). Guinea pigs in the control group received saline; while the sarin-treated group received 0.4×LD50 (16.8 μg/kg) sarin. Daily injections (a total of 14 days) were administered sc between the shoulder blades in a volume of 1.0 ml/kg body weight. At the end of the final injection, 6 animals in each group were chosen for Morris water maze test. The rest guinea pigs (n = 8 for each group) were sacrificed by decapitation, and hippocampus were dissected for analysis. Compared with the control-group, the escape latency in sarin-group was significantly (p < 0.05) longer while the crossing times were significantly decreased in the Morris water task (p < 0.05). Sarin inhibited activities of acetylcholinesterase (AChE) and neuropathy target esterase (NTE) in hippocampus. The AChE activity of hippocampus from sarin-treated groups is equivalent to 59.9 ± 6.4 %, and the NTE activity of hippocampus from sarin-groups is equivalent to 78.1 ± 8.3 % of that from control-group. Metabolites were identified and validated. A total of 14 variables were selected as potential biomarkers. Phospholipids [phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylinositol (PI), Lysophosphatidylethanolamine (LysoPE or LPE)] and sphingolipids (SPs) [sphinganine (SA), phytosphingosine (PSO) and sphinganine-1-phosphate (SA1P)] were clearly modified. In conclusion, repeated low-dose exposures to sarin disrupted the homeostasis of phospholipid and sphingolipid metabolism in guinea pig hippocampus and may lead to a neuronal-specific function disorders. Identified metabolites such as SA1P need to be studied more deeply on their biological function that against sarin lesions. In future research, we should pay more attention to characterize the physiological roles of lipid metabolism enzymes as well as their involvement in pathologies induced by repeated low-level sarin exposure.


A novel pyroptosis-related signature predicts prognosis and response to treatment in breast carcinoma.
Pyroptosis is a new form of programmed cell death (PCD), also known as cellular inflammatory necrosis. Its discovery has resulted in a novel approach to the progression and medication resistance of breast cancer (BC). However, there is still a significant gap in the investigation of pyroptosis-related genes in BC. The mRNA expression profiles and clinical data of BC patients were obtained from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. Then, using the TCGA cohort, we created a predictive multigene signature including pyroptosis-related genes and verified it using the two GEO cohorts. A pyroptosis-related gene signature was created by combining several bioinformatics and statistical methodologies to predict patient prognosis and responses to immunotherapy and chemotherapy. Furthermore, a nomogram based on the gene signature and clinicopathological markers was created to better classify the risk and quantify the risk assessment of individual patients. A pyroptosis-related gene signature consisting of 15 genes was established. The pyroptosis-related gene signature classified the patients into two groups: high-risk and low-risk. When combined with clinical variables, the risk score was discovered to be an independent predictor of overall survival (OS) in BC patients. Some immunological pathways and genes were linked to pyroptosis, according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) evaluations. Patients in the high-risk group had a worse prognosis and were not very sensitive to immunotherapy. However, several chemotherapeutic agents were predicted to have greater potential for patients in the high-risk group. Finally, a nomogram was developed that included a classifier based on the 15 pyroptosis-related genes, tumor stage, age, and histologic grade. This nomogram demonstrated good classification capacity and might help with clinical decision-making in BC.


Acute hypercalcemia-induced hypertension: the roles of calcium channel and alpha-1 adrenergic receptor.
To study the mechanism(s) of acute hypercalcemia-induced hypertension in dogs. Adult male mongrel dogs were intravenously infused with: 1) normal saline solution, 2) CaCl2 solution, 3) CaCl2 + calcium channel blocker (verapamil), 4) CaCl2 + selective alpha-1 adrenergic receptor blocker (prazosin), or 5) CaCl2 + verapamil + prazosin. Either verapamil or prazosin treatment was started at forty minutes before CaCl2 infusion and then was co-administered throughout the three-hour experimental period. Systemic and renal hemodynamics parameters were determined. Infusion of CaCl2 caused increases in mean arterial blood pressure (p < 0.01), total peripheral resistance (p < 0.001), and renal vascular resistance (p < 0.001). Prior treatment with either verapamil or prazosin lowered baseline blood pressure (p < 0.01) and could prevent hypercalcemia-induced hypertension. This occurred accompanying regaining to near normal values of abnormal systemic hemodynamics parameters. Combination of both drugs showed more profound effects, particularly on lowering renal vascular resistance. Acute hypercalemic hypertension is caused by an increase in vascular resistance mediated via the direct effect of calcium on vascular smooth muscle as well as the indirect effect of calcium induced hypercatecholaminemia. The stimulatory effect of hypercalcemia on renal vascular resistance is more prominent than that on peripheral vascular resistance.


Single-agent ibrutinib versus chemoimmunotherapy regimens for treatment-naïve patients with chronic lymphocytic leukemia: A cross-trial comparison of phase 3 studies.
Chemoimmunotherapy (CIT) and targeted therapy with single-agent ibrutinib are both recommended first-line treatments for chronic lymphocytic leukemia (CLL), although their outcomes have not been directly compared. Using ibrutinib data from the RESONATE-2 (PCYC-1115/1116) study conducted in patients ≥65 years without del(17p), we performed a cross-trial comparison with CIT data from published phase 3 studies in first-line treatment of CLL. Progression-free survival (PFS), overall survival (OS), and safety data for ibrutinib (median follow-up 35.7 months) were evaluated alongside available CIT data. CIT regimens included: fludarabine + cyclophosphamide + rituximab (CLL8, CLL10), bendamustine + rituximab (CLL10), obinutuzumab + chlorambucil and rituximab + chlorambucil (CLL11), and ofatumumab + chlorambucil (COMPLEMENT-1). Median age across studies was 61-74 years, with older populations receiving ibrutinib, obinutuzumab + chlorambucil, or rituximab + chlorambucil. Median follow-up varied across studies/regimens (range 14.5-37.4 months). Among all patients, PFS appeared longer with ibrutinib relative to CIT and OS appeared comparable. Relative to CIT studies that similarly excluded patients with del(17p) (CLL10) or enrolled older/less-fit patients (CLL11), PFS appeared favorable for ibrutinib in high-risk subgroups, including advanced disease, bulky lymph nodes, unmutated IGHV status, and presence of del(11q). Grade ≥ 3 infections ranged from 9% (ofatumumab + chlorambucil) to 40% (fludarabine + cyclophosphamide + rituximab), and was 25% with ibrutinib. Grade ≥ 3 neutropenia was 12% for ibrutinib and 26%-84% for CIT. Although definitive conclusions cannot be made due to inherent limitations of cross-trial comparisons, this report suggests that ibrutinib has a favorable benefit/risk profile and may potentially eliminate the need for chemotherapy in some patients. Randomized, comparative studies are needed to support these findings.


G-quadruplexes offer a conserved structural motif for NONO recruitment to NEAT1 architectural lncRNA.
The long non-coding RNA NEAT1 serves as a scaffold for the assembly of paraspeckles, membraneless nuclear organelles involved in gene regulation. Paraspeckle assembly requires NEAT1 recruitment of the RNA-binding protein NONO, however the NEAT1 elements responsible for recruitment are unknown. Herein we present evidence that previously unrecognized structural features of NEAT1 serve an important role in these interactions. Led by the initial observation that NONO preferentially binds the G-quadruplex conformation of G-rich C9orf72 repeat RNA, we find that G-quadruplex motifs are abundant and conserved features of NEAT1. Furthermore, we determine that NONO binds NEAT1 G-quadruplexes with structural specificity and provide evidence that G-quadruplex motifs mediate NONO-NEAT1 association, with NONO binding sites on NEAT1 corresponding largely to G-quadruplex motifs, and treatment with a G-quadruplex-disrupting small molecule causing dissociation of native NONO-NEAT1 complexes. Together, these findings position G-quadruplexes as a primary candidate for the NONO-recruiting elements of NEAT1 and provide a framework for further investigation into the role of G-quadruplexes in paraspeckle formation and function.


Managing diabetes-related infections in the elderly.
The elevated serum glucose levels of diabetics affect traditional host defenses, predisposing these individuals to infectious processes. The elderly diabetic patient is also faced with senescence of the immune system which can alter host defense mechanisms and increase the risk of infection. Infections in elderly diabetics can be severe and life-threatening, and only through the prompt recognition and treatment of these disorders can morbidity and mortality be avoided. Broad-spectrum antimicrobial agents, in conjunction with surgical intervention, are often necessary to eradicate these infections. Common sites of involvement include the skin, biliary tract, urinary tract, and the soft tissues and bones of the feet.


Targeted treatment of chronic myeloid leukemia: role of imatinib.
Chronic myeloid leukemia (CML) is a myeloproliferative disorder characterized by clonal expansion of pleuripotent hematopoetic stem cells. The incidence of CML is 1 to 2 cases per 100,000 people per year; in the Western Hemisphere, CML accounts for 15% of leukemias in adults. Discovery of the specific karyotypic abnormality of the Philadelphia (Ph) chromosome in the pathogenesis of CML has led to a better understanding of the disease and hence to an advancement of targeted therapeutics. Availability of imatinib as an accepted targeted therapy in newly diagnosed patients has changed the treatment paradigm in CML. The majority of CML patients in chronic phase achieve excellent and durable responses with standard-dose imatinib. Mechanisms of primary and secondary resistance to imatinib in CML have been extensively studied and newer tyrosine kinase inhibitors are now being evaluated for clinical use. It is important that at any time the CML treatment and response remain optimal and thus patients on imatinib require continuous monitoring for early detection of resistance. This review will discuss the treatment and guidelines for monitoring CML patients in the imatinib era.


Treatment of Chronic Plantar Heel Pain With Radiofrequency Neural Ablation of the First Branch of the Lateral Plantar Nerve and Medial Calcaneal Nerve Branches.
From March 2012 to February 2013, 37 patients experiencing plantar heel pain for ≥6 months despite treatment with physical therapy and other conservative treatment modalities were followed up. If neurogenic heel pain originating from the first branch of the lateral plantar nerve was present, with or without the medial calcaneal nerve, diagnostic nerve blocks to these nerves were performed for confirmation. If the pain was determined to be of neurogenic origin, radiofrequency neural ablation (RFNA) was applied to the corresponding sensory nerve endings. Pain was evaluated using the visual analog scale, and patients were followed for at least one year. A total of 41 feet from 37 patients (30 [81.1%] females, 7 [18.9%] males; mean age, 50.7 ± 1.6 years; mean body mass index, 30.6 ± 0.7 kg/m(2)) were included. The mean visual analog scale scores improved significantly from 1 to 6 to 12 months after the procedure relative to before the procedure, with 88% of all patients rating the treatment as either very successful or successful at 12 months postoperatively. RFNA applied to both the first branch of the lateral plantar nerve and the medial calcaneal nerve sensory branches (16 [39%] feet) and only the first branch of the lateral plantar nerve sensory branches (25 [61%] feet) showed similarly high levels of success. Of the 41 feet, 28 [68.3%] had received extracorporeal shockwave therapy, 35 [85.4%] had received steroid injections, and 22 [53.7%] had received both extracorporeal shockwave therapy and steroid injections before RFNA as an index procedure. All were unresponsive to these previous treatments. In contrast, almost all (88%) were treated successfully with RFNA. Despite a high incidence of neurologic variations, with a precise diagnosis and good application of the technique using the painful points, chronic plantar heel pain can be treated successfully with RFNA.


Reprogramming murine telomerase rapidly inhibits the growth of mouse cancer cells in vitro and in vivo.
Telomerase plays a critical role in cancer, prompting the pursuit of various telomerase-based therapeutic strategies. One such strategy, telomerase interference, exploits the high telomerase activity in cancer cells and reprograms telomerase to encode "toxic" telomeres. To date, telomerase interference has been tested in human cancer cells xenografted into mice, an approach that does not recapitulate spontaneous malignancy and offers few insights about host toxicities, because human telomerase is targeted in a mouse host. To address these limitations, we designed and validated two new gene constructs specifically targeting mouse telomerase: mutant template mouse telomerase RNA (MT-mTer) and small interfering RNA against wild-type mouse telomerase RNA (α-mTer-siRNA). Using lentiviral delivery in mouse prostate cancer cells, we achieved α-mTer-siRNA-mediated knockdown of wild-type mTer (80% depletion) and concurrent overexpression of MT-mTer (50-fold). We showed that the two constructs effectively synergize to reprogram murine telomerase to add mutant instead of wild-type telomeric repeats, resulting in rapid telomeric uncapping (5-fold increase in DNA damage foci). This, in turn, led to rapid and significant apoptosis (>90% of cells) and growth inhibition in vitro (90% reduction in viable cell mass) and in vivo (75% reduction in tumor allograft wet weight). In summary, we have shown that mouse cancer cells are vulnerable to direct telomerase interference using novel murine telomerase-targeting constructs; this approach can now be used to study the true therapeutic potential of telomerase interference in mouse spontaneous cancer models.


Occurrence and removal of micropollutants in full-scale aerobic, anaerobic and facultative wastewater treatment plants in Brazil.
This study aims to evaluate micropollutant occurrence and removal in a low-middle income country (LMIC) by investigating the occurrence of 28 chemicals from different classes (triclosan, 15 polycyclic aromatic hydrocarbons (PAHs), 4 estrogens and 8 polybrominated diphenyl ether (PBDE) congeners) in three technologically diverse full-scale Brazilian wastewater treatment plants (WWTPs). These chemicals were detected at concentrations similar to those reported in other low-middle income countries (LMICs) and high-income countries (HICs) (0.1-49 μg/L) indicating their widespread use globally and the need for more studies in LMICs that are typically characterized by relatively inadequate wastewater treatment barriers. Among the three different WWTPs investigated for removal of these chemicals, the least energy intensive system, waste stabilization ponds (WSPs), was the most effective (95-99%) compared to the activated sludge (79-94%), and Up-flow sludge blanket reactor (UASB) with trickling filters system (89-95%). These results highlight the potential of WSPs for micropollutant removal-especially in warm climates. However, the effluent from all three WWTP could pose a risk to aquatic organisms when discharged into the receiving waters as the effluent concentrations of triclosan, some estrogens, PAHs and BDE 209 were above European environmental quality standards (EQS) or predicted no effect concentration (PNEC values), indicating that receiving water bodies could benefit from further treatment. In combination, these results help to further understand prevailing concentrations of micropollutants globally and fate in current wastewater treatment systems.


Inhalation of a harmless antigen (ovalbumin) elicits immune activation but divergent immunoglobulin and cytokine activities in mice.
Exposure to aerosolized harmless antigen such as ovalbumin (OVA) has previously been shown to induce inhalation tolerance, a state characterized by inhibition of IgE synthesis and airway inflammation, upon secondary immunogenic antigen encounter. Immune events associated with this phenomenon are still poorly understood. The aim of this study was to investigate cellular and molecular mechanisms underlying this state of 'unresponsiveness'. After initial repeated OVA exposure, mice were subjected to a protocol of antigen-induced airway inflammation, encompassing two intraperitoneal injections of OVA adsorbed to aluminium hydroxide followed by airway challenge. We assessed immune events in the draining lymph nodes after sensitization, and in the lungs after challenge. In animals initially exposed to OVA, we observed, at the time of sensitization, considerable expansion of T cells, many of which expressed the activation markers CD69 and CD25, as well as increased numbers of antigen-presenting cells, particularly B cells. While these animals produced low levels of IgE, the observed elevated levels of IgG1 signified isotype switching. Splenocytes and lymph node cells from OVA-exposed mice produced low levels of IL-4, IL-5, IL-13 and IFN-gamma, indicating aborted effector function of both T helper (Th)2- and Th1-associated cytokines. Real time quantitative polymerase chain reaction (PCR) (TaqMan) analysis of costimulatory molecules in the lungs after in vivo challenge showed that B7.1, B7.2, CD28 and CTLA-4 mRNA expression was low in animals initially exposed to OVA. Ultimately, these events were associated with abrogated airway inflammation and attenuated airway hyper-responsiveness. The decreased inflammation was antigen-specific and independent of IL-10 or IFN-gamma. Initial exposure to OVA establishes a programme that prevents the generation of intact, fully functional inflammatory responses upon secondary antigen encounter. The absence of inflammation, however, is not associated with categorical immune unresponsiveness.


[Transformation of serum immunoglobulins and monoclonal antibodies into polyreactive immunoglobulins].
It was shown that polyreactive properties of natural serum immunoglobulins or some monoclonal antibodies (i.e. their ability of binding to various antigenically dissimilar antigens) can be induced by the treatment with chaotropic ions (3.5 M KSCN, 5-30 min at 20 (C). Such transformation of monoclonal antibodies or serum immunoglobulins into polyreactive immunoglobulins (PRIG) seems to result of the changes in immunoglobulin conformation but is not from their unblocking. Interaction of PRIG with some antigens immobilized on the plate is strongly dependent on temperature unlike that of monospecific antibodies.


Expression of cytochrome P450s and glutathione S-transferases in human esophagus with squamous-cell carcinomas.
In order to clarify the expression of cytochrome P450 and glutathione S-transferase in human esophagus, 41 samples of human esophagus with squamous-cell carcinoma were investigated by immunoblot analysis and enzyme assays. Cytochrome P450 1A2/1 was clearly expressed in microsomes, and the amount in samples with tumorous tissue was significantly greater than that in samples without tumourous tissues or in liver; cytochrome P450 2B6 and 3A4/3 were expressed polymorphically. Aryl hydrocarbon hydroxylase activity was detected in microsomes and was greater in samples from smokers than non-smokers. Patients who both smoked and drank alcohol, however, had activity similar to that of patients without these habits. Glutathione S-transferase M1 and A1/2 protein existed polymorphically in cytosol, and glutathione S-transferase P1-1 was detected in all samples. The frequency of expression of the glutathione S-transferase A1/2 protein was greater in patients with M1 protein than in those without; no difference in the expression was seen for glutathione S-transferase P1-1. Neither smoking nor drinking influenced the expression or activity of glutathione S-transferase. Our data support the idea that some carcinogens can be directly activated or inactivated in human esophageal epithelium.


Imaging the first trimester heart: ultrasound correlation with morphology.
First trimester sonography is a widely used technique to examine the foetus early in pregnancy. The desire to recognise complex anatomy already in early developmental stages stresses the need for a thorough knowledge of basic developmental processes as well as recognition of cardiac compartments based on their morphology. In this paper, we describe the possibilities and limitations of sonographic assessment of the foetal heart between 10 and 14 weeks of gestation and correlate this to morphology. Examples of the most commonly detected congenital anomalies are atrioventricular septal defects, transposition of the great arteries, and hypoplastic left heart, which are shown in this paper.


Concise Review: Tissue Engineering of Urinary Bladder; We Still Have a Long Way to Go?
Regenerative medicine is a new branch of medicine based on tissue engineering technology. This rapidly developing field of science offers revolutionary treatment strategy aimed at urinary bladder regeneration. Despite many promising announcements of experimental urinary bladder reconstruction, there has been a lack in commercialization of therapies based on current investigations. This is due to numerous obstacles that are slowly being identified and precisely overcome. The goal of this review is to present the current status of research on urinary bladder regeneration and highlight further challenges that need to be gradually addressed. We put an emphasis on expectations of urologists that are awaiting tissue engineering based solutions in clinical practice. This review also presents a detailed characteristic of obstacles on the road to successful urinary bladder regeneration from urological clinician perspective. A defined interdisciplinary approach might help to accelerate planning transitional research tissue engineering focused on urinary tracts. Stem Cells Translational Medicine 2017;6:2033-2043.


The spontaneously hypertensive rat as a model of human cardiovascular disease: evidence of exacerbated cardiopulmonary injury and oxidative stress from inhaled emission particulate matter.
Cardiovascular disease is considered a probable risk factor of particulate matter (PM)-related mortality and morbidity. It was hypothesized that rats with hereditary systemic hypertension and underlying cardiac disease would be more susceptible than healthy normotensive rats to pulmonary injury from inhaled residual oil fly ash (ROFA) PM. Eight spontaneously hypertensive (SH) and eight normotensive Wistar-Kyoto (WKY) rats (12-13 weeks old) were implanted with radiotelemetry transmitters on Day -10 for measurement of electrocardiographic (ECG) waveforms. These and other nonimplanted rats were exposed to filtered air or ROFA (containing leachable toxic levels of metals) on Day 0 by nose-only inhalation (ROFA, 15 mg/m(3) x 6 h/day x 3 days). ECGs were monitored during both exposure and nonexposure periods. At 0 or 18 h post-ROFA exposure, rats were assessed for airway hyperreactivity, pulmonary and cardiac histological lesions, bronchoalveolar lavage fluid (BALF) markers of lung injury, oxidative stress, and cytokine gene expression. Comparisons were made in two areas: (1) underlying cardiopulmonary complications of control SH rats in comparison to control WKY rats; and (2) ROFA-induced cardiopulmonary injury/inflammation and oxidative burden. With respect to the first area, control air-exposed SH rats had higher lung and left ventricular weights when compared to age-matched WKY rats. SH rats had hyporeactive airways to acetylcholine challenge. Lung histology revealed the presence of activated macrophages, neutrophils, and hemorrhage in control SHrats. Consistently, levels of BALF protein, macrophages, neutrophils, and red blood cells were also higher in SH rats. Thiobarbituric acid-reactive material in the BALF of air-exposed SH rats was significantly higher than that of WKY rats. Lung inflammation and lesions were mirrored in the higher basal levels of pulmonary cytokine mRNA expression. Cardiomyopathy and monocytic cell infiltration were apparent in the left ventricle of SH rats, along with increased cytokine expression. ECG demonstrated a depressed ST segment area in SH rats. With regard to the second area of comparison (ROFA-exposed rats), pulmonary histology indicated a slightly exacerbated pulmonary lesions including inflammatory response to ROFA in SH rats compared to WKY rats and ROFA-induced increases in BALF protein and albumin were significantly higher in SH rats than in WKY rats. In addition, ROFA caused an increase in BALF red blood cells in SH rats, indicating increased hemorrhage in the alveolar parenchyma. The number of alveolar macrophages increased more dramatically in SH rats following ROFA exposure, whereas neutrophils increased similarly in both strains. Despite greater pulmonary injury in SH rats, ROFA-induced increases in BALF GSH, ascorbate, and uric acid were attenuated when compared to WKY rats. ROFA inhalation exposure was associated with similar increases in pulmonary mRNA expression of IL-6, cellular fibronectin, and glucose-6-phosphate dehydrogenase (relative to that of beta-actin) in both rat strains. The expression of MIP-2 was increased in WKY but attenuated in SH rats. Thus, SH rats have underlying cardiac and pulmonary complications. When exposed to ROFA, SH rats exhibited exacerbated pulmonary injury, an attenuated antioxidant response, and acute depression in ST segment area of ECG, which is consistent with a greater susceptibility to adverse health effects of fugitive combustion PM. This study shows that the SH rat is a potentially useful model of genetically determined susceptibility with pulmonary and cardiovascular complications.


Regulating CRISPR/Cas9 Function through Conditional Guide RNA Control.
Conditional control of CRISPR/Cas9 has been developed by using a variety of different approaches, many focusing on manipulation of the Cas9 protein itself. However, more recent strategies for governing CRISPR/Cas9 function are based on guide RNA (gRNA) modifications. They include control of gRNAs by light, small molecules, proteins, and oligonucleotides. These designs have unique advantages compared to other approaches and have allowed precise regulation of gene editing and transcription. Here, we discuss strategies for conditional control of gRNA function and compare effectiveness of these methods.


[An experimental action of new injection-scleroplasty substance on eye tissues].
An experimental study was made with 8 eyes of chinchilla rabbits for the purpose of investigating the interplay between the allotransplant (administered in the Tenon's space of the eyes of experimental animals), suggested by us, on the one hand, and the neighboring ocular components, on the other hand. According to the results, the used substance is subject to necrolysis accompanied by an induced local inflammation response. As the substance dissolved, it was found to be displaced by connective tissue. At first, the tissue was granulated, then, it became more compact with its structure resembling the sclera; the tissue tightly abutted the latter intensifying its properties. It is noteworthy, that the inflammation zone was limited to the administration region. With due respect to the above, it can be stated that the suggested method can be clinically used in future as a full sclera-fixing procedure.


The next generation of rapid point-of-care testing identification tools for ventilator-associated pneumonia.
Ventilator-associated pneumonia (VAP) is a frequent issue in intensive care units (ICU), with a major impact on morbidity, mortality and cost of care. VAP diagnosis remains challenging: traditional culture-based microbiological techniques are still the gold-standard, but are too slow to enable clinicians to improve prognosis with timely antimicrobial therapy adjustment. Prolonged exposure to inappropriate antibiotics has also been shown to increase the incidence of multi-drug-resistant organisms (MDROs). Point-of-care testing (POCT) tools are diagnostic testing methods that can be used at or near the bedside, with delays ranging from a couple minutes to a few hours. The use of POCTs for VAP could allow for faster diagnosis and antimicrobial therapy adjustments. Despite uncertainty regarding their diagnostic value, C-reactive protein (CRP) and procalcitonin (PCT) can be detected using POCTs in few minutes. In VAP, CRP showed a sensitivity of 56% to 88% and specificity of 86% to 91%; PCT showed a sensitivity of 78% to 100% and a specificity between 75% and 97% using non-POCT methods. Automated microscopy could also be used in clinical ICU setting, with reported sensitivity of 100% and specificity of 97%, allowing for antibiotic susceptibility testing (AST) in less than 12 h. Multiplex polymerase chain reaction (MPCR) could allow for identification and AST approximation through the detection of drug-resistance genes in about 6 h, with reported sensitivity of 89.2% and specificity of 97.1%; although use as POCT was shown to result in test failure in about 40% of samples. Despite being at an early development stage, exhalome analysis, which allows for non-invasive fast identification, and chromogenic tests, more suited for the detection of drug-resistance enzymes, are also promising techniques for POCT diagnosis of VAP.


Nanoparticle Nucleation Is Termolecular in Metal and Involves Hydrogen: Evidence for a Kinetically Effective Nucleus of Three {Ir3H2x·P2W15Nb3O62}6- in Ir(0)n Nanoparticle Formation From [(1,5-COD)IrI·P2W15Nb3O62]8- Plus Dihydrogen.
The nucleation process yielding Ir(0)∼300 nanoparticles from (Bu4N)5Na3[(1,5-COD)Ir·P2W15Nb3O62] (abbreviated hereafter as (COD)Ir·POM8-, where POM9- = the polyoxometalate, P2W15Nb3O629-) under H2 is investigated to learn the true molecularity, and hence the associated kinetically effective nucleus (KEN), for nanoparticle formation for the first time. Recent work with this prototype transition-metal nanoparticle formation system ( J. Am. Chem. Soc. 2014 , 136 , 17601 - 17615 ) revealed that nucleation in this system is an apparent second-order in the precatalyst, A = (COD)Ir·POM8-, not the higher order implied by classic nucleation theory and its nA ⇌ An, "critical nucleus", An concept. Herein, the three most reasonable more intimate mechanisms of nucleation are tested: bimolecular nucleation, termolecular nucleation, and a mechanism termed "alternative termolecular nucleation" in which 2(COD)Ir+ and 1(COD)Ir·POM8- yield the transition state of the rate-determining step of nucleation. The results obtained definitively rule out a simple bimolecular nucleation mechanism and provide evidence for the alternative termolecular mechanism with a KEN of 3, Ir3. All higher molecularity nucleation mechanisms were also ruled out. Further insights into the KEN and its more detailed composition involving hydrogen, {Ir3H2xPOM}6-, are also obtained from the established role of H2 in the Ir(0)∼300 formation balanced reaction stoichiometry, from the p(H2) dependence of the kinetics, and from a D2/H2 kinetic isotope effect of 1.2(±0.3). Eight insights and conclusions are presented. A section covering caveats in the current work, and thus needed future studies, is also included.


How the Affordable Care Act will strengthen the nation's primary care foundation.
As the country turns toward implementation of the Patient Protection and Affordable Care Act, realizing the potential of reform will require significant transformation of the American system of health care delivery. To that end, the new law seeks to strengthen the nation's primary care foundation through enhanced reimbursement rates for providers and the use of innovative delivery models such as patient-centered medical homes. Evidence suggests that these strategies can return substantial benefits to both patients and providers by increasing access to primary care services, reducing administrative hassles and burdens, and facilitating coordination across the continuum of care. If successfully implemented, the Affordable Care Act has the potential to realign incentives within the health system and create opportunities for providers to be rewarded for delivering high value, patient-centered primary care. Such a transformation could lead to better outcomes for patients, increase job satisfaction among physicians and encourage more sustainable levels of health spending for the nation.


[Modern approach to the neuromuscular blocking agents use in pediatric patients].
Myorelaxants use decrease trend appeared since the end of 80 years of the 20th century. The trend is connected with use of the new narcotic analgesics (Remifentanil), intravenous (Propofol) and inhalation (Sevoflurane) anaesthetics. These drugs are manageable and predictable, they have not many side effects and can suppress laryngeal-pharyngeal reflex during the tracheal intubation. Furthermore there are other factors such as succinylcholines use when fast intubation is needed, the wrong myorelaxant and dosage choice. Residual curarization and side effects risk increases due to these factors. As a result the patient's activation is delayed. Nevertheless myorelaxants use refusal impairs the tracheal intubation conditions, increases the arterial hypotension and heart failure risk especially in newborns and children with severe pathology. If myorelaxants is not used, comfortable conditions of surgical manipulations impossible without big analgesics and anaesthetics doses use.


Contribution of the KCa3.1 channel-calmodulin interactions to the regulation of the KCa3.1 gating process.
The Ca(2+)-activated potassium channel of intermediate conductance, KCa3.1, is now emerging as a therapeutic target for a large variety of health disorders. The Ca(2+) sensitivity of KCa3.1 is conferred by the Ca(2+)-binding protein calmodulin (CaM), with the CaM C-lobe constitutively bound to an intracellular domain of the channel C terminus. It was proposed on the basis of the crystal structure obtained for the C-terminal region of the rat KCa2.2 channel (rSK2) with CaM that the binding of Ca(2+) to the CaM N-lobe results in CaM interlocking the C-terminal regions of two adjacent KCa3.1 subunits, leading to the formation of a dimeric structure. A study was thus undertaken to identify residues of the CaM N-lobe-KCa3.1 complex that either contribute to the channel activation process or control the channel open probability at saturating Ca(2+) (Pomax). A structural homology model of the KCa3.1-CaM complex was first generated using as template the crystal structure of the C-terminal region of the rat KCa2.2 channel with CaM. This model was confirmed by cross-bridging residues R362 of KCa3.1 and K75 of CaM. Patch-clamp experiments were next performed, demonstrating that the solvation energy of the residue at position 367 in KCa3.1 is a key determinant to the channel Pomax and deactivation time toff. Mutations of residues M368 and Q364 predicted to form anchoring points for CaM binding to KCa3.1 had little impact on either toff or Pomax. Finally, our results show that channel activation depends on electrostatic interactions involving the charged residues R362 and E363, added to a nonpolar energy contribution coming from M368. We conclude that electrostatic interactions involving residues R362 and E363 and hydrophobic effects at M368 play a prominent role in KCa3.1 activation, whereas hydrophobic interactions at S367 are determinant to the stability of the CaM-KCa3.1 complex throughout gating.


In-hospital arrhythmic burden reduction in diabetic patients with acute myocardial infarction treated with SGLT2-inhibitors: Insights from the SGLT2-I AMI PROTECT study.
Sodium-glucose co-transporter 2 inhibitors (SGLT2-i) have shown significant cardiovascular benefits in patients with and without type 2 diabetes mellitus (T2DM). They have also gained interest for their potential anti-arrhythmic role and their ability to reduce the occurrence of atrial fibrillation (AF) and ventricular arrhythmias (VAs) in T2DM and heart failure patients. To investigate in-hospital new-onset cardiac arrhythmias in a cohort of T2DM patients presenting with acute myocardial infarction (AMI) treated with SGLT2-i vs. other oral anti-diabetic agents (non-SGLT2-i users). Patients from the SGLT2-I AMI PROTECT registry (NCT05261867) were stratified according to the use of SGLT2-i before admission for AMI, divided into SGLT2-i users vs. non-SGLT2-i users. In-hospital outcomes included the occurrence of in-hospital new-onset cardiac arrhythmias (NOCAs), defined as a composite of new-onset AF and sustained new-onset ventricular tachycardia (VT) and/or ventricular fibrillation (VF) during hospitalization. The study population comprised 646 AMI patients categorized into SGLT2-i users (111 patients) and non-SGLT2-i users (535 patients). SGLT2-i users had a lower rate of NOCAs compared with non-SGLT2-i users (6.3 vs. 15.7%, p = 0.010). Moreover, SGLT2-i was associated with a lower rate of AF and VT/VF considered individually (p = 0.032). In the multivariate logistic regression model, after adjusting for all confounding factors, the use of SGLT2-i was identified as an independent predictor of the lower occurrence of NOCAs (OR = 0.35; 95%CI 0.14-0.86; p = 0.022). At multinomial logistic regression, after adjusting for potential confounders, SGLT2-i therapy remained an independent predictor of VT/VF occurrence (OR = 0.20; 95%CI 0.04-0.97; p = 0.046) but not of AF occurrence. In T2DM patients, the use of SGLT2-i was associated with a lower risk of new-onset arrhythmic events during hospitalization for AMI. In particular, the primary effect was expressed in the reduction of VAs. These findings emphasize the cardioprotective effects of SGLT2-i in the setting of AMI beyond glycemic control. Data are part of the observational international registry: SGLT2-I AMI PROTECT. ClinicalTrials.gov, identifier: NCT05261867.


Protective effects of astragaloside in rats with adriamycin nephropathy and underlying mechanism.
The present study was designed to determine the mechanism underlying the treatment of nephrotic syndrome using astragaloside by observing the effects of astragaloside on the expression of nephrin and podocin proteins and genes in kidneys of rats with adriamycin nephropathy. The rats were injected with adriamycin and, after successful model establishment, randomly divided into a model group, a Methylprednisolone (MP) group, and an astragaloside group. The 24-h complete urine samples were collected. Biochemical indicators were monitored, and kidney tissues were collected for pathological analysis using light microscopy and electron microscopy. The mRNA expression of nephrin and podocin was measured in the kidney tissues using the real-time qPCR, and the protein expression levels of nephrin and podocin were detected using Western blot analysis. At the end of 12 weeks of drug intervention, the urinary protein level was lower in the MP and astragaloside groups than that in the model group (P = 0.008 and P = 0.01, respectively). Serum albumin was higher in the MP and astragaloside groups than in the model group (P < 0.001 and P = 0.012, respectively). Podocytes in the MP group were nearly normal, and fusion of podocytes in the astragaloside group was significantly less than that in the control group. The nephrin and podocin mRNA and protein expression levels in the intervention groups were higher (P < 0.05) than that in the model group. Due to the increased expression of podocyte-related nephrin and podocin proteins, astragaloside maintained slit diaphragm integrity and decreased the level of proteinuria in rats with adriamycin nephropathy.


Left ventricular pseudoaneurysm caused by coronary spasm, myocardial infarction, and myocardial rupture.
We report a very rare case of a 47-year-old man who had coronary spasm that resulted in a silent myocardial infarction, a ruptured myocardial wall, and a nonruptured left ventricular pseudoaneurysm. The patient presented with a 6-month history of dyspnea on exertion, without evidence of fixed coronary artery stenosis. Coronary angiography showed severe coronary spasm of the left anterior descending and left circumflex arteries; the spasm was relieved promptly by nitroglycerin. Echocardiography and left ventricular angiography revealed the large left ventricular pseudoaneurysm posterolateral to the left ventricle. We performed surgical resection of the pseudoaneurysm and patch repair of the ruptured left ventricular wall, with excellent results. We present this case because of the highly unusual sequence of events. Early surgical intervention resulted in the patient's recovery.


Exercise-based motivational interviewing for female patients with fibromyalgia: a case series.
The objective of the study is to determine the effects of motivational interviewing (MI), a novel technique of behavioral counseling to promote exercise, on pain and physical function in patients with fibromyalgia (FMS). Patients who met the American College of Rheumatology criteria for FMS and had a visual analog pain score of > or =6 were enrolled in a single group intervention pilot study. Participants received two supervised exercise sessions and an exercise prescription. Thereafter, six exercise-based MI phone calls were made over a 10-week period. Assessments were done at baseline, week 12 (immediate postintervention) and week 30 (follow-up). The primary endpoints were changes from baseline in the fibromyalgia impact questionnaire (FIQ)-pain and physical impairment at week 30. Secondary measures were brief pain inventory (BPI)-pain severity and BPI-pain interference, the number of exercise minutes (NEM) per week, and the arthritis impact measurement scale (AIMS)-depression. The 19 enrolled female participants had a mean age of 52.2 +/- 9.1 years, mean disease duration of 7.5 +/- 5.0 years, and a mean FIQ-pain score of 7.7 +/- 1.4. By week 30, there was significant improvement in both FIQ-pain (-2.6 +/- 2.6, p < 0.001) and FIQ-physical impairment (-1.3 +/- 2.1, p = 0.01). Likewise, BPI-pain severity and pain interference were reduced by -2.4 +/- 2.1 (p < 0.001) and -2.4 +/- 2.0 (p < 0.001), respectively. While the median NEM per week increased from 0 to 32 min (p = 0.001) at week 30, AIMS-depression score was unchanged. In this pilot study, we conclude that telephone-delivered MI to promote exercise was associated with an improvement in patient's level of pain and physical impairment.


Comparison of serologic tests for the diagnosis and follow-up of alveolar hydatid disease.
Alveolar hydatid disease is a serious and often fatal condition caused by infection with the metacestode form of Echinococcus multilocularis. Sera of 21 patients with histologically confirmed disease were tested by an enzyme-linked immunosorbent assay (ELISA) using a semi-purified E. multilocularis antigen fraction (Em2) and by indirect hemagglutination (IHA) and double diffusion (DD5) tests using antigens prepared from E. granulosus cyst fluid. At diagnosis, sera from all 21 patients were positive by Em2 ELISA, 18 (86%) by IHA, and 5 (24%) by DD5. Em2 ELISA detected an antibody response earlier than IHA in 4 of 9 patients from whom sera were available before diagnosis. Following complete surgical resection, Em2 ELISA converted from positive to negative in serum of 2 of 3 patients, while IHA results did not change. Following incomplete resection, 14 of 15 patients tested remained positive by Em2 ELISA, while 12 remained positive by IHA. Of sera from 361 healthy persons from regions free of E. multilocularis, none were positive by Em2 ELISA, while 8% were positive by IHA. Of sera from 59 patients with non-echinococcal parasitic infections, none were positive by Em2 ELISA, while 31% were positive by IHA. Thus, in comparison with tests using E. granulosus antigens, Em2 ELISA appears to be more sensitive and specific for diagnosing AHD, useful on follow-up of resected patients, and positive earlier in the course of disease.


Development of albumin macroinitiator for polymers to use in DNA origami coating
DNA nanostructures have many advantages over polymers and lipid based drug delivery agents such as biodegradability and biocompatibility. However their transfection rates and stability still limit their widely use in nanomedicine. In this study highly versatile and straightforward albumin coating preparation method is showed for DNA nanostructures. N-methylolmaleimide was esterified with a-bromoisobutyrl bromide (BiBB) to achive bromine functional structure. Then it was attached to bovine serum albumin (BSA) via cysteine-maleimide bond further to use as macroinitiator for atom transfer radical polymerization (ATRP). Cationic polymers can be synthesized from this end further to use as binding domain for fabricated 60 Helix bundle DNA origami. Proton nuclear magnetic resonance (1H NMR) analysis used for characterization. Methyelene group hydrogens' peak in 5.0 ppm and strong peak in 1.5–2.0 ppm range showed proper methylolation of maleimide and bromine functional formation, respectively. Then BSA-macroinitiator formation is verified by 1780 Da peak shift in MALDI-TOF (Matrix-assisted laser desorption/ionization - time of flight) spectrum. Moreover electrophoretic mobility shift assay (EMSA) showed successful dense 60 Helix bundle formation. In this study, a facile method is developed to synthesize protein conjugated-ATRP initiator further can be used in polymerization and coating DNA nanostructures. It is feasible for any protein containing cysteine amino acid.


Preclinical pharmacology of the pyrrolobenzodiazepine (PBD) monomer DRH-417 (NSC 709119).
The pyrrolobenzodiazepine monomer DRH-417 is a member of the anthramycin group of anti-tumor antibiotics that bind covalently to the N2 of guanine within the minor groove of DNA. DRH-417 emerged from the EORTC-Drug Discovery Committee and NCI 60 cell line in vitro screening programs as a potent antiproliferative agent with differential sensitivity towards certain cancer types such as melanoma, breast and renal cell carcinoma (mean IC(50) = 3 nM). DRH-417 was therefore tested for in vivo activity. The maximum tolerated dose (MTD) was established as 0.5 mg/kg given i.p. Marked anti-tumor activity was seen in two human renal cell cancers, one breast cancer and a murine colon tumor model (p<0.01). A selective HPLC (LC/MS) analytical method was developed and plasma pharmacokinetics determined. At a dose of 0.5 mg kg(-1), the plasma AUC was 540 nM h (197.1 ng h ml(-1)) and the peak plasma concentration (171 nM [62.4 ng ml(-1)]) occurred at 30 min., reaching doses levels well above those needed for in vitro antiproliferative activity. Genomic profiling of in vivo sensitive tumors revealed that the latter have an activated insulin-like growth factor signaling pathway.


Use of the venture wire control catheter for the treatment of coronary artery chronic total occlusions.
The Venture catheter (St Jude, Minneapolis, MN) has a deflectable tip for facilitating wire steering and a stiff body. Both properties can be useful in percutaneous coronary interventions (PCI) of coronary chronic total occlusions (CTOs). We reviewed 26 consecutive patients in whom the Venture catheter was utilized during coronary CTO PCI at our institution between May 2008 and September 2009. Mean age was 63 ± 9 years and 96% of the patients were men. The CTO target lesion was located in the right coronary artery (35%), left anterior descending artery (27%), circumflex (27%) or a saphenous vein graft (4%). A prior attempt for CTO PCI had been done in 19%. The primary CTO PCI approach was antegrade in 92% and retrograde in 8%, but a retrograde approach was used in an additional 27% of the patients after antegrade approach failed. The Venture catheter was used to overcome vessel tortuosity (73%), for CTOs with side branch at the occlusion site (15%), to facilitate collateral branch wiring during retrograde PCI (8%), and to provide extra support (4%). The overall CTO PCI success rate was 77% and was 92% in patients with upfront Venture catheter use and in 64% of patients in whom the Venture was used after PCI attempts using other equipment failed. Procedural failure was due to inability to cross the lesion in all cases. The Venture catheter can facilitate CTO PCI, especially in patients with marked coronary tortuosity or when additional support is required.


The role of HIV and monocytes/macrophages in adipose tissue biology.
To assess the role of HIV and monocytes/macrophages in adipose tissue dysregulation. Cross-sectional study in 5 groups: HIV seronegative, HIV+ antiretroviral therapy (ART)-naive, HIV+ nonlipoatrophic on zidovudine- and/or stavudine-containing ART, HIV+ lipoatrophic on similar ART, and HIV+ on abacavir- or tenofovir-containing ART. HIV DNA in circulating monocyte subsets was quantitated by real-time polymerase chain reaction. Biopsied subcutaneous fat was examined for macrophage content by CD68 staining. Isolated adipocytes and macrophages were cultured and the supernatant assayed for secretory products by Luminex multiplex cytokine technology. Sixty-nine subjects were enrolled. Lipoatrophic subjects had higher median HIV DNA levels (270.5 copies/10 cells) in circulating peripheral CD14CD16 co-expressing monocyte subsets compared with subjects who were ART-naive (25.0 copies), nonlipoatrophic (15.0 copies), or on abacavir/tenofovir (57.5 copies), P < 0.01. Group differences in adipocytes and adipose macrophage content were marginal. Although adipocyte secretory products were similar, HIV-infected subjects had higher adipose macrophage-derived interleukin (IL)-12p40, IL-6, IL-8, and monocyte inflammatory protein 1 alpha and lower eotaxin and interferon gamma levels than HIV seronegative subjects (P < 0.05). Within HIV-infected subjects, adipose macrophage secretory products were comparable between subjects naive with ART versus those on ART. Circulating HIV-infected and proinflammatory CD14CD16 monocyte subsets contribute to the pathogenesis of HIV-associated lipoatrophy. Among HIV-infected individuals, macrophages, rather than adipocytes, are the primary source of low-grade inflammation in subcutaneous adipose tissue. HIV infection modifies these macrophages to a more proinflammatory phenotype, and these changes are not substantially mitigated by the use of ART.


Inhibitory properties of polyphenols in Malus "Winter Red" crabapple fruit on α-glucosidase and α-amylase using improved methods.
To explore the inhibitory activity of polyphenols on α-glucosidase and α-amylase, 16 polyphenols were isolated, identified, and quantified in an edible Malus "Winter Red" crabapple fruit. The limitations of two traditional methods for α-glucosidase and α-amylase activity assay in vitro were assayed. An improved method based on an HPLC assay for α-glucosidase and a colorimetric method coupled with a custom-made mini-column for α-amylase were established. Compared with positive controls, acarbose and miglito, most polyphenols, especially the four aglycones (cyanidin, quercetin, phloretin, and 3-hydroxyphloretin) showed higher inhibition rates on α-glucosidase. None of the polyphenols showed higher inhibition rates on α-amylase than acarbose, but most, especially the four aglycones, showed higher inhibition rates on α-amylase than miglito. The Malus Winter Red fruit has great potential for postprandial blood glucose management as a potential diet therapy for diabetic patients.


Controllable synthesis and formation mechanism study of homoleptic alkynyl-protected Au nanoclusters: recent advances, grand challenges, and great opportunities.
In the past decade, atomically precise coinage metal nanoclusters have been a subject of major interest in nanoscience and nanotechnology because of their determined compositions and well-defined molecular structures, which are beneficial for establishing structure-property relationships. Recently ligand engineering has been extended to alkynyl molecules. Homoleptic alkynyl-protected Au nanoclusters (Au NCs) have emerged as a hotspot of research interest, mainly due to their unique optical properties, molecular configuration, and catalytic functionalities, and more importantly, they are used as a counterpart object for fundamental study to compare with the well-established thiolate Au NCs. In this review, we first summarize the recently reported various controllable synthetic strategies for atomically precise homoleptic-alkynyl-protected Au NCs, with particular emphasis on the ligand exchange method, direct reduction of the precursor, one-pot synthesis, and the synchronous nucleation and passivation strategy. After that, we switch our focus to the formation mechanism and structure evolution process of homoleptic alkynyl-protected Au NCs, where Au144(PA)60 and Au36(PA)24 (PA = phenylacetylide) are given as examples, along with the prediction of the possible formation mechanism of some other cluster molecules. In the end of this review, the outlook and perspective of this rapidly developing field including grand challenges and great opportunities are discussed. This review can stimulate more research efforts towards developing new synthetic strategies to enrich the limited examples and unravel the formation/growth mechanism of homoleptic alkynyl-protected Au NCs.