Pooled analysis of the performance of liquid-based cytology in population-based cervical cancer screening studies in China.
Liquid-based cytology (LBC) has been widely used for cervical cancer screening. Despite numerous studies and systematic reviews, to the authors' knowledge few large studies to date have focused on biopsy-confirmed cervical lesions and controversy remains concerning its diagnostic accuracy. The objective of the current study was to assess LBC for detecting biopsy-confirmed cervical intraepithelial neoplasia (CIN) and cancer. A pooled analysis of LBC using data from 13 population-based, cross-sectional, cervical cancer screening studies performed in China from 1999 to 2008 was performed. Participants (n = 26,782) received LBC and human papillomavirus testing. Women found to be positive on screening were referred for colposcopy and biopsy. The accuracy of LBC for detecting biopsy-confirmed CIN of type 2 or worse (CIN2+) as well as CIN type 3 or worse (CIN3+) lesions was analyzed. Of 25,830 women included in the analysis, CIN2+ was found in 107 of 2612 with atypical squamous cells (4.1%), 142 of 923 with low-grade squamous intraepithelial neoplasia (15.4%), 512 of 784 with high-grade squamous intraepithelial neoplasia (65.3%), 29 of 30 with squamous cell carcinoma (96.7%), 4 of 27 with atypical glandular cells (14.8%), and 85 of 21,454 with normal cytology results (0.4%). No invasive cancers were found to have atypical squamous cells, atypical glandular cells, or cytologically normal slides. The overall sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of LBC for detecting CIN2+ were 81.0%, 95.4%, 38.3%, 99.3 %, and 94.9%, respectively. Although Hybrid Capture 2 was more sensitive than LBC, the specificity, positive predictive value, and overall accuracy of LBC were higher than those of Hybrid Capture2 at 85.2%, 18.6%, and 85.5%, respectively. The results of the current study indicate that the performance of LBC can effectively predict the risk of existing CIN2+ and may be a good screening tool for cervical cancer prevention in a developing country.


Structured illumination microscopy of a living cell.
Due to diffraction, the resolution of imaging emitted light in a fluorescence microscope is limited to about 200 nm in the lateral direction. Resolution improvement by a factor of two can be achieved using structured illumination, where a fine grating is projected onto the sample, and the final image is reconstructed from a set of images taken at different grating positions. Here we demonstrate that with the help of a spatial light modulator, this technique can be used for imaging slowly moving structures in living cells.


Potential of renin inhibition in cardiovascular disease.
Since renin catalyses the first and rate-limiting step of the renin-angiotensin system (RAS) cascade, interruption of the generation of angiotensin II (Ang II) by renin inhibitors at this highly specific initial step of the cascade has long been a therapeutic goal. The early development of renin inhibitors was hampered by problems with bioavailability and high costs of synthesis. However, more recently a potent non-peptidic inhibitor of renin, aliskiren, with acceptable oral bioavailability, has been synthesised. Aliskiren effectively reduces Ang II levels in normal volunteers and has been shown to lower blood pressure (BP) in patients with mild-to-moderate hypertension. Renin inhibitors would be expected to have similar, but not identical effects to those of the established RAS antagonists. Due to the lack of effective alternative enzyme pathways, blockade of Ang II production may be more effective with renin inhibition than with angiotensin-converting enzyme (ACE) inhibition. Furthermore, because renin has high specificity for only one substrate, angiotensinogen, side-effects would be expected to be less frequent. It is currently unclear whether blockade of Ang II type 1 (AT1) receptors, leaving other Ang II receptors (AT2, AT3 and AT4) unblocked, is preferable to the reduction in plasma and tissue Ang II levels achieved with either ACE or renin inhibition. Pharmacological suppression of the RAS, through ACE inhibition, or blockade of AT1, beta-adrenoceptor or mineralocorticoid receptors, has been proven to reduce morbidity and mortality in patients with hypertension, diabetes mellitus, atherosclerosis, heart failure and nephropathy. While, to date, aliskiren has only been shown to reduce BP, it appears likely that orally-active renin inhibitors could prove useful in the management of a wide range of cardiovascular pathologies.


Increased risk of HIV in women experiencing physical partner violence in Nairobi, Kenya.
As part of a study on etiology of sexually transmitted infections (STI) among 520 women presenting at the STI clinic in Nairobi, data on partner violence and its correlates were analyzed. Prevalence of lifetime physical violence was 26%, mainly by an intimate partner (74%). HIV seropositive women had an almost twofold increase in lifetime partner violence. Women with more risky sexual behavior such as early sexual debut, number of sex partners, history of condom use and of STI, experienced more partner violence. Parity and miscarriage were associated with a history of lifetime violence. We found an inverse association between schooling and level of violence. Six percent of the women had been raped. Gender-based violence screening and services should be integrated into voluntary counseling and testing programs as well as in reproductive health programs. Multi-sector approaches are needed to change prevailing attitudes towards violence against women.


Long-term unemployment and suicide: a systematic review and meta-analysis.
There have been a number of reviews on the association+ between unemployment and suicide, but none have investigated how this relationship is influenced by duration of unemployment. A systematic review and meta-analysis was conducted of those studies that assessed duration of unemployment as a risk factor for suicide. Studies considered as eligible for inclusion were population-based cohort or case-control designs; population-based ecological designs, or hospital based clinical cohort or case-control designs published in the year 1980 or later. The review identified 16 eligible studies, out of a possible 10,358 articles resulting from a search of four databases: PubMed, Web of Knowledge, Scopus and Proquest. While all 16 studies measured unemployment duration in different ways, a common finding was that longer duration of unemployment was related to greater risk of suicide and suicide attempt. A random effects meta-analysis on a subsample of six cohort studies indicated that the pooled relative risk of suicide in relation to average follow-up time after unemployment was 1.70 (95% CI 1.22 to 2.18). However, results also suggested a possible habituation effect to unemployment over time, with the greatest risk of suicide occurring within five years of unemployment compared to the employed population (RR = 2.50, 95% CI 1.83 to 3.17). Relative risk appeared to decline in studies of those unemployed between 12 and 16 years compared to those currently employed (RR = 1.21, 95% CI 1.10 to 1.33). Findings suggest that long-term unemployment is associated with greater incidence of suicide. Results of the meta-analysis suggest that risk is greatest in the first five years, and persists at a lower but elevated level up to 16 years after unemployment. These findings are limited by the paucity of data on this topic.


Supporting Kenyan women with advanced breast cancer through a network and assessing their needs and quality of life.
Metastatic breast cancer (MBC) patients have several unmet needs. The needs and quality of life of MBC women living in sub-Saharan Africa (SSA) are understudied. Facilitating the interaction of various caregivers is beneficial in addressing the needs. Internet-based resources play an important role in reaching out to these patients. We aimed to bring the various stakeholders into a joint network force, create a web-based portal, understand the needs of MBC patients, and assess the utilization of web-based resources for women from Kenya. A network of various stakeholders considered crucial in the care of Kenyan women with MBC was created. We conducted educational camps and assessed their needs, quality of life (QoL), and knowledge. We assessed the impact of utilizing web-based resources by MBC patients from here. We formed a network involving partners and launched the first dedicated website for MBC from Kenya. The website has received 13,944 visits and 310,379 hits in 2 years. One hundred fourteen women living with MBC were interviewed, and our findings show that psychological needs (63%), physical support needs (60%), and health care system needs (55%) are leading areas of needs that increase with rural residence (p = 0.001), less education (p = 0.003), and aggressive treatments (p = 0.008). Quality of life (QoL) confirmed better scores with urban residence (p = 0.002), internet access (p = 0.010), and stable disease (p = 0.042). Creating a network of caregivers provides opportunities for cohesive efforts in understanding the psychosocial and medical needs of patients with MBC. Internet-based resources are an effective way of reaching out to them. Kenyan patients show extremely good uptake of internet-based resources.


The prevalence of Rhesus negativity among pregnant women in Enugu, Southeast Nigeria.
Hemolytic disease of the newborn, secondary to rhesus D (Rh D) iso-immunization, contributes significantly to perinatal morbidity and mortality. Prevalence data in Nigeria, and Southeast Nigeria in particular, is very scanty. This study was carried out to provide our experience in this preventable clinical condition in Enugu, Southeast Nigeria. To determine the prevalence and trends of Rh D negativity among pregnant women in Enugu, Southeast Nigeria. A 5-year retrospective study of rhesus negative women was carried out at the University of Nigeria Teaching Hospital, Enugu, Nigeria, between 1 st January 2000 and 31 st December 2004. The prevalence rate of Rh D negative women in Enugu, Nigeria, is 4.5%. Out of 6306 women who booked for antenatal care, 282 (4.5%) were Rh D negative women. One hundred and eighty-two (182) (64.5%) of the Rh D negative women were of blood group O followed by blood group A 20%, blood group B 12.1%, and blood group AB 3.2%, respectively. There is a need for adequate counseling of pregnant women on the importance of Rh D negative factor during the antenatal period in order to prevent hemolytic disease of the newborn.


Giant-cell fibroblastoma and dermato fibro sarcoma protuberans: the same tumoral spectrum? Report of two cases of association in children.
We describe two cases of giant-cell fibroblastoma (GCF) with dermato fibro sarcoma protuberans (DFSP) component, occurring in two children in a chest wall localization. One case recurred 1 year later. The two patients were tumor-free 12 and 8 years later. GCF is a rare mesenchymal cutaneous and subcutaneous tumor reported mostly in the first two decades of life. Dermato fibro sarcoma protuberans, occurring preferentially in adults, is a rare skin tumor with a pronounced tendency to local recurrence. Some cases of association of recurrence of GFC under the form of DFSP have been reported, raising the question of a continuum between the two tumors. The treatment of choice of the two tumors is a wide local excision.


Multi-collector Inductively Coupled Plasma Mass Spectrometry: New Developments and Basic Concepts for High-precision Measurements of Mass-dependent Isotope Signatures.
Due to the development of multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS) around 25 years ago, the isotopes of a large range of elements (masses from Li to U) are now analyzed with high enough precision and accuracy to resolve subtle natural variations. These so-called 'non-traditional stable isotope systems' opened many new research avenues and are applied at an increasing rate in research and industry projects and in a broad range of different disciplines, including archeology, biology, physics, cosmochemistry and geology. Here, we briefly summarize the most basic concepts of MC-ICP-MS, introduce new technical developments and address important points on how to acquire accurate high-precision isotope measurements of non-traditional stable isotopes.


Altered polypeptide expression associated with neoplastic transformation of Syrian hamster cells by bisulfite.
Neoplastic transformation of Syrian hamster fetal cells by bisulfite is associated with qualitative and quantitative polypeptide changes. Amino acid-labeled [14C]polypeptides from neoplastic and nontransformed parental fetal cells were separated by two-dimensional gel electrophoresis and analyzed by computerized microdensitometry of autoradiographic patterns. Approximately 1000 polypeptides from parental fibroblasts at population doublings ranging from 4 to 20, and those from colony-derived malignant cell lines were compared. Most were identical. Seven malignant lines exhibited 4 qualitative polypeptide changes: 2 polypeptides had shifted slightly to the acidic side, 1 new polypeptide was observed, and 1 polypeptide was absent. The transformed bisulfite lines differed quantitatively from control cells in that 10-25% and 2-4% of the polypeptides exhibited differences in expression greater than 2- and 4-fold, respectively. Furthermore, there were 21 specific polypeptides with coordinate quantitative changes in all transformed lines. Because bisulfite at neutral pH fails to induce any significant DNA changes at concentrations that cause transformation, polypeptides expressed immediately or 48 h after bisulfite treatment were compared to those of non-treated controls, and no differences were found. Even though bisulfite does not induce detectable DNA damage or early post-treatment changes in polypeptide expression, a consistent set of qualitative and quantitative changes were observed after transformation. The qualitative polypeptide changes found in the bisulfite-induced malignant lines were similar to those seen in a benzo(a)pyrene-induced malignant line. This suggests that there is a convergence of pathways responsible for carcinogenesis independent of the nature of initiation.


Reference Values and Determinants of Spatiotemporal and Kinetic Variables in Recreational Runners.
Identifying atypical lower limb biomechanics may help prevent the occurrence or recurrence of running-related injuries. No reference values for spatiotemporal or kinetic variables in healthy recreational runners are available in the scientific literature to support clinical management. To (1) present speed- and sex-stratified reference values for spatiotemporal and kinetic variables in healthy adult recreational runners; (2) identify the determinants of these biomechanical variables; and (3) develop reference regression equations that can be used as a guide in a clinical context. Descriptive laboratory study. This study involved 860 healthy recreational runners (age, 19-65 years [38.5% women]) tested on an instrumented treadmill at their preferred running speed in randomly allocated, standardized running shoes with either hard or soft cushioning. Twelve common spatiotemporal and kinetic variables-including contact time, flight time, duty factor, vertical oscillation, step cadence, step length, vertical impact peak (VIP), time to VIP, vertical average loading rate, vertical stiffness, peak vertical ground-reaction force (GRF), and peak braking force-were derived from GRF recordings. Reference values for each biomechanical variable were calculated using descriptive statistics and stratified by sex and running speed category (≤7, 8, 9, 10, 11, 12, 13, 14, and ≥15 km/h). Correlations and multiple regression analyses were performed to identify potential determinants independently associated with each biomechanical variable and generate reference equations. The mean running speed was 10.5 ± 1.3 km/h and 9 ± 1.1 km/h in men and women, respectively. While all potential predictors were significantly correlated with many of the 12 biomechanical variables, only running speed showed high correlations (r > 0.7). The adjusted R2 of the multiple regression equations ranged from 0.19 to 0.88. This study provides reference values and equations that may guide clinicians and researchers in interpreting spatiotemporal and kinetic variables in recreational runners. The reference values can be used as targets for clinicians working with recreational runners in cases where there is a clinical suspicion of a causal relationship between atypical biomechanics and running-related injury.


Behavior of intracellular lipid droplets during cell division in HuH7 hepatoma cells.
Intracellular lipid droplets (LDs) are ubiquitous organelles found in many cell types. During mitosis, membranous organelles, including mitochondria, are divided into small pieces and transferred to daughter cells; however, the process of LD transfer to daughter cells is not fully elucidated. Herein, we investigated the behavior of LDs during mitosis in HuH7 human hepatoma cells. While fragments of the Golgi apparatus were scattered in the cytosol during mitosis, intracellular LDs retained their size and spherical morphology as they translocated to the two daughter cells. LDs were initially distributed throughout the cell during prophase but positioned outside the spindle in metaphase, aligning at the far sides of the centrioles. A similar distribution of LDs during mitosis was observed in another hepatocarcinoma HepG2 cells. When the spindle was disrupted by nocodazole treatment or never in mitosis gene A-related kinase 2A knockdown, LDs were localized in the area outside the chromosomes, suggesting that spindle formation is not necessary for LD localization at metaphase. The amount of major LD protein perilipin 2 reduced while LDs were enriched in perilipin 3 during mitosis, indicating the potential alteration of LD protein composition. Conclusively, the behavior of LDs during mitosis is distinct from that of other organelles in hepatocytes.


Development of Human Hair Reference Material Supporting the Biomonitoring of Methylmercury.
A certified reference material, NIMD-01, was developed for the analysis of mercury speciation in human hair. We collected the hair of Vietnamese males from a barbershop in Hanoi in 2016 and prepared 1200 bottles containing 3 g of sieved and blended hair powder. The certified value was given on a dry-mass basis, with the moisture content obtained by drying at 85°C for 4 h. Certified values with the expanded uncertainties (coverage factor, k = 2) were as follows: methylmercury, 0.634 ± 0.071 mg kg-1 as mercury; total mercury, 0.794 ± 0.050 mg kg-1; copper, 12.8 ± 1.4 mg kg-1; zinc, 234 ± 29 mg kg-1; selenium, 1.52 ± 0.29 mg kg-1. An indicative arsenic concentration of 0.17 ± 0.03 mg kg-1 was measured. Extended uncertainties were estimated by sample homogeneity, long- and short-term stabilities, and a characterization from measurements made by collaborating laboratories.


Ibotenic acid analogues. Synthesis and biological and in vitro activity of conformationally restricted agonists at central excitatory amino acid receptors.
A number of analogues of ibotenic acid [(RS)-3-hydroxy-5- isoxazoleglycine ] were synthesized; they were tested as excitants on neurons in the cat spinal cord, by using microelectrophoretic techniques, and as inhibitors of the binding of kainic acid (KA) in vitro, by using synaptic membranes prepared from rat brains. The excitatory effects of the 3- isoxazolol amino acids (RS)-3-hydroxy-4,5,6,7-tetrahydroisoxazolo[5, 4-c]pyridine-7-carboxylic acid (4, 7- HPCA ), (RS)-alpha-amino-3-hydroxy-5,6-dihydro-4H- cyclohept [1,2-d] isoxa zole - 8-propionic acid (8, 8- AHCP ), (RS)-alpha-amino-3- hydroxy-7,8-dihydro-6H- cyclohept [1,2-d] isoxazole -4-propionic acid (12, 4- AHCP ), and (RS)-alpha-(methylamino)-3-hydroxy-5-methyl- 4- isoxazolepropionic acid (15, N-Me-AMPA) were shown to be sensitive to (S)-glutamic acid diethyl ester (GDEE), an antagonist at quisqualic acid ( QUIS ) receptors, and insensitive to (RS)-2-amino-5-phosphonovaleric acid ( 2APV ), an antagonist at N-methyl-(R)-aspartic acid (NMDA) receptors. The compounds 4 and 12 proved to be particularly potent agonists at the former class of receptor, assumed to represent physiological glutamic acid receptors. The amino acids (RS)-beta-(2-carboxyphenyl)alanine (19), an analogue of 12, and (RS)-2-(3-carboxyphenyl) glycine were weak GDEE-sensitive excitants with potencies comparable with that of 8. All of the compounds were tested as inhibitors of KA binding. With the exception of 12 and 19, which showed very low affinity for the KA binding sites, the compounds studied were inactive in this in vitro test system.


Rapid in situ hybridization technique for the detection of ribonucleic acids in tissues using radiolabelled and fluorescein-labelled riboprobes.
In situ hybridization (ISH) is a useful diagnostic and research tool, but is also time consuming. This study was conducted to determine if a rate enhancement hybridization (REH) buffer, developed for membrane hybridization, could be used to decrease hybridization time for ISH. Tissue from swine with an enteric disease produced by a swine coronavirus, transmissible gastroenteritis virus (TGEV), was used as a model to standardize hybridization conditions for a rapid ISH technique. Small intestinal sections from pigs experimentally and naturally infected with TGEV were hybridized for various times at 52 degrees C and 70 degrees C with a radiolabelled or a fluorescein-labelled RNA probe in a standard hybridization or a REH buffer. Viral RNA was detected in intestines from as early as 30 min of hybridization by using both buffers with the radiolabelled probe; however, the signal was stronger with the REH buffer. With the fluorescein-labelled probe, viral RNA was detected in virus-infected cells of the intestines after 30 min of hybridization by using the REH buffer. Signal intensity was greater with the REH buffer than with the standard hybridization buffer when compared at each hybridization time and hybridization temperature using both radiolabelled and fluorescein-labelled probes. With the REH buffer, hybridization signal intensity was greater at 70 degrees C than at 52 degrees C for both probes. The best results were obtained when small intestinal sections were hybridized at 70 degrees C for 2 h using a radiolabelled or a fluorescein-labelled probe diluted in the REH buffer. The fluorescein-labelled RNA probe with REH buffer resulted in a minimal non-specific signal when compared with the radiolabelled probe. These studies demonstrated that the REH buffer can be used to decrease the time of ISH for the detection of viral RNA. This rapid ISH technique should have broad applications in the utilization of probe technology in diagnostics and research for the detection of target ribonucleic acids in situ


Methylomic predictors demonstrate the role of NF-κB in old-age mortality and are unrelated to the aging-associated epigenetic drift.
Changes in the DNA methylation (DNAm) landscape have been implicated in aging and cellular senescence. To unravel the role of specific DNAm patterns in late-life survival, we performed genome-wide methylation profiling in nonagenarians (n=111) and determined the performance of the methylomic predictors and conventional risk markers in a longitudinal setting. The survival model containing only the methylomic markers was superior in terms of predictive accuracy compared with the model containing only the conventional predictors or the model containing conventional predictors combined with the methylomic markers. At the 2.55-year follow-up, we identified 19 mortality-associated (false-discovery rate <0.5) CpG sites that mapped to genes functionally clustering around the nuclear factor kappa B (NF-κB) complex. Interestingly, none of the mortality-associated CpG sites overlapped with the established aging-associated DNAm sites. Our results are in line with previous findings on the role of NF-κB in controlling animal life spans and demonstrate the role of this complex in human longevity.


Socio-behavioral variables effecting oral hygiene and periodontal status of 12 year-old schoolchildren of Udaipur district.
To assess the effect of various socio-demographic in addition to oral hygiene variables on the oral hygiene and periodontal status. A combination of multi stage and cluster sampling procedure was followed to collect a representative population of 831, 12 year-old schoolchildren of Udiapur district. Oral Hygiene Index Simplified (OHI-S) and Community Periodontal Index (CPI) were used to evaluate oral hygiene and periodontal status respectively. Along with the clinical examination, each subject was interviewed to collect the socio-demographic information and oral hygiene practices. The overall mean OHI-S was 1.42 and debris component of the oral hygiene index was dominant. Unpaired 't' test showed significantly higher DI-S (p = 0.0001) and OHI-S (p = 0.007) among boys than girls. The overall prevalence of periodontal disease was 89.9% and gingival bleeding was widespread. Chi square test demonstrated significant differences (p = 0.0001) with 14.6% urban children having healthy periodontium in comparison to 5.6% rural residents. Oral hygiene scores were dependent on the father's occupation (p = 0.046), father's (p = 0.002) and mother's level of education (p = 0.0001). Oral hygiene status was poor, more among boys with debris contributing a major part for oral hygiene index. Periodontal status was poor with bleeding being most prevalent indicator. The multivariate analysis of oral hygiene status confirmed the existence of socio-behavioural determinants for oral hygiene.


Pancreatic islet cell stress induced by insulin-degrading enzyme deficiency promotes islet regeneration and protection from autoimmune diabetes.
Appropriate tuning of protein homeostasis through mobilization of the unfolded protein response (UPR) is key to the capacity of pancreatic beta cells to cope with highly variable demand for insulin synthesis. An efficient UPR ensures a sufficient beta cell mass and secretory output but can also affect beta cell resilience to autoimmune aggression. The factors regulating protein homeostasis in the face of metabolic and immune challenges are insufficiently understood. We examined beta cell adaptation to stress in mice deficient for insulin-degrading enzyme (IDE), a ubiquitous protease with high affinity for insulin and genetic association with type 2 diabetes. IDE deficiency induced a low-level UPR in both C57BL/6 and autoimmune non-obese diabetic (NOD) mice, associated with rapamycin-sensitive beta cell proliferation strongly enhanced by proteotoxic stress. Moreover, in NOD mice, IDE deficiency protected from spontaneous diabetes and triggered an additional independent pathway, conditional on the presence of islet inflammation but inhibited by proteotoxic stress, highlighted by strong upregulation of regenerating islet-derived protein 2, a protein attenuating autoimmune inflammation. Our findings establish a key role of IDE in islet cell protein homeostasis, identify a link between low-level UPR and proliferation, and reveal an UPR-independent anti-inflammatory islet cell response uncovered in the absence of IDE of potential interest in autoimmune diabetes.


Tracking the optic nervehead in OCT video using dual eigenspaces and an adaptive vascular distribution model.
Optical coherence tomography (OCT) is a new ophthalmic imaging modality generating cross sectional views of the retina. OCT systems are essentially Michelson interferometers that form images in 1.5 s by directing a superluminescent diode (SLD) beam over the retinal surface. Involuntary eye motions frequently cause incorrect locations to be imaged. This motion may leave no obvious artifacts in the scan data and can easily go undetected. For glaucoma monitoring especially, knowing the measurement path, typically a circle concentric with the nerve head, is crucial. The commercially available OCT system displays a near-infrared video of the retina showing the SLD beam. This paper presents a prototype system to detect the nerve head and SLD beam in the video, and report the true scan path relative to the nerve head. Low image contrast and limited resolution make the reliable detection of retinal features difficult. In an adaptive model construction phase, the system directly detects retinal vasculature and the nerve head and incrementally builds a model of the current subject's vascular pattern relative to the optic disk. The nerve head identification is multitiered, using a novel dual eigenspace technique and a geometric comparison of detected vessel positions and nerve head hypotheses. In its operational phase, a correspondence is achieved between the currently detected vasculature and the model. Using subjects not included in training, the system located the optic nerve head to within 5 pixels (0.07 optic disk diameters, an error well below clinical significance) in 99.75% of 2800 video fields. In current clinical practice, motions as large as 1-2 disc diameters may go undetected, so this is a vast improvement.


Effects of aging on the skin and gill microbiota of farmed seabass and seabream.
Important changes in microbial composition related to sexual maturation have been already reported in the gut of several vertebrates including mammals, amphibians and fish. Such changes in fish are linked to reproduction and growth during developmental stages, diet transitions and critical life events. We used amplicon (16S rRNA) high-throughput sequencing to characterize the skin and gill bacterial microbiota of farmed seabass and seabream belonging to three different developmental age groups: early and late juveniles and mature adults. We also assessed the impact of the surrounding estuarine water microbiota in shaping the fish skin and gill microbiota. Microbial diversity, composition and predicted metabolic functions varied across fish maturity stages. Alpha-diversity in the seabass microbiota varied significantly between age groups and was higher in older fish. Conversely, in the seabream, no significant differences were found in alpha-diversity between age groups. Microbial structure varied significantly across age groups; moreover, high structural variation was also observed within groups. Different bacterial metabolic pathways were predicted to be enriched in the microbiota of both species. Finally, we found that the water microbiota was significantly distinct from the fish microbiota across all the studied age groups, although a high percentage of ASVs was shared with the skin and gill microbiotas. We report important microbial differences in composition and potential functionality across different ages of farmed seabass and seabream. These differences may be related to somatic growth and the onset of sexual maturation. Importantly, some of the inferred metabolic pathways could enhance the fish coping mechanisms during stressful conditions. Our results provide new evidence suggesting that growth and sexual maturation have an important role in shaping the microbiota of the fish external mucosae and highlight the importance of considering different life stages in microbiota studies.


Autoantibody prevalence in active tuberculosis: reactive or pathognomonic?
To evaluate the autoantibody in patients without corresponding symptoms, whether these autoantibody are pathognomonic or not. We hypothesised that autoantibody may be reactive to chronic infection, such as tuberculosis (TB). Randomised, case-control cohort study. A tertiary centre in Taiwan. We randomly chose 100 patients out of the data bank of patients with TB in a tertiary medical centre. All patients completed the sera sampling. We chose 100 patients according to autoantibody prevalence in previous literature. We also chose 100 medical staff as control group. We tested anti-SSA, anti-SSB, anti-Sm, anti ribonucleoprotein, anti-Scl 70, anticentromere, anti-double-stranded DNA, anticardiolipin IgG and IgM in all patient and control groups. The clinical symptoms and the underlying disease were all recorded. The result of sera antibody titre was recorded. For those with specific positive serology results, following examination was carried out after a 3-month anti-TB medication. Anticardiolipin IgG titre was significantly higher in patients with TB than in control group. We compared the result with previous population study and found that anti-Scl70 is also significantly higher in patients with TB. The following up data in anti-Scl70 revealed decreased titre after treatment. No correlation between sera titre and clinical conditions was observed. In TB endemic areas, a significant proportion (32%) of patients with TB have elevated autoantibody titres, especially anticardiolipin IgG and anti-Scl-70. Mycobacterial studies should be performed in patients with elevated serum autoantibody titres but without the typical or multiple manifestations of autoimmune diseases. The study was approved by the Institutional Review Board of the hospital (NTUH REC: 9561707008) after informed consent had been obtained from the patients.


Host-Induced Genome Instability Rapidly Generates Phenotypic Variation across Candida albicans Strains and Ploidy States.
Candida albicans is an opportunistic fungal pathogen of humans that is typically diploid yet has a highly labile genome tolerant of large-scale perturbations including chromosomal aneuploidy and loss-of-heterozygosity events. The ability to rapidly generate genetic variation is crucial for C. albicans to adapt to changing or stressful environments, like those encountered in the host. Genetic variation occurs via stress-induced mutagenesis or can be generated through its parasexual cycle, in which tetraploids arise via diploid mating or stress-induced mitotic defects and undergo nonmeiotic ploidy reduction. However, it remains largely unknown how genetic background contributes to C. albicans genome instability in vitro or in the host environment. Here, we tested how genetic background, ploidy, and the host environment impacts C. albicans genome stability. We found that host association induced both loss-of-heterozygosity events and genome size changes, regardless of genetic background or ploidy. However, the magnitude and types of genome changes varied across C. albicans strain background and ploidy state. We then assessed if host-induced genomic changes resulted in fitness consequences on growth rate and nonlethal virulence phenotypes and found that many host-derived isolates significantly changed relative to their parental strain. Interestingly, diploid host-associated C. albicans predominantly decreased host reproductive fitness, whereas tetraploid host-associated C. albicans increased host reproductive fitness. Together, these results are important for understanding how host-induced genomic changes in C. albicans alter its relationship with the host.IMPORTANCECandida albicans is an opportunistic fungal pathogen of humans. The ability to generate genetic variation is essential for adaptation and is a strategy that C. albicans and other fungal pathogens use to change their genome size. Stressful environments, including the host, induce C. albicans genome instability. Here, we investigated how C. albicans genetic background and ploidy state impact genome instability, both in vitro and in a host environment. We show that the host environment induces genome instability, but the magnitude depends on C. albicans genetic background. Furthermore, we show that tetraploid C. albicans is highly unstable in host environments and rapidly reduces in genome size. These reductions in genome size often resulted in reduced virulence. In contrast, diploid C. albicans displayed modest host-induced genome size changes, yet these frequently resulted in increased virulence. Such studies are essential for understanding how opportunistic pathogens respond and potentially adapt to the host environment.


Epidemiological analysis of dyslipidemia in adults of three ethnicities in Xinjiang, China.
This study investigated the prevalence and distribution of dyslipidemia in adults of Uygur, Kazak, and Han ethnicity in Xinjiang, China. A questionnaire including general data, physical examination (blood pressure, body height, and body weight) and blood lipid [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C)] was administered to 11,506 adults in Xinjiang, China from 2009 to 2010 using a stratified sampling method. The overall prevalence rates of dyslipidemia in Uygur, Kazak, and Han adults were 42.4, 31.6, and 30.2%, respectively; they were 42.4, 31.8, and 28.2% after age standardization (P < 0.01). After standardization, the overall prevalence rates in Uygur, Kazak, and Han men were 52.6, 35.4, and 33.2%, respectively, which were significantly higher than that in women of the corresponding ethnicities (P < 0.01). In Uygur, Kazak, and Han adults, there were significant differences with respect to the standardized prevalence rates of high TG (9.3, 9.3, and 17.3%), high TC (5.2, 6.9, and 6%), low HDL-C (33.6, 20.8, and 11.1%), and high LDL-C (2.4, 2.9, and 2%) (P < 0.05). The prevalence rates of dyslipidemia in Uygur, Kazak, and Han adults in Xinjiang are higher than the average levels in China, with significant differences in ethnicity, age, and gender. Han adults exhibited the highest prevalence rate of high TG. Meanwhile, Uygur adults had the highest prevalence rate of low HDL-C. Kazak adults had high prevalence rates of high TC, low HDL-C, and high LDL-C.


Epstein-Barr Virus-Specific CD8 T Cells Selectively Infiltrate the Brain in Multiple Sclerosis and Interact Locally with Virus-Infected Cells: Clue for a Virus-Driven Immunopathological Mechanism.
Epstein-Barr virus (EBV) is a ubiquitous herpesvirus strongly associated with multiple sclerosis (MS), a chronic inflammatory disease of the central nervous system (CNS). However, the mechanisms linking EBV infection to MS pathology are uncertain. Neuropathological and immunological studies suggest that a persistent EBV infection in the CNS can stimulate a CD8 T-cell response aimed at clearing the virus but inadvertently causing CNS injury. Inasmuch as in situ demonstration of EBV-specific CD8 T cells and their effector function is missing, we searched for EBV-specific CD8 T cells in MS brain tissue using the pentamer technique. Postmortem brain samples from 12 donors with progressive MS and known HLA class I genotype were analyzed. Brain sections were stained with HLA-matched pentamers coupled with immunogenic peptides from EBV-encoded proteins, control virus (cytomegalovirus and influenza A virus) proteins, and myelin basic protein. CD8 T cells recognizing proteins expressed in the latent and lytic phases of the EBV life cycle were visualized in white matter lesions and/or meninges of 11/12 MS donors. The fraction (median value) of CD8 T cells recognizing individual EBV epitopes ranged from 0.5 to 2.5% of CNS-infiltrating CD8 T cells. Cytomegalovirus-specific CD8 T cells were detected at a lower frequency (≤0.3%) in brain sections from 4/12 MS donors. CNS-infiltrating EBV-specific CD8 T cells were CD107a positive, suggesting a cytotoxic phenotype, and stuck to EBV-infected cells. Together with local EBV dysregulation, selective enrichment of EBV-specific CD8 T cells in the MS brain supports the notion that skewed immune responses toward EBV contribute to inflammation causing CNS injury.IMPORTANCE EBV establishes a lifelong and asymptomatic infection in most individuals and more rarely causes infectious mononucleosis and malignancies, like lymphomas. The virus is also strongly associated with MS, a chronic neuroinflammatory disease with unknown etiology. Infectious mononucleosis increases the risk of developing MS, and immune reactivity toward EBV is higher in persons with MS, indicating inadequate control of the virus. Previous studies have suggested that persistent EBV infection in the CNS stimulates an immunopathological response, causing bystander neural cell damage. To verify this, we need to identify the immune culprits responsible for the detrimental antiviral response in the CNS. In this study, we analyzed postmortem brains donated by persons with MS and show that CD8 cytotoxic T cells recognizing EBV enter the brain and interact locally with the virus-infected cells. This antiviral CD8 T cell-mediated immune response likely contributes to MS pathology.


Overview of presurgical assessment and surgical treatment of epilepsy from the Italian League Against Epilepsy.
The Commission for Epilepsy Surgery of the Italian League Against Epilepsy (LICE) presents an overview of the techniques and methodologies of presurgical evaluation and of the surgical treatment of epilepsies. This overview is the result of the experience developed in the past years in the major Italian centers where programs of epilepsy surgery have been established, and it has the aim of offering a quick and easy reference tool for those involved in the treatment of patients with epilepsy. The sharing of different experiences has the additional aim of conforming and disseminating the employed techniques as well as the methods of selection and evaluation of patients. The synthetic coverage of the main issues concerning the presurgical workup and the available surgical options will hopefully provide a framework that may integrate and develop the contributions of every single center, in one of the more complex, challenging, and dynamic areas of neurological sciences.


Value choices in European COVID-19 vaccination schedules: how vaccination prioritization differs from other forms of priority setting.
With the limited initial availability of COVID-19 vaccines in the first months of 2021, decision-makers had to determine the order in which different groups were prioritized. Our aim was to find out what normative approaches to the allocation of scarce preventive resources were embedded in the national COVID-19 vaccination schedules. We systematically reviewed and compared prioritization regulations in 27 members of the European Union, the United Kingdom, and Israel. We differentiated between two types of priority categories: groups that have increased infection fatality rate (IFR) compared to the average for the general population and groups chosen because their members experience increased risk of being infected (ROI). Our findings show a clear trend: all researched schedules prioritized criteria referring to IFR (being over 65 years old and coexisting health conditions) over the ROI criteria (eg occupation and housing conditions). This is surprising since, in the context of treatment, it is common and justifiable to adopt different allocation principles (eg introducing a saving more life-year approach or prioritizing younger patients). We discuss how utilitarian, prioritarian, and egalitarian principles can be applied to interpret normative differences between the allocation of curative and preventive interventions.


A whole-genome association analysis of noncompensatory fertility in Holstein bulls.
Increasing fertility in dairy cattle is an important goal. Male infertility represents a part of the overall infertility in dairy cattle and can be partitioned into compensatory and noncompensatory components, where compensatory refers to infertility that can be overcome by increasing sperm number and noncompensatory infertility represents the remainder, presumably due to molecular and genomic defects. Through estimation of single nucleotide polymorphism (SNP) association with noncompensatory bull fertility, it is possible to identify regions of the genome influential to this trait. Use of this information in selection can allow for an increase in cattle fertility, resulting in economic benefits. In this study, high-density SNP genotypes and noncompensatory fertility data from 795 Holstein sires were used to examine SNP associations with fertility. A Bayes B analysis was performed to develop information for genomic selection and to identify genomic regions associated with noncompensatory fertility. A cross-validation approach was used to assess the effectiveness of the models within the original set of 795 bulls. Correlations of predicted and observed fertility values were approximately 0.145 in cross-validation.


Comparison of extended reality and conventional methods of basic life support training: protocol for a multinational, pragmatic, noninferiority, randomised clinical trial (XR BLS trial).
Conventional cardiopulmonary resuscitation (CPR) training for the general public involves the use of a manikin and a training video, which has limitations related to a lack of realism and immersion. To overcome these limitations, virtual reality and extended reality technologies are being used in the field of medical education. The aim of this study is to explore the efficacy and safety of extended reality (XR)-based basic life support (BLS) training. This study is a prospective, multinational, multicentre, randomised controlled study. Four institutions in 4 countries will participate in the study. A total of 154 participants will be randomly assigned to either the XR group or the conventional group stratified by institution and sex (1:1 ratio). Each participant who is allocated to either group will be sent to a separate room to receive training with an XR BLS module or conventional CPR training video. All participants will perform a test on a CPR manikin after the training. The primary outcome will be mean compression depth. The secondary outcome will be overall BLS performance, including compression rate, correct hand position, compression, and full release and hands-off time. Using virtual reality (VR) to establish a virtual educational environment can give trainees a sense of realism. In the XR environment, which combines the virtual world with the real world, trainees can more effectively learn various skills. This trial will provide evidence of the usefulness of XR in CPR education. ClinicalTrials.gov NCT04736888. Registered on 29 January 2021.


A comparison of low-molecular-weight heparin with unfractionated heparin for acute pulmonary embolism. The THESEE Study Group. Tinzaparine ou Heparine Standard: Evaluations dans l'Embolie Pulmonaire.
Low-molecular-weight heparin appears to be at least as effective and safe as standard, unfractionated heparin for the treatment of deep-vein thrombosis, but only limited data are available on the use of low-molecular-weight heparin to treat acute symptomatic pulmonary embolism. We randomly assigned 612 patients with symptomatic pulmonary embolism who did not require thrombolytic therapy or embolectomy to either subcutaneous low-molecular-weight heparin (tinzaparin) given once daily in a fixed dose or adjusted-dose, intravenous unfractionated heparin. Oral anticoagulant therapy was begun between the first and the third day and was given for at least three months. We compared the treatments at day 8 and day 90 with respect to a combined end point of recurrent thromboembolism, major bleeding, and death. In the first eight days of treatment, 9 of 308 patients assigned to receive unfractionated heparin (2.9 percent) reached at least one of the end points, as compared,with 9 of 304 patients assigned to low-molecular-weight heparin (3.0 percent; absolute difference, 0.1 percentage point; 95 percent confidence interval, -2.7 to 2.6). By day 90, 22 patients assigned to unfractionated heparin (7.1 percent) and 18 patients assigned to low-molecular-weight heparin (5.9 percent) had reached at least one end point (P=0.54; absolute difference, 1.2 percentage points; 95 percent confidence interval, -2.7 to 5.1). The risk of major bleeding was similar in the two treatment groups throughout the study. Under the conditions of this study, initial subcutaneous therapy with the low-molecular-weight heparin tinzaparin appeared to be as effective and safe as intravenous unfractionated heparin in patients with acute pulmonary embolism.


[Mechanism of the functional transformation of paralyzed intercostal muscles in rats with experimental botulism].
The rest membrane potential (RMP) and action potential (AP) of muscle fibers of external and internal intercostal muscles were examined in the local form of botulism. There was a decrease in the RMP and AP magnitudes in both groups of the muscles. This decrease was coupled with a greater proportion of the fibers with a low RMP. The excitability of the muscle fibers varied biphasically; it rose in the early period of paralysis and fell in the later one. The variations found might be accounted for by dystrophy of motoneurons innervating muscles.


Carbon-14 radiolabelling and tissue distribution evaluation of a potential anti-TB compound.
This paper describes a five-step synthesis of a carbon-14-labelled pyrazole compound (11). A total of 2.96 MBq of 11 was obtained with the specific activity of 2242.4 MBq/mmol. The radiochemical purity was >99%, and the overall radiochemical yield was 60% based on the [(14) C6 ] 4-bromoaniline starting material. Biodistribution results showed that the radiotracer (administrated orally) has a high accumulation in the small intestine, large intestine and liver of both non-infected and tuberculosis (TB)-infected mice. Therefore, this suggests that compound 11 undergoes hepatobiliary clearance. The compound under investigation has been found to be slowly released from the liver between 2 and 8 h. The study revealed that 11 has no affinity for TB cells.


Analysis of viral and defective-interfering nucleocapsids in acute and persistent infection by rhabdoviruses.
We have isolated and characterized the RNA of intracellular virus nucleocapsids recovered from a number of cell cultures persistently infected with rabies virus or vesicular stomatitis virus (VSV). VSV persistent infections in BHK21, L cells and Aedes albopictus (mosquito) cells generally showed the presence of large amounts of defective-interfering (DI) nucleocapsid RNA and much smaller amounts of standard (B) nucleocapsid RNA. Persistent infections of BHK21 cells by two rabies virus strains, challenge virus standard (CVS-11) or HEP-Flury, were followed for several months during which time the ratio of DI to B nucleocapsid RNA cycled dramatically. We also observed coordinated fluctuations in the absolute amount of incorporation of [3H]uridine into virus nucleocapsid RNA. Total incorporation was generally highest following a decrease in the relative amount of DI nucleocapsid RNA synthesis. At no time were DI nucleocapsids absent in any of the persistently infected cultures.


A polarizable force field for computing the infrared spectra of the polypeptide backbone.
The shapes of the amide bands in the infrared (IR) spectra of proteins and peptides are caused by electrostatically coupled vibrations within the polypeptide backbone and code the structures of these biopolymers. A structural decoding of the amide bands has to resort to simplified models because the huge size of these macromolecules prevents the application of accurate quantum mechanical methods such as density functional theory (DFT). Previous models employed transition-dipole coupling methods that are of limited accuracy. Here we propose a concept for the computation of protein IR spectra, which describes the molecular mechanics (MM) of polypeptide backbones by a polarizable force field of "type II". By extending the concepts of conventional polarizable MM force fields, such a PMM/II approach employs field-dependent parameters not only for the electrostatic signatures of the molecular components but also for the local potentials modeling the stiffness of chemical bonds with respect to elongations, angle deformations, and torsions. Using a PMM/II force field, the IR spectra of the polypeptide backbone can be efficiently calculated from the time dependence of the backbone's dipole moment during a short (e.g., 100 ps) MD simulation by Fourier transformation. PMM/II parameters are derived for harmonic bonding potentials of amide groups in polypeptides from a series of DFT calculations on the model molecule N-methylacetamide (NMA) exposed to homogeneous external electric fields. The amide force constants are shown to vary by as much as 20% for relevant field strengths. As a proof of principle, it is shown that the large solvatochromic effects observed in the IR spectra of NMA upon transfer from the gas phase into aqueous solution are not only excellently reproduced by DFT/MM simulations but are also nicely modeled by the PMM/II approach. The tasks remaining for a proof of practice are specified.


Increased beta 2-adrenoreceptor density in heart, kidney and lung of spontaneously hypertensive rats.
The development of beta-adrenoreceptor density and beta 1- and beta 2-adrenoreceptor distribution has been investigated in heart, kidney and lung of spontaneously hypertensive (SHR) and normotensive (WKY) rats by (-)125I-iodocyanopinolol binding at the age of 5-6, 9-10 and 19-21 weeks. At all ages beta-adrenoreceptor density was similar in hearts of both strains, while it was increased in kidney and lung of SHR compared to WKY. The beta 2-adrenoreceptor density was higher in all three tissues of SHR at all ages investigated. On the other hand, beta 1-adrenoreceptor density was decreased in heart, unchanged in lung and increased in kidney of SHR compared to WKY. Destruction of presynaptic nerve terminals by treatment of WKY with 6-hydroxydopamine produced a 24% loss of cardiac beta 2-adrenoreceptors, whereas the beta 1-adrenoreceptor density remained unchanged, suggesting that at least part of the cardiac beta 2-adrenoreceptor population is localized prejunctionally. It is suggested that beta 2-adrenoreceptors are involved in the development or maintenance of high blood pressure in SHR, possibly by facilitating noradrenaline release.


Human TrkAR649W mutation impairs nociception, sweating and cognitive abilities: a mouse model of HSAN IV.
A functional nerve growth factor NGF-Tropomyosin Receptor kinase A (TrkA) system is an essential requisite for the generation and maintenance of long-lasting thermal and mechanical hyperalgesia in adult mammals. Indeed, mutations in the gene encoding for TrkA are responsible for a rare condition, named Hereditary Sensory and Autonomic Neuropathy type IV (HSAN IV), characterized by the loss of response to noxious stimuli, anhidrosis and cognitive impairment. However, to date, there is no available mouse model to properly understand how the NGF-TrkA system can lead to pathological phenotypes that are distinctive of HSAN IV. Here, we report the generation of a knock-in mouse line carrying the HSAN IV TrkAR649W mutation. First, by in vitro biochemical and biophysical analyses, we show that the pathological R649W mutation leads to kinase-inactive TrkA also affecting its membrane dynamics and trafficking. In agreement with the HSAN IV human phenotype, TrkAR649W/m mice display a lower response to thermal and chemical noxious stimuli, correlating with reduced skin innervation, in addition to decreased sweating in comparison to TrkAh/m controls. Moreover, the R649W mutation decreases anxiety-like behavior and compromises cognitive abilities, by impairing spatial-working and social memory. Our results further uncover unexplored roles of TrkA in thermoregulation and sociability. In addition to accurately recapitulating the clinical manifestations of HSAN IV patients, our findings contribute to clarifying the involvement of the NGF-TrkA system in pain sensation.


[Principles of oral health care for children].
In this article the principles of child oral health care in the Netherlands and the consequences of cariological principals of oral health care delivery to children are described. First of all the age of the child plays a very significant role in decisions concerning prevention and restoration. In addition, the combination of a positive attitude on the part of the dentist and a pain-free treatment can prevent fear of treatment and avoidance behaviour of the children. In the treatment of carious lesions, moreover, the oral health care provider should consider to what extent the problem is related to behaviour and fits in the multi-factorial caries model. Restorative treatments should be considered as supporting prevention, reducing caries activity and eliminating the effects of neglected oral health care. Prevention has to be seen as an essential element of the treatment model and it should start early from the standpoint of habit formation and the rapid demineralization process in the deciduous dentition which results from failed preventive care.


Anaplastic astroblastoma presenting as massive, sudden-onset, intraparenchymal hemorrhage.
Astroblastoma is a rare primary glial tumor of children and young adults, typically located in the cerebral hemispheres and presenting as a well-circumscribed, nodular, often cystic enhancing mass. The existence of astroblastoma as a distinct clinicopathologic entity has long been debated but is recognized in the 2007 WHO classification of CNS tumors. The grading of these tumors remains unsettled. Currently, no grade has been established and there are no clearly defined diagnostic criteria for low- or high-grade tumors. Astroblastomas in general are thought to have a favorable outcome with low rates of recurrence following gross total resection. We report a case of astroblastoma with 'high-grade/anaplastic' histology in a 12-year-old, previously healthy girl. The patient experienced severe, sudden-onset headache, and quickly became comatose. Head CT showed a massive intraparenchymal hemorrhage in the right frontal lobe with surrounding edema. She underwent emergent posterior frontal craniotomy for decompression and hematoma evacuation. Pathologic examination revealed a sharply demarcated hemorrhagic heterogeneous glial tumor with focal papillary architecture, densely hyalinized blood vessels, and intensely GFAP-positive perivascular cells. The tumor showed unequivocal high-grade features including an elevated proliferative index. The diagnosis of anaplastic astroblastoma was rendered. The patient expired on postoperative day 30. This case illustrates the potential poor outcome of high-grade astroblastoma and highlights the morphologic heterogeneity of this rare neoplasm.


Synthesis and antiviral evaluation of the beta-L-enantiomers of some thymine 3'-deoxypentofuranonucleoside derivatives.
3'-Deoxy-beta-L-erythro- (3), 3'-deoxy-beta-L-threo- (6), 2'-fluoro- (7) and 2'-azido-2',3'-dideoxy-beta-L-erythro- (10) pentofuranonucleoside derivatives of thymine have been synthesized and their antiviral properties examined. All these derivatives were stereospecifically prepared by glycosylation of thymine with a suitable peracylated 3-deoxy-L-erythro-pentofuranose sugar (1), followed by appropriate chemical modifications. The prepared compounds were tested for their activity against HIV, but they did not show an antiviral effect.


Tracheal reconstruction by resection and end-to-end anastomosis in the horse.
A surgical technique for resection of a portion of the trachea followed by end-to-end anastomosis was developed on 4 clinically normal horses. The trachea healed without complications in 3 of the horses in which 3 tracheal cartilages were removed. Five tracheal cartilages were removed from the 4th horse. It had to be euthanatized because of excessive suture line tension and wound disruption, which occurred during recovery from anesthesia. Suture line tension was measured on 3 other clinically normal horses positioned in dorsal recumbency immediately after euthanasia after 3 and 5 tracheal cartilages had been removed and before and after flexion of the neck to 90 degrees. Suture line tension was reduced by approximately 50% when the neck was flexed. Similarly, the force required to appose the cut ends of the trachea was nearly doubled when 5 rather than 3 tracheal cartilages were removed. The surgical technique was performed on 1 horse and 1 pony with partial obstruction of the trachea. In the pony, 5 tracheal cartilages were resected. It was able to return to successful show competition. In the horse, 4 tracheal cartilages were removed. This horse died of complications resulting from bilateral hindlimb myositis, but the tracheal anastomosis was intact, sealed, and healing well.


Extended-interval dosing of aminoglycosides.
Aminoglycosides are efficacious agents. Their use has declined partly because of the development of newer, presumably less toxic agents. Research shows that aminoglycosides can be dosed differently than in the past, maintaining efficacy while reducing aminoglycoside toxicities. Aminoglycosides administered with newer agents may help overcome antibiotic bacterial resistance and thus yield safe and more effective therapy. This study focused on the efficacy and safety of the aminoglycosides extended-interval dosing regimen and was conducted by a search of the literature. Data from The Mount Sinai Hospital regarding bacterial resistance patterns were collected. A nomogram describing the administration of the extended-interval dosing regimen is provided. Extended-interval dosing of aminoglycosides is as efficacious as administering these agents every 8 hours and may result in lower rates of toxicities. Extended-interval dosing also may cost less and be easier to administer. Aminoglycosides are less susceptible to bacterial resistance than many of the newer, currently favored antibiotics. Increasing the usage of aminoglycosides is likely to be safe and beneficial in the treatment of certain bacterial infections.


Systems-Based Practice: Expert Perspectives on the Origin and Evolution of an Ambiguous Competency.
Systems-based practice (SBP) has been a core competency in graduate medical education in the United States since 1999, but it has been difficult to operationalize in residency programs due to its conceptual ambiguity. The authors explored the historical origin and subsequent development of the SBP competency from the perspective of individuals who were influential across critical phases of its implementation and ensuing development. The goal of this study was to elicit the history of SBP from the perspective of individuals who have expertise in it, and to use those findings to inform the current SBP construct. Between March and July 2021, 24 physicians, nurses, educators, and leaders in the field of SBP were individually interviewed about the origin and meaning of SBP as practiced in U.S. medical education using a semistructured guide. Individuals were selected based upon their influence on the origin or evolution of the SBP competency. Data were iteratively collected and analyzed using real-time analytic memos, regular adjudication sessions with the research team, and thematic analysis. Researchers identified themes from participants' perspectives and agreed upon the final results and quotations. Five themes were identified: SBP has many different definitions, SBP was intentionally designed to be vague, systems thinking was identified as the foundation of the SBP competency, the 6 core competencies established in the United States by the Accreditation Council for Graduate Medical Education were developed to be interdependent, and the SBP and practice-based learning and improvement competencies are uniquely related and synergistic. Interview data indicate that since its inception, SBP has been a nuanced and complex competency, resulting in a lack of mutually shared understanding among stakeholders. This deliberate historical examination of expert perspectives provides insight into specific areas for improving how SBP is taught and learned.


Effects of progesterone on chemosensitivity in normal men.
Progesterone administration increase VE in man, but its effects on ventilatory response to hypercapnia and hypoxia have not been well documented. Accordingly, VE, HVR, and HCVR were measured during placebo and MPA administration in 11 normal men. The effect of MPA (20 mg orally q 8 hr for 32 hr) on T degrees, metabolic rate (VO2 and VCO2) was also determined. With MPA, T degrees, rose 0.4 degrees C +/- 0.0008 (S.E.M.) p less than 0.0001), VE increased 0.46 +/- 0.16 L/min (p less than 0.01), and VO2 and VCO2 did not change significantly. HCVR (measured under hyperoxic conditions during rebreathing) increased significantly (P less than 0.01) from 2.9 +/- 0.33 L/min/mm Hg (placebo) to 4.0 +/- 0.29 (MPA). HVR was measured as the shape parameter A, so that when A increased, HVR was augmented. During MPA, HVR increased from A = 132 +/- 19.1 to 179 +/- 20.5 (P less than 0.02). We conclude that 60 mg of MPA daily in normal men increases VE and chemosensitivity as measured by the ventilatory response to hypercapnia and hypoxia.


The unique expression profile of FAM19A1 in the mouse brain and its association with hyperactivity, long-term memory and fear acquisition.
Neurodevelopment and mature brain function are spatiotemporally regulated by various cytokines and chemokines. The chemokine-like neuropeptide FAM19A1 is a member of family with sequence similarity 19 (FAM19), which is predominantly expressed in the brain. Its highly conserved amino acid sequence among vertebrates suggests that FAM19A1 may play important physiological roles in neurodevelopment and brain function. Here we used a LacZ reporter gene system to map the expression pattern of the FAM19A1 gene in the mouse brain. The FAM19A1 expression was observed in several brain regions starting during embryonic brain development. As the brain matured, the FAM19A1 expression was detected in the pyramidal cells of cortical layers 2/3 and 5 and in several limbic areas, including the hippocampus and the amygdala. FAM19A1-deficient mice were used to evaluate the physiological contribution of FAM19A1 to various brain functions. In behavior analysis, FAM19A1-deficient mice exhibited several abnormal behaviors, including hyperactive locomotor behavior, long-term memory deficits and fear acquisition failure. These findings provide insight into the potential contributions of FAM19A1 to neurodevelopment and mature brain function.


Genomic organization of the human NTRK1 gene.
The NTRK1 gene encodes one of the receptors for the Nerve Growth Factors and it is located at 1q21-22. Rearrangements of NTRK1 are frequently detected in human papillary thyroid carcinoma and lead to the formation of chimeric oncogenes, similarly to what observed for the other neurotrophin receptor RET. In addition, the two receptor genes are target of point mutations associated with different human diseases. RET is affected by germ line and somatic mutations in MEN2A, MEN2B tumor syndromes and in the abnormal developmental Hirschsprung disease, whereas mutations of NTRK1 have been reported very recently in patients with congenital insensitivity to pain with anidrosis (CIPA). With the aim to provide a tool for searching mutations along the whole NTRK1 gene, we have determined its genomic organization. Our results demonstrated that NTRK1 is contained within 25 Kb of DNA and is organized in 17 exons, one of which is alternatively spliced. The sequence of the 5' flanking region indicates a high content in C/G, the absence of TATA box, the presence of several putative binding sites for Sp1, AP1, AP2, AP3, ATF and GCF transcription factors.


Evidence for post-translational membrane insertion of the integral membrane protein bacterioopsin expressed in the heterologous halophilic archaeon Haloferax volcanii.
The gene coding for the integral membrane protein bacterioopsin (Bop), that is composed of seven transmembrane helices, was expressed in the halophilic archaeon Haloferax volcanii as a fusion protein with the halobacterial enzyme dihydrofolate reductase and with the cellulose binding domain of Clostridium thermocellum cellulosome. In each case, bacterioopsin was present both in the membrane and in the cytoplasmic fractions. Pulse-chase labeling experiments showed that the fusion protein in the cytoplasmic fraction is the precursor of the membrane-bound species. Bacterioopsin mutants that lack the seventh helix (BopDelta7) were found to accumulate only in the cytoplasmic fraction, whereas bacterioopsin mutants that lack either helices four and five (BopDelta4-5), or helices one and two (BopDelta1-2), were found in the cytoplasmic as well as in the membrane fractions. The seventh helix, when expressed alone, could target in trans the insertion of a separately expressed bacterioopsin mutant protein that has only the first six helices. These results support a model in which bacterioopsin is produced in H. volcanii as a soluble protein and in which its insertion into the membrane occurs post-translationally. According to this model, membrane insertion is directed by the seventh helix.


Structure-dependent all-optical switching in graphene-nanoribbon-like molecules: fully conjugated tri(perylene bisimides).
We present the structure-dependent nonlinear optical (NLO) properties of fully conjugated tri(perylene bisimides) (triPBIs) toward the understanding of the role of conformational flexibility and pi-electron conjugation in molecular NLO properties of model graphene-nanoribbon (GNR)-like molecules. In the present paper, we report the NLO absorption properties of the triPBIs in toluene excited at 532 nm with nanosecond laser pulses, where the observed transient excited state is determined to be a triplet and presented in the nonlinear process similar to the NLO properties that occur in C(60). As a result, the all-optical switching in both visible and near-infrared regions upon excitation at 532 nm was demonstrated, suggesting that the chemically synthesized model GNRs act well as smart all-optical switching devices without the need of external control. Furthermore, Raman spectral measurement was further used to characterize the conjugated structure properties of model compounds of functionalized graphene nanoribbons (F-GNRs), while the dispersion and splitting of the G-band and D-band in both frequency and intensity can help to distinguish the pi-conjugation and conformational flexibility of the two different triPBI isomers, showing the opportunity to tailor their optoelectronic properties by precisely controlling the edge orientation, edge width, and chemical termination of the edges in the synthesized F-GNRs.


Rotating electrode potentiometry: lowering the detection limits of nonequilibrium polyion-sensitive membrane electrodes.
A rotating electrode configuration is evaluated as a means to lower the detection limits of newly devised polyion-sensitive membrane electrodes (PSEs). Planar potentiometric polycation and polyanion PSEs are prepared by incorporating tridodecylmethylammonium chloride and calcium dinonylnaphthalenesulfonate, respectively, into plasticized PVC or polyurethane membranes and mounting disks of such films on an electrode body housed in a conventional rotating disk electrode apparatus. Rotation of the PSEs at 5000 rpm results in an enhancement in the detection limits toward heparin (polyanion) and protamine (polycation) of at least 1 order of magnitude (to 0.01 unit/mL for heparin; 0.02 microg/mL for protamine) over that observed when the EMF responses of the same electrodes are assessed using a stir-bar to achieve convective mass transport. A linear relationship between omega(-1/2), where omega is the rotating angular frequency, and C1/2, the polyion concentration corresponding to half the total maximum deltaEMF response toward the polyion species, is observed. It is further shown that the rotating polycation sensor can be used as an end-point detector to greatly enhance (relative to nonrotated indicator electrode) the analytical resolution and precision for measurement of low concentrations of heparin when such samples are titrated with protamine. The theoretical basis for lowering the detection limits by rotating PSEs is discussed based on the unique nonequilibrium response mechanism of such sensors.


UC2 search: using unique connectivity of uncharged compounds for metabolite annotation by database searching in mass spectrometry-based metabolomics.
For metabolite annotation in metabolomics, variations in the registered states of compounds (charged molecules and multiple components, such as salts) and their redundancy among compound databases could be the cause of misannotations and hamper immediate recognition of the uniqueness of metabolites while searching by mass values measured using mass spectrometry. We developed a search system named UC2 (Unique Connectivity of Uncharged Compounds), where compounds are tentatively neutralized into uncharged states and stored on the basis of their unique connectivity of atoms after removing their stereochemical information using the first block in the hash of the IUPAC International Chemical Identifier, by which false-positive hits are remarkably reduced, both charged and uncharged compounds are properly searched in a single query and records having a unique connectivity are compiled in a single search result. The UC2 search tool is available free of charge as a REST web service (http://webs2.kazusa.or.jp/mfsearcher) and a Java-based GUI tool. email@example.com. Supplementary data are available at Bioinformatics online.


A coaxial single fibre supercapacitor for energy storage.
An energy storage device--a coaxial single fibre supercapacitor--was developed using a dip coating method and characterised using electrochemical methods. The specific capacitance per unit area and length were calculated to be 3.18 mF cm(-2) and 0.1 mF cm(-1), respectively, for a 2.6 cm supercapacitor. A 70 cm long fibre supercapacitor was then woven into a piece of fabric. Following characterisation using cyclic voltammetry, we found that this device had the same capacitance as its original value before integration into the fabric.


Distribution of transferrin saturations in the African-American population.
To determine if transferrin saturations in African Americans may reflect the presence of a gene that influences iron metabolism, we analyzed the distribution of these values in 808 African Americans from the second National Health and Nutrition Survey. We tested for a mixture of three normal distributions consistent with population genetics for a major locus effect in which the proportion of normal homozygotes is p2; of heterozygotes, 2pq; of affected homozygotes, q2; and in which p+q = 1. Three subpopulations based on transferrin saturation were present (P < .0001) and the fit to a mixture of three normal distributions was good (P = .2). A proportion of .009 was included in a subpopulation with a mean +/- standard deviation transferrin saturation of 63.4% +/- 5.7% (postulated homozygotes for a gene that influences iron metabolism), while a proportion of .136 had an intermediate saturation of 38.0% +/- 5.7% (postulated heterozygotes) and .856 a saturation of 24.6% +/- 5.7% (postulated normal homozygotes). These proportions were consistent with population genetics because the sum of the square roots of the proportions with the lowest mean transferrin saturation (P = .925) and the highest (q = 0.093) was approximately 1 (1.018). The results are consistent with the presence in African Americans of a common locus that influences iron metabolism.


Effect of beta-adrenergic stimulation on oxygen metabolism in the fetal lamb.
Catecholamines, which are released into the circulation during stress, increase fetal metabolism. This effect appears to be related to beta-adrenoreceptor stimulation. We examined the effect of isoproterenol infusion on umbilical blood flow, oxygen delivery and consumption, and glucose and lactate uptake in late-gestation fetal lambs. Isoproterenol increased umbilical blood flow, but oxygen delivery to the fetus did not increase. Umbilical venous oxygen content fell linearly with the increase in umbilical blood flow. It is proposed that oxygen delivery to the sheep fetus is at or near a maximum and that oxygen delivery cannot be raised by increasing umbilical blood flow because oxygen diffusion at the placental site is limited. Fetal oxygen consumption increased initially but returned to control levels with an increase in infusion rate. Blood glucose concentration increased during isoproterenol infusion; this was due to release of glucose and not because of increased placental uptake. Fetal blood pH values fell in association with elevated lactate levels. It is proposed that elevated glucose concentrations resulted in increased metabolism of glucose, and because oxygen delivery could not be enhanced, increased anaerobic glycolysis caused lactate concentration to rise.


The effect of tricaine on use of the fluorescein test for detecting skin and corneal ulcers in fish.
Fluorescein has been used for rapid and sensitive detection of fish skin and corneal ulceration. Effective use of the fluorescein test requires knowledge of conditions that might cause misleading interpretations or otherwise interfere with test reliability. Examination of fish health and the clinical workup often require tricaine as one of the most commonly used anesthetics. However, tricaine may interfere with correct interpretation of the fluorescein test and might also cause significant fish injury. The effects of tricaine exposure sequence on the fidelity of the fluorescein test was studied in Pacific halibut Hippoglossus stenolepis, walleye pollock Theragra chalcogramma, and northern rock soles Lepidopsetta polyxystra by examining the fluorescence of experimentally induced epidermal wounding. Tricaine can quench fluorescence that is emitted by fluorescein retained in skin ulcers, causing a false-negative reaction. Thus, for the fluorescein test to work properly, it is important to avoid the exposure of fluorescein-treated and rinsed ulcers to tricaine. The effects of exposure to buffered versus unbuffered tricaine on epidermal and corneal integrity were studied in Nile tilapia Oreochromis niloticus and channel catfish Ictalurus punctatus subjected to the fluorescein test and histological examination. Fluorescein could detect not only ulcers but also areas with only a partial loss of epithelium (i.e., erosion). The use of unbuffered tricaine to anesthetize these fish caused serious epidermal and corneal damage. If fish are euthanized with unbuffered tricaine for clinical workup, this severe epidermal or corneal damage could be misinterpreted as an antemortem lesion, leading to misdiagnosis. Even in water with alkalinity exceeding 50 mg/L as CaCO3, it would seem prudent to always buffer tricaine with sodium bicarbonate to prevent a pH change that might lead to iatrogenic effects from unbuffered tricaine. Thus, current general recommendations suggesting that tricaine does not need to be buffered in waters with alkalinity greater than 50 mg/L might need to be modified.


Surgery of pelviureteric obstruction in 101 children over one year of age.
One hundred and one children over 1 year of age have had surgery for pelviureteric obstruction over an 11 year period. The common clinical features were abdominal pain, urinary infection or haematuria, but a number presented as an incidental finding. Less commonly, the patients presented with an abdominal mass or with hypertension. The diagnosis was usually made on intravenous pyelography (IVP) but in the latter part of the series, renal nuclide scan (RNS) and ultrasonography (US) were preferred. Ninety-three patients had a unilateral pyeloplasty, three had bilateral pyeloplasty and five had nephrectomy or heminephrectomy. Whereas initially nephrostomy drainage was used in the majority of patients after pyeloplasty, a trend away from nephrostomy evolved in the latter part of the series. With experience, the incidence of postoperative complications was also reduced and there was a reduction in the period of hospitalization. Clinical results were consistently satisfactory. Postoperative assessment after pyeloplasty was made by IVP and/or RNS and also US. A review of these investigations showed that RNS provided more factual information of the result when compared with the IVP.


The price elasticity of demand for heroin: Matched longitudinal and experimental evidence.
This paper reports estimates of the price elasticity of demand for heroin based on a newly constructed dataset. The dataset has two matched components concerning the same sample of regular heroin users: longitudinal information about real-world heroin demand (actual price and actual quantity at daily intervals for each heroin user in the sample) and experimental information about laboratory heroin demand (elicited by presenting the same heroin users with scenarios in a laboratory setting). Two empirical strategies are used to estimate the price elasticity of demand for heroin. The first strategy exploits the idiosyncratic variation in the price experienced by a heroin user over time that occurs in markets for illegal drugs. The second strategy exploits the experimentally induced variation in price experienced by a heroin user across experimental scenarios. Both empirical strategies result in the estimate that the conditional price elasticity of demand for heroin is approximately -0.80.


Reduced acetylcholine receptor density, morphological remodeling, and butyrylcholinesterase activity can sustain muscle function in acetylcholinesterase knockout mice.
Nerve-evoked contractions were studied in vitro in phrenic nerve-hemidiaphragm preparations from strain 129X1 acetylcholinesterase knockout (AChE-/-) mice and their wild-type littermates (AChE+/+). The AChE-/- mice fail to express AChE but have normal levels of butyrylcholinesterase (BChE) and can survive into adulthood. Twitch tensions elicited in diaphragms of AChE-/- mice by single supramaximal stimuli had larger amplitudes and slower rise and decay times than did those in wild-type animals. In AChE-/- preparations, repetitive stimulation at frequencies of 20 and 50 Hz and at 200 and 400 Hz produced decremental muscle tensions; however, stimulation at 70 and 100 Hz resulted in little or no loss of tension during trains. Muscles from AChE+/+ mice maintained tension at all frequencies examined but exhibited tetanic fade after exposure to the selective AChE inhibitor 1,5-bis(4-allyldimethyl-ammoniumphenyl)pentane-3-one (BW 284C51). The ability of diaphragm muscles from AChE-/- mice to maintain tension at 70 and 100 Hz suggests a partial compensation for impairment of acetylcholine (ACh) hydrolysis. Three mechanisms--including a reliance on BChE activity for termination of ACh action, downregulation of nicotinic acetylcholine receptors (nAChRs), and morphological remodeling of the endplate region--were identified. Studies of neuromuscular transmission in this model system provide an excellent opportunity to evaluate the role of AChE without complications arising from use of inhibitors.


Diffusion tensor imaging of the hippocampus and verbal memory performance: the RUN DMC study.
Cerebral small vessel disease (SVD) and hippocampal atrophy are related to verbal memory failures and may ultimately result in Alzheimer's disease. However, verbal memory failures are often present before structural changes on conventional MRI appear. Changes in microstructural integrity of the hippocampus, which cannot be detected with conventional MRI, may be the underlying pathological substrate. With diffusion tensor imaging (DTI), we investigated the relation between the microstructural integrity of the hippocampus and verbal memory performance in 503 nondemented elderly with SVD. The Radboud University Nijmegen Diffusion tensor and Magnetic resonance imaging Cohort study is a prospective cohort study among 503 nondemented elderly with cerebral SVD aged between 50 and 85 years. All participants underwent T1 MPRAGE, fluid-attenuated inversion recovery, DTI scanning and the Rey Auditory Verbal Learning Test. After manual segmentation of the hippocampi, we calculated the mean diffusivity (MD) and fractional anisotropy in both hippocampi. The relation between memory performance and hippocampal DTI parameters was adjusted for age, sex, education, depressive symptoms, hippocampal, and white-matter lesions volume and lacunar infarcts. We found inverse relations between hippocampal MD and verbal memory performance (β = -0.22; P < 0.001), immediate recall (β = -0.22; P < 0.001), delayed recall (β = -0.20; P < 0.001), and forgetting rate (β = -0.13; P = 0.025), most pronounced in participants with a normal hippocampal volume. Microstructural integrity of the hippocampus assessed by DTI is related to verbal memory performance in elderly with SVD, also in participants with an intact appearing hippocampus. Changes in hippocampal microstructure may be an early marker of underlying neurodegenerative disease, before macrostructural (i.e., volumetric) changes occur.


Conductively coupled flexible silicon electronic systems for chronic neural electrophysiology.
Materials and structures that enable long-term, intimate coupling of flexible electronic devices to biological systems are critically important to the development of advanced biomedical implants for biological research and for clinical medicine. By comparison with simple interfaces based on arrays of passive electrodes, the active electronics in such systems provide powerful and sometimes essential levels of functionality; they also demand long-lived, perfect biofluid barriers to prevent corrosive degradation of the active materials and electrical damage to the adjacent tissues. Recent reports describe strategies that enable relevant capabilities in flexible electronic systems, but only for capacitively coupled interfaces. Here, we introduce schemes that exploit patterns of highly doped silicon nanomembranes chemically bonded to thin, thermally grown layers of SiO2 as leakage-free, chronically stable, conductively coupled interfaces. The results can naturally support high-performance, flexible silicon electronic systems capable of amplified sensing and active matrix multiplexing in biopotential recording and in stimulation via Faradaic charge injection. Systematic in vitro studies highlight key considerations in the materials science and the electrical designs for high-fidelity, chronic operation. The results provide a versatile route to biointegrated forms of flexible electronics that can incorporate the most advanced silicon device technologies with broad applications in electrical interfaces to the brain and to other organ systems.


Gap acceptance behavior at unsignalized intersections: Effects of using a phone and a music player while driving.
Objectives: The present study is an attempt to analyze and compare the distraction effects caused by the use of a phone and a music player at unsignalized intersections. Method: Eighty-eight participants performed simulated driving experiments where they faced a sequence of gaps in the major road traffic at 2 unsignalized intersections. In this process, their driving behavior was evaluated in terms of gap acceptance probability, accepted lag, and maneuver completion time. These parameters were modeled with a generalized estimating equation (GEE) method by considering distraction, demographic factors, driving history, maneuver types, and driving attributes in the approach and completion zones as independent variables. Results: The results showed that gap acceptance probability decreased by 46% during the conversation task, whereas it increased by 66% during the music player task. Lower gap acceptance could be a compensatory behavior adopted by drivers during the conversation task, whereas no such measure was adapted during the music player task. The results indicate that a higher approach speed during the music player task might have led to increased gap acceptance. Further, though the effect of distraction on the accepted lag was not evident, the completion time was reduced during the conversation task. Conclusions: Overall, the results suggest that drivers are more likely to adopt a compensatory measure in complex driving situations only if they perceive a high risk. Hence, drivers are exposed to a greater risk while operating a music player, because this is not perceived as risky behavior.


Vegetable oils used as vitamin E vehicle affect the electrical activity of the rat heart.
The aim of this study is to define the possible effects of vegetable oils used as vitamin E vehicle on the electrical activity of the rat heart. To test the possible effects of vitamin E vehicles we studied the effect of i.p. injected corn oil, hazelnut oil or peanut oil on the action potential parameters recorded in both papillary and left atrial muscle strips. Four experimental groups were used. The control group was injected (i.p.) with distilled water, while the three remaining groups received injections of corn oil, hazelnut oil, or peanut oil for five weeks (in a dose of 0.4 ml/kg/day--minimum amount of oil in which vitamin E could be dissolved). We used borosilicated (15-20 megaohms) capillary electrodes and intracellular action potentials (AP) were recorded in isolated papillary and left atrium muscle strips. While administration of three different types of vegetable oil had no significant effect on AP parameters of papillary muscle, they significantly prolonged the repolarization phase of AP in atrial strips. These results show that vegetable oils used as vitamin E vehicles may alter the electrical activity of the heart in a tissue-dependent manner. The present data indicate that the possible effect of vegetable oil vehicles should be kept in mind while evaluating the possible effects of in vivo vitamin E administration.


Aberrant Amygdala-dependent Fear Memory in Corticosterone-treated Mice.
Anxiety disorder is a major psychiatric disorder characterized by fear, worry, and excessive rumination. However, the molecular mechanisms underlying neural plasticity and anxiety remain unclear. Here, we utilized a mouse model of anxiety-like behaviors induced by the chronic administration of corticosterone (CORT) to determine the exact mechanism of each region of the fear circuits in the anxiety disorders. Chronic CORT-treated mice showed a significant increase in anxiety-related behaviors as assessed by the elevated plus maze, light-dark, open-field, and marble-burying tasks. In addition, chronic CORT-treated mice exhibited abnormal amygdala-dependent tone-induced fear memory but normal hippocampus-dependent contextual memory. Consistent with amygdala hyperactivation, chronic CORT-treated mice showed significantly increased numbers of c-Fos-positive cells in the basolateral amygdala (BLA) after tone stimulation. Long-term potentiation (LTP) was markedly enhanced in the BLA of chronic CORT-treated mice compared to that of vehicle-treated mice. Immunoblot analyses revealed that autophosphorylation of Ca2+/calmodulin-dependent protein kinase (CaMK) IIα at threonine 286 and phosphorylation of cyclic-adenosine-monophosphate response-element-binding protein (CREB) at serine 133 were markedly increased in the BLA of chronic CORT-treated mice after tone stimulation. The protein and mRNA levels of brain-derived neurotrophic factor (BDNF) also significantly increased. Our findings suggest that increased CaMKII activity and synaptic plasticity in the BLA likely account for the aberrant amygdala-dependent fear memory in chronic CORT-treated mice.


An apicosome initiates self-organizing morphogenesis of human pluripotent stem cells.
Human pluripotent stem cells (hPSCs) self-organize into apicobasally polarized cysts, reminiscent of the lumenal epiblast stage, providing a model to explore key morphogenic processes in early human embryos. Here, we show that apical polarization begins on the interior of single hPSCs through the dynamic formation of a highly organized perinuclear apicosome structure. The membrane surrounding the apicosome is enriched in apical markers and displays microvilli and a primary cilium; its lumenal space is rich in Ca2+ Time-lapse imaging of isolated hPSCs reveals that the apicosome forms de novo in interphase, retains its structure during mitosis, is asymmetrically inherited after mitosis, and relocates to the recently formed cytokinetic plane, where it establishes a fully polarized lumen. In a multicellular aggregate of hPSCs, intracellular apicosomes from multiple cells are trafficked to generate a common lumenal cavity. Thus, the apicosome is a unique preassembled apical structure that can be rapidly used in single or clustered hPSCs to initiate self-organized apical polarization and lumenogenesis.


Combined cognitive and parent training interventions for adolescents with ADHD and their mothers: A randomized controlled trial.
This study examined the individual and combined effects of two nonpharmacological treatments for attention deficit/hyperactivity disorder (ADHD): Cogmed working memory training (CWMT) for adolescents and behavioral parent training (BPT) for mothers. Ninety-one adolescents (ages 11-15) and their mothers were randomized to one of four CWMT and BPT treatment and active control (placebo) group combinations of 5-week interventions. At pre- and posttest, mothers and teachers completed rating forms, and adolescents completed neuropsychological measures of working memory (WM). Individual intervention effects showed that treatment CWMT significantly improved WM spans, whereas there were no significant differences for treatment or control BPT on reports of parent-related outcomes. Combined treatment effects indicated an overall pattern of greatest improvements for the control CWMT/treatment BPT group, as compared to the other three groups, on adolescent WM deficit, behavioral regulation problems, and global executive deficit. Most significant effects for outcomes were main effects of improvements over time. A combination of CWMT and BPT did not result in increased treatment gains. However, potential effects of combined treatment may have been masked by greater perceived benefits arising from lack of struggle in the nonadaptive, CWMT active control condition. Future combined intervention research should focus on specific, theoretically driven WM deficits among individuals with ADHD, should include possible adaptations to the standard CWMT program, should examine effectiveness of cognitive treatments combined with contextual interventions and should utilize appropriate control groups to fully understand the unique and combined effects of interventions.


Do adult rat ventricular myocytes express protein kinase C-alpha?
Although calcium-insensitive protein kinase C (PKC) isoforms (PKC-epsilon and PKC-delta) are consistently detected in adult ventricular myocytes, the evidence that adult ventricular myocytes also express calcium-sensitive PKC-alpha is inconsistent. The current study used four different anti-PKC-alpha-antibodies to resolve some of the uncertainties regarding the immunodetection of PKC-alpha in adult ventricular myocytes. Three of the antibodies used in this study barely (GIBCO-BRL) or rather faintly (Transduction Laboratories and Seikagaku America) recognize PKC-alpha in crude preparations from adult ventricular myocytes. Although each of these antibodies recognizes a prominent 80-kDa band, which is similar in size to PKC-alpha, this represents nonspecific immunoreactivity and should not be confused with PKC-alpha. This conclusion is based on peptide-blocking experiments (GIBCO-BRL), the absence of the requisite sensitivity to calcium- and phorbol 12-myristate 13-acetate-induced translocation (Seikagaku America and Transduction Laboratories), and/or the failure to copurify with PKC-alpha on DEAE-Sephacel chromatography. Nevertheless, an antibody from Upstate Biotechnology clearly recognizes PKC-alpha and not other unrelated nonspecific immunoreactive species in crude preparations from adult ventricular myocytes. Each of the antisera used in this study could detect PKC-alpha immunoreactivity following chromatographic purification of the samples to enrich for PKC-alpha and remove nonspecific immunoreactive proteins. These results suggest that PKC-alpha is expressed by adult ventricular myocytes and argue that differences in the sensitivity and/or specificity of available antisera contribute to at least some of the confusion regarding PKC-alpha expression in adult ventricular myocytes.


Comparison of fibreoptic endoscopy in acute upper gastrointestinal haemorrhage in Africans and Europeans.
The results of endoscopy in acute upper gastrointestinal haemorrhage were compared in a group of 138 Africans and one of 84 Europeans. Contrary to widely held clinical opinion, the incidence of gastric and duodenal ulceration was similar in the two races. Peptic ulcers were the main source of bleeding in both groups and were surprisingly more common than varices in the Africans. Bleeding from varices, however, was far more common in the Africans than in the Europeans. Stomal ulcers were confined to Europeans. Gastric erosions, often attributed to herbal medicines, were more common in the Africans but the difference was not significant. The study was not designed to determine reduced mortality since the introduction of endoscopy, but management, especially in the Africans, was aided by early recognition of haemorrhage from oesophageal varices and acute gastric erosions.


Subterranean waterworks of biblical jerusalem: adaptation of a karst system.
Ancient Jerusalem has long been known to possess a system of subterranean waterworks by which the spring of Gihon, which issues outside the walls, could be approached from within the city, and its waters diverted to an intramural pool. Most scholars regarded these waterworks as man-made, but the techniques of underground orientation and ventilation employed by the builders, as well as the numerous anomalies and ostensible mistakes in design, mystified investigators. Geological investigation has revealed the waterworks to be part of a well-developed karst system, a network of natural dissolution channels and shafts, in the limestone and dolomite underlying the city. Thus, it was not through primary planning but by means of skillful adaptation of these pre-existing natural features that the city was ensured of a dependable water supply during both war and peace. Likewise, knowledge of the subterranean access may have played a role in David's capture of the Jebusite city.