Restriction endonuclease analysis of the pseudorabies (Aujeszky's disease) virus before and after serial passage in vivo and in vitro.
The deoxyribonucleic acid (DNA) of pseudorabies virus (PRV) was examined by restriction endonuclease analysis before and after various treatments: namely 1. plaque purification of stock virus, 2. serial passage of virus in cell culture at high (H) and low (L) multiplicity of infection (MOI), and 3. serial passage of virus in swine. The objective of the study was to determine if such treatments would either select or induce virus populations with a different predominant number or location of enzyme cleavage sites. Heterogeneity of the DNA of stock virus was revealed by differences among restriction patterns of 10 plaque-purified populations. All of these populations were then relatively stable through an additional 9 plaque purifications and passages in cell culture. Without plaque purification, heterogeneity was not evident. The predominant restriction pattern of stock virus appeared unaltered by 10 serial passages in cell culture at either HMOI or LMOI, except for the appearance in the HMOI series of several minor bands. Conversely, 10 serial passages of stock virus in swine resulted in either selection or induced changes or both. Most differences were slightly altered migration rates of relatively small (less than or equal to 5 Kbp) fragments. However, after 10 passages in swine, there were also changes in the migration rates of 2 large fragments (greater than 10 Kbp) of the viral genome.


Morphing of geometric composites via residual swelling.
Understanding and controlling the shape of thin, soft objects has been the focus of significant research efforts among physicists, biologists, and engineers in the last decade. These studies aim to utilize advanced materials in novel, adaptive ways such as fabricating smart actuators or mimicking living tissues. Here, we present the controlled growth-like morphing of 2D sheets into 3D shapes by preparing geometric composite structures that deform by residual swelling. The morphing of these geometric composites is dictated by both swelling and geometry, with diffusion controlling the swelling-induced actuation, and geometric confinement dictating the structure's deformed shape. Building on a simple mechanical analog, we present an analytical model that quantitatively describes how the Gaussian and mean curvatures of a thin disk are affected by the interplay among geometry, mechanics, and swelling. This model is in excellent agreement with our experiments and numerics. We show that the dynamics of residual swelling is dictated by a competition between two characteristic diffusive length scales governed by geometry. Our results provide the first 2D analog of Timoshenko's classical formula for the thermal bending of bimetallic beams - our generalization explains how the Gaussian curvature of a 2D geometric composite is affected by geometry and elasticity. The understanding conferred by these results suggests that the controlled shaping of geometric composites may provide a simple complement to traditional manufacturing techniques.


Gene patents related to common diseases of the eye.
Visual impairment and blindness impose substantial morbidity and premature mortality on the population. The direct costs for vision disorders have been shown to be more than the cost of coronary heart disease, stroke, arthritis or depression and were estimated to be $9.85 billion in 2004 in Australia. Hence it is important to identify the causes of common eye diseases and understand their aetiology which in turn would allow determination of better management strategies and treatment options. Age related Macular Degeneration, Cataract, Diabetic Retinopathy, Glaucoma and uncorrected refractive errors represent the majority of the visual impairment and blindness in Australia and various parts of the world. This article reviews the gene patents available for these eye conditions and highlights the important discoveries that have so far contributed to our understanding of these diseases and provides valuable information as to where research will be heading in the future.


Alternate conformations observed in catalytic serine of Bacillus subtilis lipase determined at 1.3 A resolution.
Bacillus subtilis extracellular lipase (BsL) has an exceptionally low molecular weight (19.4 kDa) for a member of the lipase family. A crystallographic study was performed on BsL in order to design and produce mutant BsL that will be more suitable for industrial uses based on analysis of the three-dimensional structure. Recently, the crystal structure of BsL has been determined at 1.5 A resolution [van Pouderoyen et al. (2001). J. Mol. Biol. 309, 215-226]. In the present study, a new crystal form of BsL which provides diffraction data to higher resolution was obtained and its structure was determined at 1.3 A using the MAD method. It was found that the active-site residue Ser77 has alternate side-chain conformations. The O(gamma) atom of the first conformer forms a hydrogen bond to the N(epsilon) atom of His155, a member of the catalytic triad. In contrast, the second conformer is constructed with a hydrogen bond to the side-chain atom of the adjacent His76. These two conformers presumably correspond to the active and inactive states, respectively. Similar alternate conformations in the catalytic serine residue have been observed in Fusarium solani cutinase determined at 1.0 A resolution and Penicillium purpurogenum acetylxylan esterase at 0.9 A resolution. In addition, a glycerol molecule, which was used as a cryoprotectant, is found to be located in the active site. On the basis of these results, a model for substrate binding in the reaction-intermediate state of BsL is proposed.


B-cell lines immortalized with an Epstein-Barr virus mutant lacking the Cp EBNA2 enhancer are biased toward utilization of the oriP-proximal EBNA gene promoter Wp1.
During Epstein-Barr virus (EBV) latent infection of B lymphocytes in vitro, six viral nuclear antigens (EBNAs) are expressed from one of two promoters, Cp or Wp, whose activities have previously been shown to be mutually exclusive in established lymphoblastoid cell lines. Initially after infection, the EBNA genes are transcribed from Wp, which is present in multiples copies within the major internal repeat of EBV. Approximately 48 to 72 h postinfection, Wp is downregulated, with a corresponding increase in transcription from Cp. An EBNA2-responsive enhancer exists upstream of Cp, and a role for EBNA2 in the induction of Cp activity during the establishment of viral latency has previously been proposed (Woisetschlaeger et al., Proc. Natl. Acad. Sci. USA 87:1725-1729, 1991). To critically assess the potential role for this enhancer region in determining relative usage of Cp and Wp, an EBNA2 enhancer deletion mutant virus was generated. Lymphoblastoid cell lines were screened by PCR and Southern blotting for the presence of mutant virus harboring the EBNA2 enhancer deletion. A quantitative S1 nuclease protection assay was developed to allow comparison of relative Cp and Wp activities for the cell lines containing mutant virus and those of the wild-type recombinants which lacked the enhancer deletion. In general, the wild-type recombinants had higher levels of Cp-initiated transcripts than Wp-initiated transcripts. In contrast, the Cp EBNA2 enhancer deletion mutants exhibited a strong bias toward Wp activity. Notably, only the first Wp (oriP-proximal Wp; Wp1) appears active in these mutants. S1 nuclease protection assays using a probe which hybridizes to the W2 exon, contained in both Cp- and Wp-initiated transcripts, indicated that the total level of transcription from Cp and Wp remained the same in wild-type and EBNA2 enhancer mutant cell lines. The presence of both Cp and Wp activity in the wild-type recombinants, as well as in newly derived lymphoblastoid cell lines established with the prototype B95.8 virus, demonstrated that Cp and Wp activities are not always mutually exclusive.


Nijmegen breakage syndrome detected by newborn screening for T cell receptor excision circles (TRECs).
Severe combined immunodeficiency (SCID) encompasses a group of disorders characterized by reduced or absent T-cell number and function and identified by newborn screening utilizing T-cell receptor excision circles (TRECs). This screening has also identified infants with T lymphopenia who lack mutations in typical SCID genes. We report an infant with low TRECs and non-SCID T lymphopenia, who proved upon whole exome sequencing to have Nijmegen breakage syndrome (NBS). Exome sequencing of DNA from the infant and his parents was performed. Genomic analysis revealed deleterious variants in the NBN gene. Confirmatory testing included Sanger sequencing and immunoblotting and radiosensitivity testing of patient lymphocytes. Two novel nonsense mutations in NBN were identified in genomic DNA from the family. Immunoblotting showed absence of nibrin protein. A colony survival assay demonstrated radiosensitivity comparable to patients with ataxia telangiectasia. Although TREC screening was developed to identify newborns with SCID, it has also identified T lymphopenic disorders that may not otherwise be diagnosed until later in life. Timely identification of an infant with T lymphopenia allowed for prompt pursuit of underlying etiology, making possible a diagnosis of NBS, genetic counseling, and early intervention to minimize complications.


Mechanistic understanding of Phenyllactic acid mediated inhibition of quorum sensing and biofilm development in Pseudomonas aeruginosa.
Pseudomonas aeruginosa depends on its quorum sensing (QS) system for its virulence factors' production and biofilm formation. Biofilms of P. aeruginosa on the surface of indwelling catheters are often resistant to antibiotic therapy. Alternative approaches that employ QS inhibitors alone or in combination with antibiotics are being developed to tackle P. aeruginosa infections. Here, we have studied the mechanism of action of 3-Phenyllactic acid (PLA), a QS inhibitory compound produced by Lactobacillus species, against P. aeruginosa PAO1. Our study revealed that PLA inhibited the expression of virulence factors such as pyocyanin, protease, and rhamnolipids that are involved in the biofilm formation of P. aeruginosa PAO1. Swarming motility, another important criterion for biofilm formation of P. aeruginosa PAO1, was also inhibited by PLA. Gene expression, mass spectrometric, functional complementation assays, and in silico data indicated that the quorum quenching and biofilm inhibitory activities of PLA are attributed to its ability to interact with P. aeruginosa QS receptors. PLA antagonistically binds to QS receptors RhlR and PqsR with a higher affinity than its cognate ligands N-butyryl-L-homoserine lactone (C4-HSL) and 2-heptyl-3,4-dihydroxyquinoline (PQS; Pseudomonas quinolone signal). Using an in vivo intraperitoneal catheter-associated medaka fish infection model, we proved that PLA inhibited the initial attachment of P. aeruginosa PAO1 on implanted catheter tubes. Our in vitro and in vivo results revealed the potential of PLA as anti-biofilm compound against P. aeruginosa.


Coccidioidomycosis Dynamics in Relation to Climate in the Southwestern United States.
Valley fever is endemic to the southwestern United States. Humans contract this fungal disease by inhaling spores of Coccidioides spp. Changes in the environment can influence the abundance and dispersal of Coccidioides spp., causing fluctuations in valley fever incidence. We combined county-level case records from state health agencies to create a regional valley fever database for the southwestern United States, including Arizona, California, Nevada, New Mexico, and Utah. We used this data set to explore how environmental factors influenced the spatial pattern and temporal dynamics of valley fever incidence during 2000-2015. We compiled climate and environmental geospatial data sets from multiple sources to compare with valley fever incidence. These variables included air temperature, precipitation, soil moisture, surface dust concentration, normalized difference vegetation index, and cropland area. We found that valley fever incidence was greater in areas with warmer air temperatures and drier soils. The mean annual cycle of incidence varied throughout the southwestern United States and peaked following periods of low precipitation and soil moisture. From year-to-year, however, autumn incidence was higher following cooler, wetter, and productive springs in the San Joaquin Valley of California. In southcentral Arizona, incidence increased significantly through time. By 2015, incidence in this region was more than double the rate in the San Joaquin Valley. Our analysis provides a framework for interpreting the influence of climate change on valley fever incidence dynamics. Our results may allow the U.S. Centers for Disease Control and Prevention to improve their estimates of the spatial pattern and intensity of valley fever endemicity.


The role of motor and nutritional individuality in childhood obesity.
Nutritional and motor individuality vary significantly among human subjects, and their mutal relationship is decisive for a desirable energy balance and turnover with regard to body composition, physical fitness level and health. Early establishment of optimal individualities, with regard to genetic, epigenetic and other factors which influence the organism early in life is desirable for a positive life-long health prognosis and life expectancy. Approaches for the evaluation of both nutritional and motor individualities have been elaborated as an important starting point for their positive development and eventual modification. This should aim to achieve not only prevention of diseases, but also to improve health prevention and achieving the status of "positive health".


Primary complications after surgery for cancer in the cardio-oesophageal region.
Results are presented from a study of 230 patients who during the past 20 years underwent radical surgery for cancer in the cardio-oesophageal region. Major postoperative complications arose in 94 patients and were lethal in more than half of them. Most of the complications, and the causes of death, were of pulmonary-pleural nature or peritonitis of varying genesis. Peritonitis occurred in half of the lethal cases and was mostly due to insufficiency of the anastomosis. The death rate and the incidence of primary complications were unrelated to the extent of the operation (total gastrectomy or upper partial gastrectomy). Oesophago-intestinal and oesophago-gastric anastomoses showed approximately equal frequency of insufficiency, but peritonitis was more common after total gastrectomy than after proximal resection of the stomach.


Phase I study of inhaled Doxorubicin for patients with metastatic tumors to the lungs.
To evaluate the toxicity profile of inhalational doxorubicin in patients with malignant disease in the lung. The OncoMyst Model CDD-2a inhalation device aerosolizes compounds to particles of 2 to 3 mum and prevents exhaled aerosol from escaping into the environment. Deposition efficiency of inhaled Technetium 99m was used to predict deposition of doxorubicin and calculate dose. Treatment was repeated every 3 weeks. No more than moderate pulmonary dysfunction was permitted (forced expiratory volume in 1 s, forced vital capacity, and diffusing capacity for carbon monoxide, all >50% predicted; resting SaO(2) >90%). Fifty-three patients were enrolled at 13 dose levels ranging from 0.4 to 9.4 mg/m(2). The most common histologic diagnoses were sarcoma (n = 19) and non-small cell lung cancer (n = 16). Dose-limiting toxicity (DLT) was observed at the 9.4 mg/m(2) dose level when two of four patients experienced pulmonary DLT. Of 11 patients treated at the 7.5 mg/m(2) dose level, only one showed DLT consisting of a decline in forced vital capacity of >20% from baseline. No significant systemic drug-related toxicity was observed. Several patients experienced declines in pulmonary function test variables, which were attributed to progressive disease. Observed activity included a partial response in a patient with metastatic soft tissue sarcoma previously treated with i.v. doxorubicin and ifosfamide. Inhaled doxorubicin is safe up to a dose of 7.5 mg/m(2) every 3 weeks in patients with cancer who had normal to moderately impaired pulmonary function.


Comparative Analysis of the Metabolic Profiles of Strains of Tribolium castaneum (Herbst) Adults with Different Levels of Phosphine Resistance Based on Direct Immersion Solid-Phase Microextraction and Gas Chromatography-Mass Spectrometry.
The management of phosphine (PH3) resistance in stored grain pests is an essential component of implementing timely and effective pest control strategies. The prevailing standard method for PH3 resistance testing involves the exposure of adult insects to a specific concentration over a fixed period. Although it is widely adopted, this method necessitates an extensive period for assay preparation and diagnosis. To address this issue, this study employed Direct Immersion Solid-Phase Microextraction (DI-SPME) coupled with Gas Chromatography-Mass Spectrometry (GC-MS) to compare and analyze the metabolic profiles of PH3-sensitive (TC-S), PH3 weak-resistant (TC-W), and PH3 strong-resistant (TC-SR) Tribolium castaneum (Herbst) adults. A total of 36 metabolites were identified from 3 different PH3-resistant strains of T. castaneum; 29 metabolites were found to present significant differences (p < 0.05) across these groups, with hydrocarbon and aromatic compounds being particularly prevalent. Seven metabolites showed no significant variations among the strains, consisting of four hydrocarbon compounds, two iodo-hydrocarbon compounds, and one alcohol compound. Further multivariate statistical analysis revealed a total of three, two, and nine differentially regulated metabolites between the TC-S versus TC-W, TC-S versus TC-SR, and TC-W versus TC-SR groups, respectively. Primarily, these metabolites comprised hydrocarbons and iodo-hydrocarbons, with the majority being associated with insect cuticle metabolism. This study demonstrates that DI-SPME technology is an effective method for studying differentially expressed metabolites in T. castaneum with different levels of PH3 resistance. This approach may help to provide a better understanding of the development of insect PH3 resistance and act as a valuable reference for the establishment of rapid diagnostic techniques for insect PH3 resistance.


Early antenatal care: does it make a difference to outcomes of pregnancy associated with syphilis? A systematic review and meta-analysis.
Despite an increase in the proportion of women who access antenatal care, mother-to-child transmission of syphilis continues to be a consequence of undiagnosed, untreated, or inadequately treated maternal syphilis. We reviewed evidence on the optimal timing of antenatal interventions to prevent mother-to-child transmission of syphilis and its associated adverse outcomes. Systematic review and meta-analysis of published literature. English-language articles were included if they (1) reported the gestational age at which the mother was screened or tested for syphilis; (2) reported on pregnancy outcome. No publication date limits were set. We identified a total of 1,199 publications, of which 84 were selected for further review and five were included. All showed a lower prevalence of any adverse outcome among women who received an intervention (to include screening and treatment) in the first and second trimesters of pregnancy compared to the third trimester. The overall odds ratio for any adverse outcome was 2.24 (95% CI 1.28, 3.93). All sub-analyses by type of outcome presented important heterogeneity between studies, except for those studies reporting an infected infant (odds ratio 2.92, 95% CI 0.66, 12.87; I(2) = 48.2%, p = 0.165). Our review has shown that the timing of antenatal care interventions makes a significant difference in the risk of having an adverse outcome due to syphilis. Women who sought care in the first two trimesters of their pregnancy, and received the appropriate intervention, were more likely to have a healthy infant, compared to women screened and treated in the third trimester. Encouraging ALL pregnant women to seek care in the first two trimesters of their pregnancy should be a priority for health programmes. For interventions to be effective within these health programmes, health systems and community engagement programmes need to be strengthened to enable pregnant women to seek antenatal care early.


Population genetics in Israel in the 1950s. The unconscious internalization of ideology.
This essay describes the effects of Zionist ideology on research into human population genetics carried out in Israel during the 1950s and early 1960s. I argue that the internalization of the dominant Zionist narrative is reflected in the articles that were written by Israeli geneticists and physicians during these years. My claim is based on a comparison of articles about human population genetics written and published by Israeli scientists between 1951 and 1963 with similar articles written by non-Israelis. The comparison reveals that during this period the Israeli human geneticists and physicians emphasized the sociological and historical aspects of their research and used their work, among other things, as a vehicle for establishing a national identity and confirming the Zionist narrative.


[Assessment of animal welfare aspects in extreme breeds of pet animals: principles, rules and other measures].
The review deals with fundamental aspects of the problems and the assessment of animal welfare aspects in extreme breeds of companion and pet animals, with legislation and with other measures to avoid breeding animals with extreme characteristics. Efforts are important in particular by breeding organisations to adapt breeding standards and to improve the education of judges and breeders. Furthermore adequate activities to correctly inform animal keepers are important.


The influence of noise levels on the perception of stress in heart disease patients.
To identify the main causes of stress in patients staying in a coronary unit and to assess the influence of noise levels on their perception of stress. This was a prospective, descriptive and quantitative study conducted between June and November 2009 in the Coronary Unit of the Hospital de Clínicas da Universidade Estadual de Campinas. The Intensive Care Unit Environmental Stressor Scale was used on the first, second and third days of hospitalization to identify stressors. The noise level was measured on the first and second nights using an Instrutherm DEC-460 decibel meter. Overall, 32 clinical heart disease patients were included. The median Intensive Care Unit Environmental Stressor Scale scores were 67.5, 60.5 and 59.5 for the first, second and third days, respectively. The differences were not statistically significant. The highest noise level (a median of 58.7 dB) was detected on the second night at 9:00 pm; the lowest level (51.5 dB) was measured on the first night at 12:00 am. In a multiple linear regression model, the first-night noise level had a 33% correlation with the second-day stress scale score, and for the second night, the correlation with the third-day stress scale score was 32.8% (p = 0.001). Patients admitted into a coronary unit have an increased perception of stress. Higher noise levels are also responsible for the perception of stress in these patients.


Transitions to mobility difficulty associated with lower extremity osteoarthritis in high functioning older women: longitudinal data from the Women's Health and Aging Study II.
To assess the impact of lower extremity osteoarthritis (OA) on transitions to mobility difficulty, and to assess the influence of pain, excess weight, and quadriceps strength on these transitions. We analyzed longitudinal data acquired from 199 participants in the Women's Health and Aging Study II (ages 70-79 years) who initially reported no lower extremity limitation (e.g., difficulty walking one-quarter mile) or difficulty in activities of daily living (ADL; e.g., transferring). Prevalent lower extremity OA was determined from validated algorithms encompassing multiple data sources. Markov transition models were created to analyze the first transition from no difficulty at baseline to lower extremity limitations, ADL difficulty, or both 18, 36, and 72 months later. Compared with women without OA (n = 140), a higher proportion of women with lower extremity OA (n = 59) initially reported pain on most days and more severe pain while walking (P < 0.05). Women with OA were also heavier, with a higher proportion being obese or overweight (P < 0.001). Lower extremity OA, higher body mass index, and lower knee extensor strength independently increased the risk of transition to combined lower extremity and ADL difficulty first over 72 months. Lower extremity OA increased the likelihood of developing difficulty in both lower extremity tasks and ADL over 72 months in a cohort of initially high functioning older women. Two modifiable factors, higher relative weight and lower knee extensor strength, substantially impacted these transitions, and therefore warrant increased attention in the management of lower extremity OA.


Epidemiology of respiratory syncytial virus-associated acute lower respiratory tract infection hospitalizations among HIV-infected and HIV-uninfected South African children, 2010-2011.
There are limited data on respiratory syncytial virus (RSV) infection among children in settings with a high prevalence of human immunodeficiency virus (HIV). We studied the epidemiology of RSV-associated acute lower respiratory tract infection (ALRTI) hospitalizations among HIV-infected and HIV-uninfected children in South Africa. Children aged <5 years admitted to sentinel surveillance hospitals with physician-diagnosed neonatal sepsis or ALRTI were enrolled. Nasopharyngeal aspirates were tested by multiplex real-time polymerase chain reaction assays for RSV and other viruses. Associations between possible risk factors and severe outcomes for RSV infection among HIV-infected and uninfected children were examined. The relative risk of hospitalization in HIV-infected and HIV-uninfected children was calculated in 1 site with population denominators. Of 4489 participants, 4293 (96%) were tested for RSV, of whom 1157 (27%) tested positive. With adjustment for age, HIV-infected children had a 3-5-fold increased risk of hospitalization with RSV-associated ALRTI (2010 relative risk, 5.6; [95% confidence interval (CI), 4.5-6.4]; 2011 relative risk, 3.1 [95% CI, 2.6-3.6]). On multivariable analysis, HIV-infected children with RSV-associated ALRTI had higher odds of death (adjusted odds ratio. 31.1; 95% CI, 5.4-179.8) and hospitalization for >5 days (adjusted odds ratio, 4.0; 95% CI, 1.5-10.6) than HIV-uninfected children. HIV-infected children have a higher risk of hospitalization with RSV-associated ALRTI and a poorer outcome than HIV-uninfected children. These children should be targeted for interventions aimed at preventing severe RSV disease.


Creative leasing strategies for medical office buildings.
The financing and ownership of medical office buildings have changed along with their design and function. Once merely a convenience for the medical staff, medical office buildings are becoming an increasingly important part of a hospital's marketing and financial strategies. Understanding the economic evolution of these buildings will help hospital financial managers approach their development in a manner that will meet the needs of the future.


[Antisera to individual histone fractions of the calf thymus. II. A study of the immunochemical specificity of the histones by an indirect immunofluorescence method].
Immune antisera to 5 fractions (H1, H2a, H2b, H3, H4) of calf thymus histone were assayed using indirect immunofluorescence (IIF). The analysis of such sera by this technique, as well as the data on complement fixation obtained previously, show that these antisera are highly active and specific for various test-objects: thymys, liver nuclei of rat, chicken, and calf, chicken erythrocytes, metaphase chromosomes of mouse fibroblasts. These antisera are of importance for the evaluation of species- and tissue-specificity of different histone fractions. Using the IIF reaction, a comparison was made between the nucleosome fraction H3, which is evolutionary stable, and fraction HI from calf thymus, rat and chicken liver, and chicken erythrocytes.


An AP-1 binding site is the predominant cis-acting regulatory element in the 1.2-kilobase early RNA promoter of human cytomegalovirus.
To extend our analysis of the regulation of human cytomegalovirus (HCMV) early gene expression, we examined a transcription unit located in the terminal repeats of the long segment of the viral genome. This region encodes a major 1.2-kb RNA which is induced at early times in infection but undergoes its largest increase in abundance after the onset of viral DNA replication. To identify the important cis-acting regulatory elements for this gene, two constructs were prepared for use in transient expression assays. One contained 413 bp of the upstream sequence and 43 bp of the leader sequence fused to the gene for chloramphenicol acetyltransferase (CAT). The second construct included 1,722 bp upstream of the start site of the 1.2-kb RNA, the entire transcribed region with an additional 166-bp insert derived from the CAT gene as an assayable marker, and 2,393 bp downstream of the polyadenylation signal. Both constructs were individually transfected into human fibroblast cells, and the cells were infected with HCMV. RNA specified by the hybrid construct was initiated at the correct position and accumulated with the same kinetics as the authentic viral transcript at early times in the infection but did not undergo the increase in abundance at late at late times. By 5'-end-deletion analysis, we determined that the promoter for the 1.2-kb RNA contains a number of cis-acting elements, the most significant of which are the TATA-like sequence CATAAA at -30 and a sequence corresponding to the binding site for the transcription factor AP-1 at -75. Using extracts prepared from HeLa cells as well as from infected and uninfected fibroblasts in gel retardation assays, we obtained evidence for the specific interaction of a cellular factor(s) with the AP-1 binding site. The pattern of binding differed in the HeLa and fibroblast cells but did not change as a function of the HCMV infection. However, the functional importance of the AP-1 binding site and its key role in the regulation of the 1.2-kb RNA was supported by analysis of constructs containing specific point mutations at this site in gel retardation and transient expression assays. Site-specific mutations in the AP-1 consensus sequence, which resulted in the complete loss of binding to cellular factors, eliminated the basal activity and reduced the inducible promoter activity by eightfold.


Antiretroviral neurotoxicity.
Combination antiretroviral therapy (CART) has proven to effectively suppress systemic HIV burden, however, poor penetration into the central nervous system (CNS) provides incomplete protection. Although the severity of HIV-associated neurocognitive disorders (HAND) has been reduced, neurological disease is expected to exert an increasing burden as HIV-infected patients live longer. Strategies to enhance penetration of antiretroviral compounds into the CNS could help to control HIV replication in this reservoir but also carries an increased risk of neurotoxicity. Efforts to target antiretroviral compounds to the CNS will have to balance these risks against the potential gain. Unfortunately, little information is available on the actions of antiretroviral compounds in the CNS, particularly at concentrations that provide effective virus suppression. The current studies evaluated the direct effects of 15 antiretroviral compounds on neurons to begin to provide basic neurotoxicity data that will serve as a foundation for the development of dosing and drug selection guidelines. Using sensitive indices of neural damage, we found a wide range of toxicities, with median toxic concentrations ranging from 2 to 10,000 ng/ml. Some toxic concentrations overlapped concentrations currently seen in the CSF but the level of toxicity was generally modest at clinically relevant concentrations. Highest neurotoxicities were associated with abacavir, efavarenz, etravirine, nevaripine, and atazanavir, while the lowest were with darunavir, emtracitabine, tenofovir, and maraviroc. No additive effects were seen with combinations used clinically. These data provide initial evidence useful for the development of treatment strategies that might reduce the risk of antiretroviral neurotoxicity.


Distribution of fiber types determined by in situ hybridization of myosin heavy chain mRNA and enzyme histochemistry in rat skeletal muscles.
We analyzed fiber types in rat skeletal muscles using a novel combination of in situ hybridization of myosin heavy chain (MyHC) mRNA, and enzyme histochemistry for succinate dehydrogenase (SD), which displayed metabolic properties. The fiber types were classified into the four major subtypes of I(beta/slow), IIA, IIX and IIB, and their intermediate types coexpressed two MyHC mRNAs: I and IIA, IIA and IIX, or IIX and IIB. The distribution of fiber types differed markedly in each skeletal muscle. The superficial region of limb muscles was composed mainly of fast-twitch fibers with oxidative-glycolytic and glycolytic activities, such as type IIX and type IIB. In contrast, the deep region was composed almost exclusively of type I and type IIA fibers both with oxidative activity. In this region, type IIA/IIX hybrid fibers were noted more frequently than type I/IIA and IIX/IIB hybrid fibers. In axial muscles, slow-twitch fibers and fast-twitch fibers composed predominantly of type IIB were distributed dispersively. The diaphragm and masseter showed a high proportion of type IIX and type IIB, respectively, to adapt to tissue-specific functional requirements and more frequently contained type IIX/IIB hybrid fibers than other observed muscles.


Evaluation of the Social Participation Questionnaire in adult patients with depressive symptoms using Rasch analysis.
The aim of this study was to explore the psychometric properties of the 22-item Social Participation Questionnaire (SPQ). The SPQ was administered to 789 adult primary care patients with depressive symptoms. As the items were intended to be summed together to provide total score, Rasch analysis (partial credit model) was applied to assess the overall fit of the model, individual item fit, differential item functioning (DIF), targeting of persons, response dependency, unidimensionality and person separation. To improve the scale's fit, it was necessary to re-score the response format. Two items demonstrated some DIF for gender and eight items showed DIF for age. To support the assumption of unidimensionality post hoc principal component analysis was performed. The analysis showed two subtests of the residuals with positive and negative loadings, but the person estimates derived from these two subtests were not statistically different to that derived from all items taken together. The response dependence between two items was identified; however, the magnitude of difficulty was very small. Although the questionnaire appeared to have insufficient items to assess the full spectrum of informal social contact, the SPQ was reasonably well targeted. The SPQ is a promising questionnaire for the measurement of social participation although it could benefit from the inclusion of further items to measure informal social contact. This study found support for the internal validity, internal consistency reliability, and unidimensionality. A future study will investigate whether targeting can be improved when additional items are included.


Ultrasonic characterization of delamination in aeronautical composites using noncontact laser generation and detection.
The characterization of delamination in composite plates with ultrasonic waves generated and detected by lasers is presented. Composite materials have become one of the most important structural materials in the aviation industry because of their excellent mechanical properties, such as high specific stiffness and antifatigue. This paper reports a new application of the laser ultrasonic technique to perform nondestructive detection of carbon-fiber-reinforced plastic (CFRP) and continuous-fiber-reinforced ceramic matrix composites (CFCCs) containing artificial internal defects, based on propagation characteristic of ultrasonic waves generated by pulse laser with a wavelength of 1064 nm and pulse duration of 10 ns. A laser interferometer based on two-wave mixing is used to measure ultrasonic wave signals. The main advantage of this technique over conventional ultrasonic testing techniques is the ability to carry out detection without using coupling agents. The research results prove that the laser ultrasonic technique is effective for the detection of internal defects in both CFRP and CFCC composite components, which should promote and expand the application of the technique in the aviation industry.


Mint3-depletion-induced energy stress sensitizes triple-negative breast cancer to chemotherapy via HSF1 inactivation.
Given the lack of therapeutic targets, the conventional approach for managing triple-negative breast cancer (TNBC) involves the utilization of cytotoxic chemotherapeutic agents. However, most TNBCs acquire resistance to chemotherapy, thereby lowering the therapeutic outcome. In addition to oncogenic mutations in TNBC, microenvironment-induced mechanisms render chemoresistance more complex and robust in vivo. Here, we aimed to analyze whether depletion of Munc18-1 interacting protein 3 (Mint3), which activates hypoxia-inducible factor 1 (HIF-1) during normoxia, sensitizes TNBC to chemotherapy. We found that Mint3 promotes the chemoresistance of TNBC in vivo. Mint3 depletion did not affect the sensitivity of human TNBC cell lines to doxorubicin and paclitaxel in vitro but sensitized tumors of these cells to chemotherapy in vivo. Transcriptome analyses revealed that the Mint3-HIF-1 axis enhanced heat shock protein 70 (HSP70) expression in tumors of TNBC cells. Administering an HSP70 inhibitor enhanced the antitumor activity of doxorubicin in TNBC tumors, similar to Mint3 depletion. Mint3 expression was also correlated with HSP70 expression in human TNBC specimens. Mechanistically, Mint3 depletion induces glycolytic maladaptation to the tumor microenvironment in TNBC tumors, resulting in energy stress. This energy stress by Mint3 depletion inactivated heat shock factor 1 (HSF-1), the master regulator of HSP expression, via the AMP-activated protein kinase/mechanistic target of the rapamycin pathway following attenuated HSP70 expression. In conclusion, Mint3 is a unique regulator of TNBC chemoresistance in vivo via metabolic adaptation to the tumor microenvironment, and a combination of Mint3 inhibition and chemotherapy may be a good strategy for TNBC treatment.


Comparative immunogenicity of two bivalent botulinum vaccines.
To compare the ability of a new single-dose botulinum vaccine containing a non-mineral oil adjuvant with a single dose of a conventional botulinum vaccine product to produce antibody to Clostridium botulinum types C and D in cattle in Northern Australia. One hundred and fifty Brahman steer weaners were randomly divided into two groups receiving either a single dose of CSL Bivalent Botulinum vaccine or Websters Singvac. Blood samples were collected at 0, 8 and 24 weeks and tested by antibody ELISA. The final samples were also tested by the toxin neutralisation test, to test titres of neutralising antibody. Six months after inoculation, cattle vaccinated with Websters Singvac had ELISA antibody response twice that of CSL conventional product. However, this difference was only evident for neutralising antibody to type C botulinum toxin. Both products produced similar titres of type D neutralising antibody after a single dose. Websters' Singvac produces a greater neutralising antibody response to type C botulism upon single inoculation than a conventional vaccine. The product produces an equivalent neutralising antibody response to type D.


Effects of short versus long bouts of aerobic exercise in sedentary women with fibromyalgia: a randomized controlled trial.
The purposes of this study were: (1) to assess the effectiveness of a 16-week progressive program of home-based, videotape-based, low-impact aerobic exercise on physical function and signs and symptoms of fibromyalgia in previously sedentary women aged 20 to 55 years and (2) to compare the effects of 1 long exercise bout versus 2 short exercise bouts per training day (fractionation) on physical function, signs and symptoms of fibromyalgia, and exercise adherence. One hundred forty-three sedentary women were randomly assigned to 1 of 3 groups: a group who trained using a long bout of exercise (LBE group, n=51), a group who trained using short bouts of exercise (SBE group, n=56), and a group who performed no exercise (NE group, n=36). The SBE group exercised twice daily, and the LBE group worked out once daily. Both groups progressed in total daily training duration from 10 to 30 minutes, 3 to 5 times a week, for 16 weeks. Physical and psychological well-being, symptoms, and self-efficacy were evaluated using a multivariate analysis of variance. Dropout rates for the NE, SBE, and LBE groups were 14%, 38%, and 29%, respectively. The NE group differed from the LBE group in disease severity, self-efficacy, and psychological well-being (midtest, efficacy analysis) and from the SBE group in disease severity and self-efficacy (posttest, efficacy analysis). Exercise adherence was greater for the LBE group than for the SBE group between weeks 5 and 8 of the training program. No other differences between exercise groups were found. Progressive, home-based, low-impact aerobics improved physical function and fibromyalgia symptoms minimally in participants who completed at least two thirds of the recommended exercise. Fractionation of exercise training provided no advantage in terms of exercise adherence, improvements in fibromyalgia symptoms or physical function. High attrition rates and problems with exercise adherence were experienced in both exercise groups.


[What is the most important factor for gastric carcinogenesis in Koreans: Helicobacter pylori, host factor or environmental factor?].
Epidemiological data including our studies demonstrated the association between Helicobacter pylori (H. pylori) infection and gastric cancer. However, this significant clinical outcome happens only in a small portion of infected person. This suggests that other contributors including host genetic and environmental factors might be involved in the disease process. Studies on the association between virulent strains of H. pylori and clinical outcomes failed to show significant results in Korea. Cytokine gene polymorphism such as interleukin-1 (IL-1) has been thought to play a role in gastric carcinogenesis. Our studies showed the controversial role of IL-1, TNF-A, IL-10 and IL-2 gene polymorphisms in the development of gastric cancer in Korea. Chronic infection and inflammation leading to tumorigenesis are mediated in part through the recognition of various stimuli by toll-like receptors (TLRs). Our studies on the polymorphisms of TLR4 and TLR2 showed no mutant form in Koreans. These discrepancies might reflect the genetic differences between Caucasians and Koreans or might be due to prevalent genetic polymorphisms with masked effect in gastric carcinogenesis in Koreans. As other candidate risk factors, there are constant or inconsistent results on the effect of dietary intake in gastric cancer. There are numerous similar risk for gastric carcinogenesis with different risk ratio including environmental factors in Caucasians and Koreans. Under the background of prevalent H. pylori infection and genetic polymorphisms, environmental factors including diet may potentiate their role in gastric carcinogenesis in Koreans.


Impact of an exercise program on adherence and fitness indicators.
Adherence to exercise is one of the most problematic health behaviors. This pilot study describes the impact of an exercise program on adherence to exercise and fitness indicators for overweight and obese adults enrolled in an insurance reimbursed exercise plan. Chart reviews were conducted retrospectively in a convenience sample of 77 subjects from a human performance lab (HPL) at a large southern university. Charts from 2004 to 2009 were reviewed for health history, fitness indicators (fitness level, weight, BMI, hip/waist ratio, % body fat, BP, HR, cholesterol), and adherence (number of exercise sessions/month). Exercise supervision was operationalized in two phases over 12 months: Phase I (3 months supervised exercise) and Phase II (9 months unsupervised exercise). Fifty-eight participants completed Phase I, and 8 completed Phase II. Six-nine percent of those completing Phase I visited the gym at least 8 times/month with significant (α=.05) improvement in all fitness indicators. Those visiting <8 times/month had improvement in fitness level, weight, BMI, and % body fat. Twenty-four subjects continued into Phase II, with only eight completing Phase II. Of those eight, only one subject visited the HPL at least 8 times/month. Health history data including co-morbidities, symptoms, habits, perceived tension, job stress, and fitness level were not associated with adherence. Symptoms of swollen, stiff, painful joints, and swollen ankles and legs were associated with decreased adherence to exercise. Supervised exercise was positively related to adherence and improved fitness indicators. Adults with joint symptoms may require more support. Based on these pilot data, designing a study with a larger sample and the inclusion of barriers and facilitators for adherence to self-directed exercise would allow additional analysis. Innovative interventions are needed that mimic the supervised environment, shifting responsibility for the exercise plan from the supervisor to those exercising.


Referencing: the cohesive use of pronouns in the written narrative of college underprepared writers, nondisabled writers, and writers with learning disabilities.
The purpose of this study was to explore whether college normally achieving writers (n = 35), writers with learning disabilities (n = 35), and underprepared writers (n = 35) demonstrated any differences in their ability to utilize the semantic roles and syntactic rules needed to apply cohesive referencing in written text. Cohesive referencing (the ability to assign roles to the speaker and addressee in written communication) is a critical aspect of written language. In addition, the study explored the relation between referential cohesion and the quality and coherence of a written text.


Neuronal Branching is Increasingly Asymmetric Near Synapses, Potentially Enabling Plasticity While Minimizing Energy Dissipation and Conduction Time.
Neurons' primary function is to encode and transmit information in the brain and body. The branching architecture of axons and dendrites must compute, respond, and make decisions while obeying the rules of the substrate in which they are enmeshed. Thus, it is important to delineate and understand the principles that govern these branching patterns. Here, we present evidence that asymmetric branching is a key factor in understanding the functional properties of neurons. First, we derive novel predictions for asymmetric scaling exponents that encapsulate branching architecture associated with crucial principles such as conduction time, power minimization, and material costs. We compare our predictions with extensive data extracted from images to associate specific principles with specific biophysical functions and cell types. Notably, we find that asymmetric branching models lead to predictions and empirical findings that correspond to different weightings of the importance of maximum, minimum, or total path lengths from the soma to the synapses. These different path lengths quantitatively and qualitatively affect energy, time, and materials. Moreover, we generally observe that higher degrees of asymmetric branching- potentially arising from extrinsic environmental cues and synaptic plasticity in response to activity- occur closer to the tips than the soma (cell body).


Toward a unifying taxonomy and definition for meditation.
One of the well-documented concerns confronting scholarly discourse about meditation is the plethora of semantic constructs and the lack of a unified definition and taxonomy. In recent years there have been several notable attempts to formulate new lexicons in order to define and categorize meditation methods. While these constructs have been useful and have encountered varying degrees of acceptance, they have also been subject to misinterpretation and debate, leaving the field devoid of a consensual paradigm. This paper attempts to influence this ongoing discussion by proposing two new models which hold the potential for enhanced scientific reliability and acceptance. Regarding the quest for a universally acceptable taxonomy, we suggest a paradigm shift away from the norm of fabricatIng new terminology from a first-person perspective. As an alternative, we propose a new taxonomic system based on the historically well-established and commonly accepted third-person paradigm of Affect and Cognition, borrowed, in part, from the psychological and cognitive sciences. With regard to the elusive definitional problem, we propose a model of meditation which clearly distinguishes "method" from "state" and is conceptualized as a dynamic process which is inclusive of six related but distinct stages. The overall goal is to provide researchers with a reliable nomenclature with which to categorize and classify diverse meditation methods, and a conceptual framework which can provide direction for their research and a theoretical basis for their findings.


Determination of sialic acid levels by using surface-enhanced Raman spectroscopy in periodontitis and gingivitis.
To compare the sialic acid (SA) levels in saliva among periodontitis-affected, gingivitis and control patients. The study involved 93 subjects. The participants were divided into three groups: (1) 30 subjects without periodontal disease (control group); (2) 30 subjects with gingivitis; and (3) 33 subjects with periodontitis. The oral parameters examined were as follows: (a) Simplified Oral Hygiene Index; (b) Calculus Index; (c) Gingival Index; (d) probing pocket depth; and (e) level of epithelial attachment. SA levels in saliva were measured by means of surface-enhanced Raman spectroscopy (SERS). This method has demonstrated the capacity to detect extremely low concentrations of molecules. The spectrum was calibrated using analytical reagent SA. The obtained median values for SA concentrations were 5.98, 7.32, and 17.12 mg/dl for control, gingivitis, and periodontitis patients, respectively. Our measurements by SERS corroborate that in periodontitis-affected patients, the SA concentration is larger than their concentrations in either control or gingivitis patients. This confirms previous reports and opens the possibility of using SERS as a diagnostic tool.


Schisandra Chinensis Baillon regulates the gene expression of phase II antioxidant/detoxifying enzymes in hepatic damage induced rats.
This study investigated the antioxidant activities and hepatoprotective effects of Schisandra chinensis Baillon extract (SCE) against tert-butyl hydroperoxide (t-BHP)-induced oxidative hepatic damage in rats. Sprague-Dawley (SD) rats were pretreated with SCE (300, 600, and 1,200 mg/kg BW) or saline once daily for 14 consecutive days. On day 14, each animal, except those belonging to the normal control group, were injected with t-BHP (0.8 mmol/kg BW/i.p.), and all of the rats were sacrificed 16 h after t-BHP injection. Although no significant differences in AST and ALT levels were observed among the TC and SCE groups, the high-dose SCE group showed a decreasing tendency compared to the TC group. However, erythrocyte SOD activity showed a significant increase in the low-dose SCE group compared with the TC group. On the other hand, no significant differences in hepatic total glutathione (GSH) level, glutathione reductase (GR), and glutathione peroxidase (GSH-Px) activities were observed among the TC and SCE groups. Hepatic histopathological evaluation revealed that pretreatment with SCE resulted in reduced t-BHP-induced incidence of lesions, such as neutrophil infiltration, swelling of liver cells, and necrosis. In particular, treatment with a high dose of SCE resulted in induction of phase II antioxidant/detoxifying enzyme expression, such as glutathione S-transferase (GST) and glutamate-cysteine ligase catalytic subunit (GCLC). Based on these results, we conclude that SCE exerts protective effects against t-BHP induced oxidative hepatic damage through the reduction of neutrophil infiltration, swelling of liver cells, and necrosis. In addition, SCE regulates the gene expression of phase II antioxidant/detoxifying enzymes independent of hepatic antioxidant enzyme activity.


Molecular tumor targeting of gelonin by fusion with F3 peptide.
Therapeutically potent macromolecular drugs have shown great promise for overcoming the limitations of small-molecule anti-cancer drugs. But tumor cell-selective intracellular delivery of the macromolecules remains a major hurdle for their successful clinical application. To overcome this challenge, we engineered a novel genetic fusion protein (F3-Gel) that composed of F3 peptide, a tumor-homing peptide, and gelonin, a plant-derived ribosome-inactivating protein (RIP), and then evaluated its anti-cancer activity in vitro and in vivo. The F3-Gel-encoding gene was synthesized by genetic recombination, and F3-Gel was successfully expressed in E coli. The anti-cancer activity of the produced F3-Gel was evaluated by various in vitro assays, which revealed that F3-Gel maintained equipotent protein synthesis inhibition activity (IC50=11 pmol/L) as unmodified gelonin (IC50=10 pmol/L). Furthermore, F3-Gel displayed enhanced cellular uptake into cancer cells (U87 MG, HeLa, LnCaP and 9L) than noncancerous cells (293 HEK and SVGp12). Compared with gelonin, F3-Gel exerted significantly higher cytotoxicity against these cancer cells. F3-Gel displayed significantly greater inhibition of protein translation in U87 MG cells: F3-Gel (0.5 μmol/L) was able to reduce the protein level to less than 50%, while gelonin (1 μmol/L) did not affect the intracellular protein level. In a U87 MG xenograft tumor-bearing mouse model, F3-Gel was accumulated in the tumor site at much higher levels and maintained for a prolonged time compared with gelonin. Administration of F3-Gel (0.5, 0.75 mol/kg, iv) caused 36% and 66%, respectively, inhibition of tumor growth in U87 MG xenograft mice, suggesting that it is a promising candidate drug for cancer treatment. Furthermore, this study demonstrates that fusion of F3 peptide to a potent macromolecule could provides an effective method for targeting tumors and eventually could improve their druggability.


Routine assessment of on-clopidogrel platelet reactivity and gene polymorphisms in predicting clinical outcome following drug-eluting stent implantation in patients with stable coronary artery disease.
This study sought to assess the usefulness of clopidogrel-pathway genotyping and on-treatment platelet reactivity (OTR) testing in predicting major adverse cardiac events (MACE) in stable coronary artery disease (CAD) patients receiving drug-eluting stents (DES) under dual antiplatelet (clopidogrel plus aspirin) therapy. The role of pharmacogenetics and OTR in predicting MACE-death, myocardial infarction, or stent thrombosis-in stable CAD patients scheduled for DES implantation is still debated. Patients with stable CAD treated by DES implantation (n = 1,432) were genotyped with a TaqMan OpenArray (Applied Biosystems, Carlsbad, California) and assessed for OTR with the VerifyNow P2Y12 test (Accumetrics Inc., San Diego, California). Genes tested were ABCB1, CYP1A2, CYP2B6*9, CYP2C8*3, CYP2C9*2, CYP2C19, CYP3A4, CYP3A5*3, P2RY12, and PON1CYP2C19. High OTR was defined as P2Y12 reaction units ≥230. The endpoint at 12-month follow-up was MACE occurring during antiplatelet therapy. All groups that were stratified for loss-of-function variants of the cytochrome P450 gene CYP2C19 had significant hazard ratios (HR) for MACE (genotypic HR: 1.41, 95% confidence interval [CI]: 1.06 to 1.89, p = 0.01; allelic HR: 1.56, 95% CI: 2.26 to 1.2, p = 0.01). Variants of other clopidogrel-pathway genes were not significantly associated with MACE. When OTR was assessed, clinical significance was found only in high-risk diabetic (HR: 2.11, 95% CI: 1.29 to 3.45, p < 0.001) and chronic kidney disease (HR: 2.03, 95% CI: 1.03 to 4.02, p = 0.04) patients. CYP2C19 metabolizer status is an independent predictor of MACE after DES implantation and can be used for prognostication in all stable CAD patients. High OTR, as assessed by the VerifyNow P2Y12 test, is an independent predictor of MACE only for high-risk subsets, that is, patients with diabetes or chronic kidney disease.


Motility changes associated with large bowel obstruction and its surgical relief.
Strain gauges and electrodes to detect intestinal motility were attached to the terminal ileum, right and left sides of the colon in Rhesus monkeys. Baseline recordings were made. An obstruction device was applied to the left colon at a laparotomy. Compromise of the bowel lumen leading to total obstruction occurred at 13 +/- 2 days. As obstruction progressed, contractions decreased in the terminal ileum and right colon but increased in the left colon proximal to the obstruction. Distention wave forms increased in the colon proximal to the obstruction. Visible intestinal movement, borborygmi and signs suggesting colic were accompanied by a motility phenomenon, simultaneous mass actions, not seen in normal unobstructed bowel. The obstructed segment was resected and an anastomosis made. The motility recordings revealed that postoperative ileus persisted for less than 48 hours and motility returned to normal significantly earlier than after a control laparotomy.


An evaluation of factors associated with sexual risk taking among Black men who have sex with men: a comparison of younger and older populations.
In the United States, rates of human immunodeficiency virus (HIV) infection are highest among Black men who have sex with men (BMSM). Prior research indicates that younger BMSM in particular (i.e., BMSM 29 years of age and younger) are most at risk for HIV infection, and that HIV incidence in this subpopulation has risen in recent years. It remains unclear, however, why younger BMSM, relative to BMSM 30 years of age and older, are at increased risk for HIV infection. For the current study, we surveyed 450 BMSM located in the Atlanta, GA metropolitan and surrounding areas. We assessed BMSM's depressive symptoms, substance use during sex, psycho-social risk factors (i.e., HIV risk perceptions, condom use self-efficacy, internalized homophobia, and perceived HIV stigmatization), and sexual risk taking (i.e., condomless anal intercourse [CAI]). We found that younger BMSM (YBMSM) and older BMSM (OBMSM) differed with respect to factors associated with CAI. In multivariable models, alcohol use before or during sex, lower educational attainment, and sexual orientation (i.e., bisexual sexual orientation) were significantly associated with increased CAI for YBMSM, while HIV risk perceptions and internalized homophobia were significantly, negatively associated with CAI among OBMSM. Rates of engaging in CAI were similar across the two age cohorts; however, factors related to CAI varied by these two groups. Findings emphasize the need to consider targeted interventions for different generational cohorts of BMSM.


Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822T.
Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822T (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104. After production and purification, the protease showed a molecular mass of 27.57 kDa and a pI of 5.8. SPFA displayed hydrolytic activity at a temperature optimum of 80 °C and a very broad pH optimum between 8.5 and 11.5, with high activity up to pH 12.5. SPFA displayed no NaCl dependence but a high NaCl tolerance, with decreasing activity up to concentrations of 5 m NaCl. The stability enhanced with increasing NaCl concentration. Based on its substrate preference for 10 synthetic peptide 4-nitroanilide substrates with three or four amino acids and its phylogenetic classification, SPFA can be assigned to the subgroup of true subtilisins. Moreover, SPFA exhibited high tolerance to 5% (w/v) SDS and 5% H2 O2 (v/v). The biochemical properties of SPFA, especially its tolerance of remarkably high pH, SDS and H2 O2 , suggest it has potential for biotechnological applications.


PAX8 Expression in Solitary Fibrous Tumor: A Potential Diagnostic Pitfall.
PAX8 is used as a diagnostic aid in classifying retroperitoneal (RP) spindle cell tumors. PAX8 positivity in a spindled RP tumor is typically associated with sarcomatoid renal cell carcinoma (SRCC). However, PAX8 expression in solitary fibrous tumor (SFT), a tumor not uncommon to the RP, has not been extensively studied. We investigated the expression of PAX8 in SFTs and other spindle cell RP tumors. We collected 30 SFT, 23 SRCC, 11 gastrointestinal stromal tumors, 2 synovial sarcomas, 6 dedifferentiated liposarcomas (DDLS), 4 well differentiated liposarcomas (WDLS), and select other tumors. We identified nuclear PAX8 expression in 13 of 30 (43%) SFT, 0 of 6 (0%) DDLS, and 1 of 4 (25%) WDLS. Twenty-eight of 30 (93%) SFT, 0 of 23 (0%) SRCC, 2 of 6 (33%) DDLS, and 1 of 4 (25%) WDLS showed nuclear STAT6 expression. All gastrointestinal stromal tumors were negative for both PAX8 and STAT6. Of the 13 SFT showing PAX8 expression, 8 showed diffuse expression and 5 expressed PAX8 focally. Extrapleural SFTs were more likely to express PAX8 compared with pleural SFTs (10/13; 77% vs. 3/17; 18%, respectively; P=0.00117). Twenty of 23 (87%) SRCC expressed PAX8; the sarcomatoid component of all 23 SRCC was negative for STAT6. Of the other spindle cell tumors studied, 1 of 2 synovial sarcomas and 1 of 2 histiocytic sarcomas showed PAX8 expression. Pathologists should be aware of the potential pitfall of the relatively frequent expression of PAX8 by SFT and STAT6 expression in liposarcoma. PAX8 expression by a spindle cell lesion of RP would not allow distinction between SFT, SRCC, or sclerosing liposarcoma by itself. A STAT6/PAX8 phenotype excludes SRCC.


International Headache Society headache diagnostic patterns in pain facility patients.
Previous studies have indicated that many patients with chronic pain (PWCP) referred to pain facilities for the treatment of neck and/or low back pain complain of associated headaches. The purpose of this study was to characterize the nature of these headaches according to International Headache Society (IHS) headache diagnostic criteria. In preparation for this study, a questionnaire that reflected IHS headache diagnostic criteria was developed. All consecutive patients admitted to our pain facility complaining of headache completed this questionnaire and received a physical and neurologic examination focused on key aspects of headache. A headache interview was also conducted, using the questionnaire as a question guide. All questionnaires were entered in a computerized database, and IHS diagnoses were arrived at for each patient. As many IHS diagnoses as possible were assigned to each PWCP as long as IHS criteria were fulfilled. In addition, a frequency distribution for headache precipitants and neck-associated symptoms was developed and evaluated by discriminant analysis to determine the diagnostic value of these factors in relation to each IHS diagnostic group. Pain facility (multidisciplinary pain center). Consecutive PWCP. Of 1,466 PWCP, 154 (10.5%) were identified as suffering from severe headache interfering with function. Of these, 55.8% indicated that their headaches were related to an injury for which they were seeking treatment and 83.7% had neck pain. Migraine headache represented the most common diagnostic group (90.3%), with cervicogenic headache representing the second most common (33.8%). Of the total group, 44.2% had more than one headache diagnosis, that is, there was overlap. Cervicogenic headache patients had the greatest percentage of overlap (94.2%), with migraine patients being second (68.3%). The most frequent headache precipitant was mental stress, followed by neck position and activity/exercise. The migraine and cervicogenic headache groups had a statistically significant greater number of neck-associated symptoms when compared with the remaining patients. Of the total headache group, 74.6% complained that they had a tender point at the back of their neck. Cervicogenic, migraine, and tension PWCP had the greatest frequency of head or neck tender points. The discriminant analysis for neck-associated symptoms yielded the following symptoms as the most common predictors of headache across IHS diagnostic groups: clues to onset were severe headache beginning at the neck or tender point and numbness in arms and legs; headache brought on by neck position and arms overhead; and neck symptoms consisting of a tender point in the neck and feeling severe headache in the neck. Headache can and should be considered a frequent comorbid condition in PWCP. Because of the overlap data, more precise diagnostic criteria may be required to separate cervicogenic headache from migraine headache. Neck-associated symptoms seem to be important even to those PWCP diagnosed with migraine headache.


Molecular dynamics simulation of composite nanochannels as nanopumps driven by symmetric temperature gradients.
In this Letter, we propose a composite nanochannel system, where half of the channel is of low surface energy, while the other half has a relatively high surface energy. Molecular dynamics simulations show that fluids in such channels can be continuously driven by a symmetric temperature gradient. In the low surface energy part, the fluid moves from high to low temperature, while the fluid migrates from low to high temperature in the high surface energy part. The mechanisms that govern the flow are explained and the conditions required to guarantee the flow and the possible applications are discussed.


Leptospirosis in Malaysia: current status, insights, and future prospects.
Among zoonotic infections, leptospirosis has a worldwide distribution and high prevalence in tropical regions. It has a broad clinical presentation from mild to severe, life-threatening infection. Leptospires, the etiological agent of leptospirosis, are found in varied ecological niches and animal species, providing a significant source of human infection. This review aims to provide the current status of leptospirosis in Malaysia and the direction for future studies. The literature search for this review was performed using PubMed, Web of Sciences, and Google Scholar databases. The incidence of leptospirosis in Malaysia from 2004 to 2020 varied; however, a large number of cases occurred during floods. Leptospira has been isolated from wild and domestic animals as well as from the environment; among them, several novel species have been identified. In Malaysia, leptospirosis infection and death were mostly associated with recreational and non-recreational water activities. Despite the endemicity of leptospirosis, the public's knowledge, attitude, and practice level are relatively low in this country. More studies are needed in Malaysia to explore the extent of leptospirosis in different settings and locations.


Residual transglutaminase in collagen - effects, detection, quantification, and removal.
In the present study, we developed an enzyme-linked immunosorbent assay (ELISA) for microbial transglutaminase (mTG) from Streptomyces mobaraensis to overcome the lack of a quantification method for mTG. We further performed a detailed follow-on-analysis of insoluble porcine collagen type I enzymatically modified with mTG primarily focusing on residuals of mTG. Repeated washing (4 ×) reduced mTG-levels in the washing fluids but did not quantitatively remove mTG from the material (p < 0.000001). Substantial amounts of up to 40% of the enzyme utilized in the crosslinking mixture remained associated with the modified collagen. Binding was non-covalent as could be demonstrated by Western blot analysis. Acidic and alkaline dialysis of mTG treated collagen material enabled complete removal the enzyme. Treatment with guanidinium chloride, urea, or sodium chloride was less effective in reducing the mTG content.


Antirecoverin autoantibodies in the patient with non-small cell lung cancer but without cancer-associated retinopathy.
The goal of the present study was to analyze serum and tumor tissue of a patient with non-small cell lung cancer (NSCLC) for the presence of autoantibodies against recoverin (anti-Rc) and recoverin expression, correspondingly. Using immunoblotting with recombinant recoverin as an antigen, we have detected anti-Rc in serum of the patient. At the same time, the patient did not manifest any signs of cancer-associated retinopathy (CAR). Polyclonal (monospecific) antibodies against recoverin used for immunohistochemical analysis of the patient's tumor revealed recoverin expression in the tumor sections. To our knowledge, this is the first case of the presence of serum anti-Rc in NSCLC patients in the absence of paraneoplastic retina degeneration.


Leptin levels are not affected by enalapril treatment after an uncomplicated myocardial infarction, but associate strongly with changes in fibrinolytic variables in men.
Leptin, an adipocyte-derived hormone, is involved in the regulation of body weight and is associated with obesity-related complications, notably cardiovascular disease (CVD). A putative link between obesity and CVD could be induction of plasminogen activator inhibitor-1 (PAI-1) synthesis by leptin. In this study, we hypothesized that the beneficial effect of the angiotensin-converting enzyme inhibitor (ACEi) enalapril on PAI-1 levels is mediated by effects on leptin levels. The association between leptin and components of the fibrinolytic system was evaluated in a non-prespecified post hoc analysis of a placebo-controlled randomized, double-blind trial where the effect of the ACEi enalapril on fibrinolysis was tested. A total of 46 men and 37 women were randomized to treatment with enalapril or placebo after (median 12 months) an uncomplicated myocardial infarction. At baseline, the participants were stable and had no signs of congestive heart failure. Leptin and fibrinolytic variables (mass concentrations of PAI-1, tissue plasminogen activator (tPA) and tPA-PAI complex) were measured at baseline, and after 10 days, 6 months and 12 months. Enalapril treatment did not change leptin levels, which increased significantly during 1 year of follow-up (p = .007). Changes in leptin levels were strongly associated with changes of tPA mass (p = .001), tPA-PAI complex (p = .003) and of PAI-1 (p = .006) in men, but not in women. Leptin levels are not influenced by treatment with an ACEi. In contrast, leptin associates strongly with changes in fibrinolytic variables notably with a sex difference, which could be of importance for obesity-related CVD.


Measurement of vasoactivity in the guinea-pig choroid.
A perfusion system for studying the vasoactive properties of the guinea-pig choroid is described. The principle of operation is that the vascular resistance of the entire vascular network of an isolated, perfused eye can be monitored by recording the pressure required to deliver a constant flow of perfusate through the network. Delivery of the pharmacological agent of interest into the perfusate stream and the subsequent determination of the magnitude of any induced pressure changes allows the vasoactive potency of various agonists to be assessed. The baseline vascular resistance was 1.35 +/- 0.16 mmHg min/microL (mean +/- SEM; n = 10) and the mean response to intraluminal delivery of 124 mmol/L K+ Krebs was an increase in resistance of 297 +/- 67%. Vasoactive responses were sustainable for more than 8 h. This system will now be used to study the vasoactive properties of the guinea-pig choroid in greater detail.


Efficacy of Mycophenolate Mofetil and Oral Cyclophosphamide on Skin Thickness: Post Hoc Analyses From Two Randomized Placebo-Controlled Trials.
To assess the efficacy of mycophenolate mofetil (MMF) and cyclophosphamide (CYC) on modified Rodnan skin score (MRSS) in participants enrolled in the Scleroderma Lung Study (SLS) I and II. SLS I participants received daily oral CYC or matching placebo for 1 year, whereas SLS II participants received daily MMF for 2 years or daily oral CYC for 1 year followed by placebo for second year. We assessed the impact of MMF and CYC on the MRSS in SLS II over a 24-month period. We also compared the change in MRSS in patients with diffuse cutaneous systemic sclerosis (dcSSc) assigned to CYC and MMF in SLS II and SLS I versus placebo in SLS I over a 24-month period using a linear mixed model. In SLS II, the baseline mean ± SD MRSS was 14.0 ± 10.6 units for CYC and 15.3 ± 10.4 units for MMF; 58.5% were classified as dcSSc. CYC and MMF were associated with statistically significant improvements in MRSS from baseline over the period of 24 months in dcSSc (P < 0.05 at each time point), but there were no differences between the 2 groups. In the dcSSc subgroup, the change in MRSS from baseline to all 6-month visits was similar in SLS II groups (MMF, CYC, pooled cohort [MMF + CYC]) and in the SLS I CYC group and showed statistically significant improvements compared to SLS I placebo at 12, 18, and 24 months (P < 0.05). In SLS II, MMF and CYC treatment resulted in improvements in MRSS in patients with dcSSc over 24 months. In addition, MMF and CYC treatment resulted in statistically significant improvements in MRSS in patients with dcSSc when compared with the SLS I placebo group.


Nuclear receptor HNF4α binding sequences are widespread in Alu repeats.
Alu repeats, which account for ~10% of the human genome, were originally considered to be junk DNA. Recent studies, however, suggest that they may contain transcription factor binding sites and hence possibly play a role in regulating gene expression. Here, we show that binding sites for a highly conserved member of the nuclear receptor superfamily of ligand-dependent transcription factors, hepatocyte nuclear factor 4alpha (HNF4α, NR2A1), are highly prevalent in Alu repeats. We employ high throughput protein binding microarrays (PBMs) to show that HNF4α binds > 66 unique sequences in Alu repeats that are present in ~1.2 million locations in the human genome. We use chromatin immunoprecipitation (ChIP) to demonstrate that HNF4α binds Alu elements in the promoters of target genes (ABCC3, APOA4, APOM, ATPIF1, CANX, FEMT1A, GSTM4, IL32, IP6K2, PRLR, PRODH2, SOCS2, TTR) and luciferase assays to show that at least some of those Alu elements can modulate HNF4α-mediated transactivation in vivo (APOM, PRODH2, TTR, APOA4). HNF4α-Alu elements are enriched in promoters of genes involved in RNA processing and a sizeable fraction are in regions of accessible chromatin. Comparative genomics analysis suggests that there may have been a gain in HNF4α binding sites in Alu elements during evolution and that non Alu repeats, such as Tiggers, also contain HNF4α sites. Our findings suggest that HNF4α, in addition to regulating gene expression via high affinity binding sites, may also modulate transcription via low affinity sites in Alu repeats.


Integration of Care in the Implementation of the Affordable Care Act: Changes in Treatment Services in a National Sample of Centers Treating Substance Use Disorders.
This study examined patterns of medicalization in substance use disorder (SUD) that are aligned with the goals of the Affordable Care Act (ACA). Using a nationally representative sample of SUD treatment programs, we examined changes in several treatment domains. While observed changes were modest, they were in directions that support the thrust of the ACA. Specifically, we found an increase in the percentage of treatment referrals from other health care providers. We found an increase in the number of physicians for programs that did have a physician on staff, and an increase in counselors certified in treating alcohol and drug addiction. There was significant growth in the availability of oral and injectable naltrexone but not of other pharmacotherapies. There was a decrease in support for the 12-step model and an increase on the emphasis of a medicalized treatment model. Finally, we found a shift away from federal block grants and other public funding, consistent with the expectations of the ACA. These data indicate that, while progress is slow, the environment of the recent past has been supportive of the goal of SUD treatment's integration into mainstream medical care.


Acute effects of whole-body proton irradiation on the immune system of the mouse.
The acute effects of proton whole-body irradiation on the distribution and function of leukocyte populations in the spleen and blood were examined and compared to the effects of photons derived from a (60)Co gamma-ray source. Adult female C57BL/6 mice were exposed to a single dose (3 Gy at 0.4 Gy/min) of protons at spread-out Bragg peak (SOBP), protons at the distal entry (E) region, or gamma rays and killed humanely at six different times thereafter. Specific differences were noted in the results, thereby suggesting that the kinetics of the response may be variable. However, the lack of significant differences in most assays at most times suggests that the RBE for both entry and peak regions of the Bragg curve was essentially 1.0 under the conditions of this study. The greatest immunodepression was observed at 4 days postexposure. Flow cytometry and mitogenic stimulation analyses of the spleen and peripheral blood demonstrated that lymphocyte populations differ in radiosensitivity, with B (CD19(+)) cells being most sensitive, T (CD3(+)) cells being moderately sensitive, and natural killer (NK1.1(+)) cells being most resistant. B lymphocytes showed the most rapid recovery. Comparison of the T-lymphocyte subsets showed that CD4(+) T helper/inducer cells were more radiosensitive than the CD8(+) T cytotoxic/suppressor cells. These findings should have an impact on future studies designed to maximize protection of normal tissue during and after proton-radiation exposure.


Styrax (Liquidambar orientalis Mill.) promotes mitochondrial function and reduces cardiac damage following myocardial ischemic injury: the role of the AMPK-PGC1α signaling pathway.
To explore the effect of extract of Styrax (ES) on myocardial ischemic injury and its molecular mechanism, indirectly providing a theoretical basis for the development of ES. In order to assess the impact of ES treatment on ischemic heart disease, both a left anterior descending ligation-induced myocardial infarction (MI) model and an ischemia/hypoxia (I/H)-induced H9c2 cell injury model have been constructed. Specifically, Sprague-Dawley rats were randomly assigned to the following groups (n = 8) and administered intragastrically once a day for seven consecutive days: Sham group, MI group, ES-L (0.2 g/kg) group, ES-M (0.4 g/kg) group, ES-H (0.8 g/kg) group, and trimetazidine (TMZ, 0.02 g/kg) group. The cardiac functions and biochemical assessment of rats were detected. Then, we validated experimentally the targets and mechanism of ES on these pathological processes in I/H-induced H9c2 cell injury model. These results showed that different doses of ES (0.2 g/kg, 0.4 g/kg, 0.8 g/kg, intragastric) significantly improved myocardial structure and function when compared to the MI group. The results of 2,3,5-triphenyltetrazolium chloride (TTC), hematoxylin-eosin, and masson staining indicated that ES could significantly reduce infarct size, inhibit myocardium apoptosis, and decrease myocardial fibrosis. Moreover, ES distinctly suppressed the serum levels of lactate dehydrogenase (LDH), cardiac troponin T (cTnT), and creatine kinase-MB (CK-MB), alleviated myocardial mitochondrial morphology, and stimulated adenosine triphosphate (ATP) production, increased the level of succinate dehydrogenase (SDH), complex I and complex V activity. Different doses of ES (5 μg/ml, 10 μg/ml, 20 μg/ml) also improved cardiomyocyte morphology and decreased the apoptosis rate in H9c2 cells that had been exposed to I/H. Furthermore, the results of western blotting and qRT-PCR indicated that ES promoted the expression of proteins and mRNA related to energy metabolism, including phosphorylated adenosine monophosphate activated protein kinase (p-AMPK), peroxisome proliferator activated receptor gamma coactivator 1 alpha (PCG-1α), nuclear respiratory factor 1, and mitochondrial transcription factor A (TFAM). Mechanically, after the administration of Compound C (dorsomorphin), an AMPK inhibitor, these effects of myocardial protection produced by ES were reversed. Collectively, these results demonstrated that ES could improve myocardial mitochondrial function and reduce ischemic injury by activating AMPK/PCG-1α signaling pathway, while indicating its potential advantages as a dietary supplement.


The RNA-Binding Landscape of HAX1 Protein Indicates Its Involvement in Translation and Ribosome Assembly.
HAX1 is a human protein with no known homologues or structural domains. Mutations in the HAX1 gene cause severe congenital neutropenia through mechanisms that are poorly understood. Previous studies reported the RNA-binding capacity of HAX1, but the role of this binding in physiology and pathology remains unexplained. Here, we report the transcriptome-wide characterization of HAX1 RNA targets using RIP-seq and CRAC, indicating that HAX1 binds transcripts involved in translation, ribosome biogenesis, and rRNA processing. Using CRISPR knockouts, we find that HAX1 RNA targets partially overlap with transcripts downregulated in HAX1 KO, implying a role in mRNA stabilization. Gene ontology analysis demonstrated that genes differentially expressed in HAX1 KO (including genes involved in ribosome biogenesis and translation) are also enriched in a subset of genes whose expression correlates with HAX1 expression in four analyzed neoplasms. The functional connection to ribosome biogenesis was also demonstrated by gradient sedimentation ribosome profiles, which revealed differences in the small subunit:monosome ratio in HAX1 WT/KO. We speculate that changes in HAX1 expression may be important for the etiology of HAX1-linked diseases through dysregulation of translation.


Validation of expert systems: examples and considerations.
The problem of a medical expert system validation is generally complex. It requires a rigorous methodology of validation and must show proof of its practical competency in order to be used currently. Validation concerns the quality of conclusions provided by the system, the quality of the deductive process leading to these conclusions as well as the validity of its utilization. In this paper, some reflections, questions, and requirements are exposed that must be addressed to proceed to the validation of a knowledge base system in the field of medicine, especially the psychiatric field.


[Prevalence and Comorbidity of Self-Reported Diagnosis of Burnout Syndrome in the General Population].
To determine the prevalence and comorbid mental disorders of self-reported diagnosis of burnout syndrome in the general population of Germany. In the German Health Interview and Examination Survey (DEGS1) self-reported diagnosis of a burnout syndrome made by a physician or psychotherapist was assessed in a standardized interview (N = 7987). For N = 4483 mental disorders were determined with the Composite International Diagnostic Interview (CIDI). Weighted lifetime and 12-month prevalences were calculated. Lifetime prevalence of diagnosed burnout syndrome was 4.2 % (women 5.2 %, men 3.3 %), 12-month prevalence was 1.5 % (women 1.9 %, men 1.1 %). Highest prevalences were found in 40 - 59 year olds, in people with middle and high socio economic status and in women with low and men with high social support. Among the 12-month cases, 70.9 % had at least one DSM-IV disorder. Associations were found for the diagnosis of burnout syndrome with somatoform, affective and anxiety disorders. The diagnosis of burnout syndrome is less frequently given and reported than expected. People with a burnout diagnosis often have a manifest mental disorder.


The Efficacy of Sunitinib Treatment of Renal Cancer Cells Is Associated with the Protein PHAX In Vitro.
Anti-angiogenic agents, such as the multi-tyrosine kinase inhibitor sunitinib, are key first line therapies for metastatic clear cell renal cell carcinoma (ccRCC), but their mechanism of action is not fully understood. Here, we take steps towards validating a computational prediction based on differential transcriptome network analysis that phosphorylated adapter RNA export protein (PHAX) is associated with sunitinib drug treatment. The regulatory impact factor differential network algorithm run on patient tissue samples suggests PHAX is likely an important regulator through changes in genome-wide network connectivity. Immunofluorescence staining of patient tumours showed strong localisation of PHAX to the microvasculature consistent with the anti-angiogenic effect of sunitinib. In normal kidney tissue, PHAX protein abundance was low but increased with tumour grade (G1 vs. G3/4; p < 0.01), consistent with a possible role in cancer progression. In organ culture, ccRCC cells had higher levels of PHAX protein expression than normal kidney cells, and sunitinib increased PHAX protein expression in a dose dependent manner (untreated vs. 100 µM; p < 0.05). PHAX knockdown in a ccRCC organ culture model impacted the ability of sunitinib to cause cancer cell death (p < 0.0001 untreated vs. treated), suggesting a role for PHAX in mediating the efficacy of sunitinib.


Skin aging.
There are two main processes that induce skin aging: intrinsic and extrinsic. A stochastic process that implies random cell damage as a result of mutations during metabolic processes due to the production of free radicals is also implicated. Extrinsic aging is caused by environmental factors such as sun exposure, air pollution, smoking, alcohol abuse, and poor nutrition. Intrinsic aging reflects the genetic background and depends on time. Various expressions of intrinsic aging include smooth, thinning skin with exaggerated expression lines. Extrinsically aged skin is characterized by photo damage as wrinkles, pigmented lesions, patchy hypopigmentations, and actinic keratoses. Timely protection including physical and chemical sunscreens, as well as avoiding exposure to intense UV irradiation, is most important. A network of antioxidants such as vitamins E and C, coenzyme Q10, alpha-lipoic acid, glutathione, and others can reduce signs of aging. Further anti-aging products are three generations of retinoids, among which the first generation is broadly accepted. A diet with lot of fruits and vegetables containing antioxidants is recommended as well as exercise two or three times a week.


Deep homology and the origins of evolutionary novelty.
Do new anatomical structures arise de novo, or do they evolve from pre-existing structures? Advances in developmental genetics, palaeontology and evolutionary developmental biology have recently shed light on the origins of some of the structures that most intrigued Charles Darwin, including animal eyes, tetrapod limbs and giant beetle horns. In each case, structures arose by the modification of pre-existing genetic regulatory circuits established in early metazoans. The deep homology of generative processes and cell-type specification mechanisms in animal development has provided the foundation for the independent evolution of a great variety of structures.


Functional outcome of surgically treated patients of ossified posterior longitudinal ligament of cervical and dorsal spine in Indian population - A single center retrospective analysis of 40 patients.
The aim of the study was to retrospectively evaluate the neurological outcome in operated patients of ossified posterior longitudinal ligament (OPLL) of cervical and/or dorsal spine using modified Japanese orthopedic association (mJOA) score and find out the factors affecting the outcome. The study design was a retrospective study. Forty operated patients of cervical and/or dorsal spine OPLL were included in the study. Neurological examination was conducted and analyzed using mJOA score pre- and postoperatively at 1, 6, and 12 months. Improvement in the mJOA score based on age, sex, type of OPLL, duration of symptoms, type of surgical procedure, and radiological parameters were calculated, analyzed, and compared with previous records of the patient. Significant improvement in mJOA scores with mean preoperative being 12.27 ± 1.95 with 1-year postoperative 13.85 ± 2.02 (P < 0.0001) noted. There is a significant difference in mean mJOA scores in posterior approach with instrumentation (P < 0.0001) as compared with laminoplasty group (P < 0.005). Patients with occupancy ratio <60% had better results (P < 0.0001) as compared to those with occupancy ratio of >60% (P = 0.003). Patients with duration of symptoms >1 year had poorer results compared to those of <1 year duration. Mean ossification kyphosis angle was 19.4° ± 5.73°. OPLL is a progressive disease which causes severe neurological deficit if left untreated. OPLL in a young patient with short duration of symptoms, low occupancy ratio, and low ossification kyphosis had better chances of neurological recovery. Type of OPLL and sex of patient do not affect the recovery. OPLL managed early and surgically has better results irrespective of anterior or posterior approach with significant improvement in mJOA score. Decompression with fusion has better results than decompression alone.


A close association of freedom from pain, migraine-related functional disability, and other outcomes: results of a post hoc analysis of randomized lasmiditan studies SAMURAI and SPARTAN.
While pain freedom at 2 h is a key primary outcome for current trials for acute treatment of migraine, the relationship between the degree of head pain and other efficacy measures at 2 h has rarely been explored. Following lasmiditan treatment of a migraine attack with moderate or severe head pain, we contrast those who achieve pain freedom with those who achieve mild pain but not pain freedom 2 h post dosing. Patient-level data were pooled across studies and treatment arms from two Phase 3 trials comparing lasmiditan and placebo, SAMURAI and SPARTAN. This post hoc analysis assessed freedom from the most bothersome symptom (MBS), freedom from migraine-related functional disability (disability), and improved patient global impression of change (PGIC) in patients who achieved 2 h pain freedom compared to those who experienced 2 h mild pain. Mild pain differs from pain relief which is defined as either mild pain or pain freedom. Patients who achieved 2 h pain freedom (N = 913), in comparison with those with 2 h mild pain (N = 864), were significantly more likely to experience MBS freedom (91.9% vs. 44.9%), disability freedom (87.1% and 13.4%), and improved PGIC (86.5% and 31.5%) (p < 0.001 for all combinations). In addition, more patients who were pain free experienced both 2 h MBS freedom and 2 h functional disability freedom (83.6%) compared to those with mild pain (10.8%; p < 0.001). The proportion of patients with pain freedom who did not achieve either MBS or disability freedom (4.6%) was lower than in patients with mild pain (52.4%). Lastly, 55.2% of patients experienced mild pain before disability freedom compared to 72.1% who experienced pain freedom and disability freedom at the same time. This study demonstrated that, at 2 h post treatment, patients who were pain free were more likely to achieve other outcomes including freedom from their MBS, freedom from migraine-related functional disability, and improved PGIC compared to those with mild pain, confirming that 2 h pain freedom is more robustly associated with other clinical outcomes than the 2 h mild pain endpoint. SAMURAI ( NCT02439320 ); SPARTAN ( NCT02605174 ).


Cognitive ability and risk for substance misuse in men: genetic and environmental correlations in a longitudinal nation-wide family study.
To investigate the association in males between cognitive ability in late adolescence and subsequent substance misuse-related events, and to study the underlying genetic and environmental correlations. A population-based longitudinal study with three different family-based designs. Cox proportional hazards models were conducted to investigate the association at the individual level. Bivariate quantitative genetic modelling in (1) full brothers and maternal half-brothers, (2) full brothers reared together and apart and (3) monozygotic and dizygotic twin brothers was used to estimate genetic and environmental correlations. Register-based study in Sweden. The full sample included 1 402 333 Swedish men born 1958-91 and conscripted at mean age 18.2 [standard deviation (SD) = 0.5] years. A total of 1 361 066 men who had no substance misuse events before cognitive assessment at mandatory military conscription were included in the Cox regression models, with a follow-up time of up to 35.6 years. Cognitive ability was assessed at conscription with the Swedish Enlistment Battery. Substance misuse events included alcohol- and drug-related court convictions, medical treatments and deaths, available from governmental registries. Lower cognitive ability in late adolescence predicted an increased risk for substance misuse events [hazard ratio (HR) for a 1-stanine unit decrease in cognitive ability: 1.29, 95% confidence interval (CI) = 1.29-1.30]. The association was somewhat attenuated within clusters of full brothers (HR = 1.21, 95% CI = 1.20-1.23). Quantitative genetic analyses indicated that the association was due primarily to genetic influences; the genetic correlations ranged between -0.39 (95% CI = -0.45, -0.34) and -0.52 (95% CI -0.55, -0.48) in the three different designs. Shared genetic influences appear to underlie the association between low cognitive ability and subsequent risk for substance misuse events among Swedish men.


Genome rearrangement distances and gene order phylogeny in gamma-Proteobacteria.
Genome rearrangements have been studied in 30 gamma-proteobacterial complete genomes by comparing the order of a reduced set of genes on the chromosome. This set included those genes fulfilling several characteristics, the main ones being that an ortholog was present in every genome and that none of them had been acquired by horizontal gene transfer. Genome rearrangement distances were estimated based on either the number of breakpoints or the minimal number of inversions separating two genomes. Breakpoint and inversion distances were highly correlated, indicating that inversions were the main type of rearrangement event in gamma-Proteobacteria. In general, the progressive increase in sequence-based distances between genome pairs was associated with the increase in their rearrangement-based distances but with several groups of distances not following this pattern. Compared with free-living enteric bacteria, the lineages of Pasteurellaceae were evolving, on average, to relatively higher rates of between 2.02 and 1.64, while the endosymbiotic bacterial lineages of Buchnera aphidicola and Wigglesworthia glossinidia were evolving at moderately higher rates of 1.38 and 1.35, respectively. Because we know that the rearrangement rate in the Bu. aphidicola lineage was close to zero during the last 100-150 Myr of evolution, we deduced that a much higher rate took place in the first period of lineage evolution after the divergence of the Escherichia coli lineage. On the other hand, the lineage of the endosymbiont Blochmannia floridanus did present an almost identical rate to free-living enteric bacteria, indicating that the increase in the genome rearrangement rate is not a general change associated with bacterial endosymbiosis. Phylogenetic reconstruction based on rearrangement distances showed a different topology from the one inferred by sequence information. This topology broke the proposed monophyly of the three endosymbiotic lineages and placed Bl. floridanus as a closer relative to E. coli than Yersinia pestis. These results indicate that the phylogeny of these insect endosymbionts is still an open question that will require the development of specific phylogenetic methods to confirm whether the sisterhood of the three endosymbiotic lineages is real or a consequence of a long-branch attraction phenomenon.