The fused SnoaL_2 domain in the Mycobacterium tuberculosis sigma factor σJ modulates promoter recognition.
Extra-cytoplasmic function (ECF) σ-factors are widespread in bacteria, linking environmental stimuli with changes in gene expression. These transcription factors span several phylogenetically distinct groups and are remarkably diverse in their activation and regulatory mechanisms. Here, we describe the structural and biochemical features of a Mycobacterium tuberculosis ECF factor σJ that suggests that the SnoaL_2 domain at the C-terminus can modulate the activity of this initiation factor in the absence of a cognate regulatory anti-σ factor. M. tuberculosis σJ can bind promoter DNA in vitro; this interaction is substantially impaired by the removal of the SnoaL_2 domain. This finding is consistent with assays to evaluate σJ-mediated gene expression. Structural similarity of the SnoaL_2 domain with epoxide hydrolases also suggests a novel functional role for this domain. The conserved sequence features between M. tuberculosis σJ and other members of the ECF41 family of σ-factors suggest that the regulatory mechanism involving the C-terminal SnoaL_2 domain is likely to be retained in this family of proteins. These studies suggest that the ECF41 family of σ-factors incorporate features of both-the σ70 family and bacterial one-component systems thereby providing a direct mechanism to implement environment-mediated transcription changes.


Risk Assessment of Ozone Fumigation Under Vacuum to Control Potential Infestation of Coffee Berry Borer and Coffee Leaf Rust in Green Coffee Beans Imported Into Hawaii.
Studies were conducted with ozone gas fumigation under vacuum as a methyl bromide alternative against life stages of coffee berry borer (CBB) Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), and the urediniospores of coffee leaf rust (CLR), Hemileia vastatrix Berkeley & Broome (Basidiomycota: Pucciniales) in green coffee, Coffea spp. L. Fumigation with 10,000 ppm O3 gas under -25.4 mm Hg vacuum1 at 13.0 ± 3.0°C for 6.0 h killed all CBB larvae, pupae, and adults, but did not kill all CBB eggs (~15% survival). Mortality of CLR urediniospores was 100% within the first hour of the 6-h fumigation. Ozone fumigation had no adverse effects on coffee quality. Results indicated that CBB adult hitchhikers may be the only target life stage of quarantine concern, and additional studies focused on this stage. CBB adult survival and reproduction decreased significantly at moisture contents ≤20%, and F1 generation survival did not occur in green coffee at moisture contents ≤15%. As the international standard for green coffee moisture content is 9-12%, adult CBB should not survive or reproduce in exported dry green coffee. Standard industry processing of harvested coffee cherries to the green coffee stage using either mechanical- or sun-drying eliminated CBB infestations from the field. A systems approach is recommended for exporting green coffee to control CBB and CLR that includes eliminating CBB life stages with standard processing methods, reducing moisture content to 9-12% to prevent egg deposition, survival or reproduction, and O3 fumigation to ensure quarantine security against potential CBB adult hitchhikers.


[Usefulness of the molecular techniques for detecting and/or identifing of parasites and fungi in humans and animals or pathogens transmitted by ticks (Part I)].
After a long period of using basic microscopic, immunological and biochemical methods for diagnosis, rapid development of nucleic acids investigation enabled introduction of specific and sensitive methods of detection of pathogenic agents on the molecular level. Among others, polymerase chain reaction (PCR), discovered in mid of 80'ies and then automatized, offered an attractive alternative to conventional testing systems. In this paper we describe reliable diagnostic tests widely used in the world, including Poland, and capable of detecting different disease agents as parasites and fungi in clinical specimens and pathogens of emerging zoonotic diseases in ticks. The possibilities of using molecular methods for determination of Plasmodium falciparum drug resistance is also discussed. Moreover, the report offers information concerning kinds of molecular tests and institutions in which there are executed.


Extracorporeal circulation in ewe's foetus: towards a reliable foetal cardiac surgery protocol. A comparison of two cases.
Foetal cardiac surgery is the ultimate goal in the treatment of congenital cardiac malformations. The aim of our research is to elucidate some of the features of the necessarily invasive experimental protocol to be used in an animal model of foetal cardiac surgery. In particular, we assessed the foetal placental reactivity to prolonged cardiac bypass in steady-flow conditions. Two cases were selected to show the outcome of prolonged (> 30 minutes) extracorporeal circulation (ECC) instituted without oxygenator under steady-flow assistance. Following the instrumentation of the animal (placement of pressure, flow and myocardial fiber length transducers) and the baseline recordings, a 60-minute bypass period was established with an axial turbopump (Hemopump 14 Fr), after systemic heparinisation and artero-venous cannulation. At the end of the circulatory assistance, the cannulae were removed and a 90 minute observation period followed. The cardiac function was assessed by means of indirectly obtained P-V loops. Case A showed a marked reduction in the end-systolic pressure-volume relationship (ESPVR) during ECC, corresponding to a rightward shift of the P-V loop, with a gradual recovery after the assisted circulation. On the contrary, case B was subjected to progressive placental dysfunction, as evidenced by haemogasanalytical data. Consequently, the haemodynamic data also outlined a negative outcome, with high ESPVR values after bypass. The present study, while confirming the possibility of cardiac intervention in the foetus, underlines the critical role of minimally invasive protocol to limit both foetal stress and placental dysfunction.


Validation of the Work Observation Method By Activity Timing (WOMBAT) method of conducting time-motion observations in critical care settings: an observational study.
Electronic documentation handling may facilitate information flows in health care settings to support better coordination of care among Health Care Providers (HCPs), but evidence is limited. Methods that accurately depict changes to the workflows of HCPs are needed to assess whether the introduction of a Critical Care clinical Information System (CCIS) to two Intensive Care Units (ICUs) represents a positive step for patient care. To evaluate a previously described method of quantifying amounts of time spent and interruptions encountered by HCPs working in two ICUs. Observers used PDAs running the Work Observation Method By Activity Timing (WOMBAT) software to record the tasks performed by HCPs in advance of the introduction of a Critical Care clinical Information System (CCIS) to quantify amounts of time spent on tasks and interruptions encountered by HCPs in ICUs. We report the percentages of time spent on each task category, and the rates of interruptions observed for physicians, nurses, respiratory therapists, and unit clerks. Compared with previously published data from Australian hospital wards, interdisciplinary information sharing and communication in ICUs explain higher proportions of time spent on professional communication and documentation by nurses and physicians, as well as more frequent interruptions which are often followed by professional communication tasks. Critical care workloads include requirements for timely information sharing and communication and explain the differences we observed between the two datasets. The data presented here further validate the WOMBAT method, and support plans to compare workflows before and after the introduction of electronic documentation methods in ICUs.


The relationship between census-derived socio-economic variables and general practice consultation rates in three town centre practices.
The relationship between socio-economic factors and consultation rates is important in determining resource allocation to general practices. To determine the relationship between general practice surgery consultation rates and census-derived socio-economic variables for patients receiving the same primary and secondary care. A retrospective analysis was taken of computerized records in three general practices in Mansfield, North Nottinghamshire, with 29,142 patients spread over 15 electoral wards (Jarman score range from -23 to +25.5). Linear regression analysis of surgery consultation rates at ward and enumeration district levels was performed against Jarman and Townsend deprivation scores and census socio-economic variables. Both the Townsend score (r2 = 59%) and the Jarman score (r2 = 39%) were associated with surgery consultation rates at ward level. The Townsend score had a stronger association than the Jarman score because all four of its component variables were individually associated with increased consultations compared with four out of eight Jarman components. Even in practices not eligible for deprivation payments there were appreciable differences in consultation rates between areas with different socio-economic characteristics. The results suggest that the variables used to determine deprivation payments should be reconsidered, and they support suggestions that payments should be introduced at a lower level of deprivation and administered on an enumeration district basis.


Bayesian prediction of winning times for elite swimming events.
To develop a statistical model of winning times for international swimming events with the aim of predicting winning time distributions and the probability of winning for the 2020 and 2024 Olympic Games. The data set included first and third place times from all individual swimming events from the Olympics and World Championships from 1990 to 2019. We compared different model formulations fitted with Bayesian inference to obtain predictive distributions; comparisons were based on mean percentage error in out-of-sample predictions of Olympics and World Championships winning swim times from 2011 to 2019. The Bayesian time series regression model, comprising auto-regressive and moving average terms and other predictors, had the smallest mean prediction error of 0.57% (CI 0.46-0.74%). For context, using the respective previous Olympics or World Championships winning time resulted in a mean prediction error of 0.70% (CI 0.59-0.82%). The Olympics were on average 0.5% (CI 0.3-0.7%) faster than World Championships over the study period. The model computes the posterior predictive distribution, which allows coaches and athletes to evaluate the probability of winning given an individual's swim time, and the probability of being faster or slower than the previous winning time or even the world record.


[A model of ecological hazard and social tension for the description of the health deterioration risk in the Russian population].
A model for quantitative evaluation of risks from long-exposure to high-intensity anthropogenic factors and chronic psychoemotional stress is presented. The model has been based on the concept of G. Seliet concerning the development of non-specific physiological reactions (the general adaptation syndrome) to chronic comparatively intensive stresses. It describes the process of time-dependent depletion of the compensatory body reserve and the probability of regulation disorders and disruption of the adaptation to single factors or their combinations. On the population level, health risks predicted with the model are growth of death probability within a year and in a delayed period (transformation of age-specific coefficients of mortality) and a magnitude of reduction of remaining life expectancy.


Renal vein cytokine release as an index of renal parenchymal inflammation in chronic experimental renal artery stenosis.
Renal parenchymal inflammation is a critical determinant of kidney injury in renal artery stenosis (RAS) but is difficult to assess in the single kidney without tissue samples. Whether renal vein (RV) levels of inflammatory markers reflect active parenchymal inflammation remains unknown. We evaluated the relationship between net RV cytokine release and tissue inflammation in the post-stenotic kidney. Pigs were studied after 10 weeks of RAS treated 4 weeks earlier with intra-renal vehicle or anti-inflammatory mesenchymal stem cells (MSCs) or normal control. Single-kidney renal blood flow was measured by fast computerized tomography. RV and inferior vena cava levels of tumor necrosis factor (TNF)-α, interferon (IF)-γ, monocyte chemoattractant protein (MCP-1) and interleukin (IL)-10 were measured by enzyme-linked immunosorbent assay, and their net release calculated. Renal expression of the same cytokines was correlated with their net release. Net release of TNF-α, IF-γ and MCP-1 was higher in RAS compared with normal and to the contralateral kidney (all P<0.05), decreased in MSC-treated pigs as was their tissue expression. Contrarily, the release of the anti-inflammatory IL-10 was lower in RAS and normalized in RAS+MSC. The net release of TNF-α, MCP-1 and IL-10 directly correlated with their tissue expression. The ratio of inflammatory-to-reparative macrophages directly correlated with the release of MCP-1, but inversely with the release of IL-10. In vitro cultured MSCs also induced a shift in the macrophage phenotype from inflammatory (M1) to reparative (M2). Our findings demonstrate that the release of inflammatory markers from the affected kidney provides an index of renal tissue inflammation in experimental RAS.


Acute and subchronic effects of methadone on the blood hormonal levels of pregnant and nonpregnant Charles River CD-1 mice.
Blood levels of ACTH, FSH, and estriol were measured throughout the estrous cycle and estriol was determined at different stages during pregnancy in Charles River CD-1 mice treated with 10 mg/kg/day of methadone or vehicle (physiological saline). Animals received one dose in a constant volume (10 ml/kg) per day subcutaneously of either methadone or saline. Blood samples of nonpregnant mice were collected 1 hour after the first dose for acute effects and 1 hour after the last dose treatment for subchronic effects. The acute administration of methadone in nonpregnant mice produced an increase in ACTH level throughout the estrous cycle whereas subchronic treatment reduced ACTH level by 51%. Acute treatment did not alter the estriol or FSH levels whereas subchronic treatment significantly lowered estriol by 17% and FSH by 79%. Methadone injected beginning on day 1 of gestation and continued through day 15 did not produce any effect on maternal body weight or food consumption but resulted in an increase in resorption sites and decrease in implantation sites. The estriol levels in control pregnant mice were 19.8, 54.8, and 109.1 ng/ml on days 1, 10, and 15 of gestation, respectively. A significant reduction of 18.8% and 35.2% in estriol was associated with methadone treatment by days 10 and 15 of gestation, respectively. Methadone, by affecting several hormonal levels in both pregnant and nonpregnant CD-1 mice, may be responsible for some of the adverse effects on reproduction encountered in this species.


[Function of transplanted heart].
Clinical and functional results of orthotopic cardiac transplantation are analyzed. It is demonstrated that case follow-up with special non-invasive methods (ECG, Holter monitoring, high-resolution ECG, EchoCG, dopler-EchoCG) is mandatory for detection of cardiac transplant function, early diagnosis of chronic rejection crisis, timely and optimal treatment tactics.


Fermented ginseng improved alcohol liver injury in association with changes in the gut microbiota of mice.
The interactions among the liver, intestine and immune system play an important role in alcoholic liver injury. In this study, C57BL/6N mice with alcoholic injury were treated with unfermented and Lactobacillus fermentum KP-3-fermented ginseng. The indicators of hepatic steatosis, inflammation and injury were evaluated. The number of beneficial and harmful bacteria in the mice ileum and colon was counted by a traditional method; moreover, the diversity analysis of the cecum flora was performed. The alcohol exposure increased the levels of ALT, AST, TNF-α and IL-6 inflammatory factors and liver steatosis. In addition, the alcohol-fed miceexhibited a lower number of Lactobacilli and Bifidobacteria in the ileum and colon; the cecum flora diversity in the mice showed that alcohol obviously enhanced the abundance of the unclassified S24-7 of the Bacteroidetes phylum and the Proteobacteria genus of the Sutterella phylum and reduced the abundance of short-chain fatty acid-producing bacteria such as Akkermansia in the Verrucomicrobia phylum and those belonging to the Allobaculum genus, the Ruminococcus genus, and the Adlercreutzia genus in the Actinobacteria phylum. All these changes were improved by fermented ginseng. Conclusively, fermented ginseng could alleviate the alcoholic liver injury and disorder of the intestine by adjusting the intestinal flora.


The development of sleep disordered breathing from 4 to 12 years and dental arch morphology.
To track the development of sleep disordered breathing (SDB) as well as dento-facial morphology in cohort of children by having them complete a questionnaire at ages 4, 6 and 12. Clinical examination, sleep studies (at ages 4 and 12) and orthodontic evaluation were carried out on all who were reported to snore regularly and children who did not snore at all. Out of the original group of 615 children, 64% (393) answered the inquiry on all three occasions. Of those, 27 snored regularly and 231 did not snore at all at the age of 12. There were differences between those groups on all answers, especially prevalence of oral breathing: 78% versus 5% (p<0.001). The prevalence of OSA decreased from 3.1% at the age of 4 to 0.8% at age 12 and the severity decreased from a mean AHI 14.8 at 4 to a mean AHI of 1.95 at age 12. The minimum prevalence of snoring regularly was estimated to 4.2% at 12 years compared to 5.3% at 4, calculated for the original cohort of 644 children. The odds for a child who snored regularly at 4 or 6 years to be snoring regularly also at age 12 was 3.7 times greater than for a not snoring child in spite of surgery (OR 3.7, 95% CI 2.4-5.7). 63 children had undergone surgery due to snoring by age 12. 14 of them never snored and 17 snored regularly at the age 12. The dental arch was narrower in the children snoring regularly at 4, 6 and 12 years compared to not snoring children. Cross-bites were more common among snoring children than among non-snoring children, at 4 and 6 as well as at 12. The prevalence of regular snoring is about the same from 4 to 12 years independent of surgery, but the prevalence of OSA decreased considerably. The children snoring regularly generally have a narrower maxilla compared to children not snoring. Surgery in young children is necessary but "cures" the snoring only temporary.


Contrast gain control in the kitten's visual system.
We have studied the effects of contrast adaptation on cortical cells from 4- and 6-wk-old kittens (49 and 47 cells, respectively) using sine-wave grating stimuli. We wished to know if the effects of adaptation to different contrast levels are more extensive than those in adult animals. Our experiments involved adapting cells to different contrasts (3.1, 12.5, and 50%) while concurrently measuring their contrast-response functions at each of these different levels. We found qualitatively that the effects of adaptation in the kitten are similar to those we have previously documented in adult animals (19). Contrast-response functions are laterally shifted along the log-contrast axis, effectively matching the response range of the cells to prevailing contrast levels. The degree to which this occurred varied from cell to cell. The average degree to which cells showed these effects, as assessed both qualitatively and quantitatively, was greater for kittens than for adult cats, and greater for 4-wk-old kittens than for those aged 6 wk. This suggests that susceptibility to adaptation varies as a function of age. Additional studies were undertaken with the intent of localizing these adaptive effects. First, lateral geniculate cells and fibers (n = 23) were studied with our standard protocol, and second, we investigated the degree to which the effects of adaptation of cortical cells transferred interocularly.(ABSTRACT TRUNCATED AT 250 WORDS)


[Biological properties of the mechanism of "marine" phospholipids containing omega-3 fatty acids].
The Laboratory mainly deals with the development of biologic technologies for producing physiologically active lipid-protein-nature compounds from marine organisms containing omega-3 fatty acids (that are membrane components) with further evaluation of their action both in the normal state and in some simulated pathologic states as well as with creation of new preparations for application in medicine and agriculture on their basis. As a result of the experiments performed, a technology for producing two biologic preparations, namely, surface active compounds (phospholipids) and a nucleopeptid-lipid complex, exhibiting a specific action, was developed. The phospholipid complex, being surface-active in composition, was characterized as a complex possessing some surfactant-type properties and displaying an antioxidant and a membrane-stabilizing effect. On the basis of the complex including marine phospholipids with omega-3 fatty acids, the preparation "Phylomek", which is the concentrate of essential marine phospholipids, and the preparation "Morephyl", which is a marine agent with surfactant-type effect, intended for animals and poultry, were created. The ingredients of the nucleopeptid-lipid complex were identified, and its effect on the increase of testosterone levels in the blood of old and sick animals was determined. A stimulating agent of genital hormones secretion was recommended for application in the geriatrics in the case of genital function disturbance and presenilation. A fraction similar in chemical content and specific activity, found in the velvet antlers, was used as the base of the biologic preparation "Pantheron". The natural complexes of marine phospholipids were shown to be able to change the composition of lipids of membranes, but the intensity of these changes differed in cells differing in function. In the study of the biochemical mechanism of correction of disturbances in the cell membranes under an oxidative stress, the interrelation between the composition of lipids of membranes, their oxidation, and the content of natural antioxidants was determined. The reparative effect of marine phospholipids on the cell membranes under progressing pathology, caused by the restoration of the composition of phospholipids, by increases in the activities of antioxidant enzymes (SOD, catalase), and by a decrease of the accumulation of LPO products, was established. Under interaction of marine phospholipids and alpha-tocopherol, synergism was noticed, the antioxidant potential of the investigated substances and their membrane-stabilizing effect increased. The phospholipids with various residues of PUFA in the molecule were found to affect the inhibition and oxidation processes, as well as the modelling of lipid membranes. This is especially true for arachidonic and docosahexaenoic acids, the ratio of which changes under the oxidative stress. At administration of phospholipids omega-3, their ratio decreases due a decrease in the level of unetherificated PUFA. The main changes of the PUFA were found to occur in phosphatidylethanolamine isolated from microsomes. The particular role of phosphatidylethanolamine and arachidonic acid in the reparation of membranes under the action of phospholipids PUFA omega-3 and alpha-tocopherol was noted.


Enhanced platelet adhesion to collagen in scleroderma. Effect of scleroderma plasma and scleroderma platelets.
The effect of plasma on platelet adhesion to collagen coated microtiter wells was investigated in 22 patients with scleroderma and 26 control subjects. In the control subjects, platelet adhesion was 38 +/- 13% (mean +/- SD) of adhesion with buffer alone; in scleroderma patients adhesion was 64 +/- 20% (p less than 0.001). No correlation was seen between the effect of plasma on platelet adhesion to collagen and the plasma levels of either FVIII/von Willebrand factor antigen or fibronectin in either scleroderma or control subjects. Furthermore, scleroderma platelets demonstrated enhanced adhesion compared to control platelets when tested in the presence of either control or scleroderma plasma.


Effect of artificial landmarks of the prefabricated auxiliary devices located at different arch positions on the accuracy of complete-arch edentulous digital implant scanning: An in-vitro study.
To examine the effect of artificial landmarks of prefabricated auxiliary devices (PAD) located at different arch positions on the accuracy of complete-arch edentulous digital implant scanning. A reference model containing four analogs and PAD were fabricated by a 3D printer (AccuFab-C1s, 3DShining). 10 digital scans were performed using an intraoral scanner (Aoralscan 3, 3DShining), sv 22.214.171.12415, with artificial landmarks located at different arch positions: group I, without any artificial landmarks; group II, with artificial landmarks at the anterior region; group III, with artificial landmarks at the posterior region. group IV: with artificial landmarks at both anterior and posterior regions. For group V: Conventional open-tray splinted impressions. The reference file and conventional stone casts were digitalized by using a dental laboratory scanner. The related files were imported into inspection software for trueness and precision assessment. Statistical analysis was performed with One-way ANOVA and Kruskal-Wallis test. The level of significance was set at α=0.05. For the global accuracy assessment, significantly higher global trueness was seen in group II (p < 0.01), III (p < 0.001), IV (p < 0.001) and V (p < 0.001) than group I. Significantly higher global precision was seen in group III (p < 0.001), IV (p < 0.001) and V (p < 0.001) than group I. For the local accuracy assessment, the PAD primarily improved accuracy on the linear deviations. Artificial landmarks of PAD at different arch positions significantly influenced the scanning accuracy. Applying the PAD in group IV could achieve comparable outcomes to conventional open-tray splinted impressions. Artificial landmarks on the posterior region may be more pivotal than those on the anterior region. Group IV could achieve comparable accuracy to conventional open-tray splinted impressions.


Obesity and orthodontic treatment: is there any direct relationship?
Obesity is a wide-spread condition directly or indirectly connected with an increase in the prevalence of a variety of human diseases. It affects over 50% of the western overall population. In 2017, a thorough analysis of 204 studies on obesity and cancer revealed that the condition increases the risk of the following types of cancer: stomach, colon, rectal, bile duct, pancreatic, esophagus, breast, endometrial, ovarian, kidney and multiple myeloma. The first study aiming at establishing a connection between obesity and the rate of induced orthodontic tooth movement was conducted by Saloom et al; however, it could not effectively nor significantly reveal any direct influence or effect. Despite being identified during the first week, differences could not be explained and treatment time remained unchanged. In spite of lack of studies in the literature on the connection between obesity and the rate of induced tooth movement, in clinical practice, courses or specialized training, we should not have protocols changed nor adopt any measures or expect significant differences between normal-weight and obese individuals. It should be emphasized that unsuccessful cases or cases of root resorption associated with treatment should not be assigned to obesity, since scientific data is insufficient to do so.


NMR solution structure of Thermotoga maritima protein TM1509 reveals a Zn-metalloprotease-like tertiary structure.
The 150-residue protein TM1509 is encoded in gene YF09_THEMA of Thermotoga maritima. TM1509 has so far no functional annotation and belongs to protein family UPF0054 (PFAM accession number: PF02130) which contains at least 146 members. The NMR structure of TM1509 reveals an alpha+beta fold comprising a four stranded beta-sheet with topology A( upward arrow), B( upward arrow), D( upward arrow), C( downward arrow) as well as five alpha-helices I-V. The structures of most members of family PF02130 can be reliably constructed using the TM1509 NMR structure, demonstrating high leverage for exploration of fold space. A multiple sequence alignment of TM1509 with homologues of family UPF0054 shows that three polypeptide segments, as well as a putative zinc-binding consensus motif HGXLHLXGYDH located at the C-terminal end of alpha-helix IV, are highly conserved. The spatial arrangement of the three His residues of this UPF0054 consensus motif is similar to the arrangement found for the His residues in the HEXXHXXGXXH zinc-binding consensus motif of matrix metallo-proteases (MMPs). Moreover, the other conserved polypeptide segments form a large cavity which encloses the putative Zn-binding pocket and might confer specificity during catalysis. However, TM1509 and the other members of the UPF0054 family do not have the crucial Glu residue in position 2 of the MMP consensus motif. Intriguingly, the TM1509 structure indicates that the Asp in the UPF0054 consensus motif (Asp 111 in TM1509) may overtake the catalytic role of the Glu. This suggests that protein family UPF0054 might contain members of a hitherto uncharacterized class of metalloproteases.


Integrative genomics of the mammalian alveolar macrophage response to intracellular mycobacteria.
Bovine TB (bTB), caused by infection with Mycobacterium bovis, is a major endemic disease affecting global cattle production. The key innate immune cell that first encounters the pathogen is the alveolar macrophage, previously shown to be substantially reprogrammed during intracellular infection by the pathogen. Here we use differential expression, and correlation- and interaction-based network approaches to analyse the host response to infection with M. bovis at the transcriptome level to identify core infection response pathways and gene modules. These outputs were then integrated with genome-wide association study (GWAS) data sets to enhance detection of genomic variants for susceptibility/resistance to M. bovis infection. The host gene expression data consisted of RNA-seq data from bovine alveolar macrophages (bAM) infected with M. bovis at 24 and 48 h post-infection (hpi) compared to non-infected control bAM. These RNA-seq data were analysed using three distinct computational pipelines to produce six separate gene sets: 1) DE genes filtered using stringent fold-change and P-value thresholds (DEG-24: 378 genes, DEG-48: 390 genes); 2) genes obtained from expression correlation networks (CON-24: 460 genes, CON-48: 416 genes); and 3) genes obtained from differential expression networks (DEN-24: 339 genes, DEN-48: 495 genes). These six gene sets were integrated with three bTB breed GWAS data sets by employing a new genomics data integration tool-gwinteR. Using GWAS summary statistics, this methodology enabled detection of 36, 102 and 921 prioritised SNPs for Charolais, Limousin and Holstein-Friesian, respectively. The results from the three parallel analyses showed that the three computational approaches could identify genes significantly enriched for SNPs associated with susceptibility/resistance to M. bovis infection. Results indicate distinct and significant overlap in SNP discovery, demonstrating that network-based integration of biologically relevant transcriptomics data can leverage substantial additional information from GWAS data sets. These analyses also demonstrated significant differences among breeds, with the Holstein-Friesian breed GWAS proving most useful for prioritising SNPS through data integration. Because the functional genomics data were generated using bAM from this population, this suggests that the genomic architecture of bTB resilience traits may be more breed-specific than previously assumed.


Postartesunate delayed hemolysis is a predictable event related to the lifesaving effect of artemisinins.
Patients with severe malaria treated with artesunate sometimes experience a delayed hemolytic episode. Artesunate (AS) induces pitting, a splenic process whereby dead parasites are expelled from their host erythrocytes. These once-infected erythrocytes then return to the circulation. We analyzed hematologic parameters in 123 travelers treated with AS for severe malaria. Among 60 nontransfused patients observed for more than 8 days, 13 (22%) had delayed hemolysis. The peak concentration of circulating once-infected erythrocytes was measured during the first week in 21 patients and was significantly higher in 9 patients with delayed hemolysis than in 12 with other patterns of anemia (0.30 vs 0.07; P = .0001). The threshold of 180 million once-infected erythrocytes per liter discriminated patients with delayed hemolysis with 89% sensitivity and 83% specificity. Once-infected erythrocyte morphology analyzed by using ImageStream in 4 patients showed an 8.9% reduction in their projected area, an alteration likely contributing to their shorter lifespan. Delayed clearance of infected erythrocytes spared by pitting during AS treatment is an original mechanism of hemolytic anemia. Our findings consolidate a disease framework for posttreatment anemia in malaria in which delayed hemolysis is a new entity. The early concentration of once-infected erythrocytes is a solid candidate marker to predict post-AS delayed hemolysis.


Insights into bacterial protein glycosylation in human microbiota.
The study of human microbiota is an emerging research topic. The past efforts have mainly centered on studying the composition and genomic landscape of bacterial species within the targeted communities. The interaction between bacteria and hosts is the pivotal event in the initiation and progression of infectious diseases. There is a great need to identify and characterize the molecules that mediate the bacteria-host interaction. Bacterial surface exposed proteins play an important role in the bacteria- host interaction. Numerous surface proteins are glycosylated, and the glycosylation is crucial for their function in mediating the bacterial interaction with hosts. Here we present an overview of surface glycoproteins from bacteria that inhabit three major mucosal environments across human body: oral, gut and skin. We describe the important enzymes involved in the process of protein glycosylation, and discuss how the process impacts the bacteria-host interaction. Emerging molecular details underlying glycosylation of bacterial surface proteins may lead to new opportunities for designing anti-infective small molecules, and developing novel vaccines in order to treat or prevent bacterial infection.


The national response for preventing healthcare-associated infections: system capacity and sustainability for improvement.
Strengthening capacity across the healthcare system for improvement is critical to ensuring that past efforts and investments establish a foundation for sustaining progress in patient safety. The objective of this analysis was to identify key system capacity issues for sustainability from evaluation of the Action Plan to prevent healthcare-associated infections, a major national initiative launched by the US Department of Health and Human Services in 2009. The analysis involves the review and synthesis of results across the components of a 3-year evaluation of the Action Plan, as described in the evaluation framework and detailed in separate analyses elsewhere in this special issue. Data collection methods included interviews with government and private stakeholders, document and literature reviews, and observations of meetings and conferences at multiple time points. Key developments in healthcare-associated infection prevention system capacity were extracted on the basis of "major activities" identified through multiple methods and organized into the level of progress based on perspectives of multiple stakeholders. Activities within each level were then examined and compared according to our evaluation's framework of 4 system functions and 5 system properties. Key system capacity and sustainability issues for the Action Plan to be addressed centered on coordination and alignment (among participating agencies, with other federal initiatives, and across levels of healthcare), infrastructure for data and accountability (including more efficient technologies and unintended consequences), cultural embedding of prevention practices, and uncertainty and variability in resources. Sustainability depends on improvements across system functions and properties and how they reinforce each other. Change is more robust if different system elements support and incentivize behavior in similar directions.


Effectiveness of phytoremediation as a secondary treatment for polycyclic aromatic hydrocarbons (PAHs) in composted soil.
A greenhouse study was conducted over a 12-month period to investigate the fate of polycyclic aromatic hydrocarbons (PAHs) in soil using phytoremediation as a secondary treatment. The soil was pretreated by composting for 12 weeks, then planted with tall fescue (Festuca arundinacea), annual ryegrass (Lolium multiflorum), and yellow sweet clover (Melilotus officinalis). Two sets of unvegetated controls also were evaluated, one fertilized and one unfertilized. Total PAH concentrations decreased in the tall fescue, annual ryegrass, and yellow sweet clover treatments by 23.9%, 15.3%, and 9.1%, respectively, whereas the control was reduced by less than 5%. The smaller two- and most of the three-ringed compounds--naphthalene, acenaphthylene, acenaphthene, fluorene, and anthracene--were not found in detectable concentrations in any of the treatments. The most probable number analysis for microbial PAH degraders did not show any statistically significant differences among treatments. There were significant differences among treatments (p < 0.05) for the residual concentrations of five of the target PAHs. Root surface area measurements indicated that tall fescue and annual ryegrass both had significantly higher root surface area than yellow sweet clover, although the two species were not significantly different from each other. The tall fescue treatment resulted in the highest root and shoot biomass, followed by annual ryegrass and yellow sweet clover, and also had the highest percent of contaminant removal after 12 months. These results imply a positive relationship between plant biomass development and PAH biodegradation.


Dimethyl sulphoxide induces a reduced growth rate, altered cell morphology and increased epidermal-growth-factor binding in Hep G2 cells.
Culture of Hep G2 cells in medium containing 2% (v/v) dimethyl sulphoxide (DMSO) resulted in a slowing of growth and a marked change in morphological appearance. By day 6, cultures containing DMSO had one-third the number of cells compared with parallel control cultures. Measurement of 125I-epidermal-growth-factor (EGF) binding to DMSO-treated cells revealed a striking time-dependent elevation in specific EGF binding to their cell surface. Increased binding was detectable within 24 h of the start of DMSO treatment, reaching, by 6 days, levels almost 25 times greater than those for control cells. Addition of EGF to DMSO-treated cells caused a rapid down-regulation of the EGF receptor, but did not alter their proliferation rate. Slowing of growth by other means, such as serum starvation, growth to confluence or culture in the presence of sodium butyrate, did not affect 125I-EGF binding, indicating a specific effect of DMSO on these cells.


Feeling Positive About Reopening? New Normal Scenarios From COVID-19 US Reopen Sentiment Analytics.
The Coronavirus pandemic has created complex challenges and adverse circumstances. This research identifies public sentiment amidst problematic socioeconomic consequences of the lockdown, and explores ensuing four potential public sentiment associated scenarios. The severity and brutality of COVID-19 have led to the development of extreme feelings, and emotional and mental healthcare challenges. This research focuses on emotional consequences - the presence of extreme fear, confusion and volatile sentiments, mixed along with trust and anticipation. It is necessary to gauge dominant public sentiment trends for effective decisions and policies. This study analyzes public sentiment using Twitter Data, time-aligned to the COVID-19 reopening debate, to identify dominant sentiment trends associated with the push to reopen the economy. Present research uses textual analytics methodologies to analyze public sentiment support for two potential divergent scenarios - an early opening and a delayed opening, and consequences of each. Present research concludes on the basis of textual data analytics, including textual data visualization and statistical validation, that tweets data from American Twitter users shows more positive sentiment support, than negative, for reopening the US economy. This research develops a novel sentiment polarity based public sentiment scenarios (PSS) framework, which will remain useful for future crises analysis, well beyond COVID-19. With additional validation, this research stream could present valuable time sensitive opportunities for state governments, the federal government, corporations and societal leaders to guide local and regional communities, and the nation into a successful new normal future.


Strong human endogenous retrovirus-specific T cell responses are associated with control of HIV-1 in chronic infection.
Eight percent of the human genome is composed of human endogenous retroviruses (HERVs), which are thought to be inactive remnants of ancient infections. Previously, we showed that individuals with early HIV-1 infection have stronger anti-HERV T cell responses than uninfected controls. In this study, we investigated whether these responses persist in chronic HIV-1 infection and whether they have a role in the control of HIV-1. Peripheral blood mononuclear cells (PBMCs) from 88 subjects diagnosed with HIV-1 infection for at least 1 year (median duration of diagnosis, 13 years) were tested for responses against HERV peptides in gamma interferon (IFN-γ) enzyme immunospot (ELISPOT) assays. Individuals who control HIV-1 viremia without highly active antiretroviral therapy (HAART) had stronger and broader HERV-specific T cell responses than HAART-suppressed patients, virologic noncontrollers, immunologic progressors, and uninfected controls (P < 0.05 for each pairwise comparison). In addition, the magnitude of the anti-HERV T cell response was inversely correlated with HIV-1 viral load (r(2) = 0.197, P = 0.0002) and associated with higher CD4(+) T cell counts (r(2) = 0.072, P = 0.027) in untreated patients. Flow cytometric analyses of an HLA-B51-restricted CD8(+) HERV response in one HIV-1-infected individual revealed a less activated and more differentiated phenotype than that stimulated by a homologous HIV-1 peptide. HLA-B51 tetramer dual staining within this individual confirmed two different T cell populations corresponding to these HERV and HIV-1 epitopes, ruling out cross-reactivity. These findings suggest a possible role for anti-HERV immunity in the control of chronic HIV-1 infection and provide support for a larger effort to design an HIV-1 vaccine that targets conserved antigens such as HERV.


Magnetization transfer in neuroimaging.
Magnetization transfer (MT) imaging has become a focus of recent basic science and clinical investigations because of its unique physical basis of contrast, and the potential to quantify aspects of the biochemical structure and composition of tissues. In this article, basic theories of MT are discussed, and several practical MT pulse and sequence considerations are addressed. The clinical utility of MT techniques in MR angiography, postgadolinium T1-weighted scanning, and spine scanning are discussed. Highlights of quantitative MT studies for pathologic tissue characterization and for studies of patients with white-matter disorders such as multiple sclerosis are included, giving a comprehensive review of the neuroimaging application of MT.


Invasion and adaptation of a warm-adapted species to montane localities: effect of acclimation potential.
Drosophila ananassae has successfully invaded the cold and dry montane localities of the Western Himalayas in recent years. The ability of this desiccation- and cold-sensitive tropical species to evolve in response to seasonal changes in montane localities is largely unknown. Here, we investigated how this sensitive species adapt to seasonally varying environmental conditions that are lethal to its survival. We observed change in the frequency of dark and light morphs of D. ananassae in five mid-altitude localities during the last decade (2000-2010). We document invasion of D. ananassae to montane localities and increase in frequency of the dark morph. The stress tolerance of morphs (dark and light) remained unaffected of developmental acclimation. However, adult acclimation has shown significant effects on tolerance to various environmental stresses in morphs and effect of this acclimation persist for long durations. Desiccation and cold stress tolerance was increased after adult acclimation for respective stress in the dark morph; while tolerance of the light morph was not affected. Further, heat tolerance of the light morph was increased after adult heat acclimation with no influence on heat tolerance of the dark morph. Our results suggest a possible role of adult acclimation in successful invasion and adaptation of D. ananassae to montane localities. Future experiments should be carried out to determine whether the survival in adverse conditions of low versus high temperature and humidity during seasonal changes is assisted by different acclimation abilities of the two morphs of D. ananassae.


Novel polysome messages and changes in translational activity appear after induction of adipogenesis in 3T3-L1 cells.
Control of translation allows for rapid adaptation of the cell to stimuli, rather than the slower transcriptional control. We presume that translational control is an essential process in the control of adipogenesis, especially in the first hours after hormonal stimulation. 3T3-L1 preadipocytes were cultured to confluency and adipogenesis was induced by standard protocols using a hormonal cocktail. Cells were harvested before and 6 hours after hormonal induction. mRNAs attached to ribosomes (polysomal mRNAs) were separated from unbound mRNAs by velocity sedimentation. Pools of polysomal and unbound mRNA fractions were analyzed by microarray analysis. Changes in relative abundance in unbound and polysomal mRNA pools were calculated to detect putative changes in translational activity. Changes of expression levels of selected genes were verified by qPCR and Western blotting. We identified 43 genes that shifted towards the polysomal fraction (up-regulated) and 2 genes that shifted towards free mRNA fraction (down-regulated). Interestingly, we found Ghrelin to be down-regulated. Up-regulated genes comprise factors that are nucleic acid binding (eIF4B, HSF1, IRF6, MYC, POLR2a, RPL18, RPL27a, RPL6, RPL7a, RPS18, RPSa, TSC22d3), form part of ribosomes (RPL18, RPL27a, RPL6, RPL7a, RPS18, RPSa), act on the regulation of translation (eIF4B) or transcription (HSF1, IRF6, MYC, TSC22d3). Others act as chaperones (BAG3, HSPA8, HSP90ab1) or in other metabolic or signals transducing processes. We conclude that a moderate reorganisation of the functionality of the ribosomal machinery and translational activity are very important steps for growth and gene expression control in the initial phase of adipogenesis.


The Impact of Principal Diagnosis on Readmission Risk among Patients Hospitalized for Community-Acquired Pneumonia.
Coding variation distorts performance/outcome statistics not eliminated by risk adjustment. Among 1596 community-acquired pneumonia patients hospitalized from 1998 to 2012 identified using an evidence-based algorithm, the authors measured the association of principal diagnosis (PD) with 30-day readmission, stratified by Pneumonia Severity Index risk class. The 152 readmitted patients were more ill (Pneumonia Severity Index class V 38.8% versus 25.8%) and less likely to have a pneumonia PD (52.6% versus 69.9%). Among patients with PDs of pneumonia, respiratory failure, sepsis, and aspiration, mortality/readmission rates were 3.9/8.5%, 28.8/14.0%, 24.7/19.6%, and 9.0/15.0%, respectively. The nonpneumonia PDs were associated with a greater risk of adjusted 30-day readmission: respiratory failure odds ratio (OR) 1.89 (95% confidence interval [CI], 1.13-3.15), sepsis OR 2.54 (95% CI, 1.52-4.26), and possibly aspiration OR 1.73 (95% CI, 0.88-3.41). With increasing use of alternative PDs among pneumonia patients, quality reporting must account for variations in condition coding practices. Rigorous risk adjustment does not eliminate the need for accurate, consistent case definition in producing valid quality measures.


Repeated human tooth allograft after a previous rejection.
A case has been presented in which a humantooth allograft was performed. It failed in a reasonably expected manner. Another tooth allograft from the same donor to the same host was then performed. It is currently functioning two years postoperatively. This seems to defy the law of second-set reaction. Various explanations for the response to this repeated tooth allograft have been discussed. It is concluded that possibility of repeating tooth allografts, even after a first allograft has failed, should be considered.


Theoretical and Experimental Gas Volume Quantification of Micro- and Nanobubble Ultrasound Contrast Agents.
The amount of gas in ultrasound contrast agents is related to their acoustic activity. Because of this relationship, gas volume has been used as a key variable in normalizing the in vitro and in vivo acoustic behavior of lipid shell-stabilized bubbles with different sizes and shell components. Despite its importance, bubble gas volume has typically only been theoretically calculated based on bubble size and concentration that is typically measured using the Coulter counter for microbubbles and nanoparticle tracking analysis (NTA) for nanoscale bubbles. However, while these methods have been validated for the analysis of liquid or solid particles, their application in bubble analysis has not been rigorously studied. We have previously shown that resonant mass measurement (RMM) may be a better-suited technique for sub-micron bubble analysis, as it can measure both buoyant and non-buoyant particle size and concentration. Here, we provide validation of RMM bubble analysis by using headspace gas chromatography/mass spectrometry (GC/MS) to experimentally measure the gas volume of the bubble samples. This measurement was then used as ground truth to test the accuracy of theoretical gas volume predictions based on RMM, NTA (for nanobubbles), and Coulter counter (for microbubbles) measurements. The results show that the headspace GC/MS gas volume measurements agreed well with the theoretical predictions for the RMM of nanobubbles but not NTA. For nanobubbles , the theoretical gas volume using RMM was 10% lower than the experimental GC/MS measurements; meanwhile, using NTA resulted in an 82% lower predicted gas volume. For microbubbles, the experimental gas volume from the GC/MS measurements was 27% lower compared to RMM and 72% less compared to the Coulter counter results. This study demonstrates that the gas volume of nanobubbles and microbubbles can be reliably measured using headspace GC/MS to validate bubble size measurement techniques. We also conclude that the accuracy of theoretical predictions is highly dependent on proper size and concentration measurements.


Effect of biomaterials on angiogenesis during vital pulp therapy.
This review intended to provide an overview of the effects of dental materials, used in dentin-pulp complex and dental pulp regeneration, on angiogenesis processes during regenerative endodontic procedures. An electronic search was performed in PubMed and MEDLINE databases via OVID using the keywords mentioned in the PubMed and MeSH headings for English language published articles from January 2005-April 2014 that evaluated the angiogenic properties of different dental materials used in regenerative endodontic procedures. Of the articles identified in an initial search, only 40 articles met the inclusion criteria set for this review. Vital pulp therapy materials might have positive effects on angiogenesis events, while most of the canal irrigating solutions and antibiotic pastes have anti-angiogenic activity except for EDTA. Future clinical studies will be helpful in defining the mechanisms of action for dental materials that promote or inhibit angiogenesis events at applied areas.


For the general internist: a review of relevant 2013 innovations in medical education.
We conducted a review of articles published in 2013 to identify high-quality research in medical education that was relevant to general medicine education practice. Our review team consisted of six general internists with expertise in medical education of varying ranks, as well as a professional medical librarian. We manually searched 15 journals in pairs, and performed an online search using the PubMed search engine for all original research articles in medical education published in 2013. From the total 4,181 citations identified, we selected 65 articles considered most relevant to general medicine educational practice. Each team member then independently reviewed and rated the quality of each selected article using the modified Medical Education Research Study Quality Instrument. We then reviewed the quality and relevance of each selected study and grouped them into categories of propensity for inclusion. Nineteen studies were felt to be of adequate quality and were of moderate to high propensity for inclusion. Team members then independently voted for studies they felt to be of the highest relevance and quality within the 19 selected studies. The ten articles with the greatest number of votes were included in the review. We categorized the studies into five general themes: Improving Clinical Skills in UME, Inpatient Clinical Teaching Methods, Advancements in Continuity Clinic, Handoffs/Transitions in Care, and Trainee Assessment. Most studies in our review of the 2013 literature in general medical education were limited to single institutions and non-randomized study designs; we identified significant limitations of each study. Selected articles may inform future research and practice of medical educators.


Linker conformation effects on the band gap in metal-organic frameworks.
In this work, we investigate how torsion in the middle aromatic ring on the terphenyldicarboxylate linker in UiO-68 affects the band gap. Furthermore, we incorporate the effect of monosubstitution on the linker (UiO-68-R; R = H, F, I, NH2, and NO2) in order to shed light on a possible route to tune the band gap by changing the torsional angle by substitutions. Our computations show that both the torsional angle and band gaps depend on the choice of the substituent, and it is, in fact, possible to tune the band gap through the substitution's effect of locking down the middle aromatic ring at different torsional angles, in combination with the substituents' electronic effects.


Monoclonal antibody-mediated modulation of parathyroid hormone secretion by dispersed parathyroid cells.
Available data suggest that ionized calcium may interact with a cell surface "sensor" or "receptor" to produce changes in one or more intracellular second messengers that ultimately regulate the release of parathyroid hormone (PTH). Recently, we developed a series of monoclonal antibodies directed toward specialized differentiation antigens expressed on endocrine cells. Since many of these monoclonal antibodies displayed exquisite specificity for cell surface molecules on the parathyroid cell, we used these reagents as probes to investigate signal recognition/transduction mechanisms associated with abnormal calcium-regulated PTH secretion. Depending on their binding site on the respective target antigen molecules, these monoclonal antibodies either stimulated or inhibited hormone secretion. Thus, defects in membrane-associated structures may contribute to deranged calcium-regulated PTH secretion in abnormal parathyroid cells.


Effects of age, sex and manual task on hand preference in wild Rhinopithecus roxellana.
Golden snub-nosed monkeys (Rhinopithecus roxellana), as typical arboreal group-living Old World monkeys, provide an appropriate animal model to research manual laterality and explore the factors affecting hand preference in nonhuman primates. This study investigated hand preference based on 63 subjects and four spontaneous manual tasks (including unimanual and bimanual feeding and grooming), and assessed the effects of age, gender and type of task on handedness in R. roxellana. A population-level left-handedness was found not only in the bimanual coordinated tasks (bimanual feeding and grooming), but also in one unimanual reaching task (unimanual feeding). There were no significant differences between the sexes in either direction or strength of hand preference among any task. However, a significant difference between adults and juveniles was found in the unimanual feeding task. This is the first report on handedness in unimanual and bimanual feeding tasks that require bipedal posture in wild R. roxellana. Furthermore, this study demonstrated spontaneous feeding tasks reported previously only in the quadrupedal posture in this species, supporting the importance of factors such as posture and task complexity in the evolution of primate manual lateralization.


Production of polyhydroxyalkanoates (PHA) using sludge from different wastewater treatment processes and the potential for medical and pharmaceutical applications.
In this study, seven strains of bacteria with polyhydroxyalkanoates (PHA)-producing ability (i.e. Bacillus cereus, Pseudomonas putida, Bacillus pumilus, Pseudomona huttiensis, Yersinia frederiksenii, Aeromonas ichthiosmia, and Sphingopyxis terrae) were isolated from various waste treatment plants in Hong Kong. Simultaneous wastewater treatment and PHA accumulation were successfully achieved in the bioreactors using isolated bacteria from different sludges. At the organic loading less than 13,000 ppm, more than 95% of chemical oxygen demand (COD) was removed by the isolated strains before the decrease of PHA accumulation. In addition, more than 95% of nitrogen removal was achieved by all isolated strains. In the bioreactors inoculated with single strains, the highest yields of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyvalerate) (PHV) were obtained in A. ichthiosmia (84 mg PHB/g) and B. cereus (69 mg/g), respectively. For the mixed culture, the highest yields of PHB and PHV were increased by 55% and 45% in the system inoculated with B. pumilus and A. ichthiosmia. The biologically synthesized PHA also showed the potential applications in drug delivery and tissue engineering. PHA-nanoparticles loaded with pyrene were successfully prepared by recombinant Escherichia coli. The results of in vitro drug release and biocompatibility tests revealed that nanoparticles could be used as safer dray carriers with high loading capacity and efficiency. After 20 days, the cells successfully grew on 90% of the PHA-aortic valve.


Biochemical markers of bone turnover and prediction of hip bone loss in older women: the study of osteoporotic fractures.
To examine the ability of commercially available biochemical markers of bone formation and resorption to predict hip bone loss, we prospectively obtained serum and timed 2-h urine specimens from 295 women age 67 years or older who were not receiving estrogen replacement therapy. Serum was assayed for two markers of bone formation: osteocalcin (OC) and bone-specific alkaline phosphatase (BALP). Urine specimens were assayed for four markers of bone resorption: N-telopeptides (NTX), free pyridinolines (Pyr), free deoxypyridinoline (Dpyr), and C-telopeptides (CTX). Measurements of hip bone mineral density were made at the time the samples were collected and then repeated an average of 3.8 years later. Higher levels of all four resorption markers were, on average, significantly associated with faster rates of bone loss at the total hip, but not at the femoral neck. Women with OC levels above the median had a significantly faster rate of bone loss than women with levels below the median, but there was no significant association between levels of BALP and hip bone loss. The sensitivity and specificity of higher marker levels for predicting rapid hip bone loss was limited, and there was considerable overlap in bone loss rates between women with high and low marker levels. We conclude that higher levels of urine NTX, CTX, Pyr, Dpyr, and serum OC are associated with faster bone loss at the hip in this population of elderly women not receiving estrogen replacement therapy, but these biochemical markers have limited value for predicting rapid hip bone loss in individuals.


A Cooperative Cobalt-Driven System for One-Carbon Extension in the Synthesis of (Z)-Silyl Enol Ethers from Aldehydes: Unlocking Regio- and Stereoselectivity.
The research presented herein explores a cobalt-based catalytic system, distinctively featuring a cooperative boron-centric element within its intricate ligand architecture. This system is strategically engineered to enable the integration of a singular carbon atom into aldehydes, a process culminating in the production of (Z)-silyl enol ethers. Beyond offering an efficient one-pot synthesis route, this method adeptly overcomes challenges inherent to conventional techniques, such as the need for large amounts of additives, restrictive functional group tolerance, and extreme reaction temperatures. Initial mechanistic studies suggest the potential role of a cobalt-carbene complex as a catalytically significant species and underscore the importance of the borane segment. Collectively, these observations highlight the potential of this system in advancing complex bond activation pursuits.


Retinal break and rhegmatogenous retinal detachment after transpupillary thermotherapy as primary or adjunct treatment of choroidal melanoma.
To report the development of retinal break or rhegmatogenous retinal detachment (RRD) after transpupillary thermotherapy (TTT) as primary or adjunct treatment of choroidal melanoma. In this noncomparative, interventional case series, the authors reviewed medical records of 13 patients who developed retinal break or RRD following TTT. The main outcome measures were clinical features and outcome of treatment of retinal break or RRD following TTT. Of 1574 patients managed on the Oncology Service at Wills Eye Institute with TTT as primary or adjunct treatment of choroidal melanoma, 13 (1%) developed retinal break with or without RRD. The mean patient age at diagnosis of choroidal melanoma was 56 years. Treatment for choroidal melanoma included combined plaque radiotherapy and TTT in 10 patients and TTT alone in 3 patients. The median number of TTT sessions before development of retinal break or RRD was 2. Retinal break or RRD developed at a median of 3 months following the last TTT. All the retinal breaks were located in the TTT-treated area. Retinal breaks were atrophic in 11 eyes and horseshoe shaped in 2 cases. The extent of retinal detachment was none in 1 eye, 1 quadrant or less in 5 eyes, 2 or 3 quadrants in 4 eyes, and 4 quadrants in 3 eyes. Seven patients underwent vitrectomy, one received cryotherapy and laser photocoagulation, and five were observed without treatment. In all eight patients who received treatment for RRD, the retina was attached after a mean follow-up period of 54 months with no intraocular or local extraocular tumor dissemination. Development of retinal break or RRD is a rare complication of TTT for treatment of choroidal melanoma. The majority of these cases develop within 6 months of TTT and most are caused by atrophic retinal holes in the TTT-treated area.


Effects of communication training on real practice performance: a role-play module versus a standardized patient module.
This study investigated the effectiveness of modules involving standardized patients and role-plays on training communication skills. The first module involved standardized patients and an Objective Structured Clinical Examination (OSCE); the second module consisted of peer role-plays and a written examination. A randomized posttest-only control group design with first-year nursing students was used. The intervention group received one-to-one communication training with direct oral feedback from the standardized patient. The control group had training with peer role-playing and mutual feedback. The posttest involved students' rating their self-efficacy, and real patients and clinical supervisors evaluated their communication skills. No significant differences were found between self-efficacy and patient ratings. However, the clinical supervisors rated the intervention group's communication skills to be significantly (p < 0.0001) superior. Assessments by clinical supervisors indicate that communication training modules including standardized patients and an OSCE are superior to communication training modules with peer role-playing.


The Genome Sequence of M228, a Chinese Isolate of Pseudomonas syringae pv. actinidiae, Illustrates Insertion Sequence Element Mobility.
We present here the complete genome sequence of M228, a Chinese biovar 3 strain of Pseudomonas syringae pv. actinidiae, a bacterial pathogen of kiwifruit. A comparison of the insertion sequence (IS) profile of M228 with that of ICMP18708, a New Zealand isolate of P. syringae pv. actinidiae, provided insight into the evolutionary history of IS elements within biovar 3.


Small-molecule activators of a bacterial signaling pathway inhibit virulence.
The Burkholderia genus encompasses multiple human pathogens, including potential bioterrorism agents, that are often extensively antibiotic resistant. The FixLJ pathway in Burkholderia is a two-component system that regulates virulence. Previous work showed that fixLJ mutations arising during chronic infection confer increased virulence while decreasing the activity of the FixLJ pathway. We hypothesized that small-molecule activators of the FixLJ pathway could serve as anti-virulence therapies. Here, we developed a high-throughput assay that screened over 28,000 compounds and identified 11 that could specifically active the FixLJ pathway. Eight of these compounds, denoted B urkholderia F ix A ctivator (BFA) 1-8, inhibited the intracellular survival of Burkholderia in THP-1-dervived macrophages in a fixLJ- dependent manner without significant toxicity. One of the compounds, BFA1, inhibited the intracellular survival in macrophages of multiple Burkholderia species. Predictive modeling of the interaction of BFA1 with Burkholderia FixL suggests that BFA1 binds to the putative ATP/ADP binding pocket in the kinase domain, indicating a potential mechanism for pathway activation. These results indicate that small-molecule FixLJ pathway activators are promising anti-virulence agents for Burkholderia and define a new paradigm for antibacterial therapeutic discovery.


Epileptic Encephalopathies: Clinical Aspects, Molecular Features and Pathogenesis, Therapeutic Targets and Translational Opportunities, and Future Research Directions.
Epileptic encephalopathies encompass a heterogeneous group of epilepsy syndromes that manifest with cognitive, behavioral, and neurologic deficits, seizures that are often intractable and multiform, aggressive electroencephalographic paroxysmal activity, and sometimes early death. As more is learned about the etiologies and manifestations of epileptic encephalopathies, progress has been made toward better treatment options. However, there is still a great need for further randomized controlled trials and research to help create clinically effective therapies. The 2015 Neurobiology of Disease in Children symposium, held in conjunction with the 44th annual meeting of the Child Neurology Society, aimed to (1) describe the clinical concerns involving diagnosis and treatment, (2) review the current status of understanding in the pathogenesis of epileptic encephalopathy, (3) discuss clinical management and therapies for epileptic encephalopathy, and (4) define future directions of research. This article summarizes the presentations and includes an edited transcript of question-and-answer sessions.


Draft Genome Sequence of Lactobacillus kefiri SGL 13, a Potential Probiotic Strain Isolated from Kefir Grains.
In this report, we present the draft genome sequence of a newly discovered potential probiotic strain of Lactobacillus kefiri, SGL 13, isolated from kefir grains. Antibiotic resistance analysis did not reveal evidence of interspecific horizontal gene transfer since the identified bacitracin resistance gene does not have mobile genetic elements.


A zinc-finger fusion protein refines Gal4-defined neural circuits.
The analysis of behavior requires that the underlying neuronal circuits are identified and genetically isolated. In several major model species-most notably Drosophila-neurogeneticists identify and isolate neural circuits with a binary heterologous expression-control system: Gal4-UASG. One limitation of Gal4-UASG is that expression patterns are often too broad to map circuits precisely. To help refine the range of Gal4 lines, we developed an intersectional genetic AND operator. Interoperable with Gal4, the new system's key component is a fusion protein in which the DNA-binding domain of Gal4 has been replaced with a zinc finger domain with a different DNA-binding specificity. In combination with its cognate binding site (UASZ) the zinc-finger-replaced Gal4 ('Zal1') was functional as a standalone transcription factor. Zal1 transgenes also refined Gal4 expression ranges when combined with UASGZ, a hybrid upstream activation sequence. In this way, combining Gal4 and Zal1 drivers captured restricted cell sets compared with single drivers and improved genetic fidelity. This intersectional genetic AND operation presumably derives from the action of a heterodimeric transcription factor: Gal4-Zal1. Configurations of Zal1-UASZ and Zal1-Gal4-UASGZ are versatile tools for defining, refining, and manipulating targeted neural expression patterns with precision.


Differentiation of catecholaminergic cells in cultures of embryonic avian sensory ganglia.
From the results of previous studies in which developing peripheral ganglia from quail embryos were transplanted into younger chicken embryo hosts, we concluded that spinal and cranial sensory ganglia contain dormant precursors with autonomic potentialities. Here we describe the differentiation of these precursors in vitro, from dorsal root and nodose ganglion cell suspensions. Dorsal root ganglia were removed from quail embryos at 9 to 15 days of incubation, dissociated to single cells, and grown in tissue culture. The differentiation of cells with autonomic features was followed by monitoring properties associated with the adrenergic phenotype (absent from quail sensory ganglia during normal embryonic development). Provided that the medium was supplemented with chicken embryo extract, numerous cells displaying tyrosine hydroxylase immunoreactivity could be detected from day 4 onward. They possessed long, multiple processes but appeared morphologically distinct from primary sensory neurons. The catalytic activity of tyrosine hydroxylase and of other enzymes required for catecholamine production was demonstrated in the cultures by glyoxylic acid-induced histofluorescence and by radiochemical measurement of the conversion of exogenous tyrosine to norepinephrine. A large proportion of tyrosine hydroxylase-positive cells were found to incorporate [3H]thymidine before and after differentiating. In contrast, recognizable sensory neurons never exhibited adrenergic properties and did not divide. Qualitatively similar results were obtained with cultures of dissociated nodose ganglia. These findings lend further weight to the assumption that latent autonomic precursors are included in the non-neuronal compartment of sensory ganglia.


Disparities in functional disability among Arab Americans by nativity, immigrant arrival cohort, and country of birth.
This study contributes to a growing literature that documents the importance of arrival cohort and country of birth for differentiating the health of U.S. immigrants. We use nationally-representative data from nine years of the American Community Survey (2008-2016) to examine if an immigrant health advantage exists among Arab Americans ages 40+ (n = 49,867) and test if differences among the foreign-born vary by arrival cohort (pre-1991, 1991-2000, and 2001+). Results from multivariate logistic regression models find that foreign-born Arab Americans have higher odds of physical and self-care disability, and this varies by immigrant arrival cohort. The post-2001 cohort had the highest odds of both disabilities, while the earlier two cohorts did not differ from the native-born after adjustments for covariates. Compositional differences in birthplace, particularly the large influx of immigrants from Iraq in the most recent cohort, explained these differences. Political instabilities globally have contributed to a growing number of U.S. immigrants with vulnerabilities that might be overlooked when arrival cohorts are not considered.


Network motif comparison rationalizes Sec1/Munc18-SNARE regulation mechanism in exocytosis.
Network motifs, recurring subnetwork patterns, provide significant insight into the biological networks which are believed to govern cellular processes. We present a comparative network motif experimental approach, which helps to explain complex biological phenomena and increases the understanding of biological functions at the molecular level by exploring evolutionary design principles of network motifs. Using this framework to analyze the SM (Sec1/Munc18)-SNARE (N-ethylmaleimide-sensitive factor activating protein receptor) system in exocytic membrane fusion in yeast and neurons, we find that the SM-SNARE network motifs of yeast and neurons show distinct dynamical behaviors. We identify the closed binding mode of neuronal SM (Munc18-1) and SNARE (syntaxin-1) as the key factor leading to mechanistic divergence of membrane fusion systems in yeast and neurons. We also predict that it underlies the conflicting observations in SM overexpression experiments. Furthermore, hypothesis-driven lipid mixing assays validated the prediction. Therefore this study provides a new method to solve the discrepancies and to generalize the functional role of SM proteins.


Reverse Genetics Systems for Filoviruses.
Filoviruses are causative agents of severe hemorrhagic fevers with high case fatality rates in humans. For studies of virus biology and the subsequent development of countermeasures, reverse genetic systems, and especially those facilitating the generation of recombinant filoviruses, are indispensable. Here, we describe the generation of recombinant filoviruses from cDNA.


Donor age and outcome of liver transplantation.
To evaluate the effect of donor age on graft and patient outcome after liver transplantation an analysis of a large-scale cohort study was performed at three tertiary referral liver transplant centers. Between April 1990 and June 1994, 772 adults underwent an initial single-organ liver transplantation. The age of the donors averaged 35 years;193 (25%) were 50 or above, the age used to define "older" donors. Groups were compared for demographic, clinical, and biochemical features. Outcome was measured using results of biochemical tests after transplantation and by graft and patient survival. Compared with younger donors, older donors were more commonly women (59% vs. 33%:P < or = .001) and died of central nervous system causes (79% vs. 28%) as opposed to trauma (13% vs. 63%:P < or = .001). The recipients of the two groups of donor livers did not differ in important respects. However, intraoperatively, livers from older donors were more likely to be assessed as either "poor" or "fair" as opposed to "good" (17% vs. 4%:P < or = .001) by the harvesting surgeon and to have initial "poor" or "fair" bile production (29% vs. 18%:P < or = .001). During the first week postoperatively, the serum aminotransferase and bilirubin levels and prothrombin times were higher in recipients of older than those of younger donor livers. During follow-up, graft survival was less for recipients of older donor livers at 3 months (81% vs.91%:P = .0001) and at 1 (76% vs. 85%:P = .007) and 2 years (71% vs. 80%:P = .005); patient survival showed similar though less marked differences. This association of donor age and poorer graft survival persisted after adjusting for many variables using bivariate and multivariate analyses. Importantly, however, the association with poor graft survival was largely among recipients of older donor livers, the quality of which was assessed as fair or poor by the harvesting surgeon; recipients of older donor livers assessed as good had a retransplant-free survival similar to that of younger donor livers (87% vs. 91% at 3 months). Thus, use of older donor livers, the quality of which are judged to be good by the harvesting surgeon, is not associated with a decrease in patient or graft survival after liver transplantation. differences. This association of donor age and poorer graft survival persisted after adjusting for many variables using bivariate and multivariate analyses. Importantly, however, the association with poorer graft survival was largely among recipients of older donor livers, the quality of which was assessed as fair or poor by the harvesting surgeon; recipients of older donor livers, the quality of which was assessed as fair or poor by the harvesting surgeon; recipients of older donor livers assessed as good had a retransplant-free survival similar to that of younger donor livers (87% vs. 91% at 3 months). Thus, use of older donor livers, the quality of which are judged to be good by the harvesting surgeon, is not associated with a decrease in patient or graft survival after liver transplantation.


Molecular, functional, and pharmacological targets for the development of gut promotility drugs.
The science of gastrointestinal motility has made phenomenal advances during the last fifty years. Yet, there is a paucity of effective promotility drugs to treat functional bowel disorders that affect 10-29% of the U.S. population. A part of the reason for the lack of effective drugs is our limited understanding of the etiology of these diseases. In the absence of this information, mostly an ad hoc approach has been used to develop the currently available drugs, which are modestly effective or effective in only a subset of the patients with functional bowel disorders. This review discusses a grounds-up approach for development of the next generation of promotility drugs. The approach is based on our current understanding of 1) the different types of contractions that produce overall motility function of mixing and orderly net distal propulsion in major gut organs, 2) the regulatory mechanisms of these contractions, 3) which receptors and intracellular signaling molecules could be targeted to stimulate specific types of contractions to accelerate or retard transit, and 4) the strengths and limitations of animal models and experimental approaches that could screen potential promotility drugs for their efficacy in human gut propulsion in functional bowel disorders.


Lovastatin Treatment Inducing Apoptosis in Human Pancreatic Cancer Cells by Inhibiting Cholesterol Rafts in Plasma Membrane and Mitochondria.
Resistance to anticancer drugs is a problem in the treatment of pancreatic ductal carcinoma (PDAC) and overcoming it is an important issue. Recently, it has been reported that statins induce apoptosis in cancer cells but the mechanism has not been completely elucidated. We investigated the antitumor mechanisms of statins against PDAC and their impact on resistance to gemcitabine (GEM). Lovastatin (LOVA) increased mitochondrial oxidative stress in PDAC cells, leading to apoptosis. LOVA reduced lipid rafts in the plasma membrane and mitochondria, suppressed the activation of epithelial growth factor receptor (EGFR) and AKT in plasma membrane rafts, and reduced B-cell lymphoma 2 (BCL2)-Bcl-2-associated X protein (BAX) binding and the translocation of F1F0 ATPase in mitochondrial rafts. In the three GEM-resistant cell lines derived from MIA and PANC1, the lipid rafts in the cell membrane and the mitochondria were increased to activate EGFR and AKT and to increase BCL2-BAX binding, which suppressed apoptosis. LOVA abrogated these anti-apoptotic effects by reducing the rafts in the resistant cells. By treating the resistant cells with LOVA, GEM sensitivity improved to the level of the parental cells. Therefore, cholesterol rafts contribute to drug resistance in PDAC. Further clinical research is warranted on overcoming anticancer drug resistance by statin-mediated intracellular cholesterol regulation.


Effectiveness of a questionnaire based intervention programme on the prevalence of arm, shoulder and neck symptoms, risk factors and sick leave in computer workers: a cluster randomised controlled trial in an occupational setting.
Arm, shoulder and neck symptoms are very prevalent among computer workers. In an attempt to reduce these symptoms, a large occupational health service in the Netherlands developed a preventive programme on exposure to risk factors, prevalence of arm, shoulder and neck symptoms, and sick leave in computer workers. The purpose of this study was to assess the effectiveness of this intervention programme. The study was a randomised controlled trial. The participants were assigned to either the intervention group or the usual care group by means of cluster randomisation. At baseline and after 12 months of follow-up, the participants completed the RSI QuickScan questionnaire on exposure to the risk factors and on the prevalence of arm, shoulder and neck symptoms. A tailor-made intervention programme was proposed to participants with a high risk profile at baseline. Examples of implemented interventions are an individual workstation check, a visit to the occupational health physician and an education programme on the prevention of arm, shoulder and neck symptoms. The primary outcome measure was the prevalence of arm, shoulder and neck symptoms. Secondary outcome measures were the scores on risk factors for arm, shoulder and neck symptoms and the number of days of sick leave. Sick leave data was obtained from the companies. Multilevel analyses were used to test the effectiveness. Of the 1,673 persons invited to participate in the study, 1,183 persons (71%) completed the baseline questionnaire and 741 persons participated at baseline as well as at 12-month follow-up. At 12-month follow-up, the intervention group showed a significant positive change (OR = 0.48) in receiving information on healthy computer use, as well as a significant positive change regarding risk indicators for work posture and movement, compared to the usual care group. There were no significant differences in changes in the prevalence of arm, shoulder and neck symptoms or sick leave between the intervention and usual care group. The effects of the RSI QuickScan intervention programme were small, possibly as a result of difficulties with the implementation process of the proposed interventions. However, some significant positive effects were found as to an increase in receiving education and a decrease in exposure to adverse postures and movements. With regard to symptoms and sick leave, only small and non-significant effects were found. Netherlands National Trial Register NTR1117.


Allogeneic bone marrow transplantation for severe aplastic anemia: the Hong Kong scenario.
Severe aplastic anemia (SAA) is a disease associated with high mortality. For young patients with HLA identical siblings, allogeneic bone marrow transplantation (BMT) offers the best chance of cure. Favourable results have also been reported using immunosuppressive therapy (IST). Transplantation is usually favoured for patients below 45 years of age. We report our experience of 11 allogeneic and one syngeneic BMT for adult Chinese patients with SAA, over a 4-year period from 1991 to 1995. Ten of the 12 (83 per cent) patients had received and failed prior IST including anti-thymocyte globulin (ATG) before being referred for BMT. Neutrophil and platelet engraftment was successful in 11 of them (92 per cent) and nine were completely transfusion independent after transplantation. Their overall 3-year survival was 67 per cent. The compromised overall result was due to a number of cases transplanted after a long time delay. No patient transplanted beyond 3 years from the initial time of diagnosis of SAA achieved long-term marrow engraftment, and they all eventually succumbed. On univariate analysis, a longer time delay and hence a larger amount of blood products exposure, were highly significantly statistically associated with inferior marrow engraftment and patient survival. Other factors including age, iron status, infused cell dose and the conditioning protocol were not found to significantly affect engraftment and survival. Graft versus host disease was clinically mild or absent in most patients. This may be related to ethnicity or previous ATG exposure. In conclusion, early allogeneic BMT was a safe and effective treatment in our small series of patients with SAA failing IST.


Diseases and disorders associated with excess body weight.
Excess body weight is a very serious problem, especially in North America and Europe. It has been referred to as a "pandemic" since it has progressively increased over the past several decades. Moreover, excess body weight significantly increases the risk of numerous diseases and clinical disorders, including all-cause mortality, coronary and cerebrovascular diseases, various cancers, type 2 diabetes mellitus, hypertension, liver disease and asthma, as well as psychopathology, among others. Unfortunately, overweight and obesity are now common in both young children and adolescents. Although the causes of excess body weight are multi-factorial, the most important factors are excess caloric intake coupled with limited energy expenditure. Therefore, lifestyle modification can significantly reduce the risk of morbidity and mortality and thereby increase longevity and improve the quality of life.


Use of synthetic peptides in the study of the antibody response to Plasmodium vivax sporozoites.
Synthetic peptides reproducing 4 DRADGQPAG (D4) and a sequential array of DRADGQPAG and DRAAGQPAG repeats (DDAAD) of the Plasmodium vivax circumsporozoite (CS) protein were investigated for their potential use in the detection of P. vivax sporozoite antibodies in human sera. These peptides specifically inhibited the binding of monoclonal antibodies to the P. vivax CS protein in Western blots. However, when D4 and DDAAD peptides were used in an enzyme-linked immunosorbent assay (ELISA) for the detection of human antibodies, more sera bound to the DDAAD (61%) than to the D4 peptide (22%). This binding was specific, and suggested that the DDAAD peptide contained epitopes constituted by the sequential array of DRADGQPAG and DRAAGQPAG repeat variants and absent in the D4 peptide. The ELISA using the DDAAD peptide was applied to the detection of P. vivax CS protein antibodies in a large number of sera from Kataragama, an endemic area in Sri Lanka. The prevalence of these antibodies increased with age, reaching 40% in adults greater than 50 years old. The ELISA employing the DDAAD peptide represents a simple and useful tool for the analysis of the antibody response to P. vivax sporozoites in naturally exposed individuals.


Adsorption of Cd(II) ions on magnetic graphene oxide/cellulose modified with β-cyclodextrin: Analytical interpretation via statistical physics modeling and fractal like kinetic approach.
In the present study, β-cyclodextrin modified magnetic graphene oxide/cellulose (CN/IGO/Cel) was fabricated for removal of Cd(II) ions. The material was characterized through various analytical techniques like FTIR, XRD, TGA/DTA, SEM, TEM, and XPS. The point of zero charge of the material was obtained as 5.38. The controllable factors were optimized by Taguchi design and optimum values were: adsorbent dose-16 mg, equilibrium time-40 min, and initial concentration of Cd(II) ions-40 mg/L. The material shows high adsorption capacity (303.98 mg/g) under optimal conditions. The good fitting of Langmuir model to adsorption data (R2 = 0.9918-0.9936) revealed the monolayer coverage on adsorbent surface. Statistical physics model M 2 showed best fitting to adsorption data (R2 > 0.997), suggesting the binding of Cd(II) ions occurred on two different receptor sites (n). Stereographically n > 1 confirming vertical multi-molecular mechanisms of Cd(II) ions adsorption on CN/IGO/Cel surface. The adsorption energies (E1 = 23.71-28.95 kJ/mol; E2 = 22.69-29.38 kJ/mol) concluded the involvement of physical forces for Cd(II) ions adsorption. Kinetic data fitted well to fractal-like pseudo first-order model (R2 > 0.9952), concluding the adsorption of Cd(II) ions occurred on energetically heterogeneous surface. The kinetic analysis shows that the both the film-diffusion and pore-diffusion were responsible for Cd(II) ions uptake. XPS analysis was utilized to explain the adsorption mechanism of Cd(II) ions onto CN/IGO/Cel.


Salmonella Typhi acquires diverse plasmids from other Enterobacteriaceae to develop cephalosporin resistance.
Recent reports have established the emergence and dissemination of extensively drug resistant (XDR) H58 Salmonella Typhi clone in Pakistan. In India where typhoid fever is endemic, only sporadic cases of ceftriaxone resistant S. Typhi are reported. This study aimed at elucidating the phylogenetic evolutionary framework of ceftriaxone resistant S. Typhi isolates from India to predict their potential dissemination. Five ceftriaxone resistant S. Typhi isolates from three tertiary care hospitals in India were sequenced on an Ion Torrent Personal Genome Machine (PGM). A core genome single-nucleotide-polymorphism (SNP) based phylogeny of the isolates in comparison to the global collection of MDR and XDR S. Typhi isolates was built. Two of five isolates were additionally sequenced using Oxford Nanopore MinION to completely characterize the plasmid and understand its transmission dynamics within Enterobacteriaceae. Comparative genomic analysis and detailed plasmid characterization indicate that while in Pakistan (4.3.1 lineage I) the XDR trait is associated with blaCTX-M-15 gene on IncY plasmid, in India (4.3.1 lineage II), the ceftriaxone resistance is due to short term persistence of resistance plasmids such as IncX3 (blaSHV-12) or IncN (blaTEM-1B + blaDHA-1). Considering the selection pressure exerted by the extensive use of ceftriaxone in India, there are potential risks for the occurrence of plasmid transmission events in the predominant H58 lineages. Therefore, continuous monitoring of S. Typhi lineages carrying plasmid-mediated cephalosporin resistant genes is vital not just for India but also globally.


Effects of a drought period on physico-chemical surface water quality in a regional catchment area.
Hydrological drought periods are expected to become more severe in North-Western Europe as a result of climate change. This may have implications for water quality, as demonstrated by declining water quality of large rivers (e.g. Rhine, Meuse) during droughts. However, similar investigations in regional catchment areas are lacking to date. In the present study, we investigated the effects of a drought period on the water quality of the Dommel River, a tributary of the Meuse river in the Netherlands. Water quality during the drought of 2003 was compared to that in reference years (2004-2006) for 18 physical/chemical parameters using ANOVA analysis. It was demonstrated that the drought period of 2003 did not significantly affect water quality, although the origin of river flow during the drought shifted from mainly overland flow to deep groundwater flow and (treated) communal effluents. Significant differences in water quality were noted for some monitoring stations during the study period, which could be related to operational water management such as cleaning of sediment traps in the river and improvements in communal effluent treatment. The results of this study are interesting to water managers in Western Europe as they contribute to understanding the potential impact of climate change on water quality/quantity patterns in regional water systems.


Perioperative beta-blockers for major noncardiac surgery: Primum Non Nocere.
Recent studies have called into question the benefit of perioperative beta blockade, especially in patients at low to moderate risk of cardiac events. Once considered standard of care, the role of beta-blocker therapy now lies mired in conflicting data that are difficult to apply to the at-risk patient. We provide an overview of the evolution of perioperative beta blockade, beginning with the physiology of the adrenergic system, with emphasis on the biologic rationale for the perioperative implementation of beta-blockers. Although initial studies were small in size and statistically limited, early data showed cardiac benefit with the use of perioperative beta-blockers. However, larger, more recent studies now suggest a lack of benefit and potential harm from this practice. This paradigm holds true especially in those at low-to-moderate cardiovascular risk profiles. Potential explanations for these paradoxical results are discussed, stressing the key differences between earlier and current studies that may explain these divergent outcomes. We conclude by commenting on performance measures as they relate to perioperative beta-blockers and make recommendations for the continued safe implementation of this practice.


Ontogeny of D1 and DARPP-32 gene expression in the rat striatum: an in situ hybridization study.
D1 dopamine receptor (D1R) and DARPP-32 (a dopamine and adenosine 3',5'-monophosphate regulated phosphoprotein), gene expression was studied in the rat striatum in adults and during ontogeny by in situ hybridization. D1R mRNA was first detected in the striatal primordium at day 17 of gestation. At day 18, D1R mRNA was found throughout the striatum. Before birth, the striatal neurons had neuroblastic aspect and were close together, giving homogeneous and compact labelling. After birth, the topography and aspect of the neurons containing D1R mRNA and DARPP-32 mRNA were similar. The two mRNAs were detectable in the caudate-putamen, accumbens nucleus and olfactory tubercle. The microautoradiographic analysis demonstrated that D1R and DARPP-32 genes are massively expressed by the medium-sized striatal neurons. The proportion of medium-sized neurons containing the DARPP-32 mRNA was however higher than that of the neurons containing the D1R mRNA. Furthermore, an unexpected proportion of large-sized neurons express these genes. This proportion varies with development. Comparison between the appearance, topography and frequency of choline-acetyltransferase immunoreactive neurons and large-sized neurons containing D1R or DARPP-32 mRNA suggest that these large-sized neurons containing D1R and DARPP-32 mRNAs are cholinergic ones.


Pimavanserin for the treatment of Parkinson's disease psychosis: number needed to treat, number needed to harm, and likelihood to be helped or harmed.
Our aim was to describe the efficacy and tolerability of pimavanserin, a highly selective serotonin 5-HT2A receptor inverse agonist/antagonist indicated for the treatment of hallucinations and delusions associated with Parkinson's disease psychosis (PDP), using the metrics of number needed to treat (NNT) and number needed to harm (NNH). Categorical efficacy and tolerability data were extracted from the clinical trial databases of the double-blind placebo-controlled studies of pimavanserin in persons with PDP. NNT and NNH values were calculated with their respective 95% confidence intervals. The likelihood to be helped or harmed (LHH) was then calculated contrasting therapeutic response versus discontinuation because of an adverse event. NNT values for pimavanserin 34 mg/d versus placebo for several definitions of clinical response are 10, and/or are not statistically significant, and/or show an advantage for pimavanserin over placebo (such as for postural hypotension). In terms of LHH, pimavanserin 34 mg/d is about five times more likely to result in clinical response (as measured by a ≥3 point decrease from baseline on the Scale for the Assessment of Positive Symptoms adapted for Parkinson's disease) versus discontinuation due to an adverse event. Using the metrics of NNT, NNH, and LHH, pimavanserin 34 mg/d for the treatment of PDP appears to have a compelling benefit/risk profile.