Genetics of coronary artery disease in the post-GWAS era.
During the past decade, genome-wide association studies (GWAS) have transformed our understanding of many heritable traits. Three recent large-scale GWAS meta-analyses now further markedly expand the knowledge on coronary artery disease (CAD) genetics in doubling the number of loci with genome-wide significant signals. Here, we review the unprecedented discoveries of CAD GWAS on low-frequency variants, underrepresented populations, sex differences and integrated polygenic risk. We present the milestones of CAD GWAS and post-GWAS studies from 2007 to 2021, and the trend in identification of variants with smaller odds ratio by year due to the increasing sample size. We compile the 321 CAD loci discovered thus far and classify candidate genes as well as distinct functional pathways on the road to indepth biological investigation and identification of novel treatment targets. We draw attention to systems genetics in integrating these loci into gene regulatory networks within and across tissues. We review the traits, biomarkers and diseases scrutinized by Mendelian randomization studies for CAD. Finally, we discuss the potentials and concerns of polygenic scores in predicting CAD risk in patient care as well as future directions of GWAS and post-GWAS studies in the field of precision medicine.


[Application of biosurfactant in composting of agricultural waste].
The biosurfactant of rhamonolipid (Rh) obtained from Pseudomonas aeruginosa was used in co-composting of rice straw and wheat bran, the efficiency of the promoter was studied under temperature control during the first stage of composting. Composting was done in an aerobic static bed of composting during which moisture content of mixture materials was controlled between 60% 70%, and the control of aeration adopted the timer by turns of 0.18m3/h for 20 minutes and pause for 40 minutes. Changes were studied including pH, organic matter, dissolved organic carbon (DOC), the biomass of microorganism, hemicellulase activity, carboxymethyl cellulose enzyme (CMCase) activity, hemi-cellulose and cellulose contents. The experimental results show that the degradation rate of organic matter is 13.4% higher than that of the control, and the average of DOC is improved by 2.2g/kg. The degradation contents of hemi-cellulose and cellulose are increased by 5.7%, 10.7% compared with the control. This indicate that the addition of Rh can improve microenvironment, strengthen polymers hydration, accelerate the co-composting process and improve the quality of composting production.


INVASIVE FUNGAL INFECTIONS IN CHILDREN TREATED FOR HEMATOLOGIC MALIGNANCIES - A FIVE-YEAR SINGLE CENTER EXPERIENCE.
Invasive fungal infections (IFI) are life-threatening complications of intensive chemotherapy treatment, with the incidence in pediatric patients ranging from 2% to 21%. In this article, we describe our 5-year experience of IFI in pediatric oncology patients and its clinical manifestations with radiological findings, treatment and outcome. A retrospective and descriptive survey of IFI in children with hematologic neoplasms was conducted at the Department of Oncology and Hematology, Zagreb Children's Hospital. Medical charts of children 0-17 years of age, of both sexes, treated for leukemias and lymphomas from January 2016 to December 2020 were reviewed. In a 5-year period, 60 patients were treated for hematologic malignancy, acute lymphoblastic leukemia (ALL) being the most prevalent diagnosis. IFI was verified in 9 (15%) children, predominantly in patients with ALL (75%). The specific causative agent was detected in one child, whereas other infections were classified as probable pulmonary aspergillosis. All the patients received standard prophylaxis with fluconazole and treatment with liposomal amphotericin B and voriconazole. The majority of our patients achieved recovery. IFI prevention, diagnosis and treatment remain a challenge. Uniform prophylaxis and therapy protocols, as well as environmental control are of vital importance for the development of better strategies in the prevention, early detection and treatment of IFI in pediatric hematology patients.


Predicting environmental mitigation requirements for hydropower projects through the integration of biophysical and socio-political geographies.
Uncertainty about environmental mitigation needs at existing and proposed hydropower projects makes it difficult for stakeholders to minimize environmental impacts. Hydropower developers and operators desire tools to better anticipate mitigation requirements, while natural resource managers and regulators need tools to evaluate different mitigation scenarios and order effective mitigation. Here we sought to examine the feasibility of using a suite of multi-faceted explanatory variables within a spatially explicit modeling framework to fit predictive models for future environmental mitigation requirements at hydropower projects across the conterminous U.S. Using a database comprised of mitigation requirements from more than 300 hydropower project licenses, we were able to successfully fit models for nearly 50 types of environmental mitigation and to apply the predictive models to a set of more than 500 non-powered dams identified as having hydropower potential. The results demonstrate that mitigation requirements are functions of a range of factors, from biophysical to socio-political. Project developers can use these models to inform cost projections and design considerations, while regulators can use the models to more quickly identify likely environmental issues and potential solutions, hopefully resulting in more timely and more effective decisions on environmental mitigation.


Biological interaction of thiamine with lysozyme using binding capacity concept and molecular docking.
The binding of thiamine (vitamin B1) on lysozyme has been examined at various ionic strengths of phosphate buffer (pH 6.9), various pH values, and various protein concentrations at 25°C using thiamine selective membrane electrode. This method is faster and more precise than equilibrium dialysis technique which can obtain sufficient and accurate data for binding analysis. The values of Hill equation parameters were estimated for each set using binding capacity concept and used for calculation of intrinsic binding affinity. The results represent two binding sets for thiamine on lysozyme at various experimental conditions.


Effects of proxigermanium on interferon production and 2',5'-oligoadenylate synthetase activity in the lung of influenza virus-infected mice and in virus-infected human peripheral blood mononuclear cell cultures.
Proxigermanium (SK-818) is a synthesized organic germanium compound having various biological activities. The effects of proxigermanium on interferon (IFN) production in mice infected with influenza virus and virus-infected human peripheral blood mononuclear cells (hPBMC) were investigated. Proxigermanium alone did not induce IFN production in normal mice or in hPBMC without viral infection. On the other hand, proxigermanium enhanced alpha/beta IFN production in viral-infected mice and hPBMC. Since proxigermanium is known to have antiviral activity in vivo but not in vitro, it is likely that the IFN production augumenting activity of proxigermanium is involved in its antiviral activities.


Team Sports and Sexual Violence: Examining Perpetration by and Victimization of Adolescent Males and Females.
The authors examined whether team athletic involvement is related to sexual violence victimization and perpetration among male and female high school students. A secondary data analysis was conducted with Minnesota Student Survey data gathered from 122,501 Minnesota eighth-grade, ninth-grade, and 11th grade students. Multivariable logistic regression indicates that sport-involved youth are more likely to be involved in sexual violence than peers not involved in sports. For example, highly sport-involved males are significantly more likely to coerce a partner into sex (odds ratio [OR] = 1.35, 95% confidence interval [CI] = [1.01, 1.67]) and be coerced (OR = 1.19, 95% CI = [1.02, 1.38]) than nonsport peers. The results suggest high school sport programs could be an important site to implement comprehensive sexual violence prevention programs targeted at individual actions as well as harmful cultural norms and systematic inequities.


Large Ganglion Cyst with Unusual Location on the Back-A Case Report.
A ganglion cyst is a soft tissue tumor-like lesion filled with colloid material commonly located on the hand and wrist. We report a case of a large ganglion cyst with an unusual location on the back. The patient presented with a mass growing over 2 months measuring 11.2 × 4.7 × 7.2 cm on magnetic resonance imaging. Ultrasound and puncture was attempted twice without achieving drainage. After surgical removal, histologic examination diagnosed the tumor as a ganglion cyst. We conclude that when evaluating a subcutaneous soft tissue mass, regardless of localization, a ganglion cyst may be a differential diagnosis.


Association of human leukocyte antigen polymorphism with hepatitis B virus infection and genotypes.
Associations were studied between the polymorphism of northern Han Chinese leukocyte antigen (HLA) alleles and the outcomes of hepatitis B virus (HBV) infection and HBV genotypes. HLA-A, B, and DRB1 alleles in peripheral blood mononuclear cells (PBMCs) were detected by polymerase chain reaction (PCR) with sequence-specific primers. The PBMCs were collected from 61 persons who tested positive for hepatitis B surface antigen (HBsAg) for more than 6 months (Persistent group), 32 persons who tested negative for both HBsAg and HBV DNA but positive for both anti-HBc and anti-HBs (Recovered group), and 40 persons who tested negative for all serologic markers of HBV infection (Uninfected group). HBV genotypes in serum specimens from 56 of 61 patients with persistent HBV infection were determined by nested PCR with 6 pairs of HBV genotype-specific primers (A to F). The frequency of HLA-DRB1*12 was significantly higher in the Persistent group than in the Recovered group (P=0.004). HLA-A*02 was significantly higher in the Recovered group than in the Persistent group (P=0.044). HLA-DRB1*15 was significantly higher in the HBV genotype B group than in the C group (P=0.013). These findings suggested that there were associations not only between HLA polymorphisms and outcomes of HBV infection but also between HLA polymorphisms and the infected HBV genotypes.


Depopulation of lymphocyte migration sites in the lymph node by irradiation and colloidal carbon.
The purpose of the present investigation was to examine, in the light of recent histological findings, whether irradiation and colloidal carbon can have a lymphocyte depopulating effect on preferentially particular structures of the rat lymph nodes. Normal eight-week-old Sprague-Dawley rats received a 500 R whole body irradiation or a subcutaneous injection of 0.02 ml of India ink. The animals were then sacrificed at various time-intervals. The histological analysis of the irradiated and draining nodes revealed that both treatments almost completely eliminated small lymphocytes from the affected nodal structures, except in the center of the deep cortex units. The affected structures had been predominantly populated by recirculating lymphocytes.Thus, the treatment had a rather preferential depleting effect on a node population of recirculating lymphocytes. This finding provides another possible explanation for the carbon-induced augmentation of a GVH reaction in nodes. This augmentation had previously been attributed to a stimulation by the carbon of host macrophages, which would mediate the proliferation of antigen-reactive donor cells. From our present findings, it appears that carbon, like irradiation, could act by depleting a node of recirculating lymphocytes, thereby weakening its immunological potential against the inoculated lymphocytes.


Epiretinal membranes in vitro.
Monolayers of cells were grown from epiretinal membranes using standard tissue culture techniques. The growing cells were filmed by time-lapse cinephotomicrography to record their locomotory characteristics and cell-to-cell interactions. The established layers of cells were examined in the scanning electron microscope. In addition, the cultured cells were studied immunohistochemically to identify specific cell types and the distribution of contractile elements (e.g. actin). The locomotory patterns, the topography and the immunohistochemistry of cells from epiretinal membranes were compared to the movement, appearance and staining characteristics of a wide range of normal cells in culture. From the growth characteristics in tissue culture, early membranes appeared more hypercellular than longer standing membranes. Cell outgrowth was established more quickly and was more widespread in the young epiretinal membranes than in older, more fibrous, specimens. The locomotory and immunohistochemical studies demonstrated that a wide variety of cell types may contribute to epiretinal membranes. A remarkably wide range of cell forms were observed in cultures of individual membranes. Epithelial and glial cells were found in many of the monolayers. However, they did not constitute the majority of cells in the cultures. The predominating cell types had the morphological and locomotory characteristics of fibroblasts or macrophages.


Quantification of levetiracetam in plasma and urine and its application to a pharmacokinetic study of traumatic brain injury patients.
Background: Levetiracetam is an antiepileptic drug used to prevent or treat seizure in patients with severe traumatic brain injury. This study aimed to develop and validate methodology suitable for measuring levetiracetam concentrations in human plasma and urine. Methods: Plasma or urine (10 μl) samples were spiked with [2H6]-levetiracetam and processed using an acetonitrile precipitation. ESI-LC-MS/MS was employed for analyte detection. Results: The levetiracetam calibration was linear from 0.1 to 50 mg/l in a combined matrix of plasma and urine. Intra- and inter-assay imprecision and accuracy in plasma were <7.7 and 109%, and in urine were <7.9 and 108%, respectively. Conclusion: The validated method was applied to a pharmacokinetic study of levetiracetam in critically ill patients with severe traumatic brain injury.


Web of science: a unique method of cited reference searching.
The number of times an article is acknowledged as a reference in another article reflects its scientific impact. Citation analysis is one of the parameters for assessing the quality of research published in scientific, technology and social science journals. Web of Science enables users to search current and retrospective multidisciplinary information. Parameters and practical applications evaluating journal and article citation characteristics available through the Science Citation Index are summarized.


Short communication: Effect of washing method, grinding size, and the determination of an indigestible fraction on in situ degradation of starch in mature corn grain.
The objectives of this study were to determine (1) the effect of grinding size (1, 2, 4, and 6 mm) to determine effective ruminal disappearance (ERD); (2) the most adequate method to estimate the rapidly degradable fraction (A); (3) a time point to measure the indigestible fraction (C); and (4) the viability of using fewer time points to estimate starch fractional disappearance rate (kd) of mature corn grain. Fraction A was determined by rinsing in a bucket or washing machine, rumen immersion followed by bucket or washing machine, and water immersion for 30 min followed by bucket or washing machine. Ruminal in situ incubations were performed at 48, 72, 96, and 120 h to determine fraction C, and at 0 (washing machine), 3, 6, 12, 18, 24, and 48 h to determine the kinetics of starch disappearance. Models were used with either 2 or 3 pools and kd was determined by the linear slope of the log-transformed bag residues as a proportion of incubated samples over time. The ERD was calculated as A + B [kd/(kd + kp)], where kp is the ruminal fractional passage rate = 16.0% h-1. Data were analyzed using PROC MIXED of SAS (SAS Institute Inc., Cary, NC) with the fixed effects of run (for fraction A analysis only) method (either washing or model), grinding size, and method by grinding size interaction, with cow as a random effect. Correlation between estimates calculated using all time points or combinations of 2 and 3 time points were determined using PROC CORR. Fraction A was reduced as grinding size increased, but was not altered by washing method. Samples ground at 6 mm had greater fraction C than other grinding sizes at 48, 72, or 96 h, but not at 120 h. Model affected the slowly degradable fraction (B) values solely, but the difference was minor (0.5 percentage units). Greater fractions B and C but reduced kd and ERD were observed as grinding size increased. Based on correlation analysis the 2-pool model, incubation times of 0, 3, and 48 h were suitable to evaluate ruminal starch degradation kinetics in mature corn. Ruminal in situ incubation at 120 h highlighted the lack of a fraction C of starch (0.13% of starch). Washing method did not affect determination of fraction A of starch. Ruminal in situ incubations of 0, 3, and 48 h for starch degradation kinetics using a 2-pool model were adequate for mature ground corn, but 120 h of incubation is suggested to confirm the existence or absence of a fraction C. Grinding size affected starch degradation kinetics and fraction A determination.


Surgical management of lesions of the internal carotid artery using a modified Fisch type A infratemporal approach.
To report seven cases of vascular repair of the internal carotid artery (ICA) using a modified Fisch type A infratemporal approach and a venous grafting. Retrospective case review. Tertiary care center. We have analyzed the clinical presentation, paraclinical assessment, and postoperative results regarding the vascular repair and the facial and auditory function from seven consecutive patients. All patients have been operated on by a multidisciplinary team of ENT and vascular surgeons. The study includes four men and three women, aged from 21 to 62 years old. Six patients suffered from vascular traumatic injury after motor vehicle accident (n = 5) or cervical manipulation (n = 1) and one patient presented an atheromatous stenosis. All benefited from a vascular repair with a venous grafting through a modified Fisch Type A infratemporal approach. No death and no new stroke were noted (mean follow-up, 34 mo). The postoperative angiographies showed six functional grafts and one asymptomatic thrombosis. Six immediate postoperative facial palsy occurred but recovered to Grade I or II within 6 months. There was one traumatic injury of the facial nerve and one postoperative anacusis. For the six other patients, the reconstitution of the external auditory canal and ossicular chain allowed to limit the hearing loss to a mean air-bone gap of 22.5 dB (range, 15-35 dB). The lesions of the intrapetrous aspects of the ICA remain the subject of debates regarding the indication for a vascular repair. For young or in good health patients, the infratemporal approach provides a safe and reliable access to the horizontal segment of the ICA, offering to the vascular surgeons optimal conditions for the vascular repair.


Persistence of rabbit haemorrhagic disease virus genome in vaccinated rabbits after experimental infection.
Rabbit haemorrhagic disease (RHD) is usually a fatal disease in rabbits which has spread rapidly across the continents. While previous studies suggested persistence in rabbits to be an important factor in the epidemiology, the relevance of field virus infection of immune rabbits has not been investigated in experimentally infected animals before. This report describes for the first time the persistence of rabbit haemorrhagic disease virus (RHDV) genome for at least 15 weeks in rabbits immunized with an inactivated vaccine as well as a subunit vaccine and subsequently challenged with virulent RHDV. The viral RNA loads were determined by real-time reverse transcription-polymerase chain reaction. No conspicuous association of the detectable amount of RHDV RNA with the type of vaccine, the time after infection and--with one exception--the level of RHDV-specific antibodies in the immunized animals was observed. The results presented in this study are an urgent evidence for the existence of carrier animals as an important factor in the epidemiology of RHD.


Compatibility of immunogenicity guidance by the EMA and the US FDA.
The guidelines for immunogenicity studies by the European Medicines Agency and the US FDA are based on different legislations and regulatory philosophies. In spite of the different background, the main guidelines are compatible on the scientific level, especially for new innovative therapeutic protein products. The importance of sensitive and drug-tolerant antidrug antibody assays and multidisciplinary approach to development and assessment are highlighted by both agencies. The main differences are in the field of biosimilars. The nonclinical in vivo immunogenicity studies are seen more useful by the FDA than by the European Medicines Agency. The draft FDA guidance on interchangeability will complicate global biosimilar development by requiring clinical switch studies with US sourced reference product.


High Spatial Resolution Chemical Imaging of Implant-Associated Infections with X-ray Excited Luminescence Chemical Imaging Through Tissue.
Microbial infections associated with implantable medical devices are a major concern in fracture fixation failure. Early diagnosis of such infection will allow successful eradication with antibiotics without an extra cost for a second surgery. Herein, we describe XELCI as a technique with high X-ray resolution, implant specificity, and chemical sensitivity to noninvasively image chemical concentrations near the surface of implanted medical devices. The devices are coated with chemically reporting surfaces. This chemically responsive surface consists of two layers coated on an implantable medical device; a pH-sensitive layer (bromothymol blue or bromocresol green incorporated hydrogel) which is coated over a red-light emitting scintillator (Gd2O2S: Eu) layer for monitoring. A focused X-ray beam irradiates a spot on the implant, and the red light generated by the scintillator (with 620 nm and 700 nm peaks) is transmitted through the sensing layer which alters the spectral ratio depending on the pH. An image is generated by scanning the X-ray beam across the implant and measuring the spectral ratio of light passing through the tissue point-by-point. We used this imaging technique for monitoring implant-associated infections previously on the bone surface of the femur with a modified implantable plate sensor. Now we are studying pH changes that occur from tibial intramedullary rod infections. Two different types of intramedullary rod designs are used in pre-pilot rabbit studies, and we learned that the XELCI technique could be used to monitor any chemical changes that occur not only on the bone surface but also inside the bone. Thus, this enables noninvasive, high spatial resolution, low background local pH imaging to study implant-associated infection biochemistry.


Anthropometric body fat composition reference values in Spanish adolescents. The AVENA Study.
To determine reference values for body mass index (BMI), sum of six skinfolds (sigma6 skinfolds) and body fat percentage (BF%) in Spanish adolescents aged 13-18 years, included in the AVENA Study (Alimentación y Valoración del Estado Nutricional en Adolescentes: Food and Assessment of the Nutritional Status of Adolescents). Multicentre cross-sectional study. Representative sample of Spanish adolescents. The population was selected by means of a multiplestep, simple random sampling. The final number of subjects included in the AVENA Study was 2859 adolescents; 2160 adolescents had a complete set of anthropometric measurements and were then included in this study (1109 males and 1051 females). Weight, height and six skinfold thicknesses were measured. As indices of total adiposity, we calculated BMI, summation sigma6 skinfolds and BF% with the formulas described by Slaughter et al. Sigma6 skinfolds and BF% in each age group were significantly higher in females than in males. In males, age showed a significant effect for BMI, sigma6 skinfolds and BF%; however, in females, the effect was only significant for BF%. The percentile distribution was more disperse towards higher sigma6 skinfolds and BF% values in males when compared with females. The presented percentile values will help us to classify adolescents in comparison with a well-established reference population, and to estimate the proportion of adolescents with high or low adiposity amounts. The AVENA-Study was supported by the Spanish Ministry of Health (FIS 00/0015), and grants from Panrico SA, Madaus SA and Procter and Gamble SA. This study was also supported by Instituto de Salud Carlos III (Spain), RCESP (C03/09) and Spanish Ministry of Education (AP2003-2128).


Association of knowledge of HIV and other factors with individuals' attitudes toward HIV infection: a national cross-sectional survey among the Japanese non-medical working population.
The stigma of and discrimination because of HIV has been described as the most important obstacle to prevention and treatment efforts. The purpose of this study was to investigate negative attitudes and prejudice toward HIV among the Japanese non-medical working population and to explore contributing factors. An online anonymous nationwide survey involving approximately 3,000 individuals was conducted in Japan. Questions ranged from background information and HIV knowledge to individuals' attitudes towards HIV infection in the workplace. Descriptive statistics and logistic regression were applied for analysis. Thirty-three percent of participants feared transmission of HIV from infected colleagues, 34% tended to avoid contact with them and 40% had prejudiced opinions about HIV infection. Despite a relatively high level of knowledge of HIV/AIDS overall (11.9 ± 3.3 from 15 points), only 50% of individuals were aware of some issues. Greater knowledge was associated with less negative attitudes towards HIV infection (OR 0.39, 95% CI 0.31-0.48 for prejudiced opinion, high compared with low level of knowledge), whereas greater health consciousness was inversely related to attitude (OR 1.97, 95% CI 1.50-2.58 for prejudiced opinion, high compared with low health consciousness). Knowledge neutralizes peoples' negative attitudes towards HIV infection, whereas greater health consciousness may worsen them. Educational programs should balance knowledge with health consciousness to improve the efficacy of HIV interventions.


Individual tree segmentation of airborne and UAV LiDAR point clouds based on the watershed and optimized connection center evolution clustering.
Light detection and ranging (LiDAR) data can provide 3D structural information of objects and are ideal for extracting individual tree parameters, and individual tree segmentation (ITS) is a vital step for this purpose. Various ITS methods have been emerging from airborne LiDAR scanning (ALS) or unmanned aerial vehicle LiDAR scanning (ULS) data. Here, we propose a new individual tree segmentation method, which couples the classical and efficient watershed algorithm (WS) and the newly developed connection center evolution (CCE) clustering algorithm in pattern recognition. The CCE is first used in ITS and comprehensively optimized by considering tree structure and point cloud characteristics. Firstly, the amount of data is greatly reduced by mean shift voxelization. Then, the optimal clustering scale is automatically determined by the shapes in the projection of three different directions. We select five forest plots in Saihanba, China and 14 public plots in Alpine region, Europe with ULS or ALS point cloud densities from 11 to 3295 pts/m2. Eleven ITS methods were used for comparison. The accuracy of tree top detection and tree height extraction is estimated by five and two metrics, respectively. The results show that the matching rate (R match) of tree tops is up to 0.92, the coefficient of determination (R 2) of tree height estimation is up to .94, and the minimum root mean square error (RMSE) is 0.6 m. Our method outperforms the other methods especially in the broadleaf forests plot on slopes, where the five evaluation metrics for tree top detection outperformed the other algorithms by at least 11% on average. Our ITS method is both robust and efficient and has the potential to be used especially in coniferous forests to extract the structural parameters of individual trees for forest management, carbon stock estimation, and habitat mapping.


Hyperlipidaemia and aortic valve disease.
Degenerative aortic valve stenosis is a common disease in the elderly, and traditional risk factors for atherosclerotic disease including hyperlipidaemia have been associated with the condition in several studies. This review addresses the role of the various risk factors and the potential for intervention. The association of lipid abnormalities such as high lipoprotein(a) levels and the presence of the apolipoprotein E4 allele with aortic stenosis, as well as the presence of several inflammatory markers both in plasma and in surgically excised valves, suggest that the stenotic process is driven by many of the same factors behind atherosclerosis. The aortic valves of animals fed a cholesterol-rich diet exhibit many characteristics in common with the early stages of aortic stenosis. This opens up the potential of retarding the process through intervention strategies. Hyperlipidaemia is associated with degenerative aortic valve stenosis, and the disease resembles the inflammatory process of atherosclerosis. Randomized controlled clinical trials will be needed to demonstrate the role of lipid intervention in patients with this condition.


Can bovine in vitro-matured oocytes selectively process X- or Y-sorted sperm differentially?
It has been reported that the mammalian female could have a preconceptual influence on the sex of her offspring, and it has been hypothesized that this influence could go some way toward accounting for the reported lower fertility following insemination with sex-sorted sperm. To test whether in vitro matured oocytes are able to select X- or Y-bearing spermatozoa following in vitro fertilization (IVF), we fertilized in vitro 1788 oocytes with X-sorted semen, Y-sorted semen, a mix of X- and Y-sorted semen, and unsorted semen from the same bull, and cultured until Day 9. Fertility was assessed by recording cleavage rate at 48 h postinsemination (hpi) and blastocyst development until Day 9. Embryos were sexed at the two- to four-cell stage and the blastocyst stage. The proportion of zygotes cleaving at 48 hpi was not different between X- and Y-sorted groups and the mix of X- and Y-sorted semen group; however, all were significantly lower than the unsorted group (P < 0.001). Blastocyst yield on Day 6 was significantly higher (P < or = 0.01) in the control group compared with the rest of the groups. Cumulative blastocyst yields on Days 7, 8, and 9 were also significantly higher (P < or = 0.01) in the unsorted group compared with the sorted groups. The proportion of female and male two- to four-cell embryos obtained following IVF with X- and Y-sorted sperm was 88% and 89%, respectively and the sex ratio at the two- to four-cell stage was not different following IVF with unsorted or sorted/recombined sperm (56.9% males vs. 57% males, respectively). At the blastocyst stage, similar percentages were obtained. In conclusion, the differences in cleavage and blastocyst development using sorted versus unsorted sperm are not due to the oocyte preferentially selecting sperm of one sex over another, but are more likely due to spermatic damage caused by the sorting procedure.


Bionomics of malaria vectors in Lao PDR, 2018-2020: entomological surveillance as a key tool for malaria elimination.
The Lao PDR National Strategic Plan for malaria control and elimination for year 2021-2025 emphasizes the importance of routine entomological surveillance being conducted in areas with high transmission and in active malaria foci in elimination targeted areas. The collection of entomological surveillance data that is closely linked to recent epidemiological data is crucial for improving impact, as it contributes to the evidence package that supports operational and strategic decision-making of national malaria programmes, as they accelerate their last mile of elimination. The Center for Malariology Parasitology and Epidemiology (CMPE) entomology team conducted entomological surveillance activities at 13 sentinel sites in 8 provinces and at active transmission foci sites from 2018 to 2020. The techniques used for the mosquito collection were indoor and outdoor human landing collections (from houses and from cultivation areas) and cattle baited net trap collections. There were 5601 Anopheles mosquito females captured and identified throughout the study, on both human and cow bait. They represented 15 different species or species complexes. The primary malaria vectors as well as the secondary vectors were present in all collection sites in the south, indicating that people living in these rural areas with high malaria incidence are exposed to the vectors. The vectors were highly zoophilic, but they still bite humans throughout the night with a high peak of activity before midnight, both indoors and outdoors. Overall, 17% of the malaria vectors were collected indoors when the people are sleeping. This confirms the importance of bed net use during the night. Thirty-two percent of primary and secondary vectors were collected outdoors at times when people are usually awake and outdoors, which shows that people are exposed to potentially infectious mosquitoes and the importance of personal protection at these times. The findings showed that residual transmission may occur outdoors in the villages, and outside the villages in cultivation fields and forested areas. Epidemiological data showed that transmission was higher in surveillance sites which were targeted as part of a malaria response rather than sentinel sites. Understanding where and how transmission is persisting, monitoring and mapping vector species distribution in areas with active transmission, monitoring biting trends, and designing evidence based and effective vector control interventions are critical to accelerating progress toward malaria elimination. In this context, the role of entomological surveillance combined with epidemiological data should be considered as a cornerstone in achieving malaria elimination.


[Investigation of seropositivity of Bordetella pertussis in adults in a university hospital].
Whooping cough is a vaccine-preventable infectious diseases caused by Bordetella pertussis/parapertussis. Despite of routine immunization programs in the world, pertussis still remains endemic. Recently unvaccinated or partially immunized infants have infected with this pathogen and also increase of incidence was observed in adolescents and adults. The source of pertussis in newborns are attributed to household, especially due to the family members. Theaim of this study was to determine B.pertussis IgG antibody positivity by ELISA method in adults. Eighty-four of the total study population (39.6%) were anti-pertussis IgG positive, 128 (60.4%) were negative. The samples included in the study (n= 212) were divided into three groups according to ages: 19-35 years (Group 1, n= 61), 36-50 years (Group 2, n= 58), 51-65 years (Group 3, n= 93). Anti-pertussis IgG antibody positivity was 26.2% (n= 22) in Group 1, 26.2% (n= 22) in Group 2 and 47.6% (n= 40) in Group 3. According to the anti-pertussis IgG positivity results, no significant difference was observed between genders. The results were obtained as absorbance values by ELISA test, then transformed into semi-quantitative values as NovaTec Test Unit (NTU). NTU positive values were between 11.01-39.4. Ninety four percent of NTU values were in the range of 11.01 to 28.01 and 6% were between 28.01 to 39.4. It was observed that seropositivity rates peaked at ages of 27, 55 and 65. The highest NTU values were observed in the age of 32 in females and in the age of 24 in males. Relatively, low seropositivity values (NT= 15-20) were observed in both females and males to the age of 45 in Group 2. However a slight increase was observed in females after the age of 45. An increase (NTU ≥ 20) was determined in the age of 49 in males. The seropositivity rates were generally low (NT ≤ 20) in females in Group 3, but the highest values (NTU ≥ 30) were observed in 55 and 65 years of age in males in this group. The highest seropositivity (NTI ≥ 30) were in 55 and 65 years of age (NTI ≥ 30) in males and 27 years of age (NTU ≥ 30) in females while in 36-50 age range, it was relatively low (NTI ≤ 20). Routine pertussis vaccination program is not yet implemented for adults in our country. However, the causes more clearly demonstrate the need for adult pertussis vaccination since adults may be incompletely vaccinated or not vaccinated in the childhood, current vaccinations are not available in the childhood of adults, adults become more susceptible to infections as the age increases, life expectancy increases and the likelihood of encountering infections, and childhood vaccination antibodies diminish over time. According to the results of this study, a single dose of pertussis vaccine is recommended to implement for adults in our country.


Characterization and management of neurological adverse events during immune-checkpoint inhibitors treatment: an Italian multicentric experience.
Neurological immune-related adverse events (nirAEs) are rare toxicities of immune-checkpoint inhibitors (ICI). With the increase of ICI oncological indications, their incidence is growing. Their recognition and management remain nevertheless challenging. A national, web-based database was built to collect cases of neurological symptoms in patients receiving ICI and not attributable to other causes after an adequate workup. We identified 27 patients who developed nirAEs (20 males, median age 69 years). Patients received anti-PD1/PDL1 (78%), anti-CTLA4 (4%), or both (19%). Most common cancers were melanoma (30%) and non-small cell lung cancer (26%). Peripheral nervous system was mostly affected (78%). Median time to onset was 43.5 days and was shorter for peripheral versus central nervous system toxicities (36 versus 144.5 days, p = 0.045). Common manifestations were myositis (33%), inflammatory polyradiculoneuropathies (33%), and myasthenia gravis (19%), alone or in combination, but the spectrum of diagnoses was broad. Most patients received first-line glucocorticoids (85%) or IVIg (15%). Seven patients (26%) needed second-line treatments. At last follow-up, four (15%) patients were deceased (encephalitis, 1; myositis/myasthenia with concomitant myocarditis, 2; acute polyradiculoneuropathy, 1), while seven (26%) had a complete remission, eight (30%) partial improvement, and six (22%) stable/progressing symptoms. ICI treatment was discontinued in most patients (78%). Neurological irAEs are rare but potentially fatal. They primarily affect neuromuscular structures but encompass a broad range of presentations. A prompt recognition is mandatory to timely withheld immunotherapy and administrate glucocorticoids. In corticoresistant or severely affected patients, second-line treatments with IVIg or plasmapheresis may result in additional benefit.


The immune responses of common carp, Cyprinus carpio L., injected with carp interleukin-1beta gene.
The function of the common carp, Cyprinus carpio L., interleukin-1beta (IL-1beta) gene was studied by DNA injection. To investigate the immune responses to IL-1beta, a plasmid construct of cytomegalovirus (CMV)-driven carp IL-1beta was injected into the epaxial muscle of carp. IL-1beta protein expressed in serum on 1, 3, and 5 days after plasmid injection was quantified by ELISA and immunohistochemistry. IL1-beta gene injection increased proliferation of the lymphocytes by phytohemagglutinin (PHA). Macrophage functions, such as production of superoxide anion and phagocytosis, also were stimulated by IL-1beta gene injection. Moreover, an increase in resistance to Aeromonas hydrophila infection was recorded in IL-1beta-injected fish compared with control fish. Thus, the cloned homolog of IL-1beta from carp has all the functional similarities to the mammalian IL-1beta gene.


Further observations on cystic dilatation of the intrahepatic biliary system in biliary atresia after hepatic portoenterostomy: report on 10 cases.
This is a report on ten patients with cystic dilatation of the intrahepatic biliary system (CDIB) after hepatic portoenterostomy. They were five girls and five boys and the diagnosis of CDIB was made at ages 6 months to 11 years (mean age: 2.8 +/- 3.3 years). Follow-up ranged from one month to 15 years (mean: 5.5 +/- 4.9 years). In order to elucidate the factors which affect the clinical outcome of such patients, the types of CDIB (Type A: noncommunicating solitary cyst, Type B: communicating solitary cyst, Type C: multi-cystic dilatation), clinical symptoms at onset of CDIB and the method for the treatment were reviewed in relation to the outcome. For the purpose of understanding pathogenesis of CDIB, immunohistochemical study on hepatobiliary system was done with monoclonal antibody for cytokeratin. Outcome of the patients of Type C was poor, whereas the outcome of patients with type A and B was good. The outcome of preoperatively jaundiced patients was poor, but jaundice-free patients showed good outcome. Method of treatment was not related to the outcome. As epithelium of CDIB was positive for monoclonal antibody of cytokeratin, it was suspected that pathogenesis of CDIB might be related to peribiliary gland which originated from ductal plate.


A demand-offer critical analysis of current drug development. Phase I drugs versus TCGA sequencing data.
Phase I clinical trials have become increasingly critical to regulatory approvals of novel agents. In phase I drug development, a global problem of unknown magnitude is the multiplicity of similar drugs being investigated against the same target, colloquially known as the 'me too' phenomenon. Ranging from December 2020 to December 2022 we annotated phase I clinical trials present on clinicaltrials.gov. Public databases were queried for annotation of investigational agents (IAs). Extensive literature research and data mining were performed to annotate agents not present in public databases. The Cancer Genome Atlas (TCGA) pan-cancer sequencing cohort was used to perform second-level analyses to evaluate tumour types with a higher number of IA matches. A total of 1054 unique drug targets were identified. The most frequent IA classes were: cell products (1223), small-molecule inhibitors (1110), antibodies (733), and vaccines (346). Only a minority (8.9%) of phase I IAs were explored against a target without a competitive agent; 7% of agents shared targets with 2-3 other agents. Unfortunately, the majority (84%) shared targets with at least four other agents. Using data from the TCGA pan-cancer sequencing potentially underserved histologies were identified. Analysis of alteration-IA matches revealed potentially frequent and unexplored alterations in many tumour types. The majority of IAs (86%) shared targets with at least three other agents. We argue that these duplicative efforts could be redirected toward unmet needs instead.


Ticks of Poland. Review of contemporary issues and latest research.
The paper presents current knowledge of ticks occurring in Poland, their medical importance, and a review of recent studies implemented in the Polish research centres on ticks and their significance in the epidemiology of transmissible diseases. In the Polish fauna there are 19 species of ticks (Ixodida) recognized as existing permanently in our country: Argas reflexus, Argas polonicus, Carios vespertilionis, Ixodes trianguliceps, Ixodes arboricola, Ixodes crenulatus, Ixodes hexagonus, Ixodes lividus, Ixodes rugicollis, Ixodes caledonicus, Ixodes frontalis, Ixodes simplex, Ixodes vespertilionis, Ixodes apronophorus, Ixodes persulcatus, Ixodes ricinus, Haemaphysalis punctata, Haemaphysalis concinna, Dermacentor reticulatus. Occasionally, alien species of ticks transferred to the territory of Poland are recorded: Amblyomma sphenodonti, Amblyomma exornatum, Amblyomma flavomaculatum, Amblyomma latum, Amblyomma nuttalli, Amblyomma quadricavum, Amblyomma transversale, Amblyomma varanensis, Amblyomma spp., Dermacentor marginatus, Hyalomma aegyptium, Hyalomma marginatum, Ixodes eldaricus, Ixodes festai, Rhipicephalus rossicus, Rhipicephalus sanguineus. The most common species of the highest medical and veterinary importance in Poland is invariably Ixodes ricinus. The review also sets out information on the risks of tickborne diseases in recreational areas of large cities in Poland, ticks as the cause of occupational diseases and dangerous species of ticks attacking people outside the Polish borders. Selected problems of the biology of ticks, the spread of alien species transferred on hosts and prevention of tick attacks have also been presented. The Polish studies on ticks are a valuable contribution to global research on the Ixodida.


True and false memory in the absence of perceptual identification.
Two experiments are reported in which, after attempting to identify a briefly flashed, masked test word, participants were asked to rate the likelihood that it had been presented in an earlier study list. Even when people were unable to identify such items, they demonstrated an ability to discriminate between those that were studied and those that were not studied; ratings given to studied items were significantly higher than ratings given to nonstudied items. This effect does not appear to be a data-driven phenomenon. In Experiment 1 it was found when the presentation modality was changed from study to test. In Experiment 2 false memory for unidentified items that were related to studied items was shown.


[Traumatic unilateral renal artery thrombosis and protein C deficiency. A case report].
Post traumatic renal artery thrombosis is rarely described in the literature. This pathology can result from stretch injury to inelastic intima of the renal artery, or by the direct flow to the abdomen causing compression injury to the renal artery against the vertebral column. However, the association of this pathology with hematologic diseases (in particular protein C deficit) was never described. We report an observation of a 28-year-old man with an uneventful history who was admitted to the intensive care unit for traumatic head injury associated with post traumatic renal artery thrombosis requiring nephrectomy. The etiologic investigation of this thrombo-embolic complication reveals a protein C deficit. Our patient was improved under treatment. This original observation confirms that post traumatic renal artery thrombosis can be associated with hematologic diseases (in particular protein C deficit).


Theoretical and practical aspects of receptor hormone assay in breast cancer.
The Authors have discussed the importance of ER/PR assay in breast cancer care. 162 patients were studied. The dextran-coated charcoal technique was used as the standard most readily available. The Authors point out the importance of specimen handling as is shown by time-decay studies, and the differences in ER/PR level at the time of the biopsy and mastectomy cut-section, and its eventual influence to better understanding assay results. The establishment of a center of control between different labs would also help in the development of more exact regimens for cancer care.


Claudins 1, 4, 5, 7 and occludin in ameloblastomas and developing human teeth.
To analyze the distribution pattern of claudins 1, 4, 5, 7 and occludin in benign and malignant ameloblastomas and developing human teeth. Paraffin-embedded tissue specimens of 25 benign and four malignant ameloblastomas and two developing human teeth were examined immunohistochemically using antibodies against claudins 1, 4, 5, 7 and occludin. In ameloblastomas strongest expression was seen for claudins 1 and 7 while claudin 4 was expressed less frequently. Claudin 5 and occludin were seen only in a minority of cases. There were no evident differences in the expression of claudins or occludin neither between different histologic subtypes of ameloblastomas nor between benign or malignant cases. The strongest expression for claudins was present in the central stellatum reticulum-like cells surrounding the microcysts and in the areas with squamous differentiation of the ameloblastomas. In developing teeth both claudin 1 and 7 stained strongly in the enamel epithelium, ameloblasts, and enamel matrix, but staining for claudin 4 was relatively weak. Claudin 5 was preferentially expressed only in vessels, and occludin staining ranged from negative to weak in ameloblastomas and teeth germs. There were no clear differences in the expression levels between benign and malignant ameloblastic tumors. The overexpression of claudins in the areas with microcyst formation may indicate their attempt to maintain the interepithelial cohesion of the cells. The strong immunoreactivity of ameloblasts and newly synthesized enamel matrix for claudins 1 and 7 indicates that they may be involved in cell signaling influencing enamel formation.


Physiologically based pharmacokinetic modeling of polyethylene glycol-coated polyacrylamide nanoparticles in rats.
Nanoparticles' health risks depend on their biodistribution in the body. Phagocytosis may greatly affect this distribution but has not yet explicitly accounted for in whole body pharmacokinetic models. Here, we present a physiologically based pharmacokinetic model that includes phagocytosis of nanoparticles to explore the biodistribution of intravenously injected polyethylene glycol-coated polyacrylamide nanoparticles in rats. The model explains 97% of the observed variation in nanoparticles amounts across organs. According to the model, phagocytizing cells quickly capture nanoparticles until their saturation and thereby constitute a major reservoir in richly perfused organs (spleen, liver, bone marrow, lungs, heart and kidneys), storing 83% of the nanoparticles found in these organs 120 h after injection. Key determinants of the nanoparticles biodistribution are the uptake capacities of phagocytizing cells in organs, the partitioning between tissue and blood, and the permeability between capillary blood and tissues. This framework can be extended to other types of nanoparticles by adapting these determinants.


An automated process for supporting decisions in clustering-based data analysis.
Metrics are commonly used by biomedical researchers and practitioners to measure and evaluate properties of individuals, instruments, models, methods, or datasets. Due to the lack of a standardized validation procedure for a metric, it is assumed that if a metric is appropriate for analyzing a dataset in a certain domain, then it will be appropriate for other datasets in the same domain. However, such generalizability cannot be taken for granted, since the behavior of a metric can vary in different scenarios. The study of such behavior of a metric is the objective of this paper, since it would allow for assessing its reliability before drawing any conclusion about biomedical datasets. We present a method to support in evaluating the behavior of quantitative metrics on datasets. Our approach assesses a metric by using clustering-based data analysis, and enhancing the decision-making process in the optimal classification. Our method assesses the metrics by applying two important criteria of the unsupervised classification validation that are calculated on the clusterings generated by the metric, namely stability and goodness of the clusters. The application of our method is facilitated to biomedical researchers by our evaluomeR tool. The analytical power of our methods is shown in the results of the application of our method to analyze (1) the behavior of the impact factor metric for a series of journal categories; (2) which structural metrics provide a better partitioning of the content of a repository of biomedical ontologies, and (3) the heterogeneity sources in effect size metrics of biomedical primary studies. The use of statistical properties such as stability and goodness of classifications allows for a useful analysis of the behavior of quantitative metrics, which can be used for supporting decisions about which metrics to apply on a certain dataset.


Job satisfaction among obstetrician-gynecologists: a comparison between private practice physicians and academic physicians.
Physician job satisfaction has been the subject of much research. However, no studies have been conducted comparing academic and private practice physician satisfaction in obstetrics and gynecology. This study was undertaken to measure satisfaction levels for academic and private practice obstetrician-gynecologists and compare different aspects of their practice that contributed to their satisfaction. A survey was mailed to randomly selected obstetrician-gynecologists in Memphis, TN; Birmingham, AL; Little Rock, AR; and Jackson, MS. Physicians were asked to respond to questions concerning demographics and career satisfaction. They were also asked to assess the contribution of 13 different aspects of their practice in contributing to their job selection and satisfaction using a Likert scale. A score of 1 meant the physician completely disagreed with a statement regarding a factor's contribution or was completely dissatisfied; a score of 5 meant the physician completely agreed with a factor's contribution or was completely satisfied. Simple descriptive statistics, as well as the 2-sample t test, were used. Likert scale values were assumed to be interval measurements. Of the 297 questionnaires mailed, 129 (43%) physicians responded. Ninety-five (74%) respondents rated their overall satisfaction as 4 or 5. No significant difference was found between academic and private physicians when comparing overall job satisfaction (P = .25). When compared to private practice physicians, the aspects most likely contributing to overall job satisfaction for academic physicians were the ability to teach, conduct research, and practice variety (P = .0001, P = .0001, and P = .007, respectively). When compared with academic physicians, the aspects most likely contributing to job satisfaction for private practice physicians were autonomy, physician-patient relationship, and insurance reimbursement (P = .0058, P = .0001, and P = .0098, respectively). When choosing a practice setting, academic physicians found variety, teaching, and research to be more important (P = .0027, P = .0001, and P = .0001, respectively). In contrast, private practice physicians found autonomy, physician-patient relationship, coworkers, and geographic location to be more important (P = .0005, P = .0001, P = .0035, and P = .0016, respectively). Academic and private practice physicians are equally satisfied with their careers. However, teaching, research and variety contribute more to academic satisfaction, whereas autonomy, physician-patient relationship, and coworkers contribute more to satisfaction for the physician in private practice. This study may be used when counseling residents concerning their career options.


Evaluation of Quantitative PET/MR Enterography Biomarkers for Discrimination of Inflammatory Strictures from Fibrotic Strictures in Crohn Disease.
To retrospectively evaluate positron emission tomography (PET)/magnetic resonance (MR) enterography for the differentiation of fibrotic strictures from inflammatory strictures in patients with Crohn disease. This HIPAA-compliant retrospective study was approved by the institutional review board. Patients gave their written informed consent for study enrollment. PET/MR enterography images were evaluated in 19 patients with Crohn disease who had strictures that underwent surgical resection with pathologic confirmation. Two radiologists and a nuclear medicine physician in consensus evaluated the following bowel wall PET/MR enterography biomarkers: signal intensity (SI) on T2-weighted images, apparent diffusion coefficient (ADC), PET maximum standardized uptake value (SUVmax), SI on T2-weighted images × SUVmax, and ADC × SUVmax values at levels that corresponded to pathologic specimens. MR, PET, and hybrid PET/MR biomarkers were compared, and the performance for differentiation of inflammatory strictures from fibrotic strictures was assessed. Mixed-model regression analysis was used to compare the mean imaging parameters between groups; the P values were corrected for the five comparisons by using the Bonferroni method. Three of the PET/MR enterography biomarkers, SUVmax, SI on T2-weighted images × SUVmax, and ADC × SUVmax, showed significant differences in the fibrosis group compared with the fibrosis with active inflammation group and the active inflammation only group. The best discriminator between fibrosis and active inflammation was the combined PET/MR enterography biomarker ADC × SUVmax cutoff of less than 3000, which was associated with accuracy, sensitivity, and specificity values of 0.71, 0.67, and 0.73, respectively. PET/MR enterography offers a potential noninvasive technique for the differentiation of purely fibrotic strictures from mixed or inflammatory strictures. A hybrid biomarker that incorporates both MR and PET information performed better for stricture evaluation than either modality alone.


[Forced downward and convergent deviation of the eyes and periodic ocular movements in aneurysmal hemorrhage of the mesencephalic tegmentum].
The authors report a case of forced convergent and downward deviation of the eyes with paralysis of upward gaze and, at times, with lateral periodic alternating movements. Pathological examination disclosed a subarachnoid aneurysm behind the midbrain with a fresh hematoma within the mesencephalic tegmentum at the level of the superior colliculi. The clinical and pathological data are discussed.


KAT6A mutations in Arboleda-Tham syndrome drive epigenetic regulation of posterior HOXC cluster.
Arboleda-Tham Syndrome (ARTHS) is a rare genetic disorder caused by heterozygous, de novo mutations in Lysine(K) acetyltransferase 6A (KAT6A). ARTHS is clinically heterogeneous and characterized by several common features, including intellectual disability, developmental and speech delay, and hypotonia, and affects multiple organ systems. KAT6A is the enzymatic core of a histone-acetylation protein complex; however, the direct histone targets and gene regulatory effects remain unknown. In this study, we use ARTHS patient (n = 8) and control (n = 14) dermal fibroblasts and perform comprehensive profiling of the epigenome and transcriptome caused by KAT6A mutations. We identified differential chromatin accessibility within the promoter or gene body of 23% (14/60) of genes that were differentially expressed between ARTHS and controls. Within fibroblasts, we show a distinct set of genes from the posterior HOXC gene cluster (HOXC10, HOXC11, HOXC-AS3, HOXC-AS2, and HOTAIR) that are overexpressed in ARTHS and are transcription factors critical for early development body segment patterning. The genomic loci harboring HOXC genes are epigenetically regulated with increased chromatin accessibility, high levels of H3K23ac, and increased gene-body DNA methylation compared to controls, all of which are consistent with transcriptomic overexpression. Finally, we used unbiased proteomic mass spectrometry and identified two new histone post-translational modifications (PTMs) that are disrupted in ARTHS: H2A and H3K56 acetylation. Our multi-omics assays have identified novel histone and gene regulatory roles of KAT6A in a large group of ARTHS patients harboring diverse pathogenic mutations. This work provides insight into the role of KAT6A on the epigenomic regulation in somatic cell types.


Results of a modified Duhamel Operation for Hirschsprung's disease using the GIA stapler.
Thirty-six patients with Hirschsprung's disease underwent surgery using modified Duhamel operation with the GIA (gastrointestinal autosuture) stapler over a 12-year period at our institution. We now present our experience from the perspective of postoperative complications and anorectal function. Postoperative complications included minimal anal bleeding due to the GIA stapler in 2 patients, and mild enterocolitis in 6 patients. Severe complications such as anastomotic leakage or stricture were not observed in our series. Overall, postoperative anorectal function was good in patients without Down's syndrome or cerebral palsy. Follow-up of our patients showed that a good quality of life was achieved during childhood. We conclude, therefore, that the modified Duhamel operation, with the use of the GIA stapler, is a safe and easy procedure for correction of Hirschsprung's disease.


Genetic liability to fractures in the elderly.
The genetic impact on the causation of osteoporotic fractures is unclear. A large twin study is ideally suited to determine the genetic liability to categories of fracture at various ages. A cohort of all 33 432 Swedish twins born from 1896 to 1944 was used to evaluate the genetic liability to fracture occurrence in the elderly. The Swedish Inpatient Registry and computer-assisted telephone interviews enabled us to identify 6021 twins with any fracture, 3599 with an osteoporotic fracture, and 1055 with a hip fracture after the age of 50 years. Genetic variation in liability to fracture differed considerably by type of fracture and age. Less than 20% of the overall age-adjusted fracture variance was explained by genetic variation. The age-adjusted heritability of any osteoporotic fracture was slightly greater (0.27; 95% confidence interval [CI], 0.09-0.28), and for hip fracture alone, it was 0.48 (95% CI, 0.28-0.57). Heritability was not attenuated after further adjustment for several known osteoporotic covariates but was considerably greater for first hip fractures before the age of 69 years (0.68; 95% CI, 0.41-0.78) and between 69 and 79 years (0.47; 95% CI, 0.04-0.62) than for hip fractures after 79 years of age (0.03; 95% CI, 0.00-0.26). The importance of genetic factors in propensity to fractures depends on fracture site and age. The search for susceptibility genes and environmental factors that may modulate expression of these genes in younger elderly patients with hip fracture, the most devastating osteoporotic fracture, should be encouraged. Prevention of fractures in the oldest elderly should focus on lifestyle interventions.


Involvement of adrenal and thyroid activity in the annual testicular regression of red headed bunting (Emberiza bruniceps).
Thyroid and adrenal activities are closely associated with reproductive cycle and any alteration in these endocrine functions may cause changes in the pituitary-gonadal axis. To understand this interrelationship during testicular senstive phase (month of April) male birds were injected with metapyrone(corticosterone synthesis blocks 1 microg/bird/day & 10 microg/bird/day) and newmercazol (thyroxine synthesis blocks 10 microg/bird/day) over a period of 12 weeks. During late breeding phase (month of August) two sets of birds having large gonad (photosensitive) and regressed gonad (photorefractory) were injected with metapyrone (10 microg/bird/alternate day). Results indicate that decreased activity of both adrenal and thyroid, extended the breeding phase but rate of regression decreased only in the case of bird receiving higher level of metapyrone.However,in the second group rate of gonadal regression was slow only in those bird where treatment was started during photosensitive phase. It may be suggested that optimum level of activity of adrenal and thyroid function are essential for termination of reproduction and any alteration in these function may alter seasonal pattern of neuroendocrine gonadal axis.


Culex pipiens pipiens: characterization of immune peptides and the influence of immune activation on development of Wuchereria bancrofti.
Stimulating or augmenting the innate immune response of insect vectors has been shown to impede or disrupt the development and transmission of eukaryotic pathogens; however, the majority of such studies have utilized model systems and not natural parasite-vector systems. The Culex pipiens complex of mosquitoes functions as a primary urban vector of Wuchereria bancrofti, a causative agent of lymphatic filariasis. To test the effects of immune activation on this vector-parasite interaction, Culex pipiens pipiens from the filariasis-endemic Nile Delta were subjected to bacteria inoculation and subsequently fed a blood meal containing W. bancrofti. No difference was seen between parasite development in these mosquitoes as compared to non-inoculated controls. A set of expressed sequence tags from blood-fed midgut and bacteria-inoculated Cx. p. pipiens reveals transcripts for the immune peptides cecropin, gambicin and defensin--all of which have been reported to have antiparasitic effects. Sequences and transcriptional profiles for these peptides are reported. The discrepancy between these results and those reported for the model parasite, Brugia malayi, in the mosquito Aedes aegypti are discussed.


The Surgical Biology of Cancer.
The surgical principles in the treatment of cancer were first clearly enunciated in 1906 by Halsted in the treatment of carcinoma of the breast, namely wide surgical removal of the breast together with the regional lymph nodes to which it spreads. The hope was that the regional nodes would serve as filters and prevent the spread of malignant cells into the blood stream, enabling the surgeon to excise every last cancer cell from the patient, thereby curing the cancer.


Identification of cytochrome P450 2C2 protein complexes in mouse liver.
Interactions of microsomal cytochromes P450 (CYPs) with other proteins in the microsomal membrane are important for their function. In addition to their redox partners, CYPs have been reported to interact with other proteins not directly involved in their enzymatic function. In this study, proteins were identified that interact with CYP2C2 in vivo in mouse liver. Flag-tagged CYP2C2 was expressed exogenously in mouse liver and was affinity purified, along with associated proteins which were identified by MS and confirmed by Western blotting. Over 20 proteins reproducibly copurified with CYP2C2. The heterogeneous sedimentation velocity of CYP2C2 and associated proteins by centrifugation in sucrose gradients and sequential immunoprecipitation analysis were consistent with multiple CYP2C2 complexes of differing composition. The abundance of CYPs and other drug metabolizing enzymes and NAD/NADP requiring enzymes associated with CYP2C2 suggest that complexes of these proteins may improve enzymatic efficiency or facilitate sequential metabolic steps. Chaperones, which may be important for maintaining CYP function, and reticulons, endoplasmic reticulum proteins that shape the morphology of the endoplasmic reticulum and are potential endoplasmic reticulum retention proteins for CYPs, were also associated with CYP2C2.


Long-term follow-up of drusen-like lesions in patients with type II mesangiocapillary glomerulonephritis.
Drusen-like lesions beneath the retina in patients with partial lipodystrophy and type II mesangiocapillary glomerulonephritis (MCGN) were first reported in 1989. This study reports the long-term follow-up of this original cohort of patients more than 10 years later. Three patients had undergone renal transplantation. Retinopathy was graded semiquantitatively using an international classification and grading system. Progression was assessed by comparing the visual acuities and the colour, red-free and fluorescein angiographic photographs at baseline and review. The visual acuity in all four patients was unchanged, as were the bilateral drusen-like lesions. The retinal pigment hypertrophy at the posterior pole of two of the patients was also unchanged. There were no signs of choroidal neovascularisation or central serous retinopathy in any patient. There was no progression of retinopathy over 10 years in any patient. This study suggests that in patients with type II MCGN, (a) factors other than drusen may contribute to choroidal neovascularisation and (b) renal transplantation does not appear to increase the risk of progression of retinopathy.


Eicosapentaenoic- and arachidonic acid metabolism in isolated liver cells.
The oxidation and esterification of 14C labelled eicosapentaenoic acid (20:5,n-3) and arachidonic acid (20:4,n-6) in isolated liver cells has been studied. The bioconversion of C18 and C22 polyunsaturated fatty acids to 20:5(n-3) and 20:4(n-6) is also discussed. Adrenic acid (22:4,n-6) and docosahexaenoic acid (22:6,n-3) are retroconverted to 20:4(n-6) and 20:5(n-3) respectively by peroxisomal beta-oxidation. 20:4(n-6) and 20:5(n-3) are both mainly oxidized in the mitochondria. The peroxisomal contribution to the oxidation of 20:5,n-3 is however larger than with 18:2(n-6) and 18:1(n-9) which are predominantly oxidized by mitochondrial beta-oxidation. Isolated liver cells oxidize more 20:5(n-3) and esterify less than observed with 20:4(n-6) as substrate. In liver cells from essential fatty acid deficient animals 20:5(n-3) and 20:4(n-6) are both efficiently directed to the phospholipids. In hepatocytes from animals fed a diet rich in 18:2(n-6) and 18:3(n-3) arachidonic acid is still to a large extent esterified in the phospholipids while 20:5(n-3) only to a small extent is esterified in this lipid fraction. 18:2(n-6) is much more efficiently esterified in the phospholipids than is 18:3(n-3) which is also rapidly removed from the phospholipids. The delta 4 desaturase activity is increased in essential fatty acid deficiency similar to delta 6 desaturase. The competition between 18:3(n-3) and 18:2(n-6) for delta 6 desaturation in combination with a much higher dietary intake of 18:2(n-6) than of 18:3(n-3) may limit the conversion of dietary 18:3(n-3) to 20:5(n-3).(ABSTRACT TRUNCATED AT 250 WORDS)


Right lower lobe sleeve resection: bronchial flap to correct caliber disparity.
Sleeve resection of the right lower lobe is performed infrequently. Caliber disparity between the truncus intermedius and the middle lobe bronchus is a major problem. We report a case of lung cancer completely resected by sleeve resection of the right lower lobe. A bronchial flap constructed from the distal bronchial end was used for correction of the caliber disparity, and interlobar dissection between the upper and middle lobes effectively reduced the tension on the anastomotic site. These procedures are useful for sleeve resection of the right lower lobe.


The influence of charge trapping on the electrochromic performance of poly(3,4-alkylenedioxythiophene) derivatives.
This paper describes the electrochromic properties of a series of poly(3,4-alkylenedioxythiophene) (PXDOT) derivatives featuring various ring sizes and substitutions. The presence of a bulky group on the monomer resulted in a polymer possessing a more-open morphology, which promoted reversible ionic transfer. We used an electrochemical quartz crystal microbalance and cyclic voltammetry to investigate the properties of these polymers. We found that both cations and anions were involved in the charge compensation process. Furthermore, PXDOT derivatives possessing larger ring sizes and/or longer alkyl substituents exhibited less trapping of ions within the polymer during the redox process. The long-term electrochromic stability of these PXDOTs depended strongly on the number of trapped ions. Although the transmittance attenuation of poly(3,4-ethylenedioxythiophene) (PEDOT) decreased from 53 to 42%, we observed no significant decay for poly(diethyl-3,4-dihydro-2H-thieno[3,4-b]-[1,4]dioxepine) (PProDOT-Et(2)) after 400 cycles.


Effectiveness of various methods of educating children and adolescents for the maintenance of oral health: A systematic review of randomized controlled trials.
School-based oral health education has emerged as one of the most effective methods to instill a positive attitude toward oral health in children. This systematic review (SR) aimed to assess the effectiveness of different methods of oral health education in children and adolescents. Systematic search was conducted in PubMed, Cochrane, Web of Science, LILACS, Scopus, and EMBASE on January 29, 2023. Review included only randomized controlled trials (RCTs), and their risk of bias (ROB) was assessed by Cochrane RoB-2. Data were extracted and analyzed by expert group. The GRADE approach was used to assess the quality of evidence for each outcome. Every step was conducted in duplicate, and disagreements were resolved by consulting the third reviewer. The SR included 10 RCTs with majority showing a high ROB. They included 12 methods with the commonest being the use of leaflets/flash cards. Most interventions were effective in improving oral hygiene, gingival health, and knowledge except conventional teaching methods, lecture by using PowerPoint by dentists, and e-learning interventions. The grade of evidence in the majority of outcomes was found to be low or very low. Interventions using the interactive methods such as games, motivational-experiential learning, and audiovisual methods were found to be more effective than controls and other interventions. Variabilities in study methods and outcome variables must be addressed in the future.


Synergistic bombesin and insulin stimulation of DNA synthesis in human fetal kidney in serum-free culture.
The respective influences of growth factors during kidney development can be directly evaluated using the chemically-defined serum-free culture system perfected in our laboratory. Since, in this culture model, conditions are minimal for growth and differentiation, DNA synthesis sharply decreases during the first 48 h. The addition of epidermal growth factor (EGF, 100 ng/ml), insulin (5 micrograms/ml) and transferrin (5 micrograms/ml) significantly restores this important cellular function. The objective of the present study was to determine the influence of bombesin, a potent mitogen, supplemented alone or in combination with insulin, transferrin and/or EGF. Cortical explants of human fetal kidneys (17-20 weeks) were maintained during 5 days in culture. When compared with 5 day controls (L-15 medium only), bombesin generated a maximal though weak effect on DNA synthesis at a concentration of 0.3 nM, corresponding to a stimulation index (SI) of 22%. When combined with either transferrin or EGF, or with transferrin plus EGF, bombesin did not alter the SI of individual factors. Insulin, in turn, greatly increased DNA synthesis (SI = 169%), while bombesin strongly potentiated this effect (SI = 275%). Transferrin also enhanced insulin SI from 169 to 240%. When added as a third factor, bombesin further potentiated the effectiveness (SI = 338%) of the combination insulin plus transferrin. These results indicate that bombesin controls cell proliferation in synergism with other regulators and hence may act as a competence growth factor during nephrogenesis.


[Role of apoptosis and mitochondrial apoptotic pathway in glucolipotoxicity-induced islet beta-cell dysfunction].
To investigate the mechanism of beta-cell dysfunction induced by glucolipotoxicity in high fat-fed obese rats. Eighteen high-fat obese male Wistar rats were assigned into 3 groups and underwent 48-hour infusion through the jugular vein with normal saline (n=6), 20% intralipid + heparin (FFA group, n=6), or 25%glucose +20% intralipid + heparin (GS-FFA group, n=6). The plasma beta-hydroxybutyric acid (beta-HBA) was measured before and at the end of the infusion. After the infusion, the rats were sacrificed following an intravenous glucose tolerance test (IVGTT) to remove the tail of the pancreas for detection of apoptotic islet cells using TUNEL method. Immunohistochemical staining was performed to detect the expression of cytochrome c (cyt c), apoptosis-inducing factor (AIF), caspase-9 and caspase-3 in the islet cells. At the end of the infusion, all the rats exhibited increased plasma beta-HBA levels, which was the highest in the GS-FFA group (P<0.05). IVGTT performed after the infusion showed a significantly lower insulinogenic index in GS-FFA group than that in NS and FFA groups. Greater number of apoptotic islet cells was found in the GS-FFA group than in the FFA and NS groups (P<0.05), and the islets had significantly higher levels of cyt c, AIF, caspase-9 and caspase-3 in the former group than in the latter two groups (P<0.05). Hyperglycemia and high free fatty acid level synergistically impair insulin secretions to cause ketone overproduction in high fat-fed obese rats. The beta-cell dysfunction due to glucolipotoxicity is associated with increased beta-cell apoptosis and activation of mitochondrial apoptotic pathway.


First-Principles Study of B16N16 Cluster-Assembled Porous Nanomaterials.
Owing to the similar valence electron structures between the B-N bond and the C-C bond, boron nitride, similar to carbon, can form abundant polymorphs with different frameworks, which possess rich mechanical and electronic properties. Using the hollow, cage-like B16N16 cluster as building blocks, here, we established three new BN polymorphs with low-density porous structures, termed Cub-B16N16, Tet-B16N16, and Ort-B16N16, which have cubic (P4¯3m), tetragonal (P4/nbm), and orthomorphic (Imma) symmetries, respectively. Our density functional theory (DFT) calculations indicated that the existence of porous structure Cub-B16N16, Tet-B16N16, and Ort-B16N16 were not only energetically, dynamically, thermally and mechanically stable, they were even more stable than some known phases, such as sc-B12N12 and Hp-BN. The obtained Pugh's ratio showed that the Cub-B16N16 and Tet-B16N16 structures were brittle materials, but Ort-B16N16 was ductile. The analysis of ideal strength, Young's moduli, and shear moduli revealed that the proposed new phases all exhibited sizable mechanical anisotropy. Additionally, the calculation of electronic band structures and density of states showed that they were all semiconducting with a wide, indirect band gap (~3 eV). The results obtained in this work not only identified three stable BN polymorphs, they also highlighted a bottom-up way to obtain the desired materials with the clusters serving as building blocks.


In vitro activity of artemether against African isolates (Senegal) of Plasmodium falciparum in comparison with standard antimalarial drugs.
The in vitro activity of artemether against 56 African isolates of Plasmodium falciparum from Senegal was evaluated using an isotope-based drug susceptibility semi-microtest. The 50% inhibitory concentration (IC50) values for artemether were in a narrow range from 0.8 to 15.2 nM (mean IC50 = 3.43 nM) and the 95% confidence interval (CI) was 2.50-4.36 nM. Artemether was equally effective on chloroquine-sensitive and chloroquine-resistant isolates (mean IC50 = 346 nM, 95% CI = 2.08-4.84 nM versus mean IC50 = 2.80 nM, 95% CI = 2.00-3.60 nM). There was a significant positive correlation between responses to artemether and mefloquine (r2 = 0.36, P < 0.001), artemether and quinine (r2 = 0.085, P < 0.05), artemether and halofantrine (r2 = 0.075, P < 0.05), quinine and mefloquine (r2 = 0.205, P < 0.01), quinine and halofantrine (r2 = 0.124, P < 0.05), and mefloquine and halofantrine (r2 = 0.801, P < 0.001). A positive correlation between these drugs suggests in vitro cross-resistance or at least in vitro cross-susceptibility.


Management of Hyperthyroidism in Pregnancy: Results of a Survey among Members of the European Thyroid Association.
An optimal management of maternal hyperthyroidism is important for positive pregnancy outcome, and to this end, the Endocrine Society published their guidelines in 2007. This survey aimed to investigate to what extent the clinical practice relating to the management of hyperthyroidism during pregnancy in Europe is uniform and consistent with the guidelines. We e-mailed an online questionnaire survey based on clinical case scenarios to 605 members of the European Thyroid Association. We analysed 190 responses from 28 European countries. For a woman with newly diagnosed Graves' disease (GD) and wishing pregnancy, 78% of the responders would initiate antithyroid drugs (ATDs), while 22% would recommend definitive treatment with radioiodine or surgery. In case of a relapsed GD before pregnancy, 80% preferred definitive treatment. For a woman with newly diagnosed GD during pregnancy, 53% would treat with propylthiouracil, 12% with methimazole, and 34% with propylthiouracil initially and switch to methimazole after the first trimester. Responders used several combinations of tests to monitor the dose of ATDs, and the thyroid test results they targeted were inconsistent. For a lactating woman with GD, 68% would give ATDs without stopping lactation. Variation in the clinical practices surrounding the management of hyperthyroid pregnant women in Europe still exists.


Rift Valley fever knowledge, mitigation strategies and communication preferences among male and female livestock farmers in Eastern Province, Rwanda.
The Government of Rwanda reported an outbreak of Rift Valley fever (RVF) in the Eastern Province in 2018. To respond to the outbreak, vaccination and education campaigns about the disease were carried out. Because RVF cases continue to be detected in Rwanda and the disease impacts livelihoods and health, accurate knowledge and communication are imperative. The objectives of this study were to evaluate knowledge and risk perceptions of RVF transmission among livestock farmers in Nyagatare District, Eastern Province, Rwanda, and to compare RVF knowledge, risk perceptions, and farming practices between male and female livestock farmers. This cross-sectional, quantitative study was conducted in selected sectors of Nyagatare District in the Eastern Province of Rwanda in June 2019. A 34-question survey was used to ask about demographics, livestock ownership, risk perceptions about zoonotic diseases and livestock management, RVF knowledge, preferred communication sources and information sharing strategies, and protective strategies for RVF mitigation while working with livestock. Livestock farmers were interviewed at three milk collection centers, two village meeting points, a farm cooperative meeting, and during door-to-door visits in villages. In total, 123 livestock farmers were interviewed. The survey found that most livestock farmers lacked knowledge about epizootic and zoonotic transmission of RVF, more male livestock farmers were familiar with RVF and risk mitigation strategies, and female livestock farmers are not viewed as reliable sources of information. Additionally, most livestock farmers had not vaccinated their animals against RVF despite past vaccination campaigns. Radio was the most popular communication channel. These findings show that RVF knowledge and information sharing are inadequate among livestock farmers in Eastern Province. Therefore, vaccination and education campaigns may need to be reevaluated within the context of these trends in order to prepare for future RVF outbreaks.


Unexpected electronic process of H2 activation by a new nickel borane complex: comparison with the usual homolytic and heterolytic activations.
H-H σ-bond activation promoted by Ni[MesB(o-Ph2PC6H4)2] (1(Mes)) was theoretically investigated with the density functional theory method. In 1(Mes), the nickel 3d, 4s, and 4p orbital populations are similar to those of the typical nickel(II) complex. First, one H2 molecule coordinates with the nickel center to form a dihydrogen complex, 2, which induces an increase in the nickel 3d and 4p orbital populations and thus a decrease in the nickel oxidation state. Then, the H-H σ-bond is cleaved under the unusual cooperation of the electron-rich nickel center and the electron-deficient borane ligand in a polarized manner, leading to an unprecedented trans-nickel(II) hydridoborohydrido complex, 3. In the transition state, charge transfer (CT) occuring from the H2 moiety to the 1(Mes) moiety (0.683 e) is much larger than the reverse CT (0.284 e). As a result, cleavage of the H-H σ-bond affords two positively charged hydrogen atoms. In this process, the boron atomic population and the nickel 4p orbital population increase, but the nickel 3d orbital population decreases. After cleavage of the H-H σ-bond, CT from the nickel 4p orbital to these positively charged hydrogen atoms occurs to afford 3, where the oxidation state of the nickel center increases to +2. These electronic processes are different from those of the usual homolytic and heterolytic H-H σ-bond activations. Regeneration of 1(Mes) and the role of the borane ligand in these reactions are also discussed in detail.


Intercalated chemotherapy and erlotinib for non-small cell lung cancer (NSCLC) with activating epidermal growth factor receptor (EGFR) mutations.
Among attempts to delay development of resistance to tyrosine kinase inhibitors (TKIs) in patients with advanced non-small cell lung cancer (NSCLC) with activating mutations of epidermal growth factor receptor (EGFR), intercalated therapy has not been properly evaluated. In a phase II trial, 38 patients with EGFR mutated NSCLC in advanced stage were treated with 4 to 6 3-weekly cycles of intercalated schedule with gemcitabine (1250 mg/m2, days 1 and 4), cisplatin (75 mg/m2, day 2) and erlotinib (150 mg, days 5 - 15), followed by continuous erlotinib as maintenance. In addition to standard radiologic evaluation according to RECIST, PET/CT was done prior to treatment and at 6 months, using PERCIST as a method for assessment of response. The primary endpoint was progression-free survival (PFS). In general, tolerance to treatment was good, even among 8 patients with performance status 2-3 and 13 patients with brain metastases; grade 4 toxicity included 2 cases of neutropenia and 4 thrombo-embolic events. Complete response (CR) or partial response (PR) were seen in 15 (39.5%) and 17 (44.7%) cases, respectively. All cases of CR were confirmed also by PET/CT. Median PFS was 23.4 months and median overall survival (OS) was 38.3 months. After a median follow-up of 35 months, 8 patients are still in CR and on maintenance erlotinib. In conclusion, intercalated treatment for treatment-naive patients with EGFR activating mutations leads to excellent response rate and prolonged PFS and survival. Comparison of the intercalated schedule to monotherapy with TKIs in a randomized trial is warranted.


Inhibition of MRP1/ABCC1, MRP2/ABCC2, and MRP3/ABCC3 by nucleoside, nucleotide, and non-nucleoside reverse transcriptase inhibitors.
Many drug interactions with drugs used for the therapy of human immunodeficiency virus (HIV) occur at the level of different cytochrome P450 isozymes. Increasing evidence suggests that antiretrovirals may also modify activity and expression of active drug transport systems. Such interactions may alter drug absorption, elimination, and also drug distribution and reach clinical importance if thereby access to the target site is affected. Beyond P-glycoprotein, the family of multidrug resistance-related proteins (MRP/ABCC) substantially contributes to the elimination of numerous drugs and their metabolites. Because the interaction of MRPs with non-HIV protease inhibitor antiretrovirals has not been studied thoroughly, we investigated whether important non-nucleoside reverse transcriptase inhibitors (NNRTI) (delavirdine, efavirenz, and nevirapine), nucleoside reverse transcriptase inhibitors (NRTI) (abacavir, emtricitabine, and lamivudine), and tenofovir as a nonnucleotide reverse transcriptase inhibitor can interact with MRP1, MRP2, and MRP3 in vitro. Inhibition of these ABC transporters was quantified by confocal laser-scanning microscopy using the 5-chloromethylfluorescein diacetate assay. With the exception of abacavir, which had no effect on MRP3, all the test compounds increased intracellular 5-chloromethylfluorescein fluorescence in a concentration-dependent manner, and this effect was observed in all the overexpressing cell lines but not in the parental cell line, indicating inhibition of MRP1, MRP2, and MRP3. In conclusion, the present study provides the first evidence for a significant and concentration-dependent inhibition of MRPs by NNRTI, NRTI, and tenofovir, which was most pronounced for delavirdine, efavirenz, and emtricitabine, suggesting that this might contribute to some of the known drug interactions impairing HIV therapy and also to the superior effectiveness of combination pharmacotherapy.


[Effect of Electroacupuncture at "Neiguan"(PC 6) on Serum and Myocardial Metabolites in Rats with Myocardial Ischemia Reperfusion Injury Based on Nuclear Magnetic Resonance Spectroscopy].
We have repeatedly demonstrated that electroacupuncture (EA) of "Neiguan"(PC 6) can improve myocardial ischemia in rats. The present study was designed to investigate the metabolomic profile of peripheral blood se-rum and myocardium involving EA-induced improvement of myocardial ischemia-reperfusion injury (MIRI) in rats by using nuclear magnetic resonance spectroscopy. Thirty male SD rats were equally randomized into blank control, model and EA groups. Rats of the control group were only banded for 20 min, once a day for 7 days. The MIRI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 60 min, and rats of the model group were banded as those in the control group. EA (10 Hz/50 Hz, 1 mA) was applied to bilateral PC 6 for 20 min, once daily for 7 days. The blood samples and left ventricular myocardial tissues were collected for assaying the profiles of differential metabolites using 1H nuclear magnetic resonance (1H NMR) spectroscopy and multivariate statistical analysis such as the principal components analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and orthogonal PLS-DA (O-PLS-DA) with SIMCA-P software 12.0. A total of 19 differential metabolites (17 down-regulated, 2 up-regulated) in the serum and 14 differential metabolites (13 down-regulated and 1 up-regulated) in the ischemic left myocardium were identified after MIRI. Of the 19 serum differential metabolites, amino acids (leucine, isoleucine, valine,alanine, lysine, glycine, glutamine), 3-hydroxy butyric acid (3-HB), lactic acid, acetate, N-acetyl glycoprotein (NAc), acetone, acetoacetate, succinate, polyunsaturated fatty acids (PUFA), creatine, glycerophosphocholine (GPC) were down-regulated; while low density lipoprotein (LDL), LDL/very low density lipoprotein(LDL/VLDL)and glucose obviously up-regulated. Of the 14 myocardial differential metabolites, amino acids (alanine, lysine, glutamate, glutamine, aspartate, taurine, glycine, threonine), GPC, creatine, lactic acid, adenosine monophosphate (AMP), nicotinamide adenine dinucleotide (NAD+) were significantly decreased, and glucose was up-regulated. Following EA treatment, most of the decreased serum differential metabolites except acetone, acetoacetate and PUFA, and the increased serum LDL, LDL/VLDL and glucose recovered, basically close to the control level; and the decreased myocardial creatine, GPC and NAD+ were also apparently up-regulated and the increased myocardial glucose was down-regulated. But, myocardial threonine and AMP still presented a decreasing state. Although the pattern of myocardial differential metabolites of the EA group had a trend to be close to the control group, the significant difference still existed, while the metabolic pattern of serum metabolites in the EA group was close to that of the control group. EA stimulation of PC 6 can regulate serum or/and myocardial metabolites as amino acids, carbohydrates, lipids, etc. in MIRI rats, of which both serum and myocardial creatine, GPC and glucose may be jointly confer a favorable potential for EA-induced improvement of MIRI.


Adult pedestrian behavior when accompanying children on the route to school.
Pedestrian injuries are a major cause of morbidity and mortality to children, especially boys. Adults serve as pedestrian role models and provide learning opportunities for children when walking to school. The research objectives were to investigate adult pedestrian behavior when accompanying boys and girls. Behavioral observation of 140 adult pedestrians accompanying 4- to 9-year-old children was done in British residential locations. Observations took place at light-controlled crossings, speed-restricted school safety zones, and mid-block unmarked crossing sites. Behaviors observed included stopping at the curb, waiting at the curb, looking left and right before and during road crossing, holding hands, talking, and walking straight across. In general, adults modeled safe road crossing behaviors. Adult safe behavior scores were higher when accompanying girls than when accompanying boys. No statistically significant differences were found by child age group. The fewest safe pedestrian behaviors were observed at light-controlled crossings. Adult pedestrians behave differently when with boys and girls and at different types of road crossing site. Interventions aimed at reducing pedestrian injuries to children may need to take these different everyday experiences into consideration.


A specific receptor for retinol binding protein as detected by the binding of human and bovine retinol binding protein to pigment epithelial cells.
By means of autoradiographic techniques the specific plasma carrier of retinol, namely retinol binding protein (RBP) in a radioactive form (retinol-125I.RBP), bound specifically in vivo to the choroidal surface of intact, isolated bovine pigment epithelial cells. The retinol-125I.RBP did not bind to the retinal surface of the pigment epithelial cells nor did not bind to photoreceptors. Retinol is normally provided to the retina from the blood via a specific complex formation with a receptor on the chordial surface of the pigment epithelial cells. Retinol metabolism might be deranged in some diseases through a defect in the pigment epithelial receptor for RBP.


A Recombinant Porcine Reproductive and Respiratory Syndrome Virus Stably Expressing DsRed Protein Based on Bacterial Artificial Chromosome System.
Recombinant viruses possessing reporter proteins as tools are widely applied in investigating viral biology because of the convenience for observation. Previously, we generated a recombinant pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) with enhanced green fluorescent protein (EGFP) reporter for monitoring virus spread and screening of neutralizing antibodies. PRRSV with different kinds of reporters can support more application scenarios. Here, we described a new genetically stable infectious clones of a highly pathogenic PRRSV (HP-PRRSV) harboring the DsRed (a red fluorescent protein isolated from the coral Discosoma) gene. In the recombinant infectious clone, the transcription regulatory sequence 2 (TRS2) of PRRSV was inserted between the open reading frame 7 (ORF7) and 3'UTR to drive the transcription of DsRed gene, which makes it a separate transcription unit in the viral genome. Using the bacterial artificial chromosome (BAC) system and cytomegalovirus (CMV) promoter, the recombinant HP-PRRSV with the DsRed insertion was successfully rescued and showed similar growth and replication patterns compared with the wild-type virus in the MARC-145 cells. In addition, the DsRed protein was stably expressed in the recombinant virus for at least 10 passages with consistent fluorescence intensity and density. Using the recombinant HP-PRRSV with DsRed protein, the virus tracking in MARC-145 was observed by live-cell imaging. Meanwhile, quantification of the DsRed fluorescence positive cells by flow cytometry provides an alternative to standard methods for testing the level of PRRSV infection. This recombinant PRRSV with DsRed fluorescence protein expression could be a useful tool for fundamental research on the viral biology and shows the new design for stable expression of foreign genes in PRRSV.


[Studies on the chemical constituents of the roots of Anemone altaica].
To investigate the chemical constituents of the roots of Anemone altaica Fisch. ex C. A. May. The constituents of n-BuOH-soluble portion were isolated and purified by means of chromatography. Compounds were identified by their physical characteristics and spectral features. Six compounds were isolated and identified as cimigenol-3-O-beta-D-xylopyranoside (1), cimigenol-3-O-beta-D-xylopyranol (1 -->3)-beta-D-xylopyranoside (2), isolariciresinol-9-O-beta-D-glucopyranoside (3), adenosine (4), uridine (5) and methyl-beta-D-glucopyranoside (6). All compounds are isolated from this genus for the first time.


Addition of an androgen-free epididymal protein extract increases the ability of immature hamster spermatozoa to fertilize in vivo and in vitro.
The fertility of spermatozoa from the different epididymal segments of hamsters was tested by in-vivo insemination. Caput and proximal corpus spermatozoa were non-fertile; spermatozoa from the distal corpus epididymidis fertilized 13% (38/290) oocytes and those from the proximal and distal cauda epididymidis 71 and 87%, respectively. When tested by in-vitro insemination, distal corpus spermatozoa penetrated 44% of oocytes while those from the distal cauda fertilized 87% of oocytes. Spermatozoa from the distal corpus recovered in Medium BMOC fertilized 13% (28/219) of oocytes in vivo, while those mixed with an epididymal protein preparation (0.8 mg protein/ml) fertilized 24% (49/204; P less than 0.01) of oocytes. When distal corpus spermatozoa were inseminated in vivo with 0.8 mg epididymal protein preparation 34% (31/90) oocytes were fertilized and only 22% (23/103; P less than 0.05) oocytes were fertilized when the proteins were obtained from epididymides of animals castrated for 30 days. When distal corpus spermatozoa were preincubated for 5 h in medium without (control) or with protein preparation (0.8 or 1.6 mg protein/ml), a significant increase in in-vitro oocyte penetration was found (25 compared with 45%; P less than 0.05) when the protein was present at 1.6 mg/ml. These results confirm and extend previous observations suggesting a role for androgen-dependent glycoproteins secreted by the epididymis in the acquisition of fertilizing ability that occurs during sperm maturation.