Subtle memory decline over 12 months in mild cognitive impairment.
Screening of normal older persons for progressive memory decline is a worthwhile strategy in the pursuit of the earliest possible stages of pre-clinical Alzheimer's disease (AD) or mild cognitive impairment (MCI). Reliable tests are needed to both detect MCI and measure the natural history of decline over months rather than years. We aimed to detect memory decline over 1 year in a group of older individuals with well-characterised amnestic MCI. The continuous learning task (CLT) from the CogState test battery was administered 8 times in 12 months to 15 individuals with MCI and 35 controls matched for age, education, IQ and gender. All subjects were recruited from an ongoing aging study. The rate of change in CLT performance over the year was compared between groups and also compared to that detected with a word list learning task and a computerised paired associate learning task. At baseline, memory performance in the amnestic MCI group was significantly worse than controls on all memory tests. However, at 12 months the magnitude of the difference between the groups had increased significantly on the CLT due to decline in memory accuracy in the MCI group. No decline over 12 months was detectable on the routine memory tests. Subtle memory decline is detectable in amnestic MCI using reliable and sensitive tests of memory. Such measures may assist in the early identification of AD and also in trials of putative disease-modifying therapies to be conducted over as little as 12 months.


CHST7 Methylation Status Related to the Proliferation and Differentiation of Pituitary Adenomas.
Pituitary adenomas (PAs) are the second most common primary brain tumor and may develop from any of the cell lineages responsible for producing the different pituitary hormones. DNA methylation is one of the essential epigenetic mechanisms in cancers, including PAs. In this study, we measured the expression profile and promoter methylation status of carbohydrate sulfotransferase 7 (CHST7) in patients with PA; then, we investigated the effect of the CHST7 methylation status on the proliferation and differentiation of PAs. The volcano map and Metascape results showed that the levels of CHST7 were related to the lineages' differentiation and the cell adhesion of PAs, and patients with low CHST7 had greater chances of having an SF-1 lineage (p = 0.002) and optic chiasm compression (p = 0.007). Reactome pathway analysis revealed that most of the DEGs involved in the regulation of TP53 regulated the transcription of cell cycle genes (HSA-6791312 and HSA6804116) in patients with high CHST7. Correlation analysis showed that CHST7 was significantly correlated with the eIF2/ATF4 pathway and mitochondrion-related genes. The AUC of ROC showed that CHST7 (0.288; 95% CI: 0.187-0.388) was superior to SF-1 (0.555; 95% CI: 0.440-0.671) and inferior to FSHB (0.804; 95% CI: 0.704-0.903) in forecasting the SF-1 lineage (p < 0.001). The SF-1 lineage showed a higher methylation frequency for CHST7 than the Pit-1 and TBX19 lineages (p = 0.009). Furthermore, as the key molecule of the hypothalamic-pituitary-gonadal axis, inhibin βE (INHBE) was positively correlated with the levels of CHST7 (r = 0.685, p < 0.001). In summary, CHST7 is a novel pituitary gland specific protein in SF-1 lineage adenomas with a potential role in gonadotroph cell proliferation and lineage differentiation in PAs.


Recombination contributes to population diversification in the polyploid intestinal symbiont Epulopiscium sp. type B.
Epulopiscium sp. type B (Lachnospiraceae) is an exceptionally large, highly polyploid, intestinal symbiont of the coral reef dwelling surgeonfish Naso tonganus. These obligate anaerobes do not form mature endospores and reproduce solely through the production of multiple intracellular offspring. This likely makes them dependent on immediate transfer to a receptive host for dispersal. During reproduction, only a small proportion of Epulopiscium mother-cell DNA is inherited. To explore the impact of this unusual viviparous lifestyle on symbiont population dynamics, we investigated Epulopiscium sp. type B and their fish hosts collected over the course of two decades, at island and reef habitats near Lizard Island, Australia. Using multi-locus sequence analysis, we found that recombination plays an important role in maintaining diversity of these symbionts and yet populations exhibit linkage disequilibrium (LD). Symbiont populations showed spatial but not temporal partitioning. Surgeonfish are long-lived and capable of traveling long distances, yet the population structures of Epulopiscium suggest that adult fish tend to not roam beyond a limited locale. Codiversification analyses and traits of this partnership suggest that while symbionts are obligately dependent on their host, the host has a facultative association with Epulopiscium. We suggest that congression of unlinked markers contributes to LD estimates in this and other recombinant populations of bacteria. The findings here inform our understanding of evolutionary processes within intestinal Lachnospiraceae populations.


The upper airway response to pollen is enhanced by exposure to combustion particulates: a pilot human experimental challenge study.
Although human experimental studies have shown that gaseous pollutants enhance the inflammatory response to allergens, human data on whether combustion particulates enhance the inflammatory response to allergen are limited. Therefore, we conducted a human experimental study to investigate whether combustion particulates enhance the inflammatory response to aeroallergens. "Enhancement" refers to a greater-than-additive response when combustion particulates are delivered with allergen, compared with the responses when particulates and allergen are delivered alone. Eight subjects, five atopic and three nonatopic, participated in three randomized exposure-challenge sessions at least 2 weeks apart (i.e., clean air followed by allergen, particles followed by no allergen, or particles followed by allergen). Each session consisted of nasal exposure to combustion particles (target concentration of 1.0 mg/m3) or clean air for 1 hr, followed 3 hr later by challenge with whole pollen grains or placebo. Nasal lavage was performed immediately before particle or clean air exposure, immediately after exposure, and 4, 18 and 42 hr after pollen challenge. Cell counts, differentials, and measurement of cytokines were performed on each nasal lavage. In atopic but not in nonatopic subjects, when allergen was preceded by particulates, there was a significant enhancement immediately after pollen challenge in nasal lavage leukocytes and neutrophils (29.7 X 10(3) cells/mL and 25.4 X 10(3) cells/mL, respectively). This represents a 143% and 130% enhancement, respectively. The enhanced response for interleukin-4 was 3.23 pg/mL (p = 0.06), a 395% enhancement. In atopic subjects there was evidence of an enhanced response when particulates, as compared to clean air, preceded the allergen challenge.


Two-magnon bound state causes ultrafast thermally induced magnetisation switching.
There has been much interest recently in the discovery of thermally induced magnetisation switching using femtosecond laser excitation, where a ferrimagnetic system can be switched deterministically without an applied magnetic field. Experimental results suggest that the reversal occurs due to intrinsic material properties, but so far the microscopic mechanism responsible for reversal has not been identified. Using computational and analytic methods we show that the switching is caused by the excitation of two-magnon bound states, the properties of which are dependent on material factors. This discovery allows us to accurately predict the onset of switching and the identification of this mechanism will allow new classes of materials to be identified or designed for memory devices in the THz regime.


Delineation of genetic zones in the European Norway spruce natural range: preliminary evidence.
We applied geostatistics to previously reported chloroplast simple sequence repeats (cpSSR) haplotype frequency data from 95 European Norway spruce populations to provide preliminary evidence about the following issues: (i) delineation of genetically homogeneous regions ('genetic zones'); (ii) prediction of their haplotype frequencies and definition of related criteria to be applied for provenance identification and certification of seedlots; (iii) identification of putative allochthonous stands; and (iv) construction of a continental-scale 'availability map' of the intraspecific biodiversity for Norway spruce. Direct evidence of large-scale geographical structure over the European natural range was obtained, detecting both geographical clines and stationary patterns. The increase of the mean genetic divergence by geographical distance (up to approximately 1800 km apart) provided a strong hint that geographical distance is a major factor of differentiation in Norway spruce. Haplotype frequency surfaces were obtained by applying ordinary kriging to sampling frequency data. Cluster analysis carried out on haplotype frequency surfaces revealed a fair discrimination among 16 genetic zones, with an accuracy of 0.916. Dendrogram analysis carried out on the predicted mean haplotype frequency confirmed a fairly good separability of the genetic zones detected. Reclassification of populations by discriminant analysis confirmed the robustness of the genetic subdivision obtained. Combining the information from discriminant analysis and cross-validation of sampling points, three populations likely to be of nonlocal origin were identified. The application of geostatistical analysis to genetic marker data is discussed in relation to breeding activities and to the formulation of appropriate conservation strategies.


Managing flowering time in Miscanthus and sugarcane to facilitate intra- and intergeneric crosses.
Miscanthus is a close relative of Saccharum and a potentially valuable genetic resource for improving sugarcane. Differences in flowering time within and between Miscanthus and Saccharum hinders intra- and interspecific hybridizations. A series of greenhouse experiments were conducted over three years to determine how to synchronize flowering time of Saccharum and Miscanthus genotypes. We found that day length was an important factor influencing when Miscanthus and Saccharum flowered. Sugarcane could be induced to flower in a central Illinois greenhouse using supplemental lighting to reduce the rate at which days shortened during the autumn and winter to 1 min d-1, which allowed us to synchronize the flowering of some sugarcane genotypes with Miscanthus genotypes primarily from low latitudes. In a complementary growth chamber experiment, we evaluated 33 Miscanthus genotypes, including 28 M. sinensis, 2 M. floridulus, and 3 M. ×giganteus collected from 20.9° S to 44.9° N for response to three day lengths (10 h, 12.5 h, and 15 h). High latitude-adapted M. sinensis flowered mainly under 15 h days, but unexpectedly, short days resulted in short, stocky plants that did not flower; in some cases, flag leaves developed under short days but heading did not occur. In contrast, for M. sinensis and M. floridulus from low latitudes, shorter day lengths typically resulted in earlier flowering, and for some low latitude genotypes, 15 h days resulted in no flowering. However, the highest ratio of reproductive shoots to total number of culms was typically observed for 12.5 h or 15 h days. Latitude of origin was significantly associated with culm length, and the shorter the days, the stronger the relationship. Nearly all entries achieved maximal culm length under the 15 h treatment, but the nearer to the equator an accession originated, the less of a difference in culm length between the short-day treatments and the 15 h day treatment. Under short days, short culms for high-latitude accessions was achieved by different physiological mechanisms for M. sinensis genetic groups from the mainland in comparison to those from Japan; for mainland accessions, the mechanism was reduced internode length, whereas for Japanese accessions the phyllochron under short days was greater than under long days. Thus, for M. sinensis, short days typically hastened floral induction, consistent with the expectations for a facultative short-day plant. However, for high latitude accessions of M. sinensis, days less than 12.5 h also signaled that plants should prepare for winter by producing many short culms with limited elongation and development; moreover, this response was also epistatic to flowering. Thus, to flower M. sinensis that originates from high latitudes synchronously with sugarcane, the former needs day lengths >12.5 h (perhaps as high as 15 h), whereas that the latter needs day lengths <12.5 h.


Diagnosis and classification of ileal pouch disorders: consensus guidelines from the International Ileal Pouch Consortium.
Restorative proctocolectomy with ileal pouch-anal anastomosis is an option for most patients with ulcerative colitis or familial adenomatous polyposis who require colectomy. Although the construction of an ileal pouch substantially improves patients' health-related quality of life, the surgery is, directly or indirectly, associated with various structural, inflammatory, and functional adverse sequelae. Furthermore, the surgical procedure does not completely abolish the risk for neoplasia. Patients with ileal pouches often present with extraintestinal, systemic inflammatory conditions. The International Ileal Pouch Consortium was established to create this consensus document on the diagnosis and classification of ileal pouch disorders using available evidence and the panellists' expertise. In a given individual, the condition of the pouch can change over time. Therefore, close monitoring of the activity and progression of the disease is essential to make accurate modifications in the diagnosis and classification in a timely manner.


Homogenization of Heterogeneously Coupled Bistable ODE's-Applied to Excitation Waves in Pancreatic Islets of Langerhans.
We consider a lattice of coupled identical differential equations. The coupling is between nearest neighbors and of resistance type, but the strength of coupling varies from site to site. Such a lattice can, for example, model an islet of Langerhans, where the sites in the lattice model individual but identical β-cells, and the coupling between cells is made of gap junctions.By using a homogenization technique we approximate the coupled discrete equations by a PDE, basically a nonlinear heat equation (a Fisher equation). For appropriate parameters this equation is known to have wave-solutions. Of importance is the fact, that the resulting diffusion coefficient does not only depend on the average of the coupling, but also on the variance of the strength. This means that the heterogeneity of the coupling strength influences the wave velocity-the greater the variance, the slower is the wave. This result is illustrated by simulations, both of a simple prototype equation, and for a full model of coupled beta-cells in both one and two dimensions, and implies that the natural heterogeneity in the islets of Langerhans should be taken into account.


A survey on the usage of supplements in canadian patients living with HIV.
To assess the usage, knowledge and attitudes of Patients Living with HIV with respect to supplements. A questionnaire was mailed to people living with HIV via HIV/AIDS organizations in Ontario and distributed to those attending the HIV-clinic of the University Health Network. The survey was completed by 312 subjects (95 female, 207 male). Self-rated health status was considered fair/good in 77.5% and excellent in 15.4% of participants. Vitamin/mineral supplement was used by 75.6%. Main reasons to take supplements were to: prolong life (56.1%); treat HIV-related conditions (19.9%); increase energy level (42.6%) and to boost immunity (36.5%). Among participants, 54.2% were somewhat familiar with supplements, 44.7% trusted the information on the labels and 28.2% felt that if a supplement is available for sale, it is safe. Supplements were mostly purchased at pharmacies (45.5%) and health food stores (30.1%). Only 25.9% and 27.9% of participants discussed their use of supplements with their HIV or family doctor respectively. Supplements are frequently used by people living with HIV without consulting their doctors. Current research has not shown a clear benefit from micronutrient supplementation and with the possible potential drug interactions, people living with HIV will need nutrition education regarding supplement usage and should report their use to their physicians.


Absorption of nutrients is only slightly reduced by supplementing enteral formulas with viscous fiber in miniature pigs.
Viscous polysaccharides reduce intestinal absorption of glucose and diminish postprandial hyperglycemia. However, it is unknown whether viscous fiber also inhibits absorption of nutrients under conditions of enteric feeding. Therefore, we measured the absorption rates of nutrients in miniature pigs by perfusing a 150-cm length of jejunum with 8.37 kJ/min of the three following enteral diets: an isoosmotic oligomeric diet (1670 kJ/L), a hyperosmotic oligomeric diet and an isoosmotic polymeric diet (both 3350 kJ/L). The diets were supplemented with guar gum from 0 to 4.4 g/L. With the three guar-free diets, the mean absorption rate of energy was 5.2 +/- 0.32 kJ/min, corresponding to 62% of the energy infused. Absorption rates of carbohydrate, protein, fat and energy linearly declined as concentrations of guar or the logarithm of chyme viscosity increased. Due to modulations in viscosity, the inhibitory effects of guar were significantly different among the three diets. With the isoosmotic and hyperosmotic oligomeric and the polymeric diets, the addition of 1 g guar/L diminished the absorption of energy by 9.7, 6. 6 and 3.7%, respectively. The strong inhibitory effect on nutrient absorption with the isoosmotic oligomeric diet was caused by an increase in chyme viscosity due to water absorption. With the hyperosmotic oligomeric and the polymeric diets, the chyme viscosity and thus inhibitory effects on absorption were diminished by water secretion and the concomitant infusion of pancreatic enzymes. Results indicate that the addition of small amounts of guar gum to enteral diets of high energy density exerts only small effects on absorption of nutrients.


Effects of oral motor therapy in children with cerebral palsy.
Oral motor dysfunction is a common issue in children with cerebral palsy (CP). Drooling, difficulties with sucking, swallowing, and chewing are some of the problems often seen. In this study, we aimed to research the effect of oral motor therapy on pediatric CP patients with feeding problems. Included in this single centered, randomized, prospective study were 81 children aged 12-42 months who had been diagnosed with CP, had oral motor dysfunction and were observed at the Pediatric Neurology outpatient clinic of the Children's Health and Diseases Department, Istanbul Medical Faculty, Istanbul University. Patients were randomized into two groups: The training group and the control group. One patient from the training group dropped out of the study because of not participating regularly. Following initial evaluation of all patients by a blinded physiotherapist and pedagogue, patients in the training group participated in 1 h oral motor training sessions with a different physiotherapist once a week for 6 months. All patients kept on routine physiotherapy by their own physiotherapists. Oral motor assessment form, functional feeding assessment (FFA) subscale of the multidisciplinary feeding profile (MFP) and the Bayley scales of infant development (BSID-II) were used to evaluate oral motor function, swallowing, chewing, the gag reflex, the asymmetrical tonic neck reflex, tongue, jaw, and mouth function, severity of drooling, aspiration, choking, independent feeding and tolerated food texture during the initial examination and 6 months later. When the initial and post-therapy FFA and BSID-II scores received by patients in the training and the study group were compared, the training group showed a statistically significant improvement (P < 0.05). Oral motor therapy has a beneficial effect on feeding problems in children with CP.


Why Women Request Labiaplasty.
In recent years, labiaplasty has jumped in popularity, despite opposition to the procedure. In 2007, the American College of Obstetricians and Gynecologists declared the recommendation of cosmetic vaginal procedures to be "untenable," although in 2016 they allowed consideration of labiaplasty in adolescents if symptoms persist. The reasons for labiaplasty requests are not yet fully understood, and physician opposition limits patient access to surgical relief. In this prospective study, 50 consecutive patients consulting about labiaplasty were given a questionnaire assessing 11 physical and appearance-related symptoms associated with elongated labia. The mean patient age was 33.5 years (range, 17 to 51 years). Fifty-eight percent of women had given birth, 52 percent noticed that their labia had become elongated as they got older, and 93 percent had bilateral elongation. When asked about physical symptoms, over half of patients experienced tugging during intercourse, found tight pants uncomfortable, experienced twisting of the labia, and noted labia visibility in yoga pants. Nearly half experienced pain during intercourse, and 40 percent said their labia could become exposed in bathing suits. Regarding appearance, almost all patients were self-conscious and over half felt less attractive to their partner, experienced restricted clothing choice, and noted a negative impact on self-esteem and intimacy. Nearly all patients experienced at least four symptoms. Most patients requesting labiaplasty experience both physical and appearance-related symptoms. Understanding this patient perspective is crucial in assessing surgical outcomes. Furthermore, the better all physicians understand labia symptomatology, the better supported patients will be in seeking surgical relief.


Clinical immunotherapy experiences in the Southeastern Cancer Study Group.
The Southeastern Cancer Study Group has been particularly interested in the use of immunological adjuvants in the treatment of melanoma and acute myelogenous leukemia. A study of chemotherapy versus chemoimmunotherapy in randomly selected patients with metastatic malignant melanoma revealed no significant increase in either complete remissions or overall survival in the group receiving chemoimmunotherapy. Preliminary results in a study involving maintenance of patients with acute myelogenous leukemia (chemotherapy versus chemoimmunotherapy) revealed suggestive but not statistically significant increases in remission duration and survival in the group receiving immunotherapy. Data from these ongoing studies and plans for new group studies are presented in this paper.


Trisomy 21 and Rett syndrome: a double burden.
Rett syndrome is a severe neurodevelopmental disorder generally affecting girls. Affected individuals are apparently normal at birth but later pass through a period of regression with loss of hand and communication skills and the development of hand stereotypies and dyspraxia. Mutations in the methyl-CpG binding protein 2 (MECP2) gene, have now been found to cause Rett syndrome in up to 80% of classical cases. We report a girl with Down syndrome, one of three children with birth defects in a family of five. From the age of 18 months she developed symptomatology considered by her primary physician to be very characteristic of Rett syndrome. However, this remained a clinical diagnosis till the age of 12 years. Laboratory confirmation of the dual diagnosis, which includes a R168X mutation in the MECP2 gene in addition to trisomy 21, has now been possible. The presence of one neurological or developmental disorder does not necessarily preclude a diagnosis of Rett syndrome.


Road rule enforcement and where to find it: An investigation of applications used to avoid detection when violating traffic rules.
One of the primary countermeasures in place to prevent road rule violations is legal enforcement, yet there are numerous applications that can undermine such efforts by notifying drivers of enforcement locations. However, the capabilities of these applications and how they can impact offending behavior is currently unknown. Two studies were conducted to understand which of these applications are being used by drivers and how these applications are impacting road rule violations. Study 1 consisted of a content analysis that involved searching the Google Play Store and Apple iTunes Store for applications that could be used to avoid road rule violations using pre-determined keywords. Meanwhile, Study 2 consisted of 468 licensed Australian drivers (54.5% males) over the age of 17 years (Mage = 35 years) who completed a survey. A total of 73 applications were identified for Study 1, with most of the applications displaying speed camera locations. It was found that applications that notify drivers of traffic enforcement locations are widely prevalent, can be used on a variety of interfaces and include numerous additional features. Study 2 found that those who use the applications were more willing to speed than those who do not use the applications, while there was no difference in phone use while driving between those who do and do not use the applications. The findings have important implications for stakeholders, policy, and future research. For example, it is suggested that specific functions of these applications need to be regulated to reduce road rule violations and crash risk. Meanwhile, enforcement initiatives need to evolve at a faster rate to keep up to date with the changing technology that can undermine them.


BK channels: multiple sensors, one activation gate.
Ion transport across cell membranes is essential to cell communication and signaling. Passive ion transport is mediated by ion channels, membrane proteins that create ion conducting pores across cell membrane to allow ion flux down electrochemical gradient. Under physiological conditions, majority of ion channel pores are not constitutively open. Instead, structural region(s) within these pores breaks the continuity of the aqueous ion pathway, thereby serves as activation gate(s) to control ions flow in and out. To achieve spatially and temporally regulated ion flux in cells, many ion channels have evolved sensors to detect various environmental stimuli or the metabolic states of the cell and trigger global conformational changes, thereby dynamically operate the opening and closing of their activation gate. The sensors of ion channels can be broadly categorized as chemical sensors and physical sensors to respond to chemical (such as neural transmitters, nucleotides and ions) and physical (such as voltage, mechanical force and temperature) signals, respectively. With the rapidly growing structural and functional information of different types of ion channels, it is now critical to understand how ion channel sensors dynamically control their gates at molecular and atomic level. The voltage and Ca(2+) activated BK channels, a K(+) channel with an electrical sensor and multiple chemical sensors, provide a unique model system for us to understand how physical and chemical energy synergistically operate its activation gate.


The impact of surgical treatment for penile cancer -- patients' perspectives.
Penile cancer is a rare but highly treatable condition. Whilst over 80% survive for over five years, treatment can have a significant impact on quality of life. There has been little research conducted to date on men's experiences of treatment for penile cancer. The Patients Experiences of Penile Cancer study (PEPC) aimed to redress this shortfall by exploring men's experiences of surgical treatment for penile cancer. The study used a narrative history design in which data were collected using one-on-one semi-structured interviews. Maximum variation sampling was used to acquire the widest possible range of experiences. Twenty-seven interviews of around one hour were conducted with men with an average age of 63 years at diagnosis (range = 41-82). The data were analysed using constant comparison analysis. The physical impact of surgery was inter-connected with broader events in the lives of the men experiencing treatment. These experiences cover urinary function, sexual function and sexual relationships, healing and recovery, masculinity, mental well-being, coping and support. A key area for the development of care is to devise and evaluate procedures for ensuring that men are well-informed about the extent and potential consequences of their treatment. Men's experiences of penile cancer surgery will be informed by a complex web interlaced with their broader lives, making it difficult for health professionals to judge how surgery will impact on a men presenting to them. Further research is required to ascertain the most appropriate strategies for rehabilitation of men experiencing penile cancer surgery.


Psychosocial risk screening during pregnancy: additional risks identified during a second interview.
The Prenatal Risk Overview (PRO) screens for 13 psychosocial risk factors associated with poor birth outcomes. This study assessed the extent to which risk factors unreported during an intake interview were identified during a subsequent interview. A total of 708 pregnant women were screened and re-screened at three urban community health care centers between July 2007 and April 2010. Study participants were predominantly young (mean age 23.5 years), unmarried (75.1%) women of color (92.5%); 38.4% were foreign-born. The proportional increase in participants identified as being at risk for individual domains at the second interview ranged from 5.6% to 49.0% for the combined Moderate/High Risk classification and from 5.6% to 73.0% for the High Risk only classification. For women whose health and well-being are challenged by poverty, violence, social isolation, and other stressors, both initial screening and repeat screening offer opportunities to alleviate identified risks.


miR-146a and miR-146b in the diagnosis and prognosis of papillary thyroid carcinoma.
The present study investigated the relationship between the expression of miR-146a and miR‑146b with the occurrence and prognosis of papillary thyroid carcinoma. Experiments in vitro were also used to explore the effect of the knocked down expression of the miRNAs on growth and migration of papillary thyroid carcinoma cells. A total of 73 patients with papillary thyroid carcinoma admitted to Yidu Central Hospital of Weifang from September 2013 to September 2015 were enrolled in the study. Carcinoma samples were obtained from each patient, and adjacent tissues were used as control samples to determine expression levels of miR-146a and miR146b by semi-quantitative RT-PCR. An analysis was conducted to find possible correlations between the miRNAs expression levels and clinicopathological features in the patients followed up for one year after diagnosis. Additionally, to examine the function of miR-146a and miR‑146b on TPC-1 cells, the expression of miRNAs was knocked down using specific siRNAs. MTT and Transwell assays were used to evaluate cell proliferation and migration, respectively, in the miRNA cell lines. Finally, western blot analysis was used to analyze the expression of IRAK1 in PTC cancer cells. Our results showed that the expression levels of miR-146a and miR-146b in carcinoma tissues were significantly higher than the levels in cancer-free tissues (p<0.01). The relative expression levels of miR-146a and miR-146b in cancerous tissues could be associated with the pathological type and presence or absence of lymph node metastasis (p<0.05). Compared with the siRNA-control cell, MTT and Transwell assays showed that the cell growth and migration of TPC-1 cells were decreased in miR-146a and miR-146b low expression cells (p<0.01). Western blot analysis showed that the expression of IRAK1 in papillary thyroid carcinoma was higher than in adjacent tissue (p<0.01). Based on our findings, the expression of miR-146a and miR-146b correlates with the occurrence and prognosis of papillary thyroid carcinoma, and the expression levels of miR-146a and miR-146b seem to affect the cell proliferation and migration and regulate the expression of IRAK1 protein in cancer cells. Further studies are needed to validate our results to provide new targets for prevention and treatment of papillary thyroid carcinoma.


Interdisciplinary approach towards a systems medicine toolbox using the example of inflammatory diseases.
Electronic access to multiple data types, from generic information on biological systems at different functional and cellular levels to high-throughput molecular data from human patients, is a prerequisite of successful systems medicine research. However, scientists often encounter technical and conceptual difficulties that forestall the efficient and effective use of these resources. We summarize and discuss some of these obstacles, and suggest ways to avoid or evade them.The methodological gap between data capturing and data analysis is huge in human medical research. Primary data producers often do not fully apprehend the scientific value of their data, whereas data analysts maybe ignorant of the circumstances under which the data were collected. Therefore, the provision of easy-to-use data access tools not only helps to improve data quality on the part of the data producers but also is likely to foster an informed dialogue with the data analysts.We propose a means to integrate phenotypic data, questionnaire data and microbiome data with a user-friendly Systems Medicine toolbox embedded into i2b2/tranSMART. Our approach is exemplified by the integration of a basic outlier detection tool and a more advanced microbiome analysis (alpha diversity) script. Continuous discussion with clinicians, data managers, biostatisticians and systems medicine experts should serve to enrich even further the functionality of toolboxes like ours, being geared to be used by 'informed non-experts' but at the same time attuned to existing, more sophisticated analysis tools.


Phaeohyphomycotic soft tissue disease caused by Pleurophomopsis lignicola in a kidney transplant patient.
A 44-year-old immunocompromised man presented with multiple tissue abscesses, covering the entire left limb. A dematiaceous fungus compatible with Pleurophomopsis lignicola Petrak was isolated from the diseased tissue in pure culture. This is the second reported isolation of this fungus from man and the first report of fatal soft tissue infection. A detailed morphological description of the isolate is provided.


Evolution of physical and electronic structures of bilayer graphene upon chemical functionalization.
The chemical behavior of bilayer graphene under strong covalent and noncovalent functionalization is relatively unknown compared to monolayer graphene, which has been far more widely studied. Bilayer graphene is significantly less chemically reactive than monolayer graphene, making it more challenging to study its chemistry in detail. However, bilayer graphene is increasingly attractive for electronic applications rather than monolayer graphene because of its electric-field-controllable band gap, and there is a need for a greater understanding of its chemical functionalization. In this paper, we study the covalent and noncovalent functionalization of bilayer graphene using an electrochemical process with aryl diazonium salts in the high conversion regime (D/G ratio >1), and we use Raman spectroscopic mapping and conductive atomic force microscopy (cAFM) to study the resulting changes in the physical and electronic structures. Covalent functionalization at high chemical conversion induces distinct changes in the Raman spectrum of bilayer graphene including the broadening and shift in position of the split G peak. Also, the D peak becomes active with four components. We report for the first time that the broadening of the 2D22 and 2D21 components is a distinct indicator of covalent functionalization, whereas the decrease in intensity of the 2D11 and 2D12 peaks corresponds to doping. Conductive AFM imaging shows physisorbed species from noncovalent functionalization can be removed by mechanical and electrical influence from the AFM tip, and that changes in conductivity due to functionalization are inhomogeneous. These results allow one to distinguish covalent from noncovalent chemistry as a guide for further studies of the chemistry of bilayer graphene.


Adjunctive armodafinil for major depressive episodes associated with bipolar I disorder: a randomized, multicenter, double-blind, placebo-controlled, proof-of-concept study.
To evaluate the efficacy and safety of armodafinil, the longer-lasting isomer of modafinil, when used adjunctively in patients with bipolar depression. In this 8-week, multicenter, randomized, double-blind, placebo-controlled study conducted between June 2007 and December 2008, patients who were experiencing a major depressive episode associated with bipolar I disorder (according to DSM-IV-TR criteria) despite treatment with lithium, olanzapine, or valproic acid were randomly assigned to adjunctive armodafinil 150 mg/d (n = 128) or placebo (n = 129) administered once daily in the morning. The primary outcome measure was change from baseline in the total 30-item Inventory of Depressive Symptomatology, Clinician-Rated (IDS-C₃₀) score. Secondary outcomes included changes from baseline in scores on the Montgomery-Åsberg Depression Rating Scale, among other psychological symptom scales. Statistical analyses were performed using analysis of covariance (ANCOVA), with study drug and concurrent mood stabilizer treatment for bipolar disorder as factors and the corresponding baseline value as a covariate. A prespecified sensitivity analysis was done using analysis of variance (ANOVA) if a statistically significant treatment-by-baseline interaction was found. Tolerability was also assessed. A significant baseline-by-treatment interaction in the total IDS-C₃₀ score (P = .08) was found. Patients administered adjunctive armodafinil showed greater improvement in depressive symptoms as seen in the greater mean ± SD change on the total IDS-C₃₀ score (-15.8 ± 11.57) compared with the placebo group (-12.8 ± 12.54) (ANOVA: P = .044; ANCOVA: P = .074). No differences between treatment groups were observed in secondary outcomes. Adverse events reported more frequently in patients receiving adjunctive armodafinil were headache, diarrhea, and insomnia. Armodafinil was not associated with an increased incidence and/or severity of suicidality, depression, or mania or with changes in metabolic profile measurements. In this proof-of-concept study, adjunctive armodafinil 150 mg/d appeared to improve depressive symptoms according to some, but not all, measures and was generally well tolerated in patients with bipolar depression. clinicaltrials.gov Identifier: NCT00481195.


Respiratory muscle pressure development during breath holding in apnea divers.
We sought to characterize the patterns of active pressure development of the inspiratory and expiratory rib cage muscles (P(rcm,i) and P(rcm,e)), the diaphragm (P(di,i)), and the expiratory abdominal muscles (P(abm,e)) during maximal "dry" breath holding in trained apnea divers (n = 8). Respiratory contractions were assessed via esophageal and gastric manometry. It was expected that inspiratory/expiratory pressures would progressively increase in both magnitude and frequency during the struggle phase, and that inspiratory rib cage muscle pressures would rise at a rate exceeding that of the diaphragm by the break point. P(rcm,i), P(di,i), P(rcm,e), and P(abm,e) significantly increased from the beginning until the end of the struggle phase (P < 0.05). Moreover, P(di,i)/P(rcm,i) and P(abm,e)/P(rcm,e) ratios had declined by the break point (P < 0.05), indicating that rib cage muscles increased their contribution to net inspiratory/expiratory pressure development by the end of the breath hold, relative to that contributed by the diaphragm and abdominal muscles. The pressure-time indices of the diaphragm and inspiratory rib cage muscles continuously increased over the struggle phase (P < 0.05). The "extradiaphragmatic" shift in inspiratory muscle recruitment, commensurate with increasing P(rcm,e) and P(abm,e), may reflect an extreme loading response to breathing against a heavy elastance (i.e., closed glottis). In addition, the relative intensity of diaphragmatic and inspiratory rib cage muscle contractions approaches potentially "fatiguing" levels by the break point of maximal breath holding.


Twelve-year experience with the Thow long intestinal tube: a means of preventing postoperative bowel obstruction.
The purpose of this study was to determine the effectiveness of the Thow long intestinal tube (LIT) for prevention of postoperative adhesive small bowel obstruction (ASBO) and to compare the Thow tube with other LITs. The charts of all patients who had placement of a Thow tube between January 1986 and November 1998 were reviewed. Thirty-four patients ranging in age from 9 to 86 years (mean 57.9) were included in the study. Twenty-five were contacted by phone for long-term follow-up. Twenty-nine patients had undergone previous abdominal surgery, and in 11 of 29 the previous surgery was for ASBO. Indications for surgery and Thow tube placement included: bowel obstruction (25), perforated viscus (five), carcinomatosis (two), colitis (one), and atonic bowel (one). Review of the operative notes revealed no difficulty in advancing the Thow tube in 32 of 34 patients (94%). Thow tube-related complications occurred in nine patients (25%). All complications were associated with the gastrostomy site, and only one patient required surgery for the complication. Two (5.9%) patients developed recurrent obstruction during a mean follow-up of 52 months. In one patient the obstruction was caused by adhesions and in another it was the result of an intra-abdominal abscess. Of 23 patients treated for ASBO at the time of Thow tube placement no patient (0%) developed recurrent ASBO during the follow-up period (total 110.5 patient-years). This study along with a review of the literature suggests that LITs decrease the risk of recurrent ASBO. The Thow tube, however, is easily placed and is associated with fewer and less severe complications than other LITs.


Utility of CSF Cytokine/Chemokines as Markers of Active Intrathecal Inflammation: Comparison of Demyelinating, Anti-NMDAR and Enteroviral Encephalitis.
Despite the discovery of CSF and serum diagnostic autoantibodies in autoimmune encephalitis, there are still very limited CSF biomarkers for diagnostic and monitoring purposes in children with inflammatory or autoimmune brain disease. The cause of encephalitis is unknown in up to a third of encephalitis cohorts, and it is important to differentiate infective from autoimmune encephalitis given the therapeutic implications. To study CSF cytokines and chemokines as diagnostic biomarkers of active neuroinflammation, and assess their role in differentiating demyelinating, autoimmune, and viral encephalitis. We measured and compared 32 cytokine/chemokines using multiplex immunoassay and APRIL and BAFF using ELISA in CSF collected prior to commencing treatment from paediatric patients with confirmed acute disseminated encephalomyelitis (ADEM, n = 16), anti-NMDAR encephalitis (anti-NMDAR E, n = 11), and enteroviral encephalitis (EVE, n = 16). We generated normative data using CSF from 20 non-inflammatory neurological controls. The sensitivity of CSF cytokine/chemokines to diagnose encephalitis cases was calculated using 95th centile of control values as cut off. We correlated CSF cytokine/chemokines with disease severity and follow up outcome based on modified Rankin scale. One-way hierarchical correlational cluster analysis of molecules was performed in different encephalitis and outcome groups. In descending order, CSF TNF-α, IL-10, IFN-α, IL-6, CXCL13 and CXCL10 had the best sensitivity (>79.1%) when all encephalitis patients were included. The combination of IL-6 and IFN-α was most predictive of inflammation on multiple logistic regression with area under the ROC curve 0.99 (CI 0.97-1.00). There were no differences in CSF cytokine concentrations between EVE and anti-NMDAR E, whereas ADEM showed more pronounced elevation of Th17 related (IL-17, IL-21) and Th2 (IL-4, CCL17) related cytokine/chemokines. Unlike EVE, heat map analysis showed similar clustering of cytokine/chemokine molecules in immune mediated encephalitis (ADEM and anti-NMDAR E). Th1 and B cell (CXCL13 and CXCL10) molecules clustered together in patients with severe encephalopathy at admission and worse disability at follow up in all encephalitis. There was no correlation between CSF neopterin and IFN-γ or IFN-α. A combination panel of cytokine/chemokines consisting of CSF TNF-α, IL-10, IFN-α, IL-6, CXCL13 and CXCL10 measured using multiplex immunoassay may be used to diagnose and monitor intrathecal inflammation in the brain. Given their association with worse outcome, certain key chemokines (CXCL13, CXCL10) could represent potential therapeutic targets in encephalitis.


℮-conome: an automated tissue counting platform of cone photoreceptors for rodent models of retinitis pigmentosa.
Retinitis pigmentosa is characterized by the sequential loss of rod and cone photoreceptors. The preservation of cones would prevent blindness due to their essential role in human vision. Rod-derived Cone Viability Factor is a thioredoxin-like protein that is secreted by rods and is involved in cone survival. To validate the activity of Rod-derived Cone Viability Factors (RdCVFs) as therapeutic agents for treating retinitis Pigmentosa, we have developed e-conome, an automated cell counting platform for retinal flat mounts of rodent models of cone degeneration. This automated quantification method allows for faster data analysis thereby accelerating translational research. An inverted fluorescent microscope, motorized and coupled to a CCD camera records images of cones labeled with fluorescent peanut agglutinin lectin on flat-mounted retinas. In an average of 300 fields per retina, nine Z-planes at magnification X40 are acquired after two-stage autofocus individually for each field. The projection of the stack of 9 images is subject to a threshold, filtered to exclude aberrant images based on preset variables. The cones are identified by treating the resulting image using 13 variables empirically determined. The cone density is calculated over the 300 fields. The method was validated by comparison to the conventional stereological counting. The decrease in cone density in rd1 mouse was found to be equivalent to the decrease determined by stereological counting. We also studied the spatiotemporal pattern of the degeneration of cones in the rd1 mouse and show that while the reduction in cone density starts in the central part of the retina, cone degeneration progresses at the same speed over the whole retinal surface. We finally show that for mice with an inactivation of the Nucleoredoxin-like genes Nxnl1 or Nxnl2 encoding RdCVFs, the loss of cones is more pronounced in the ventral retina. The automated platform ℮-conome used here for retinal disease is a tool that can broadly accelerate translational research for neurodegenerative diseases.


Anticoagulation combined with antiplatelet therapy in patients with left ventricular thrombus after first acute myocardial infarction.
There are limited data about the optimal anti-thrombotic therapy for preventing embolism while minimizing bleeding events in patients with first acute myocardial infarction (AMI) complicated by left ventricular thrombus (LVT). Among 2301 consecutive patients with AMI hospitalized between 2001 and 2014, we studied 1850 patients with first AMI who discharged alive to investigate clinical characteristics, incidence of systemic embolism (SE), and association between anticoagulation and embolic or bleeding events. Left ventricular thrombus was diagnosed by echocardiography, left ventriculography, or cardiac magnetic resonance imaging in 92 (5.0%) patients (62 ± 12 years). During a median follow-up period of 5.4 years (interquartile range 2.1-9.1 years), SE occurred in 15 of 92 patients with LVT (16.3%) and 51 of 1758 patients without LVT (2.9%), respectively. Kaplan-Meier analysis showed a significantly higher incidence of SE in the LVT group (log-rank test, P < 0.001). Multivariate analysis showed that LVT was an independent predictor of SE. Among the LVT patients treated with vitamin K antagonists (n = 84), we compared the patients with therapeutic range (TTR) ≥50% (n = 34) and those with TTR <50% (n = 50). Only one embolic event developed in the TTR ≥50% group and nine embolic events developed in the TTR <50% group (2.9% vs. 19%, P = 0.036). There was no difference in major bleeding events (TTR ≥50%; 9% vs. TTR <50%; 8%, P = 0.89). Appropriate anticoagulation therapy may decrease the incidence of embolic events without increasing the incidence of bleeding events in patients with first AMI complicated by LV thrombus.


Cloning, overexpression, and mutagenesis of the gene for homoprotocatechuate 2,3-dioxygenase from Brevibacterium fuscum.
Homoprotocatechuate (hpca, 3,4-dihydroxyphenylacetate) is a central intermediate for the bacterial degradation of aromatic compounds. Homoprotocatechuate 2,3-dioxygenase (HPCD) catalyzes the key ring cleavage step in the metabolism of hpca by the Gram (+) bacterium Brevibacterium fuscum to yield alpha-hydroxy-delta-carboxymethyl cis-muconic semialdehyde. A genomic DNA library of B. fuscum was constructed in Escherichia coli using a cosmid vector and screened by spraying the cells with hpca. One clone was found to contain the gene for HPCD based on its ability to convert hpca into the yellow-colored product. This cosmid clone was further subcloned and the gene for HPCD was localized and sequenced. The open reading frame codes for a protein with 365 amino acids and M(r) = 41,699, in accord with the characteristics of the previously purified wild-type enzyme. The gene for HPCD was overexpressed in E. coli to approximately 30% of the total soluble protein, and purification of the recombinant enzyme to apparent homogeneity was achieved by a two-step procedure. Iron was the only abundant metal found in the purified recombinant enzyme, and the specific activity per iron was comparable to that observed for the wild-type enzyme. The deduced amino acid sequence of HPCD has a very high level of homology (78.6% identity in the 337-aa overlap) to the manganese-dependent homoprotocatechuate 2,3-dioxygenase (MndD) from Arthrobacter globiformis CM-2. The basis for the difference in metal selection by HPCD and MndD was investigated by mutagenesis of a 50-base-pair region of the HPCD gene containing three frame shifts relative to the MndD gene. The purified triple mutant of HPCD did not exhibit a significant change in the metal content; therefore, other factors must contribute to the selection of the active site metal.


Comparison of the Contact Force Exerted on Teeth by Conventional Macintosh Laryngoscope Versus Video Laryngoscopes.
During laryngoscopy, the laryngoscope blade sometimes comes in contact with the teeth, fracturing or dislocating them. However, no studies have compared the effects of newly marketed video laryngoscopes and the Macintosh laryngoscope (Mac) on teeth. In this study, we measured and compared the force exerted on the teeth of an intubating manikin by the Mac, the Airway Scope (Pentax), and the McGrath MAC (Covidien). The mean force exerted was 141.1 ± 15.7 kg by the Mac, 39.2 ± 10.3 kg by the Airway Scope, and 48.7 ± 6.7 kg by the McGrath MAC. No significant difference was observed between the Airway Scope and the McGrath MAC. When the Mac is inserted, the glottis has to be visually located from outside the oral cavity. However, a significant force is not necessary when inserting video laryngoscopes because a camera is mounted on the blade tip. In this laboratory model, the lower force exerted by the video laryngoscopes should contribute to a reduction in their impact on fracture or dislocation of teeth.


Sclerotherapy for lymphatic malformations in children: a scoping review.
This scoping review assesses the literature and summarizes the current evidence on sclerotherapy for the treatment of lymphatic malformations in pediatric patients. A comprehensive search of published and unpublished literature was conducted using multiple databases. Title, abstract, and full-text screening was conducted by 2 independent clinicians. All discrepancies were resolved during consensus meetings. A total of 182 articles were retrieved. Forty-four articles were removed as duplicates, and 11 articles were added after reviewing prominent studies. After full-text abstraction, 44 articles and 2 conference proceedings (N = 882 patients) were included in the final results. Twelve articles were classified as level II and 34 articles as level IV evidence. Picibanil (OK-432) was the primary agent used in most included studies. Postinjection symptoms with OK-432 were primarily fever, swelling, and erythema at the site. Life-threatening complications were uncommon and involved postinjection swelling of cervical lesions causing airway compromise. The literature regarding sclerotherapy for lymphatic malformations is of a low level of evidence and suffers from a lack of standardization. Randomized clinical trials focused on OK-432, bleomycin, or alcoholic solution of zein; standardized dosing protocols; and consistent and reliable outcome reporting will be necessary for further development of treatment guidelines.


[Place of reflexotherapy and some other methods of alternative medicine in modern medical practice].
Assessment of the role and place of nontraditional methods of treatment and reflexotherapy, widely applied in hospitals is presented in the article. Besides, we become alerted regarding not serious approach of some scientists and health service managers to reflexotherapy as a whole and to one of its methods--acupuncture. An analysis of the situation developed in the legislation concerning training of reflexotherapy specialists for last 15-20 years not only in Ukraine, but also abroad was done. The article presents a historical parallel between the use of medicamentous and nonmedicamentous methods of treatment.


An overview of benefit/risk of disease modifying treatment of rheumatoid arthritis as of today.
In chronic rheumatoid arthritis (RA), disease modifying drugs are used in an attempt to suppress the progressive damage to tissues and joints that is associated with active disease. Their success in achieving this goal is variable; responses vary from complete suppression of all signs and symptoms of RA to continued active disease, with progressive disability, despite prolonged therapy. Because disease activity almost always recurs after the therapy is stopped, early interruption of an effective therapy for any reason will make its benefit insignificant in a lifelong disease such as RA. Similarly, short-term sequential use of multiple disease modifying therapies is unlikely to be beneficial. The immediate problems with these therapies are substantial. In general, fewer than 50% of patients are able to continue a particular drug for more than one year. Since it takes three to 12 months or longer to achieve maximum effects, those patients who are unable to continue the drug receive little benefit from it. Inevitable delayed side-effects, such as those associated with chronic corticosteroid therapy, may make the benefit/risk ratio unacceptable. Potential late lethal adverse effects, such as malignancy, weight the benefit/risk ratio to varying extents for individual patients, depending on the relative probability that the adverse effect will occur during the remainder of the patient's anticipated life span, and are of greater importance in younger patients. In that minority of patients who achieve remission or near remission and are able to tolerate a disease modifying treatment for many years, it is of truly significant benefit. We are still searching for a therapy that will reliably achieve this goal for most patients with RA.


Detecting Audio Adversarial Examples in Automatic Speech Recognition Systems Using Decision Boundary Patterns.
Automatic Speech Recognition (ASR) systems are ubiquitous in various commercial applications. These systems typically rely on machine learning techniques for transcribing voice commands into text for further processing. Despite their success in many applications, audio Adversarial Examples (AEs) have emerged as a major security threat to ASR systems. This is because audio AEs are able to fool ASR models into producing incorrect results. While researchers have investigated methods for defending against audio AEs, the intrinsic properties of AEs and benign audio are not well studied. The work in this paper shows that the machine learning decision boundary patterns around audio AEs and benign audio are fundamentally different. Using dimensionality-reduction techniques, this work shows that these different patterns can be visually distinguished in two-dimensional (2D) space. This in turn allows for the detection of audio AEs using anomal- detection methods.


Would integrated Western and traditional Chinese medicine have more benefits for stroke rehabilitation? A systematic review and meta-analysis.
Stroke is a major cause of death or long-term disability worldwide. Many patients with stroke receive integrative therapy consisting of Western medicine (WM) and routine rehabilitation in conjunction with Chinese medicine (CM), such as acupuncture and Chinese herbal medicine. However, there is no available evidence on the effectiveness of the combined use of WM and CM interventions in stroke rehabilitation. The purpose of this meta-analysis is to evaluate the results of all individual studies to assess the combined use of CM and WM in stroke rehabilitation compared with WM only. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines were followed. MEDLINE, EMBASE, Cochrane and China National Knowledge Infrastructure (CNKI) were searched. The included outcomes were dependency, motor function, depression and swallowing function. Subgroup analysis was performed, and publication bias was assessed using funnel plots. 58 studies and 6339 patients were included in the meta-analysis. Subgroup analysis revealed that combined therapy comprising both acupuncture and WM had a superior effect on improving dependency and swallowing function compared with standard WM therapy alone. Potential superiority of combined therapy comprising CM and WM in improving depression compared with standard WM therapy was also found. Our results indicate that the combined use of CM and WM could be more efficacious in stroke rehabilitation compared with the use of WM therapy alone. However, most studies were short in duration (2 to 4 weeks) and prone to different types of biases, which prevents making any conclusion regarding the long-term effects and raises concerns regarding true efficacy in context of high likelihood of Hawthorn bias. So, more randomised controlled trials with more rigorous design and longer duration of treatment and follow-up need to be conducted to compare WM alone versus WM and CM combined. CRD42020152050.


Interleukin-26 is associated with the level of systemic inflammation and lung functions in obese and non-obese moderate-to-severe asthmatic patients.
Obese asthma is a complex syndrome, which includes different phenotypes of disease. At present, these phenotypes only have started to acquire a sufficient understanding. It was suggested that IL-26 is a potential biomarker of disease severity in asthma without signs of Th2-mediated inflammation. In this study, we investigated the serum and exhaled levels of IL-26 and its associations with the level of systemic inflammation, lung functions, and body weight in obese and non-obese moderate-to-severe asthmatic patients MATERIAL AND METHODS: The study included 10 healthy subjects, 10 obese subjects without lung pathologies, 10 non-obese asthmatics (NOA) (BMI 18.5-24.9 kg/m²), and 40 obese asthmatics (OA) (BMI 25.0-49.9 kg/m²). During the visit, patients' examination and spirometry with the bronchodilator reversibility test were conducted, the exhaled breath condensate (EBC) was obtained, and the blood samples were collected. The level of IL-26, interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), TNF-α, interleukin-10 (IL-10), total and specific immunoglobulin E (IgE), and high sensitive C reactive protein (hs-CRP) were measured using the ELISA kits. Statistical comparison between 2 groups was analyzed using the Mann-Whitney rank-sum test. Chi-square with Yates' correction was used to compare frequencies. Spearman's rank test was used for correlating nonparametric variables. The Receiver Operating Characteristic (ROC) curve and the area under ROC curve (AUC) were used for evaluating the diagnostic power of IL-26 as a possible biomarker. NOA had a reversible airway obstruction with reduced FEV1, FEV1/FVC, FVC 25/75, and positive post-bronchodilator test (PBT), significantly increased serum levels of IL-10, IL-4, and slightly increased IL-26. NOA had significantly increased exhaled IL-26 in comparison with healthy subjects. The obese subjects had a normal ventilatory pattern without airway obstruction, and differences in serum IL-26, IL-10, and IL-4 concentrations in comparison with healthy subjects. Obese subjects had a significant escalation of hs-CRP and no differences in the levels of exhaled IL-26, IL-10, and hs-CRP as compared with healthy subjects. OA had reduced FEV1, FEV1/FVC, and FEV25-75 in comparison with non-obese asthmatics. OA had elevated IL-26, IL-10, IL-4, and hs-CRP concentrations as compared with healthy subjects. These patients had a partial similarity with both non-obese asthmatics (elevated IL-26, IL-10, and IL-4) and obese subjects (elevated, IL-1β, IL-6, TNF-α, hs-CRP). OA had a reduced concentration of exhaled IL-26 in comparison with NOA and elevated exhaled IL-10 in comparison with obese subjects. Furthermore, OA had an increased concentration of IL-1β and TNF-α in comparison with healthy individuals and NOA. Exhaled IL-26 concentration distinguished non-obese asthmatics from healthy subjects, asthmatic patients from non-asthmatics (healthy and obese subjects), all asthmatic patients from non-asthmatics (healthy and obese subjects). Exhaled IL-26 elevated in obese and non-obese moderate-to-severe asthmatic patients. Exhaled IL-26 might be a perspective biomarker in non-obese and obese asthmatics. The obese asthmatic phenotype comprised the combined systemic and local airway inflammation.


Leptin and leptin binding activity in the preovulatory follicle of polycystic ovary syndrome patients.
To investigate the clinical importance of leptin's intraovarian effects, we studied the concentration of leptin and leptin binding activity in the plasma and in the follicular fluid of PCOS patients (n=20; median BMI: 27.1 kg/m2, range 19.7-36.3) undergoing controlled ovarian stimulation with long-term GnRH agonist, recombinant FSH, and in vitro fertilization. Follicular fluid and blood samples were collected during follicle aspiration for IVF. Total leptin concentration was measured by radioimmunoassay, and specific leptin binding activity was accessed by a gel filtration column assay. Follicular fluid and plasma leptin levels were similar (median 1135 pmol/l vs. 1409 pmol/l; p=0.81). Follicular fluid to plasma leptin ratio was independently associated with cumulative FSH dose (r=0.63; p=0.006) and insulin resistance index (r=-0.45; p=0.04). Specific leptin binding activity was higher in the plasma than in the follicular fluid [median 7.94% vs. 3.49%; p<0.001]. When multivariate analysis was used to predict FSH consumption, only follicular fluid leptin levels were significantly associated with cumulative FSH dose (r=0.46; p=0.04). We infer that at least in part by increased intrafollicular leptin levels, obesity directly affects ovarian function in PCOS, and may induce a relative resistance to gonadotropin stimulation. This intraovarian effect of leptin can be even more profound because of low leptin binding activity in the preovulatory follicle of obese patients.


Recombinant infectious hematopoietic necrosis viruses induce protection for rainbow trout Oncorhynchus mykiss.
Infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) are rhabdoviruses that infect salmonids, producing serious economic losses. Two recombinant IHN viruses were generated by reverse genetics. For one (rIHNV GFP) the IHNV NV gene was replaced with the green fluorescent protein (GFP) gene. In the other (rIHNV-Gvhsv GFP) the G gene was also exchanged for that of VHSV. No mortalities, external signs or histological lesions were observed in experimental infections conducted with the recombinant viruses. Neither the rIHNV GFP nor rIHNV-Gvhsv GFP was detected by RT-PCR in any of the examined tissues from experimentally infected fish. In order to assess their potential as vaccines against the wild type viruses, rainbow trout were vaccinated with the recombinant viruses by intraperitoneal injection and challenged 30 d later with virulent IHNV or VHSV. The GFP viruses provided protection against both wild type viruses. None of the recombinant viruses induced antibody production, and the expression of interferon (IFNalpha4) and interferon induced genes such as Mx protein and ISG-15 was not different to that of controls. The rIHNV-Gvhsv GFP did not inhibit cellular apoptosis as it was observed in an IHNV inoculated fish cell line. These studies suggest that the recombinant rIHNV-Gvhsv GFP is a promising candidate as a live recombinant vaccine and also provides a good model to further study viral pathogenicity and the molecular basis of protection against these viral infections.


USP8 mutations in corticotroph adenomas determine a distinct gene expression profile irrespective of functional tumour status.
Pituitary corticotroph adenomas commonly cause Cushing's disease (CD) but part of these tumours are hormonally inactive (silent corticotroph adenomas, SCA). USP8 mutations are well-known driver mutations in corticotrophinomas. Differences in transcriptomic profiles between functioning and silent tumours or tumours with different USP8 status have not been investigated. Forty-eight patients (28 CD, 20 SCA) were screened for USP8 mutations with Sanger sequencing. Twenty-four patients were included in transcriptomic profiling with Ampliseq Transcriptome Human Gene Expression Core Panel. The entire patients group was included in qRT-PCR analysis of selected genes expression. Immunohistochemistry was used for visualization of selected protein. We found USP8 mutation in 15 patients with CD and 4 SCAs. USP8 mutations determine molecular profile of the tumours as showed by hierarchical clustering and identification of 1648 genes differentially expressed in USP8-mutated and USP8-wild-type tumours. Mutations affect many molecular pathways as observed in Gene Set Enrichment analysis. USP8-mutated adenomas showed higher level of POMC, CDC25A, MAPK4 but lower level of CCND2, CDK6, CDKN1B than USP8-wt tumours. Eighty-seven genes differentially expressed between CD-related adenomas and SCAs were found, including those involved in cell signalling (GLI2, DLC1, TBX2, RASSF6), cell adhesion (GJA1, CDH6), ion transport (KCNN4, KCNJ5) and GABA signalling (GABBR2, GABRD). USP8 mutations occur in functioning and silent corticotrophinomas. They have pleiotropic effect, not limited to EGFR signalling, and affect expression levels of many genes involved in different pathways. Expression of GABA-related genes GABBR2, GNAL, GABARD and KCNJ5 correspond to functional status of the tumours.


Therapeutic developments in Friedreich ataxia.
Friedreich ataxia is an inherited, severe, progressive neuro- and cardiodegenerative disorder for which there currently is no approved therapy. Friedreich ataxia is caused by the decreased expression and/or function of frataxin, a mitochondrial matrix protein that binds iron and is involved in the formation of iron-sulfur clusters. Decreased frataxin function leads to decreased iron-sulfur cluster formation, mitochondrial iron accumulation, cytosolic iron depletion, oxidative stress, and mitochondrial dysfunction. Cloning of the disease gene for Friedreich ataxia and elucidation of many aspects of the biochemical defects underlying the disorder have led to several major therapeutic initiatives aimed at increasing frataxin expression, reversing mitochondrial iron accumulation, and alleviating oxidative stress. These initiatives are in preclinical and clinical development and are reviewed herein.


Treatment outcome of patients with carcinoma of vulva: experience from a tertiary cancer center of India.
The aim of our retrospective study was to analyze and report the clinical outcome of patients with vulvar carcinoma (VC) treated at our center. We retrieved the information regarding patients' clinical details, treatment given, survival and complications from the case records of all VC patients who were treated at our center during the year 1998-2005. Overall survival (OS) was determined with respect to age, histopathological grade, stage of disease, treatment group, pathological lymph node status, etc. A total of 60 case records were retrieved for this retrospective analysis. Age ranged from 24 to 92 years (median 63 years). International Federation of Gynecology and Obstetrics (FIGO) stage distribution was as follows: stage I: 2 patients; stage II: 17 patients; stage III: 31 patients; stage IV: 9 patients; and unknown stage: 1 patient. Thirty-three patients underwent surgery (wide local excision 3, radical vulvectomy 30). Eleven patients received postoperative radiation therapy (PORT), 12 received palliative radiation therapy (RT) and 15 underwent definitive RT (5 of them received concurrent chemotherapy). Median follow-up period was 23 months (range 2-144 months). The 5-year OS for all stages was 41%. FIGO stage and pathological node positivity were found to be statistically significant prognostic factors for survival. Despite the majority of patients presenting in advanced stage, the 5-year OS of 41% in our series reflects a decent therapeutic outcome. The results have shown FIGO stage and pathological node positivity to be significant prognostic factors for survival. The use of preoperative chemotherapy/RT needs to be studied in our setup.


The specificity of the Eating Disorder Inventory.
A self-report measure, the Eating Disorder Inventory (EDI), has recently been published. An examination of the responses on the EDI of a sample of psychiatric out-patients who did not have eating problems revealed that many of its subscales may be non-specific in character reflecting the general level of psychological disturbance.


TRP channels in oxygen physiology: distinctive functional properties and roles of TRPA1 in O2 sensing.
Transient Receptor Potential (TRP) proteins form cation channels characterized by a wide variety of activation triggers. Here, we overview a group of TRP channels that respond to reactive redox species to transduce physiological signals, with a focus on TRPA1 and its role in oxygen physiology. Our systematic evaluation of oxidation sensitivity using cysteine-selective reactive disulphides with different redox potentials reveals that TRPA1 has the highest sensitivity to oxidants/electrophiles among the TRP channels, which enables it to sense O2. Proline hydroxylation by O2-dependent hydroxylases also regulates the O2-sensing function by inhibiting TRPA1 in normoxia; TRPA1 is activated by hypoxia through relief from the inhibition and by hyperoxia through cysteine oxidation that overrides the inhibition. TRPA1 enhances neuronal discharges induced by hyperoxia and hypoxia in the vagus to underlie respiratory adaptation to changes in O2 availability. This importance of TRPA1 in non-carotid body O2 sensors can be extended to the universal significance of redox-sensitive TRP channels in O2 adaptation.


Repair of the immature and mature craniofacial skeleton with a carbonated calcium phosphate cement: assessment of biocompatibility, osteoconductivity, and remodeling capacity.
The apatite compounds used most commonly in craniofacial reconstruction are highly crystalline and biologically inert ceramics. Because their capacity to be replaced by native bone is limited, they have found little application in repair of the growing craniofacial skeleton. Carbonated calcium phosphate cements more closely resemble the mineral phase of bone, thereby offering enhanced bioresorption and osteoconductivity, but their fate in the immature and mature craniofacial skeleton has not been investigated. The authors hypothesized that the capacity for cell-mediated remodeling of carbonated calcium phosphate cements is based on (1) their crystallographic and compositional similarity to the mineral phase of bone and (2) the osteogenic capacity of the host. Four noncritical-sized calvarial defects were created in six 3-week-old and six 16-week-old Yorkshire pigs. The defects were repaired with autologous bone, sintered carbonated calcium phosphate cement disks with a higher crystal order, or carbonated calcium phosphate cement (Norian CRS; Synthes Maxillofacial, West Chester, Pa.). The fourth defect was left empty as a control. Specimens were harvested at 30 and 90 days postoperatively. Empty defects healed with dense fibroconnective tissue in all groups. Autologous bone grafts underwent complete remodeling and replacement with woven bone at both time points. Sintered carbonated calcium phosphate disks demonstrated no bony ingrowth or remodeling. In immature animals, carbonated calcium phosphate cement implants were progressively replaced with woven bone through osteoclast-mediated resorption and osteoblast-mediated bone formation. Only minimal remodeling of the carbonated calcium phosphate cement implants was observed in skeletally mature animals. The results of these experiments suggest that the extent of remodeling of carbonated calcium phosphate cement is dependent on both the composition of the implant itself and the osteogenic capacity of the host and that carbonated calcium phosphate cement may be used successfully for inlay applications in the immature craniofacial skeleton.


Two-dimensional gel electrophoresis (2D-GE) image analysis based on CellProfiler: Pseudomonas aeruginosa AG1 as model.
Two-dimensional gel electrophoresis (2D-GE) is an indispensable technique for the study of proteomes of biological systems, providing an assessment of changes in protein abundance under various experimental conditions. However, due to the complexity of 2D-GE gels, there is no systematic, automatic, and reproducible protocol for image analysis and specific implementations are required for each context. In addition, practically all available solutions are commercial, which implies high cost and little flexibility to modulate the parameters of the algorithms. Using the bacterial strain, Pseudomonas aeruginosaAG1 as a model, we obtained images from 2D-GE of periplasmic protein profiles when the strain was exposed to multiple conditions, including antibiotics. Then, we proceeded to implement and evaluate an image analysis protocol with open-source software, CellProfiler. First, a preprocessing step included a bUnwarpJ-Image pipeline for aligning 2D-GE images. Then, using CellProfiler, we standardized two pipelines for spots identification. Total spots recognition was achieved using segmentation by intensity, whose performance was evaluated when compared with a reference protocol. In a second pipeline with the same program, differential identification of spots was addressed when comparing pairs of protein profiles. Due to the characteristics of the programs used, our workflow can automatically analyze a large number of images and it is parallelizable, which is an advantage with respect to other implementations. Finally, we compared six experimental conditions of bacterial strain in the presence or absence of antibiotics, determining protein profiles relationships by applying clustering algorithms PCA (Principal Components Analysis) and HC (Hierarchical Clustering).


Coronary image quality of 320-MDCT in patients with heart rates above 65 beats per minute: preliminary experience.
High heart rate may negatively influence the image quality of cardiac CT. The technical advances of 320-MDCT may overcome issues with poor image quality associated with high heart rate. This study aimed to evaluate the coronary image quality of 320-MDCT in patients with heart rates above 65 beats/min. Patients who presented for cardiac CT were divided into two groups according to heart rate, either greater than 65 beats/min or less than or equal to 65 beats/min. Two radiologists were blinded to the patient groups and evaluated images of 15 coronary artery segments per patient using 320-MDCT with consensus agreement. The image quality was scored subjectively as 1 or 2 (diagnostic quality) or 3 (poor quality and nondiagnostic). There were no statistically significant differences between the two groups in terms of age, sex, and body mass index (p > 0.05). The median heart rate was 70 beats/min (range, 67-110 beats/min) for the group with heart rate greater than 65 beats/min and 60 beats/min (range, 48-65 beats/min) for the group with heart rate less than or equal to 65 beats/min (p < 0.001). In patients with heart rates greater than 65 beats/min, diagnostic quality images (scores of 1 or 2) were obtained in 95.6% of the analyzed segments, compared with 96.9% in the group with heart rate less than or equal to 65 beats/min (p = 0.7). Our initial evaluation suggests that coronary artery images of diagnostic quality can be obtained using 320-MDCT in most patients with heart rates greater than 65 beats/min, in percentages similar to those for patients with heart rates less than or equal to 65 beats/min. This finding may be the result of the inherent image acquisition and reconstruction technique of 320-MDCT.


Prenatal diagnosis and elective termination of neural tube defects in Hawaii, 1986-1997.
To determine the influence of various factors on the prenatal diagnosis and elective termination of neural tube defects (NTDs) in Hawaii in 1986-1997. Data from a birth defects registry were analyzed and included 245 cases. 74% of the cases were prenatally diagnosed and 48% were terminated. Anencephaly was more likely to be prenatally diagnosed and electively terminated than spina bifida or encephalocele. Other factors such as maternal age, race/ethnicity, place of residence, and elevated maternal serum alpha-fetoprotein influenced the prenatal diagnosis and/or elective termination of NTD-affected pregnancies. However, a given factor may influence prenatal diagnosis and elective termination in different ways. Various diagnostic and demographic factors can influence the prenatal diagnosis and the subsequent termination of NTD-affected pregnancies.


Increased apoptosis, tumor necrosis factor-α, and DNA damage attenuated by 3',4'-dihydroxyflavonol in rats with brain İschemia-reperfusion.
This research was aimed to find out the effects of 3',4'-dihydroxyflavonol (DiOHF) on apoptosis, DNA damage, and tumor necrosis factor-α (TNF-α) levels in the frontal cortex of rats with induced experimental brain ischemi reperfusion. A total of 38 Wistar albino male rats were used. Groups were created as 1-Sham; 2-Ischemia-reperfusion (I/R); 3-I/R + DiOHF (10 mg/kg); 4-Ischemia + DiOHF + reperfusion; 5-DiOHF + I/R. I/R was performed by carotid artery ligation for 30 min in anesthesized animals. Following experimental applications, blood samples were taken from anesthetized rats to obtain erythrocyte and plasma. Later, the rats were killed by cervical dislocation, and frontal cortex samples were taken and stored at - 80oC for the analysis. In the ischemic frontal cortex tissue sections degenerate neuron numbers, Terminal deoxynucleotidyl transferase-dUTP nick end labeling (TUNEL) positive cell ratio and caspase-3 positive cell ratio increased. Malondialdehyde, TNF-α, and 8-OHdG levels were increased in both plasma and tissue in ischemia group, whereas tissue and erythrocyte glutathione levels were significantly suppressed. However, these values were significantly reversed by DiOHF treatment. The results of the study showed that I/R significantly increased apoptosis, TNF-α, and DNA damage in rats with brain I/R. However, 10 mg/kg intraperitoneal DiOHF treatment improved deterioted parameters.


Flow cytometric analysis of thiazole orange uptake by platelets: a diagnostic aid in the evaluation of thrombocytopenic disorders.
Thiazole orange (TO), a fluorescent dye originally synthesized for reticulocyte analysis, is characterized by a large fluorescence enhancement and high quantum yield on binding to nucleic acids, particularly RNA. In addition, the dye readily permeates live cell membranes. We applied TO staining, followed by fluorescence-activated flow cytometric analysis, to platelets in whole blood samples from hematologically normal subjects and patients with various quantitative platelet disorders. The percentage of TO-positive platelets in 50 control subjects was 8.6 +/- 2.8% (mean +/- SD) ranging from 2.8% to 15.8%. In 21 thrombocytopenic patients whose bone marrow contained normal to increased numbers of megakaryocytes, the percentage of fluorescently labeled platelets was significantly elevated (P less than .0001) to 26.9 +/- 10.9% (range, 13.3% to 57.1%). In contrast, the proportion of positively stained platelets in 23 patients with thrombocytopenia due to impaired platelet production (various conditions with reduced marrow megakaryocytes) did not significantly differ from the controls, whereas the absolute counts of TO-positive platelets were significantly lowered (P less than .0001). Differences in the distributions of the percentage values as well as of the absolute counts for TO-positive platelets between the two patient groups were again highly significant (P less than .0001). Both the sensitivity and the specificity of this method in distinguishing between these categories of thrombocytopenia were greater than or equal to 95%. We conclude that flow cytometric analysis of platelets after staining with TO is a sensitive and specific test that rapidly provides information on the thrombopoietic activity in thrombocytopenic disorders. Our data further suggest that increased amounts of residual RNA characterize platelets released under conditions of "stress thrombopoiesis."


Smad3-dependent and -independent pathways are involved in peritoneal membrane injury.
Transition of peritoneal mesothelial cells to a mesenchymal phenotype plays an integral role in the angiogenic and fibrotic changes seen in the peritoneum of patients receiving long-term peritoneal dialysis. While signaling by transforming growth factor (TGF)-beta through Smad proteins likely causes these changes, it is possible that non-Smad pathways may also play a role. Here, we found that Smad3-deficient mice were protected from peritoneal fibrosis and angiogenesis caused by adenovirus-mediated gene transfer of active TGF-beta1 to mesothelial cells; however, mesothelial transition occurred in this setting, suggesting involvement of non-Smad mechanisms. The phosphatidyl inositol 3 kinase (PI3K) target, Akt, was upregulated in both Smad-deficient and wild-type mice after exposure to TGF-beta1. In vivo inhibition of the mammalian target of rapamycin (mTOR) by rapamycin completely abrogated the transition response in Smad3-deficient but not in wild-type mice. Rapamycin blocked nuclear localization of beta-catenin independent of glycogen synthase kinase 3beta activity. Further, in Smad3-deficient mice rapamycin reduced the expression of alpha-smooth muscle actin, which is an epithelial-to-mesenchymal transition-associated gene. Hence, we conclude that TGF-beta1 causes peritoneal injury through Smad-dependent and Smad-independent pathways; the latter involves redundant mechanisms inhibited by rapamycin, suggesting that suppression of both pathways may be necessary to abrogate mesothelial transition.


What Works for Adolescent Black Males at Risk of Suicide: A Review.
We reviewed the controlled studies that report outcome findings for Black adolescent males 24 years of age and younger at risk of suicide. Our review identified 48 articles published from 2000 to 2015, 33 that met our initial criteria for full-text articles review, resulting in 6 that met all inclusion criteria. We sought to understand what works for Black males experiencing suicide ideation or engaging in suicidal behaviors (e.g., attempts). We identified crossover effects for multisystemic therapy for reducing the risk for suicide ideation and attempts. Attachment-based family therapy was salient for use as a component of clinical practice for Black males being treated for suicide ideation. While remaining randomized control trials did involve Black youth, dis-aggregated data based on ethnicity and gender were not reported. Overall, the located studies are too few to provide unequivocal guidance for practice.


Adventures in Rhodococcus - from steroids to explosives.
Rhodococcus is a genus of mycolic-acid-containing actinomycetes that utilize a remarkable variety of organic compounds as growth substrates. This degradation helps maintain the global carbon cycle and has increasing applications ranging from the biodegradation of pollutants to the biocatalytic production of drugs and hormones. We have been using Rhodococcus jostii RHA1 as a model organism to understand the catabolic versatility of Rhodococcus and related bacteria. Our approach is exemplified by the discovery of a cluster of genes specifying the catabolism of cholesterol. This degradation proceeds via β-oxidative degradation of the side chain and O2-dependent cleavage of steroid ring A in a process similar to bacterial degradation of aromatic compounds. The pathway is widespread in Actinobacteria and is critical to the pathogenesis of Mycobacterium tuberculosis, arguably the world's most successful pathogen. The close similarity of some of these enzymes with biphenyl- and polychlorinated-biphenyl-degrading enzymes that we have characterized is facilitating inhibitor design. Our studies in RHA1 have also provided important insights into a number of novel metalloenzymes and their biosynthesis, such as acetonitrile hydratase (ANHase), a cobalt-containing enzyme with no significant sequence identity with characterized nitrile hydratases. Molecular genetic and biochemical studies have identified AnhE as a dimeric metallochaperone that delivers cobalt to ANHase, enabling its maturation in vivo. Other metalloenzymes we are characterizing include N-acetylmuramic acid hydroxylase, which catalyzes an unusual hydroxylation of the rhodococcal and mycobacterial peptidoglycan, and 2 RHA1 dye-decolorizing peroxidases. Using molecular genetic and biochemical approaches, we have demonstrated that one of these enzymes is involved in the degradation of lignin. Overall, our studies are providing fundamental insights into a range of catabolic processes that have a wide variety of applications.


Comparison of immune responses and protective efficacy of intranasal prime-boost immunization regimens using adenovirus-based and CpG/HH2 adjuvanted-subunit vaccines against genital Chlamydia muridarum infection.
An efficacious Chlamydia vaccine is urgently needed to control Chlamydia infections. Heterologous prime-boost vaccination regimens are emerging as a promising strategy for preventing intracellular viral and bacterial infections. However, it remains to be determined if this regimen would be a feasible and effective approach for Chlamydia infection. In this study, we examined the immune response and the protective efficacy induced by various vaccination regimens using a recombinant adenovirus vector expressing the Chlamydia antigen CPAF (AdCPAF) and recombinant CPAF (rCPAF) subunit vaccines formulated with CpG oligodeoxynucleotides and/or a synthetic immunomodulatory peptide HH2 as adjuvants. A single dose of AdCPAF stimulated potent antibody production but weak cellular immune responses in mice. A booster rCPAF vaccine formulated with both CpG and HH2, but not CpG alone or HH2 alone, showed robust adjuvant effects on induction of Th1-biased cellular immune responses in mice primed with AdCPAF. In contrast, a homologous regimen using rCPAF/CpG/HH2 subunit vaccine for both priming and boosting induced a weak antibody response, but potent cellular immunity with a mixed Th1/Th17 profile. Despite the disparities observed in humoral and cellular immune responses, both the heterologous and homologous prime-boost regimens conferred significant immune protection against genital Chlamydia muridarum challenge in C3H/HeN and BALB/c mice.


Plasmid Dissemination and Selection of a Multidrug-Resistant Klebsiella pneumoniae Strain during Transplant-Associated Antibiotic Therapy.
Antibiotics, which are used both to prevent and to treat infections, are a mainstay therapy for lifesaving procedures such as transplantation. For this reason, and many others, increased antibiotic resistance among human-associated pathogens, such as the carbapenem-resistant Enterobacteriaceae species, is of grave concern. In this study, we report on a hematopoietic stem cell transplant recipient in whom cultures detected the emergence of carbapenem resistance and spread across five strains of bacteria that persisted for over a year. Carbapenem resistance in Citrobacter freundii, Enterobacter cloacae, Klebsiella aerogenes, and Klebsiella pneumoniae was linked to a pair of plasmids, each carrying the Klebsiella pneumoniae carbapenemase gene (blaKPC). Surveillance cultures identified a carbapenem-susceptible strain of Citrobacter freundii that may have become resistant through horizontal gene transfer of these plasmids. Selection of a multidrug-resistant Klebsiella pneumoniae strain was also detected following combination antibiotic therapy. Here we report a plasmid carrying the blaKPC gene with broad host range that poses the additional threat of spreading to endogenous members of the human gut microbiome.IMPORTANCE Antibiotic-resistant bacteria are a serious threat to medically fragile patient populations. The spread of antibiotic resistance through plasmid-mediated mechanisms is of grave concern as it can lead to the conversion of endogenous patient-associated strains to difficult-to-treat pathogens.


Repurpose terbutaline sulfate for amyotrophic lateral sclerosis using electronic medical records.
Prediction of new disease indications for approved drugs by computational methods has been based largely on the genomics signatures of drugs and diseases. We propose a method for drug repositioning that uses the clinical signatures extracted from over 13 years of electronic medical records from a tertiary hospital, including >9.4 M laboratory tests from >530,000 patients, in addition to diverse genomics signatures. Cross-validation using over 17,000 known drug-disease associations shows this approach outperforms various predictive models based on genomics signatures and a well-known "guilt-by-association" method. Interestingly, the prediction suggests that terbutaline sulfate, which is widely used for asthma, is a promising candidate for amyotrophic lateral sclerosis for which there are few therapeutic options. In vivo tests using zebrafish models found that terbutaline sulfate prevents defects in axons and neuromuscular junction degeneration in a dose-dependent manner. A therapeutic potential of terbutaline sulfate was also observed when axonal and neuromuscular junction degeneration have already occurred in zebrafish model. Cotreatment with a β2-adrenergic receptor antagonist, butoxamine, suggests that the effect of terbutaline is mediated by activation of β2-adrenergic receptors.


Estimating confidence intervals for cerebral autoregulation: a parametric bootstrap approach.
Cerebral autoregulation (CA) refers to the ability of the brain vasculature to control blood flow in the face of changing blood pressure. One of the methods commonly used to assess cerebral autoregulation, especially in participants at rest, is the analysis of phase derived from transfer function analysis (TFA), relating arterial blood pressure (ABP) to cerebral blood flow (CBF). This and other indexes of CA can provide consistent results when comparing groups of subjects (e.g. patients and healthy controls or normocapnia and hypercapnia) but can be quite variable within and between individuals. The objective of this paper is to present a novel parametric bootstrap method, used to estimate the sampling distribution and hence confidence intervals (CIs) of the mean phase estimate in the low-frequency band, in order to optimise estimation of measures of CA function and allow more robust inferences on the status of CA from individual recordings. A set of simulations was used to verify the proposed method under controlled conditions. In 20 healthy adult volunteers (age 25.53.5 years), ABP and CBF velocity (CBFV) were measured at rest, using a Finometer device and Transcranial Doppler (applied to the middle cerebral artery), respectively. For each volunteer, five individual recordings were taken on different days, each approximately 18 min long. Phase was estimated using TFA. Analysis of recorded data showed widely changing CIs over the duration of recordings, which could be reduced when noisy data and frequencies with low coherence were excluded from the analysis (Wilcoxon signed rank testp= 0.0065). The TFA window-lengths of 50s gave smaller CIs than lengths of 100s (p< 0.001) or 20s (p< 0.001), challenging the usual recommendation of 100s. The method adds a much needed flexible statistical tool for CA analysis in individual recordings.


CD8+ T Cells Keep Their (K+)urrency for Function.
CD8+ T-cell responses are influenced by ion abundance, which can widely vary within the tumor microenvironment. In this issue, Collier and colleagues investigated how intracellular versus extracellular potassium ion (K+) regulates intratumoral CD8+ T cells. They show that, while excessive extracellular K+ induces exhaustion, intracellular K+ is needed for protection from dysfunction. This work shows additional evidence that the regulation of CD8+ T-cell responses depends on a fine balance between intracellular and extracellular metabolite levels. See related article by Collier et al. (2) .


Morphological and functional impairment in the gut in a partial body irradiation minipig model of GI-ARS.
Purpose: Göttingen minipig (G-MP) displays classic gastrointestinal acute radiation syndrome (GI-ARS) following total body irradiation (TBI) at GI doses which are lethal by 10-14 days. In collaboration with BARDA, we are developing a hemi-body/partial body irradiation (PBI) model by exposing only the abdomen and lower extremities to study GI structure/function impairment, natural history of injury and recovery, as well as correlative biomarkers out to 30 days.Materials and methods: Twenty-four G-MP were exposed to either 12 or 16 Gy (LINAC Elekta); head, forelimbs, and thorax were outside the irradiation field, sparing ∼50% of the bone marrow. Animals were followed for 30 days with euthanasia scheduled at pre-set intervals to study the time course of GI injury and recovery. Hematological profiles, clinical symptoms, gross- and histo-pathology including markers of proliferation and apoptosis in the small intestines, gut function parameters (food tolerance, digestion, absorption, citrulline production), and levels of two biomarkers, CRP and IGF-1, were evaluated.Results: PBI at 16 Gy yielded higher lethality than 12 Gy. Unlike TBI, PBI did not cause severe pancytopenia or external hemorrhage, as expected, and allowed to focus the injury on GI organs while sparing the radiation sensitive heart and lung. Compromised animals showed inactivity, anorexia, vomiting, diarrhea, and weight loss. Histology revealed that in 12 Gy irradiated animals, lesions recovered overtime. In 16 Gy irradiated animals, lesions were more pronounced and persistent. BrdU and Ki67 labelling demonstrated dose-dependent loss of crypts and subsequent mucosal ulceration which recovered over time. Minimal apoptosis was observed at both doses. Reductions in food tolerance, digestion, absorption, and citrulline production were time and dose-dependent. Loss of citrulline reached a nadir between 6-12 days and then recovered partially. CRP and IGF-1 were upregulated following PBI at GI doses.Conclusions: This lower hemi-body irradiation model allowed for extended survival at GI-specific ARS doses and development of a well-controlled GI syndrome with minimal hematopoietic injury or confounding mortality from cardiopulmonary damage. A dose-dependent impairment in the intestinal structure resulted in overall decreased gut functionality followed by a partial recovery. However, while the structure appeared to be recovered, not all functionality was attained. PBI induced systemic inflammation and altered the IGF-1 hormone indicating that these can be used as biomarkers in the minipig even under partial body conditions. This PBI model aligns with other minipig models under BARDA's large animal consortium to test medical countermeasure efficacy against a less complex GI-specific ARS injury.