Graded müllerectomy for correction of graves upper eyelid retraction: effect on eyelid movements.
To measure the effect of müllerectomy from posterior approach on the amplitudes of spontaneous blinking and downward eyelid saccades. Spontaneous blinks and downward upper eyelid saccadic movements of 16 patients (23 eyelids) with Graves orbitopathy were measured before and after müllerectomy from posterior approach. A new video system was used to continuously register the blinking activity while subjects viewed a commercial movie for 5 minutes. Downward eyelid saccades (30° of downgaze) were also measured with the video system. Müllerectomy had no effect on the amplitude of the blink. However, as the eyelid margins were significantly lowered by the surgery, the amplitude of the blink movements relative to the pupil center increased substantially. The number of movements occluding the pupil center increased from 0% to 13%. Due to the increased efficiency of blinking, the blink rate decreased. Surgery induced a mean increase of 1.1 mm of downward saccades. The effects of müllerectomy on the blinks are indirect and related to correction of eyelid retraction. The relative amplitude of blink movements increases and blink rate decreases. Müllerectomy does affect the downward eyelid saccades increasing the ability of the upper eyelid to relax on downgaze.


Association of Autologous Costal Cartilage Harvesting Technique With Donor-Site Pain in Patients Undergoing Rhinoplasty.
Postoperative pain at the donor site is a common morbidity following autologous costal cartilage grafting. To evaluate postoperative pain at the donor site after the use of a muscle-sparing costal cartilage harvesting technique compared with a muscle-cutting technique using electrocautery. Designed as a controlled trial without randomization, this prospective, comparative cohort study was conducted between January 1, 2016, and March 31, 2017. Participants included 20 patients who underwent rhinoplasty for various cosmetic and functional complaints from January 1, 2016, to February 28, 2017. Of the 20 patients, 1 was excluded owing to an infection that developed on postoperative day (POD) 7. Patients were grouped by the rib harvesting technique used that was either a muscle-sparing technique (n = 11) or a muscle-cutting technique (n = 8). Skin incisions for both groups were carried out with a blade. Transection of muscle fascia and muscle fibers was performed with monopolar electrocautery in the muscle-cutting technique group. Blunt dissection with a hemostat was performed in the muscle-sparing technique group. All other surgical techniques were identical. Postoperative pain was assessed with visual analog scale scores for resting pain and movement pain. Eight pain measurements were noted at the sixth postoperative hour and on PODs 1, 2, 3, 7, 15, 30, and 45. During the hospital stay, the postoperative need for analgesics was recorded daily as the number of analgesic infusion vials used. The 19 patients in the study included 11 women and 8 men whose mean age (SD) was 33.2 (10.3) years The mean (SD) visual pain analog scale scores for resting pain and movement pain were consistently higher in the muscle-cutting technique group than in the muscle-sparing technique group. This difference was statistically significant on PODs 2, 3, and 15 for resting pain and on PODs 2, 3, 7, 15, 30, and 45 for movement pain. The mean postoperative need for analgesic infusion vials during hospital stay was higher in the muscle-cutting technique group, and the difference was statistically significant on POD 2 (1.9 [0.6] vials vs 1.0 [0.9] vials; P = .02). Both resting and movement pain at the donor site was significantly reduced in the muscle-sparing technique group during the postoperative period, findings that align with anecdotal reports in the literature. Routine use of the muscle-sparing technique in autologous costal cartilage harvesting is recommended to reduce postoperative pain. 2.


[Pharmacological analysis of the role of dorsal hippocampus adrenergic effects in the central mechanisms of ovulation and compensatory ovarian hypertrophy in rats].
In experiments on mature female rats it was shown that L-DOPA administration restores normal ovulation in rats after the dorsal hippocampus destruction and related partial ovulation blockade. Pharmacological analysis after microinjections of alpha- and beta-adrenoblockers into the dorsal hippocampus region has shown that alpha- and beta-adrenoreception of the dorsal hippocampus plays a definite role in the regulation of gonadotropin secretion. However, the effect of phentolamine injection on the rat ovulatory reaction was less pronounced than under conditions of this hippocampus region destruction. The beta-adrenoblocker propranolol injected into the dorsal hippocampus inhibited compensatory hypertrophy of the ovaries in hemicastrated rats, decreased the hypophyseal FSH content and did not influence ovulation.


Structural basis of flocculin-mediated social behavior in yeast.
In the budding yeast Saccharomyces cerevisiae, self-recognition and the thereby promoted aggregation of thousands of cells into protective flocs is mediated by a family of cell-surface adhesins, the flocculins (Flo). Based on this social behavior FLO genes fulfill the definition of "greenbeard" genes, which direct cooperation toward other carriers of the same gene. The process of flocculation plays an eminent role in the food industry for the production of beer and wine. However, the precise mode of flocculin-mediated surface recognition and the exact structure of cognate ligands have remained elusive. Here, we present structures of the adhesion domain of a flocculin complexed to its cognate ligands derived from yeast high-mannose oligosaccharides at resolutions up to 0.95 Å. Besides a PA14-like architecture, the Flo5A domain reveals a previously undescribed lectin fold that utilizes a unique DcisD calcium-binding motif for carbohydrate binding and that is widely spread among pro- and eukaryotes. Given the high abundance of high-mannose oligosaccharides in yeast cell walls, the Flo5A structure suggests a model for recognition, where social non-self- instead of unsocial self-interactions are favored.


Emotion regulation dynamics predict substance use in high-risk adolescents.
Emotional factors such as stress and anxiety contribute to risk of substance use in adolescents. Descriptive measures of within-person affect variability are often used to predict morbidity, but indices derived from theoretical models may provide more interpretable alternatives. A continuous-time state-space model of emotion regulation as closed-loop feedback control was used to estimate the homeostatic tendency of affect in each of 94 adolescent participants. The resulting indices of emotion regulation were then compared to within-person affect sum score means and standard deviations in predicting total counts of nicotine, alcohol, and cannabis use. Model-based emotion regulation was significantly associated with lower frequencies of nicotine, alcohol, and cannabis use, while mean negative affect sum score was associated with higher frequencies. Model comparisons revealed that while model-based predictors and descriptive statistics explained similar amounts of variance in substance use, the explained variance proportions were independent between the approaches. The greatest predictive value was achieved by a combined model with both sets of affect indices. We conclude that theoretically defined and model-estimated individual characteristics may serve an important role in conceptualizing and predicting substance use behavior.


Contribution of Patient Interviews as Part of a Comprehensive Approach to the Identification of Drug-Related Problems on Geriatric Wards.
Inappropriate prescribing is linked to increased risks for adverse drug reactions and hospitalisation. Combining explicit and implicit criteria of inappropriate prescribing with the information obtained in patient interviews seems beneficial with regard to the identification of drug-related problems (DRPs) in hospitalised patients. We aimed to investigate the inclusion of pharmacist interviews as part of medication reviews (including the use of explicit and implicit criteria of inappropriate prescribing) to identify DRPs in older inpatients. Clinical medication reviews were performed on geriatric and associated physical and neurological rehabilitation wards in a regional secondary care hospital. Data from electronic medical records, laboratory data, and current treatment regimens were complemented with a novel structured patient interview performed by a clinical pharmacist. The structured interview questioned patients on administration issues, prescribed medication, self-medication, and allergies. The reviews included the use of current treatment guidelines, the Medication Appropriateness Index, the Screening Tool of Older People's Prescriptions (STOPP, v2), and the Screening Tool to Alert to Right Treatment (START, v2). The potential relevance of the DRPs was estimated using the German version of the CLEO tool. In 110 patients, 595 DRPs were identified, averaging 5.4 per patient (range 0-17). The structured interviews identified 249 DRPs (41.8%), of which 227 were not identified by any other source of information. The majority of DRPs (213/249, i.e. 85.5%) identified by patient interview were estimated to be of minor clinical relevance (i.e. limited adherence, knowledge, quality of life, or satisfaction). We demonstrated that structured patient interviews identified additional DRPs that other sources did not identify. Embedded within a comprehensive approach, the structured patient interviews were needed as data resource for over one-third of all DRPs.


The Mediating Role of Sense of Control in the Associations Between Remote Contacts and Loneliness Among Older Adults.
This study explored whether a sense of control over social life mediated the associations between using remote contact (phone calls, letters/emails, social media) and loneliness for socially isolated older adults. We used path analysis with the 2014 and 2016 Health and Retirement Study datasets (N = 3767). Results showed that more frequent phone calls and letters/emails were associated with lower levels of loneliness through sense of control. However, sense of control did not mediate the association between social media and loneliness. Findings suggest that promoting sense of control over social life by remote contact, particularly phone calls and letters/emails, may be effective in alleviating loneliness for isolated older adults.


Assessment of diclofenac LC50 reference values in juvenile and embryonic stages of the zebrafish (Danio rerio).
The aim of the study was to compare the acute toxicity of diclofenac to juvenile and embryonic stages of the zebrafish (Danio rerio). Acute toxicity tests were performed on the aquarium fish Danio rerio, which is one of the model organisms most commonly used in toxicity testing. The tests were performed using a semi-static method according to OECD guideline No. 203 (Fish, acute toxicity test). Embryo toxicity tests were performed in zebrafish embryos (Danio rerio) in compliance with OECD No. 212 methodology (Fish, short-term toxicity test on embryo and sac-fry stages). The results were subjected to a probit analysis using the EKO-TOX 5.2 programme to determine 96hLC50 and 144hLC50 (median lethal concentration, 50% mortality after a 96 h or 144 h interval, respectively) values of diclofenac. The statistical significance of the difference between LC50 values in juvenile and embryonic stages of Danio rerio was tested using the Mann-Whitney non-parametric test implemented in the Unistat 5.1 programme. The LC50 mean value of diclofenac was 166.6 +/- 9.8 mg/L in juvenile Danio rerio, and 6.11 +/- 2.48 mg/L in embryonic stages of Danio rerio. The study demonstrated a statistically higher sensitivity to diclofenac (P < 0.05) in embryonic stages compared to the juvenile fish.


Evaluation of radiation dose and image quality for dental cone-beam computed tomography in pediatric patients.
Children are sensitive to radiation; therefore, it is necessary to reduce radiation dose as much as possible in pediatric patients. In addition, it is crucial to investigate the optimal imaging conditions as they considerably affect the radiation dose. In this study, we investigated the effect of different imaging conditions on image quality and optimized the imaging conditions for dental cone-beam computed tomography (CBCT) examinations to diagnose ectopic eruptions and impacted teeth in children. To achieve our aims, we evaluated radiation doses and subjective and objective image quality. The CBCT scans were performed using 3D Accuitomo F17. All combinations of a tube voltage (90 kV), tube currents (1, 2, 3 mA), fields of view (FOVs) (4 × 4, 6 × 6 cm), and rotation angles (360°, 180°) were used. Dose-area product values were measured. SedentexCT IQ cylindrical phantom was used to physically evaluate the image quality. We used the modulation transfer function as an index of resolution, the noise power spectrum as an index of noise characteristics, and the system performance function as an overall evaluation index of the image. Five dentists visually evaluated the images from the head-neck phantom. The results showed that the image quality tended to worsen, and scores for visual evaluation decreased as tube currents, FOVs and rotation angles decreased. In particular, image noise negatively affected the delineation of the periodontal ligament space. The optimal imaging conditions were 90 kV, 2 mA, 4 × 4 cm FOV and 180° rotation. These results suggest that CBCT radiation doses can be significantly reduced by optimizing the imaging conditions.


Decreased prevalence of moderate to severe COPD over 15 years in northern Sweden.
The burden of COPD in terms of mortality, morbidity, costs and prevalence has increased worldwide. Recent results on prevalence in Western Europe are conflicting. In Sweden smoking prevalence has steadily decreased over the past 30 years. The aim was to study changes in prevalence and risk factor patterns of COPD in the same area and within the same age-span 15 years apart. Two population-based cross-sectional samples in ages 23-72 years participating at examinations in 1994 and 2009, respectively, were compared in terms of COPD prevalence, severity and risk factor patterns. Two different definitions of COPD were used; FEV1/FVC < LLN and FEV1/FVC < 0.7. The severity of COPD was assessed by FEV1, both as % of predicted and in relation to the LLN. The prevalence of COPD decreased significantly from 9.5% to 6.3% (p = 0.030) according to the FEV1/FVC < LLN criterion, while the decrease based on the FEV1/FVC < 0.7 criterion from 10.5% to 8.5% was non-significant. The prevalence of moderate to severe COPD decreased substantially and significantly, and the risk factor pattern was altered in 2009 when, beside age and smoking, also socio-economic status based on occupation was significantly associated with COPD. Changes in both prevalence and risk factor patterns of COPD were observed between surveys. Following a continuing decrease in smoking habits over several decades, a decrease in the prevalence of moderate to severe COPD was observed from 1994 to 2009 in northern Sweden.


In vivo characterization of the effects of abscisic acid and drying protocols associated with the acquisition of desiccation tolerance in alfalfa (Medicago sativa L.) somatic embryos.
Although somatic embryos of alfalfa (Medicago sativa L.) had acquired some tolerance to desiccation at the cotyledonary stage of development (22 d after plating), additional culturing in 20 microm abscisic acid (ABA) for 8 d induced greater desiccation tolerance, as determined by increased germination. Compared with fast drying, slow drying of the ABA-treated embryos improved desiccation tolerance. However, slow drying of non-ABA-treated embryos led to the complete loss of germination capacity, while some fast-dried embryos survived. An electron paramagnetic resonance spin probe technique and in vivo Fourier transform infrared microspectroscopy revealed that cellular membrane integrity and a-helical protein secondary structure were maintained during drying in embryos cultured in media enriched with 20 microM ABA, but not in embryos cultured in the absence of ABA. Slow-dried, non-ABA-treated embryos had low oligosaccharide to sucrose ratios, an increased proportion of beta-sheet protein secondary structures and broad membrane phase transitions extending over a temperature range of more than 60 degrees C, suggestive of irreversible phase separations. The spin probe study showed evidence of imbibitional damage, which could be alleviated by prehydration in humid air. These observations emphasize the importance of appropriate drying and prehydration protocols for the survival and storage of somatic embryos. It is suggested that ABA also plays a role in suppressing metabolism, thus increasing the level of desiccation tolerance; this is particularly evident under stressful conditions such as slow drying.


Diversity of genomic breakpoints in TFG-ALK translocations in anaplastic large cell lymphomas: identification of a new TFG-ALK(XL) chimeric gene with transforming activity.
Anaplastic large cell lymphomas are associated with chromosomal aberrations involving the anaplastic lymphoma kinase (ALK) gene at 2p23 that result in the expression of novel chimeric ALK proteins with transforming properties. In most of these tumors, the t(2;5)(p23;q35) generates the NPM-ALK fusion gene. However, several studies have now demonstrated that genes other than NPM may be fused to the ALK gene. We have recently described two different ALK rearrangements involving the TRK-fused gene (TFG) in which the same portion of ALK was fused to different length fragments of the 5' TFG region. These two rearrangements encoded chimeric proteins of 85 kd (TFG-ALK(S)) and 97 kd (TFG-ALK(L)), respectively. In this study, we have identified a new ALK rearrangement in which the catalytic domain of ALK was fused to a larger fragment of the TFG gene (TFG-ALK(XL)), encoding for a fusion protein of 113 kd. Genomic analysis of these three TFG-ALK rearrangements revealed that the TFG breakpoints occur at introns 3, 4, and 5, respectively, whereas the ALK breakpoints always occur in the same intron. No homologous regions or known recombination sequences were found in these regions. Transfection experiments using NIH-3T3 fibroblasts showed a similar transforming efficiency of TFG-ALK variants compared with NPM-ALK. In addition, in common with NPM-ALK, the TFG-ALK proteins formed stable complexes with the signaling proteins Grb2, Shc, and PLC-gamma. In conclusion, these findings indicate that the TFG may use a variety of intronic breakpoints in ALK rearrangements generating fusion proteins of different molecular weights, but with similar transforming potential than NPM-ALK.


How to Handle Concomitant Asymptomatic Prosthetic Joints During an Episode of Hematogenous Periprosthetic Joint Infection, a Multicenter Analysis.
Prosthetic joints are at risk of becoming infected during an episode of bacteremia, especially during Staphylocococcus aureus bacteremia. However, it is unclear how often asymptomatic periprosthetic joint infection (PJI) occurs, and whether additional diagnostics should be considered. In this multicenter study, we retrospectively analyzed a cohort of patients with a late acute (hematogenous) PJI between 2005-2015 who had concomitant prosthetic joints in situ. Patients without at least 1 year of follow-up were excluded. We included 91 patients with a hematogenous PJI and 108 concomitant prosthetic joints. The incident PJI was most frequently caused by Staphylococcus aureus (43%), followed by streptococci (26%) and Gram-negative rods (18%). Of 108 concomitant prosthetic joints, 13 were symptomatic, of which 10 were subsequently diagnosed as a second PJI. Of the 95 asymptomatic prosthetic joints, 1 PJI developed during the follow-up period and was classified as a "missed" PJI at the time of bacteremia with S. aureus (1.1%). Infected prosthetic joints were younger than the noninfected ones in 67% of cases, and prosthetic knees were affected more often than prosthetic hips (78%). During an episode of hematogenous PJI, concomitant asymptomatic prosthetic joints have a very low risk of being infected, and additional diagnostic work-up for these joints is not necessary.


SARS-CoV-2 seroprevalence and respiratory disease disability claims in Mexico City Metropolitan Area.
To characterize the impact of SARS-CoV-2 infection in workers from an essential large-scale company in the Greater Mexico City Metropolitan Area using point prevalence of acute infection, point prevalence of past infection through serum antibodies and respiratory disease short-term disability claims (RD-STDC). Four randomized surveys, three during 2020 before and one after (December 2021) vaccines' availability. point prevalence of acute infection through saliva PCR (polymerase chain reaction) testing, point prevalence of past infection through serum antibodies against Covid-19, RD-STDC and prevalence of symptoms during the previous six months. Prevalence of SARS-CoV-2 cases was 1.29-4.88%, on average, a quarter of participants pre-vaccination were seropositive; over half of participants with a RD-STDC had antibodies. The odds of having antibodies were 6-7 times more among workers with an RD-STDC. High antibody levels against Covid-19 in this study population reflects that coverage is high among workers in this industry. STDCs are a useful tool to track workplace epidemics.


[A new paradigm for the treatment of sternomediastinitis].
To analyze the results of simultaneous surgical treatment of sternomediastinitis compared to stage-by-stage approach. The study included 27 patients between October 2022 and March 2023. All patients underwent heart surgery through median sternotomy. There were 11 (40.7%) women and 16 (59.3%) men. Mean age of patients was 68.4±9.8 years. All patients were divided into 2 groups: 12 patients underwent partial necrectomy and vacuum wound therapy (or long-term dressings) before surgery (group 1), 15 patients underwent surgery without prior conservative treatment (group 2). The most common causative agent of infection was staphylococcus (48.1%). In all patients, we found histological signs of osteomyelitis. Preoperative clinical status was similar in both groups. There were differences in the following indicators: C-reactive protein upon admission - 74.9±18.6 versus 94.8±23.8 mg/l, procalcitonin - 0.13 [0.02; 1.43] versus 0.21 [0.02; 0.94] ng/ml. Postoperative outcomes were similar in both groups. Mortality was 8.3% (n=1) and 13.3% (n=2), respectively. Simultaneous combined surgical treatment (sequestrectomy + muscle flap grafting) demonstrates optimal results in the treatment of sternomediastinitis.


Preliminary study on determining stereotypical motor movements.
Stereotypical motor movements are one of the most common and least understood behaviors occurring in individuals with Autism Spectrum Disorder (ASD). To overcome problems with traditional methods for measuring stereotypical motor movements in persons with ASD, the Kinect sensor from Microsoft and gesture recognition algorithms were used to automatically detect hand flapping. This study provides a valuable tool to monitor stereotypes in order to understand and to cope with this problem. At the end it facilitates to identify behavioral patterns especially relevant when studying interaction skills in children with ASD.


Subchronic, Low-Level Intraperitoneal Injections of Manganese (IV) Oxide and Manganese (II) Chloride Affect Rat Brain Neurochemistry.
Manganese (Mn) is neurotoxic and can induce manganism, a Parkinson-like disease categorized as being a serious central nervous system irreversible neurodegenerative disease. An increased risk of developing symptoms of Parkinson disease has been linked to work-related exposure, for example, for workers in agriculture, horticulture, and people living near areas with frequent use of Mn-containing pesticides. In this study, the focus was placed on neurochemical effects of Mn. Rats were dosed intraperitoneally with 0.9% NaCl (control), 1.22 mg Mn (as MnO2)/kg bodyweight (bw)/day, or 2.5 mg Mn (as MnCl2)/kg bw/day for 7 d/wk for 8 or 12 weeks. This dosing regimen adds relevant new knowledge about Mn neurotoxicity as a consequence of low-dose subchronic Mn dosing. Manganese concentrations increased in the striatum, the rest of the brain, and in plasma, and regional brain neurotransmitter concentrations, including noradrenaline, dopamine (DA), 5-hydroxytrytamine, glutamate, taurine, and γ-amino butyric acid, and the activity of acetylcholinesterase changed. Importantly, a target parameter for Parkinson disease and manganism, the striatal DA concentration, was reduced after 12 weeks of dosing with MnCl2. Plasma prolactin concentration was not significantly affected due to a potentially reduced dopaminergic inhibition of the prolactin release from the anterior hypophysis. No effects on the striatal α-synuclein and synaptophysin protein levels were detected.


Idiopathic granulomatous angiitis of the central nervous system. Diagnostic challenges.
Granulomatous angiitis of the central nervous system (CNS) is a rare cause of vasculitis involving the brain and spinal cord and is included in lists of disorders causing strokes. To determine the frequency of strokes (eg, sudden onset of focal symptoms) as a presenting complication and to determine the sensitivity of angiography and other technologies in detecting vasculitis in histologically proved cases of idiopathic granulomatous angiitis of the CNS (IGANS), we reviewed the published literature. A biopsy-proved case of IGANS in a patient presenting without strokes and with a normal angiogram is reported. Additional cases of pathologically proved IGANS where results of angiography or other neuroimaging procedures were available were found by search of MEDLINE and bibliographies of pertinent articles and books. We compared our case with 39 reported cases of histologically proved granulomatous angiitis of the CNS not associated with Hodgkin's disease, herpes zoster, sarcoidosis, or other proximate cause. We included only those cases that had been evaluated with angiography or other neuroimaging techniques before death or biopsy. Analysis of these cases shows that strokes as presenting complications are rare in IGANS. Most patients present with a diffuse encephalopathy and, when focal symptoms develop, they tend to develop gradually. Including our case, 56% of 41 angiograms performed in 31 reported patients with histologically proved IGANS were abnormal, but only 27% were diagnostic for vasculitis. We conclude that stroke is uncommon as a presenting complaint in IGANS and angiography is insensitive as a screening test for these patients.


Spirituality of Science: Implications for Meaning, Well-Being, and Learning.
Scientists often refer to spiritual experiences with science. This research addresses this unique component of science attitudes-spirituality of science: feelings of meaning, awe, and connection derived through scientific ideas. Three studies (N = 1,197) examined individual differences in Spirituality of Science (SoS) and its benefits for well-being, meaning, and learning. Spirituality of Science was related to belief in science, but unlike other science attitudes, spirituality of science was also associated with trait awe and general spirituality (Study 1). spirituality of science also predicted meaning in life and emotional well-being in a group of atheists and agnostics, showing that scientific sources of spirituality can provide similar psychological benefits as religious spirituality (Study 2). Finally, Spirituality of Science predicted stronger engagement and recall of scientific information (Study 3). Results provide support for an experience of spirituality related to science, with benefits for meaning, well-being, and learning.


Does fat in milk, butter and cheese affect blood lipids and cholesterol differently?
To compare the effects of isoenergetic amounts of milk, cheese and butter (adjusted to the same content of lactose and casein) on fasting and postprandial blood lipids and lipoproteins, and on postprandial glucose and insulin response. The experiments were designed to provide 20% of total energy from dairy fat, as either whole milk, mean (+/-SD) 2164 (+/-97) g, butter 93 (+/-4) g, and hard cheese 305 (+/-45) g, which were served to 14 healthy young men for three periods of three weeks each, separated by washout periods, in a randomized, cross-over study with strictly controlled dietary intake. Fasting blood samples were taken at the end of the study periods. Measurements of the postprandial effect of the three different dairy test products (0.7 g of milk fat/kg body weight) were carried out on day 4 of each intervention period. Blood samples were taken before and at 2, 4, 6 and 8 hours following intake of the meals. Fasting LDL cholesterol concentration was significantly higher after butter than cheese diet (p = 0.037), with a borderline significant difference in total cholesterol (p = 0.054) after the experimental periods of three weeks. Postprandial glucose showed a higher response after cheese diet than after milk diet (p = 0.010, diet x time interaction). A different effect of fat in milk and butter could not be confirmed in this study. The moderately lower LDL cholesterol after cheese diet compared to butter diet should be investigated further.


Oxytocin Reduces Alcohol Cue-Reactivity in Alcohol-Dependent Rats and Humans.
Approved pharmacological treatments for alcohol use disorder are limited in their effectiveness, and new drugs that can easily be translated into the clinic are warranted. One of those candidates is oxytocin because of its interaction with several alcohol-induced effects. Alcohol-dependent rats as well as post-mortem brains of human alcoholics and controls were analyzed for the expression of the oxytocin system by qRT-PCR, in situ hybridization, receptor autoradiography ([125I]OVTA binding), and immunohistochemistry. Alcohol self-administration and cue-induced reinstatement behavior was measured after intracerebroventricular injection of 10 nM oxytocin in dependent rats. Here we show a pronounced upregulation of oxytocin receptors in brain tissues of alcohol-dependent rats and deceased alcoholics, primarily in frontal and striatal areas. This upregulation stems most likely from reduced oxytocin expression in hypothalamic nuclei. Pharmacological validation showed that oxytocin reduced cue-induced reinstatement response in dependent rats-an effect that was not observed in non-dependent rats. Finally, a clinical pilot study (German clinical trial number DRKS00009253) using functional magnetic resonance imaging in heavy social male drinkers showed that intranasal oxytocin (24 IU) decreased neural cue-reactivity in brain networks similar to those detected in dependent rats and humans with increased oxytocin receptor expression. These studies suggest that oxytocin might be used as an anticraving medication and thus may positively affect treatment outcomes in alcoholics.


Rugby-related adult maxillofacial trauma injuries: a NEISS database study.
The primary objective of this study is to delineate the data on maxillofacial trauma in rugby utilizing the National Electronic Injury Surveillance System (NEISS) database. Specifically, we want to establish the prevalence of facial rugby injuries in terms of age, mechanism of injury, and degree of injury in order to develop ways to limit facial trauma in the future. The NEISS database was accessed in February 2020 in order to identify adult patients (> 19 years of age) presenting to the emergence department (ED) for rugby-related head and facial injuries from the previous 10 years (2009-2018). Descriptive statistics were organized and presented. Chi-squared testing (χ2) was performed to compare categorical variables, and ANOVA was performed to compare continuous variables. A total of 507 patients (national estimate = 18,952) from 2009 to 2018 were identified as appropriate for study inclusion. The most common injuries were those to the facial region including the eyelid, eye area, and nose (59.4%). The most frequently encountered facial fracture while playing rugby was the nasal bone (58.6%). Overall, 98.4% of patients who presented to the ED with rugby injuries were treated and released, 1.2% were admitted or observed, and 0.4% left against medical advice. When evaluating a patient with a rugby-related injury, one should expect injuries to the eyelid, eye area, or nose. The most common fracture pattern will most likely be nasal bone. Despite these injuries, the vast majority of patients will be treated and released.


A Target-Detecting Visual Neuron in the Dragonfly Locks on to Selectively Attended Targets.
The visual world projects a complex and rapidly changing image onto the retina of many animal species. This presents computational challenges for those animals reliant on visual processing to provide an accurate representation of the world. One such challenge is parsing a visual scene for the most salient targets, such as the selection of prey amid a swarm. The ability to selectively prioritize processing of some stimuli over others is known as 'selective attention'. We recently identified a dragonfly visual neuron called 'Centrifugal Small Target Motion Detector 1' (CSTMD1) that exhibits selective attention when presented with multiple, equally salient targets. Here we conducted in vivo, electrophysiological recordings from CSTMD1 in wild-caught male dragonflies (Hemicordulia tau), while presenting visual stimuli on an LCD monitor. To identify the target selected in any given trial, we uniquely modulated the intensity of the moving targets (frequency tagging). We found that the frequency information of the selected target is preserved in the neuronal response, while the distracter is completely ignored. We also show that the competitive system that underlies selection in this neuron can be biased by the presentation of a preceding target on the same trajectory, even when it is of lower contrast than an abrupt, novel distracter. With this improved method for identifying and biasing target selection in CSTMD1, the dragonfly provides an ideal animal model system to probe the neuronal mechanisms underlying selective attention.SIGNIFICANCE STATEMENT We present the first application of frequency tagging to intracellular neuronal recordings, demonstrating that the frequency component of a stimulus is encoded in the spiking response of an individual neuron. Using this technique as an identifier, we demonstrate that CSTMD1 'locks on' to a selected target and encodes the absolute strength of this target, even in the presence of abruptly appearing, high-contrast distracters. The underlying mechanism also permits the selection mechanism to switch between targets mid-trial, even among equivalent targets. Together, these results demonstrate greater complexity in this selective attention system than would be expected in a winner-takes-all network. These results are in contrast to typical findings in the primate and avian brain, but display intriguing resemblance to observations in human psychophysics.


A vasoconstrictor cardiogenic chemoreflex induced by prostaglandin F2 alpha.
Systemic and skeletal muscle vascular responses to intracardiac prostaglandin F2 alpha (PGF2 alpha) were studied in anesthetized dogs. Resistance changes in a vascularly isolated but innervated reservoir-perfused gracilis muscle could occur only neurogenically. Injection of PGF2 alpha or serotonin (5-HT) raised systemic arterial blood pressure (BP) and caused reflex increases in gracilis muscle perfusion pressure (GMPP) that were abolished by alpha-adrenergic blockade. These changes in GMPP are opposite to the expected baroreceptor responses to an increase in BP. Subepicardial local anesthesia attenuated both delta BP and delta GMPP to 5-HT but only the delta GMPP to PGF2 alpha. Changes in BP and GMPP to 5-HT were reduced by vagotomy but not by stellate ganglionectomy. Conversely, stellate ganglionectomy significantly reduced the delta GMPP to PGF2 alpha but not the delta BP. Vagotomy affected neither delta BP nor delta GMPP following PGF2 alpha. Thus, while PGF2 alpha and 5-HT produce reflex vasoconstriction in gracilis muscle that is mediated through sympathetic efferent nerves and initiated by cardiogenic reflexes, apparently PGF2 alpha does so through sympathetic afferent and 5-HT through vagal afferent pathways. Further, since the systemic pressor response to PGF2 alpha persisted after stellatectomy, vagotomy, or local anesthesia, it is independent of the sympathetic cardiogenic chemoreflex.


Lack of humoral immune response to the tetracycline (Tet) activator in rats injected intracranially with Tet-off rAAV vectors.
The ability to safely control transgene expression from viral vectors is a long-term goal in the gene therapy field. We have previously reported tight regulation of GFP expression in rat brain using a self-regulating tet-off rAAV vector. The immune responses against tet regulatory elements observed by other groups in nonhuman primates after intramuscular injection of tet-on encoding vectors raise concerns about the clinical value of tet-regulated vectors. However, previous studies have not examined immune responses following injection of AAV vectors into brain. Therefore, rat striatum was injected with tet-off rAAV harboring a therapeutic gene for Parkinson's disease, either hAADC or hGDNF. The expression of each gene was tightly controlled by the tet-off regulatory system. Using an ELISA developed with purified GST-tTA protein, no detectable immunogenicity against tTA was observed in sera of rats that received an intrastriatal injection of either vector. In contrast, sera from rats intradermally injected with an adenovirus containing either tTA or rtTA, as positive controls, had readily detectable antibodies. These observations suggest that tet-off rAAV vectors do not elicit an immune response when injected into rat brain and that these may offer safer vectors for Parkinson's disease than vectors with constitutive expression.


Alterations in lectin binding accompany increased permeability in the dystrophic rat model for proliferative retinopathy.
In the dystrophic Royal College of Surgeons (RCS) rat, migration of vessels from the inner retina into the retinal pigment epithelium (RPE) is associated with neovascular proliferation and formation of vitreo-retinal membranes (VRMs), (Caldwell et al., 1988; Frank and Das, 1988). We studied permeability and luminal membrane glycoconjugates in these vessels using horseradish peroxidase (HRP) and lectin-ferritin (Fe) techniques. RCS and genetic control rats were injected with HRP, their retinas were fixed, incubated in Fe conjugates of wheat germ agglutinin (WGA-Fe) or concanavalin-A (ConA-Fe), reacted to demonstrate HRP, and prepared for electron microscopy. The RPE and VRM vessels in RCS retinas were compared with the normal inner retina and choriocapillaris vessels in RCS and genetic control rats. In both groups inner retinal vessels formed a barrier to HRP, while fenestrated choriocapillaris (CE) vessels were permeable to the tracer. In both of these vascular beds plasma membrane WGA-Fe binding was dense and uniform, while ConA-Fe binding was sparse and patchy. Studies with competitive sugars showed that WGA-Fe binding was primarily to N-acetylglucosamine (NAG) and that ConA-Fe was to mannose. In both RPE and VRM vessels tight junctions appeared intact, but both vessel types were permeable to HRP with the RPE vessels often containing fenestrae and channels. As compared with binding in the inner retina and CE vessels, WGA-Fe binding was lower in VRM vessels and normal in RPE vessels, while ConA-Fe binding was higher in both RPE and VRM vessels. Thus, increased permeability is accompanied by alterations in both NAG and mannose residues in the VRM vessels and with alterations in mannose residues and the presence of fenestrations and channels in the RPE vessels.


The time course of miniature endplate currents and its modification by receptor blockade and ethanol.
Miniature endplate currents (MEPCs) recorded from mouse diaphragms with a point voltage clamp, without inhibition of acetylcholinesterase (AChE) and in the absence of any drug, showed in their decay phase consistent deviations from an exponential time course, consisting of (a) "curvature," a progressive increase of decay rate during most of the decay phase, followed by (b) "late" tails. Both phenomena persisted when MEPCs (and channel lifetime) were prolonged by ethanol. Curvature was increased by muscle fiber depolarization and decreased by hyperpolarization. Receptor blockade by (+)-tubocurarine, alpha-bungarotoxin, hexamethonium, or myasthenic IgG accelerated the decay of the main part of MEPCs and eliminated curvature; the time constant of MEPCs became close to the channel time constant. We conclude that curvature arises from repeated action of ACh with cooperativity in ACh-receptor interaction; the voltage sensitivity of curvature follows from the voltage sensitivity of channel closing. Ethanol, in addition to its effect to prolong channel lifetime, enhances the tendency of ACh to act more than once to open channels before being lost to the system. Analysis of the rising phase of the MEPC, in terms of driving functions, also indicated that ethanol promotes channel opening by ACh; this action can account for a substantial increase of MEPC height by ethanol when MEPCs are made small by receptor blockade. Driving functions were also voltage sensitive, in a manner indicating acceleration of channel opening, but reduction of channel conductance, with hyperpolarization. Poisoning or inhibition of AChE prolonged MEPCs without altering the duration of ionic channels. Since ethanol caused further prolongation of MEPCs after poisoning of AChE, with little change in MEPC height, we conclude that the extension of mean channel lifetime by ethanol is accompanied by a similar extension of ACh binding to receptors. After poisoning of AChE, MEPCs became very variable in time course and the decay rate (tau-1) was correlated with MEPC height with a slope of log tau vs. log height of 0.77 for MEPCs of greater than 60% mean size. This slope is larger than expected from cooperativity in ACh-receptor interaction. Correlation of tau and height of MEPCs also exists when AChE is intact; the slope of log tau vs. log height was 0.12 with or without prolongation of MEPCs by ethanol.


Oral administration of the 5-HT2Creceptor agonist, mCPP, reduces body weight gain in rats over 28 days as a result of maintained hypophagia.
The 5-HT(2C) receptor subtype has been implicated extensively in the regulation of ingestive behaviour. To assess whether chronic administration of the preferential 5-HT(2C) receptor agonist, mCPP, reduces rat body weight gain and to determine if this effect is wholly or partially attributable to the effect of the drug on daily food intake. Animals were orally dosed with mCPP (10 mg/kg P.O., b.i.d.) or d-fenfluramine (2.5 mg/kg P.O., b.i.d.) for 28 days. Further groups of animals received drug treatments for the first 14 days and then received vehicle for the remainder of the experiment. Locomotor activity was assessed on days 2, 14, and 28. In a second study, animals received mCPP or d-fenfluramine for a 14-day period (dose and route were identical to the previous study). A group of pair-fed controls were included to determine whether the effects on body weight gain were attributable entirely to drug-induced hypophagia. Both mCPP and d-fenfluramine reduced body weight relative to vehicle-treated controls over the 28-day period. Withdrawal of the drugs on day 14 resulted in a significant rebound weight gain. Neither mCPP nor d-fenfluramine induced significant changes in locomotor activity compared to controls on any of the days tested (2, 14 or 28). In the second, 14-day study, changes in the body weights of pair-fed controls closely paralleled those of their drug-treated counterparts. These data indicate that chronic oral treatment with mCPP and d-fenfluramine significantly reduces rat body weight gain, an effect that is reversible upon withdrawal and wholly attributable to maintained hypophagia.


Anticoagulation practices for continuous renal replacement therapy: a survey of physicians from the United States.
During continuous renal replacement therapy (CRRT), anticoagulants are recommended for patients at low risk of bleeding and not already receiving systemic anticoagulants. Current anticoagulants used in CRRT in the US are systemic heparins or regional citrate. To better understand use of anticoagulants for CRRT in the US, we surveyed nephrologists and critical care medicine (CCM) specialists. The survey contained 30 questions. Respondents were board certified and worked in intensive care units of academic medical centers or community hospitals. 150 physicians (70 nephrologists and 80 CCM) completed the survey. Mean number of CRRT machines in use increased ∼30% from the pre-pandemic era to 2022. Unfractionated heparin was the most used anticoagulant (43% of estimated patients) followed by citrate (28%). Respondents reported 29% of patients received no anticoagulant. Risk of hypocalcemia (52%) and citrate safety (42%) were the predominant reasons given for using no anticoagulant instead of citrate in heparin-intolerant patients. 84% said filter clogging was a problem when no anticoagulant was used, and almost 25% said increased transfusions were necessary. Respondents using heparin (n = 131) considered it inexpensive and easily obtainable, although of moderate safety, citing concerns of heparin-induced thrombocytopenia and bleeding. Anticoagulant citrate dextrose solution was the most used citrate. Respondents estimated that 37% of patients receiving citrate develop hypocalcemia and 17% citrate lock. Given the increased use of CRRT and the lack of approved, safe, and effective anticoagulant choices for CRRT in the US, effective use of current and other anticoagulant options needs to be evaluated.


Fluorescent-antibody reagents for the identification of Clostridium botulinum.
Fluorescent-antibody reagents were prepared against vegetative cells of representative strains of each physiological group and toxin type of Clostridium botulinum known to have caused botulism in humans. A fluorescent-antibody reagent was also prepared for C. botulinum type G, which has been isolated from autopsy specimens but which has not clearly been implicated in botulism. These fluorescent-antibody reagents were evaluated against 200 strains of C. botulinum and 64 strains of other clostridia. Each reagent reacted with at least a 2+ intensity with all of the strains in its same toxin type and physiological group. Ninety-seven percent of the strains gave at least a 3+ reaction with the homologous group or toxin type reagent. Some cross-reactions occurred with reagents against different toxin type strains within a physiological group; there was less cross-reaction between physiological groups and very little reactivity of C. botulinum reagents with nontoxigenic organisms. Absorption of cross-reacting antibodies was not successful. Certain reagents could be used for presumptive laboratory identification of C. botulinum strains causing botulism, especially in infants. The type G reagent provided a good means of identifying C. botulinum type G, which lacks the lipase marker and whose toxigenicity may be more difficult to demonstrate in mixed cultures. There was a serological relationship between C. botulinum type G and some strains of Clostridium subterminale. This relationship provided evidence of differences between strains of C. botulinum type G isolated in two different countries.


Body proportions in Hyaenidae.
Indexes of skeletal measurements are used to compare body proportions in Hyaenidae, Canidae and Felidae. Hyaena and Crocuta are characterized by a long neck and long forelimbs. The humerus is relatively short and the radius and metacarpal bones are long. In the the os femoris is long and the tibia short. The proportions are adapted to carry large and heavy prey and provide maximum static stability. Proteles cristatus which mainly feeds on termites and insects has the long neck and forelimb in common with the other Hyaenidae, but its hindlimbs are longer and the length of the limbsegments reveals more canid proportions.


Regulation of lipoprotein metabolism by adrenergic mechanisms.
The available data on the adrenergic influences on lipoprotein metabolism are reviewed. It is clear that the influences are complex. It appears that adrenergic stimuli act at multiple sites in the lipoprotein metabolic pathways. This explains in part, the confusion in interpreting the literature. Based on the effects of alpha- and beta-adrenergic blocker administration in animals and humans, it is clear that the net effect of alpha 1-receptor stimulation is a decrease in HDL cholesterol and an increase in LDL cholesterol and VLDL triglyceride. On the other hand, beta-receptor stimulation will result in a net reduction of LDL and an increase in HDL. Further studies to elucidate the exact mechanism of action by the adrenergic stimuli on lipoprotein metabolism should provide insights into the relationship between the adrenergic nervous system and atherosclerosis and an enhanced understanding of the mechanisms by which antihypertensive-adrenergic mechanisms--alpha 1-Inhibition--Atherosclerosis--Serum lipids.


Getting to zero: Impact of a device to reduce blood culture contamination and false-positive central-line-associated bloodstream infections.
To assess the impact of initial specimen diversion device (ISDD) on inpatient and emergency department blood culture contamination (BCC), central-line-associated bloodstream infection (CLABSI) standardized infection ratios (SIRs), and antibiotic administration. Single-center quasi-experimental prospective cohort study wherein phlebotomists used traditional venipuncture with or without the ISDD while registered nurses (RNs) used traditional venipuncture. BCC events among phlebotomists and RNs were observed and compared from March 17, 2019, through January 21, 2020, defined by contaminant detection in 1 of 4 bottles for matched sets or 1 of 2 bottles in both subsets for coagulase negative staphylococci. CLABSIs throughout this period were recorded and SIRs were calculated. Enhanced oversight took place through July 21, 2019, with chart review assessing antibiotic use for patients with possible BCC. Overall, 24% of blood cultures obtained were from patients in intensive care. Phlebotomists using traditional venipuncture (n = 4,759) had a 2.3% BCC rate; phlebotomists using the ISDD (n = 11,202) had a 0% BCC rate. RNs drew 7,411 BCs with a 0.8% BCC rate. The CLABSI SIR was decreased from 1.103 in 2017 and 0.658 in 2018 to 0.439 in 2019. The CLABSI incidence was 33%-64% of predicted value for each 2019 quarter. This range fell to 18%-37% after the exclusion of likely false-positive results. Among 42 patients with possible BCC under enhanced oversight, 2 patients were treated with prolonged antibiotic courses. ISDD use by phlebotomists was associated with BCC reduction and reduced false-positive CLABSI results. This patient-care quality improvement could constitute sustainable antibiotic stewardship expansion.


Anatomic correlates of changing aqueous outflow facility in excised human eyes.
Five pairs of excised human eyes were examined for anatomic correlates of changes in aqueous outflow facility that result from changing the intraocular pressure (IOP) and from placing mechanical tension on the irido-corneal angle (lens depression). An increase in IOP from 0 to 40 mm Hg tended inconsistently to compress trabecular meshwork as a whole and to distend the juxtacanalicular tissue. The most constant and significant effect of increasing IOP, however, was to compress Schlemm's canal, thereby progressively diminishing its volume to its virtual collapse at 40 mm Hg IOP. Lens depression increased Schlemm's canal volume and partially prevented its collapse at high levels of IOP. The mean frequency of endothelial vacuoles was remarkably constant at IOPs from 2.5 to 40.0 mm Hg, with and without lens depression, but was significantly lower when the IOP was reduced to 0 mm Hg. When correlated with existing physiologic data, these anatomic findings suggest that Schlemm's canal collapse, but not vacuole frequency, is an important contributor to pressure-dependent changes in aqueous outflow facility.


Phosphorylated CRMP1, axon guidance protein, is a component of spheroids and is involved in axonal pathology in amyotrophic lateral sclerosis.
In amyotrophic lateral sclerosis (ALS), neurodegeneration is characterized by distal axonopathy that begins at the distal axons, including the neuromuscular junctions, and progresses proximally in a "dying back" manner prior to the degeneration of cell bodies. However, the molecular mechanism for distal axonopathy in ALS has not been fully addressed. Semaphorin 3A (Sema3A), a repulsive axon guidance molecule that phosphorylates collapsin response mediator proteins (CRMPs), is known to be highly expressed in Schwann cells near distal axons in a mouse model of ALS. To clarify the involvement of Sema3A-CRMP signaling in the axonal pathogenesis of ALS, we investigated the expression of phosphorylated CRMP1 (pCRMP1) in the spinal cords of 35 patients with sporadic ALS and seven disease controls. In ALS patients, we found that pCRMP1 accumulated in the proximal axons and co-localized with phosphorylated neurofilaments (pNFs), which are a major protein constituent of spheroids. Interestingly, the pCRMP1:pNF ratio of the fluorescence signal in spheroid immunostaining was inversely correlated with disease duration in 18 evaluable ALS patients, indicating that the accumulation of pCRMP1 may precede that of pNFs in spheroids or promote ALS progression. In addition, overexpression of a phospho-mimicking CRMP1 mutant inhibited axonal outgrowth in Neuro2A cells. Taken together, these results indicate that pCRMP1 may be involved in the pathogenesis of axonopathy in ALS, leading to spheroid formation through the proximal progression of axonopathy.


Methylation of nuclear DNA in Physarum polycephalum.
The restriction endonucleases HpaII and HhaI, whose action is inhibited by the presence of methylated base analogues at the recognition sequences in the DNA substrate, were used to investigate the distribution of 5-methylcytosine in nuclear DNA from Physarum polycephalum. Physarum DNA is digested into two fractions by these enzymes: a low-molecular-weight (M--) compartment comprising 80% of the DNA, and a high-molecular-weight (M+) compartment containing 20% of the DNA. The DNA fraction showing resistance to digestion by restriction endonuclease HpaII is cleaved by its isoschizomer MspI, indicating that methylated endonuclease-HpaII-specific sites are present in M + DNA. Additional properties of sequences in the M+ compartment were investigated.


From physician intent to the pharmacy label: prevalence and description of discrepancies from a cross-sectional evaluation of electronic prescriptions.
The objectives of this cross-sectional study were to estimate the prevalence of unintended discrepancies between three sources of prescription information and to describe the types of electronic prescribing system vulnerabilities identified. Staff from community pharmacies identified approximately 200 new prescriptions written at three participating ambulatory care clinics (2 adult, 1 paediatric). Unintended discrepancies were identified by comparing three sources of prescription information: (1) the prescriber's note as documented in the patient's chart; (2) the electronic prescription (e-prescription) entered into the clinic's electronic prescribing software; (3) the medication that was ultimately dispensed by the pharmacy as indicated on the prescription label. The discrepancy rate was calculated by dividing the number of discrepancies identified by the number of prescriptions evaluated. A total of 602 prescriptions written by 33 prescribers were evaluated from the 3 ambulatory care clinics. The discrepancy rate between the prescriber's note and the e-prescription was 1.7%, 0.6% and 3.9% for the three clinics. The discrepancy rate between the e-prescription (clinic) and the prescription label (pharmacy) was 4.2%, 0.9% and 1.5%. Differences between directions for administration was the most common type of discrepancy identified. Discrepancy rates between the prescriber's note and the e-prescription were similar to the discrepancy rates between the e-prescription and pharmacy label. To reduce outpatient medication errors, a better understanding is needed of the sources of discrepancies that occur within the prescriber's clinic, and those that occur between the clinic and pharmacy.


Grb7 upregulation is a molecular adaptation to HER2 signaling inhibition due to removal of Akt-mediated gene repression.
The efficacy of anti-HER2 therapeutics, such as lapatinib and trastuzumab, is limited by primary and acquired resistance. Cellular adaptations that allow breast cancer cell to survive prolonged HER2 inhibition include de-repression of the transcription factor FOXO3A with consequent estrogen receptor activation, and/or increased HER3 signaling. Here, we used low-density arrays, quantitative PCR, and western blotting to determine how HER2 signaling inhibition with lapatinib or PI3K inhibitors affects the expression of genes involved in breast cancer metastatic spread and overall prognosis. Retroviral transgenesis was used to express constitutively active forms of Akt in the HER2(+) breast cancer cell line SKBR3, and Grb7 in MCF7 cells. Specific gene silencing was obtained by siRNAs transfection. A murine BT474 xenograft cancer model was used to assess the effect of lapatinib on gene expression in vivo. We found that lapatinib induces upregulation of Grb7, an adaptor protein involved in receptor tyrosine kinase signaling and promoting cell survival and cell migration. Grb7 upregulation induced by lapatinib was found to occur in cancer cells in vitro and in vivo. We demonstrate that Grb7 upregulation is recreated by PI3K inhibitors while being prevented by constitutively active Akt. Thus, Grb7 is repressed by PI3K signaling and lapatinib-mediated Akt inhibition is responsible for Grb7 de-repression. Finally, we show that Grb7 removal by RNA-interference reduces breast cancer cell viability and increases the activity of lapatinib. In conclusion, Grb7 upregulation is a potentially adverse consequence of HER2 signaling inhibition. Preventing Grb7 accumulation and/or its interaction with receptor tyrosine kinases may increase the benefit of HER2-targeting drugs.


Ultraviolet stellar spectroscopy on gemini 11.
Objective-prism and objective-grating spectrograms were obtained in six star fields during the Gemini 11 mission. The grating spectra show absorption lines in the 2000 to 3000 angstrom wavelength region of Canopus and Sirius and provide ultraviolet-energy distribution data for approximately 50 stars. Many prism spectra show the Balmer discontinuity due to hydrogen and two absorption features probably due to ultraviolet iron multiplets.


β-Catenin mutations as determinants of hepatoblastoma phenotypes in mice.
Hepatoblastoma (HB) is the most common pediatric liver cancer. Although long-term survival of HB is generally favorable, it depends on clinical stage, tumor histology, and a variety of biochemical and molecular features. HB appears almost exclusively before the age of 3 years, is represented by seven histological subtypes, and is usually associated with highly heterogeneous somatic mutations in the catenin β1 (CTNNB1) gene, which encodes β-catenin, a Wnt ligand-responsive transcriptional co-factor. Numerous recurring β-catenin mutations, not previously documented in HB, have also been identified in various other pediatric and adult cancer types. Little is known about the underlying factors that determine the above HB features and behaviors or whether non-HB-associated β-catenin mutations are tumorigenic when expressed in hepatocytes. Here, we investigated the oncogenic properties of 14 different HB- and non-HB-associated β-catenin mutants encoded by Sleeping Beauty vectors following their delivery into the mouse liver by hydrodynamic tail-vein injection. We show that all β-catenin mutations, as well as WT β-catenin, are tumorigenic when co-expressed with a mutant form of yes-associated protein (YAP). However, tumor growth rates, histologies, nuclear-to-cytoplasmic partitioning, and metabolic and transcriptional landscapes were strongly influenced by the identities of the β-catenin mutations. These findings provide a context for understanding at the molecular level the notable biological diversity of HB.


Diagnosis and treatment of iron-deficiency anaemia in pregnancy and postpartum.
Iron deficiency occurs frequently in pregnancy and can be diagnosed by serum ferritin-level measurement (threshold value < 30 μg/L). Screening for iron-deficiency anemia is recommended in every pregnant women, and should be done by serum ferritin-level screening in the first trimester and regular hemoglobin checks at least once per trimester. In the case of iron deficiency with or without anaemia in pregnancy, oral iron therapy should be given as first-line treatment. In the case of severe iron-deficiency anemia, intolerance of oral iron, lack of response to oral iron, or in the case of a clinical need for rapid and efficient treatment of anaemia (e.g., advanced pregnancy), intravenous iron therapy should be administered. In the postpartum period, oral iron therapy should be administered for mild iron-deficiency anemia (haemorrhagic anemia), and intravenous iron therapy for moderately severe-to-severe anemia (Hb < 95 g/L). If there is an indication for intravenous iron therapy in pregnancy or postpartum, iron-containing drugs which have been studied in well-controlled clinical trials in pregnancy and postpartum such as ferric carboxymaltose must be preferred for safety reasons. While anaphylactic reactions are extremely are with non-dextrane products, close surveillance during administration is recommended for all intravenous iron products.


Genotypes, nt 1858 variants, and geographic origin of hepatitis B virus--large-scale analysis using a new genotyping method.
The nucleotide at position 1858 of hepatitis B virus has importance in chronic hepatitis B (HB) because a cytosine at nt 1858 effectively prevents virus escape through the precore TAG stop codon mutation. The relatedness between nt 1858 and genotypes was analyzed using a new genotyping method based on restriction fragment length polymorphism (RFLP) analysis of an S gene amplicon. Seventy-three gene bank sequences were analyzed by phylogenetic tree construction and RFLP prediction. A tree supporting the existence of 6 genotypes (A-F) was obtained, with C-1858 found in 9 of 9 A genotypes, 0 of 7 B, 0 of 19 C, 1 of 10 D, 0 of 2 E, and 3 of 3 F. Serum samples from 187 HB e antigen-positive chronic carriers were analyzed: Genotypes in northern Europeans were 60% A, 31% D; in southern Europeans and Middle Easterners 96% D; in Africans 53% A, 27% D, 20% E; and in East Asians 14% A, 43% B, 43% C. Cytosine at nt 1858 was found in 36 of 44 A, 0 of 32 B, 8 of 34 C, 0 of 59 D, 0 of 3 E, and 1 of 1 F genotype samples.


[Three-dimensional morphological modeling and visualization of wheat root system].
Crop three-dimensional (3D) morphological modeling and visualization is an important part of digital plant study. This paper aimed to develop a 3D morphological model of wheat root system based on the parameters of wheat root morphological features, and to realize the visualization of wheat root growth. According to the framework of visualization technology for wheat root growth, a 3D visualization model of wheat root axis, including root axis growth model, branch geometric model, and root axis curve model, was developed firstly. Then, by integrating root topology, the corresponding pixel was determined, and the whole wheat root system was three-dimensionally re-constructed by using the morphological feature parameters in the root morphological model. Finally, based on the platform of OpenGL, and by integrating the technologies of texture mapping, lighting rendering, and collision detection, the 3D visualization of wheat root growth was realized. The 3D output of wheat root system from the model was vivid, which could realize the 3D root system visualization of different wheat cultivars under different water regimes and nitrogen application rates. This study could lay a technical foundation for further development of an integral visualization system of wheat plant.


Prevalence of borrelia burgdorferi sensu lato infection among rodents and host-seeking ticks in South Carolina.
Tissues of rodents and host-seeking adult ticks collected in the Piedmont, Sandhills, Coastal Plain, and Coastal Zone of South Carolina were cultured in attempts to isolate Borrelia burgdorferi (Johnson, Schmid, Hyde, Steigerwalt & Brenner), the etiologic agent of Lyme disease. An exploratory, tree-based statistical analysis was used to identify ecological variables that were associated with spirochete infection among rodents and ticks. Spirochetes were isolated from tissues of 71 rodents: 22 (69%) of 32 eastern woodrats, 39 (53%) of 74 cotton mice, and 11(25%) of 44 hispid cotton rats. Rodent infection prevalences were significantly higher in the Coastal Zone than in other regions. Spirochetes were also cultured from 31 (2.6%) of 1,193 questing ticks. Prevalence of spirochetes in Ixodes affinis Neumann (19/74, 26%) was significantly higher than in I. scapularis Say (12/864, 1.3%) and other species (0/255) of ticks tested. In addition, two (9%) of 23 adult I. minor Neumann removed from woodrats contained spirochetes. Isolates from rodents and ticks were analyzed immunologically by indirect immunofluorescence and Western blots, and further characterized by polymerase chain reaction assays and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All were determined to be B. burgdorferi sensu lato. Results of this study confirmed that B. burgdorferi is endemic in South Carolina, and that enzootic transmission cycles exist at foci in the Coastal Zone. These findings add additional evidence that I. affinis and I. minor are potentially significant maintenance vectors of the spirochete.


LRP16 integrates into NF-κB transcriptional complex and is required for its functional activation.
Nuclear factor κB (NF-κB)-mediated pathways have been widely implicated in cell survival, development and tumor progression. Although the molecular events of determining NF-κB translocation from cytoplasm to nucleus have been extensively documented, the regulatory mechanisms of NF-κB activity inside the nucleus are still poorly understood. Being a special member of macro domain proteins, LRP16 was previously identified as a coactivator of both estrogen receptor and androgen receptor, and as an interactor of NF-κB coactivator UXT. Here, we investigated the regulatory role of LRP16 on NF-κB activation. GST pull-down and coimmunoprecipitation (CoIP) assays assessed protein-protein interactions. The functional activity of NF-κB was assessed by luciferase assays, changes in expression of its target genes, and its DNA binding ability. Annexin V staining and flow cytometry analysis were used to evaluate cell apoptosis. Immunohistochemical staining of LRP16 and enzyme-linked immunosorbent assay-based evaluation of active NF-κB were performed on primary human gastric carcinoma samples. We demonstrate that LRP16 integrates into NF-κB transcriptional complex through associating with its p65 component. RNA interference knockdown of the endogenous LRP16 in cells leads to impaired NF-κB activity and significantly attenuated NF-κB-dependent gene expression. Mechanistic analysis revealed that knockdown of LRP16 did not affect tumor necrosis factor α (TNF-α)-induced nuclear translocation of NF-κB, but blunted the formation or stabilization of functional NF-κB/p300/CREB-binding protein transcription complex in the nucleus. In addition, knockdown of LRP16 also sensitizes cells to apoptosis induced by TNF-α. Finally, a positive link between LRP16 expression intensity in nuclei of tumor cells and NF-κB activity was preliminarily established in human gastric carcinoma specimens. Our findings not only indicate that LRP16 is a crucial regulator for NF-κB activation inside the nucleus, but also suggest that LRP16 may be an important contributor to the aberrant activation of NF-κB in tumors.


Complementary peptide epitopes and anti-idiotypic antibodies in autoimmunity.
Application of complementary B and T cell epitopes in inducing anti-idiotypic and anti-clonotypic antibodies capable of regulating or suppressing the autoimmune responses in experimental autoimmune myasthenia gravis (EAMG), allergic neuritis (EAN) and allergic encephalomyelitis (EAE) has been the stimulus of many research efforts. Studies on the idiotypic/anti-idiotypic network of anti-La/SSB positive sera from patients with Sjogren's Syndrome (SS) and Systemic Lupus Erythematosus (SLE) and on animals immunized with the complementary epitopes are presented.


Fleck (Mouchetée) dystrophy of the cornea.
Five families, four Caucasian and one Negro (14 patients) with fleck (speckled or Mouchetée) dystrophy of the cornea are presented. In each, the typical presentation of fine scattered fleck-like dystrophic lesions was found throughout all layers of the central and peripheral corneal stroma but not affecting the epithelium, Bowman's membrane, Descemet's membrane or the endothelium. Clear stroma was noted between each lesion. Visual acuity in all patients was normal or only slightly affected. Except for an occasional patient with minor photophobia, almost all patients found affected with this dystrophy were asymptomatic in regard to their corneal condition. There was no decrease in corneal sensitivity noted in any patient. All families displayed evidence of an autosomal dominant hereditary pattern and progression of the dystrophy is slow and benign in character. With the exception of one family with atopic disease, no systemic organic illness was noted by histroy in all of the families. Laboratory screening of 2 of the 5 families showed no abnormalities of any systemic metabolic disorders. The characteristic clinical picture and favorable prognosis of this condition enables one to easily differentiate this condition from other known parenchymatous corneal dystrophies. The incidence of this condition is probably much more common than the reported cases in the literature might indicate.


Identification of an unintended consequence of Nrf2-directed cytoprotection against a key tobacco carcinogen plus a counteracting chemopreventive intervention.
NF-E2-related factor 2 (Nrf2) is a major cytoprotective gene and is a key chemopreventive target against cancer and other diseases. Here we show that Nrf2 faces a dilemma in defense against 4-aminobiphenyl (ABP), a major human bladder carcinogen from tobacco smoke and other environmental sources. Although Nrf2 protected mouse liver against ABP (which is metabolically activated in liver), the bladder level of N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG-C8-ABP), the predominant ABP-DNA adduct formed in bladder cells and tissues, was markedly higher in Nrf2(+/+) mice than in Nrf2(-/-) mice after ABP exposure. Notably, Nrf2 protected bladder cells against ABP in vitro. Mechanistic investigations showed that the dichotomous effects of Nrf2 could be explained at least partly by upregulation of UDP-glucuronosyltransferase (UGT). Nrf2 promoted conjugation of ABP with glucuronic acid in the liver, increasing urinary excretion of the conjugate. Although glucuronidation of ABP and its metabolites is a detoxification process, these conjugates, which are excreted in urine, are known to be unstable in acidic urine, leading to delivery of the parent compounds to bladder. Hence, although higher liver UGT activity may protect the liver against ABP, it increases bladder exposure to ABP. These findings raise concerns of potential bladder toxicity when Nrf2-activating chemopreventive agents are used in humans exposed to ABP, especially in smokers. We further show that 5,6-dihydrocyclopenta[c][1,2]-dithiole-3(4H)-thione (CPDT) significantly inhibits dG-C8-ABP formation in bladder cells and tissues but does not seem to significantly modulate ABP-catalyzing UGT in liver. Thus, CPDT exemplifies a counteracting solution to the dilemma posed by Nrf2.


Modeling evolutionary growth of a microRNA-mediated regulation system.
Gene duplication plays a crucial role in the development of complex biosystems, but the evolutionary forces behind the growth of biosystems are poorly understood. In this work, we introduce a model for such a growth through gene duplication. Plant microRNAs (miRNAs) are considered as a model. miRNAs are one of the non-coding small RNAs (19-25 nucleotides), which are involved in the post-transcriptional gene regulation. A single kind of miRNAs can be encoded by multiple genomic regions called miRNA genes, and can regulate multiple kinds of functional gene families. It is assumed that a single miRNA system involves all these genes, miRNA genes and their target gene families. We are interested in how duplication of miRNA genes affects the evolution of the miRNA system by focusing on the numbers of miRNA genes and their target gene families, denoted by x and y, respectively. We here theoretically explore the evolutionary growth of (x,y); the former increases by duplication of the miRNA gene while the latter increases when an independent gene family acquires a novel binding site of the miRNA by mutations. We first investigate the evolutionary patterns of (x,y) under three commonly assumed scenarios for the evolution of duplicated genes, that is, the positive and negative dosage and neofunctionalization scenarios. The results indicate that under the three scenarios, the transient process of (x,y) is unidirectional, although the direction is different depending on the model. This pattern is not consistent with the observation in the Arabidopsis thaliana genome, suggesting that a model that incorporates at least two directional evolutionary forces is needed to explain the observation. Then, such a model called the "complexity growth model" is introduced, in which we assume that duplication of miRNA genes is evolutionary advantageous in that the system can encode a complex and sophisticated pattern of regulation because multiple miRNA genes can have different expression patterns. This is helpful to optimize the regulation of a few particular functional gene families, but there is a cost; once the system is optimized for one purpose, it could be difficult for other purposes to use it. That is, duplication of miRNA genes would narrow down the potential gene families that can join the system. Our theoretical analysis revealed that this model can explain the observation of Arabidopsis miRNAs. Although we consider plant miRNAs as an example in this work, the model can be readily applied to other regulation systems with some modifications. Further development of such models would provide insights into the evolutionary growth of the complexity of biosystems.


Lactobacillus paralimentarius sp. nov., isolated from sourdough.
Six strains of lactic acid bacteria isolated from sourdough were characterized taxonomically. They were Gram-positive, catalase-negative, facultatively anaerobic rods that did not produce gas from glucose. Morphological and physiological data indicated that the strains belong to the genus Lactobacillus and they were similar to Lactobacillus alimentarius in phenotypic characteristics. These strains shared the same phenotypic characteristics and exhibited intragroup DNA homology values of over 89.8%, indicating that they comprised a single species. The G + C content of the DNA for the strains was 37.2-38.0 mol%. The 16S rRNA sequence of representative strain TB 1T was determined and aligned with that of other Lactobacillus species. This strain was placed in the genus Lactobacillus on the basis of phylogenetic analysis. L. alimentarius was the most closely related species in the phylogenetic tree and this species also showed the highest sequence homology value (96%) with strain TB 1T. DNA-DNA hybridization indicated that strain TB 1T did not belong to L. alimentarius. It is proposed that these strains are placed in the genus Lactobacillus as a new species, Lactobacillus paralimentarius sp. nov. The type strain of L. paralimentarius is TB 1T, which has been deposited in the Japan Collection of Microorganisms (JCM) as strain JCM 10415T.


Nonlinear performance of multi-granularity orthogonal transmission systems with frequency division multiplexing.
Orthogonal transmission with frequency division multiplexing technique is investigated for next generation optical communication systems. Coherent optical orthogonal frequency division multiplexing (OFDM) and single-carrier frequency division multiplexing (SCFDM) schemes are compared in combination with polarization-division multiplexing quadrature phase shift keying (QPSK) or 16-QAM (quadrature amplitude modulation) formats. Multi-granularity transmission with flexible bandwidth can be realized through ultra-dense wavelength division multiplexing (UDWDM) based on the orthogonal technique. The system performance is numerically studied with special emphasis on transmission degradations due to fiber Kerr nonlinearity. The maximum reach and fiber capacity for different spectral efficiencies are investigated for systems with nonlinear propagation over uncompensated standard single-mode fiber (SSMF) links with lumped amplification.


Genetic dissection of honeybee (Apis mellifera L.) foraging behavior.
We demonstrate the effects of a new quantitative trait locus (QTL), designated pln3, that was mapped in a backcross population derived from strains of bees selected for the amount of pollen they store in combs. We independently confirmed pln3 by demonstrating its effects on individual foraging behavior, as we did previously for QTLs pln1 and pln2 (Hunt et al. 1995). QTL pln2 is very robust in its effects on foraging behavior. In this study, pln2 was again shown to affect individual foraging behavior of workers derived from a hybrid backcross of the selected strains. In addition, pln2 was shown to affect the amount of pollen stored in combs of colonies derived from a wide cross of European and Africanized honeybees. This is noteworthy because it demonstrates that we can map QTLs for behavior in interstrain crosses derived from selective breeding and study their effects in unselected, natural populations. The results we present also demonstrate the repeatability of finding QTLs with measurable effects, even after outcrossing selected strains, suggesting that there is a relatively small subset of QTLs with major effects segregating in the population from which we selected our founding breeding populations. The different QTLs, pln1, pln2, and pln3, appear to have different effects, revealing the complex genetic architecture of honeybee foraging behavior.


Why Hypothesis Testers Should Spend Less Time Testing Hypotheses.
For almost half a century, Paul Meehl educated psychologists about how the mindless use of null-hypothesis significance tests made research on theories in the social sciences basically uninterpretable. In response to the replication crisis, reforms in psychology have focused on formalizing procedures for testing hypotheses. These reforms were necessary and influential. However, as an unexpected consequence, psychological scientists have begun to realize that they may not be ready to test hypotheses. Forcing researchers to prematurely test hypotheses before they have established a sound "derivation chain" between test and theory is counterproductive. Instead, various nonconfirmatory research activities should be used to obtain the inputs necessary to make hypothesis tests informative. Before testing hypotheses, researchers should spend more time forming concepts, developing valid measures, establishing the causal relationships between concepts and the functional form of those relationships, and identifying boundary conditions and auxiliary assumptions. Providing these inputs should be recognized and incentivized as a crucial goal in itself. In this article, we discuss how shifting the focus to nonconfirmatory research can tie together many loose ends of psychology's reform movement and help us to develop strong, testable theories, as Paul Meehl urged.


The immunopathological spectrum of crescentic glomerulonephritis: a survey of 106 patients in a single Chinese center.
Crescentic glomerulonephritis (CrGN) is a severe form of glomerular injury. We retrospectively analyzed data from Chinese patients with CrGN in our center to characterize the immunopathological spectrum of CrGN. A total of 106 consecutive patients with biopsy-proven CrGN were recruited. CrGN was classified into 3 types according to findings on immunofluorescence microscopy; type I was defined as a linear deposition of immunoglobulins along the glomerular basement membrane (GBM), type II as a glomerular deposition of immune-complex and type III as a pauci-immune deposition. A total of 17/106 (16.0%) patients were classified as type I, 43/106 (40.6%) as type II and 46/106 (43.4%) as type III. Serum antineutrophil cytoplasmic antibodies (ANCA) could be detected in 1/17 (5.9%) patient with type I, 11/43 (25.6%) patients with type II and 29/46 (63.0%) patients with type III CrGN (p < 0.001). Serum anti-GBM antibodies could be detected in 14/17 (82.4%) patients with type I, 2/43 (4.7%) patients with type II and 3/46 (6.5%) patients with type III. In type II CrGN, ANCA-positive patients were older (p < 0.001), had more multi-system involvement (p < 0.001) and more fibrinoid necrosis in renal histopathology than ANCA-negative patients (p = 0.003). Pauci-immune CrGN might be the most common type of CrGN in Northern China. There was much heterogeneity within each type of CrGN.


Effect of gender and preoperative diagnosis on results of revision total knee arthroplasty.
Recent studies question an effect of gender on outcome of primary TKA. We questioned whether the results of revision TKA were affected by gender. We separated 67 revision TKAs by gender and preoperative diagnosis into four groups (arthrofibrosis, infection, instability, and wear and loosening). Each revision TKA was individually matched by age and gender to two primary TKAs. Postoperative Knee Society pain and function scores after revision TKA were lower than for primary TKA for both females and males. However, postoperative Knee Society pain and function scores were similar in males and females. Postoperative pain and function scores were lower for all revision groups compared with primary TKA, except for pain and function scores after revision for instability. Postoperative pain and function scores were higher for instability and wear or loosening than for arthrofibrosis. Our data suggest the results of revision TKA are affected by preoperative diagnosis but not gender. Level III, retrospective matched cohort study. See Guidelines for Authors for a complete description of levels of evidence.


A 36 kg Giant Ovarian Fibroma with Meigs Syndrome: A Case Report and Literature Review of Extremely Giant Ovarian Tumor.
Ovarian tumors can get extremely giant to occupy the whole abdominal cavity. We report a case of 36 kg solid ovarian tumor, which was the largest ovarian solid tumor that have been ever reported. A 54-year-old woman presented to our hospital with a chief complaint of markedly distended abdominal wall. Preoperative imaging examinations revealed that most of the tumor was uniform and its density was like that of subcutaneous fat. Pleural effusion was detected in the right thoracic region. We organized a multidisciplinary team and successfully resected the right adnexa. The patient had an uneventful postoperative course, and she was discharged on the 7th postoperative day and diagnosed with a fibroma of the ovary with Meigs syndrome. A comprehensive literature search revealed 48 cases of extremely giant ovarian tumor in these 20 years. Six out of 48 cases are solid. Twelve out of 48 cases are malignant or borderline malignant, and patients' age and tumor size/weight were not related to the frequency of malignancy/borderline malignancy. As many as 4 out of 48 patients died before their first hospital visit or early after surgery. Clinicians should consider a considerable high mortality and frequency of severe surgical complications when planning the treatment strategy for extremely giant ovarian tumors.


Differential Effects of Physiological Arousal Following Acute Stress on Police Officer Performance in a Simulated Critical Incident.
Background: Police officer response in a critical incident is often a life-or-death scenario for the officer, the suspect, and the public. Efficient and accurate decisions are necessary to ensure the safety of all involved. Under these conditions, it is important to understand the effects of physiological arousal in response to acute stress on police officer performance in critical and dangerous incidents. Prior research suggests that physiological arousal following a stressor differentially affects police performance - communication may be impaired, whereas well-rehearsed, tactical behaviors may be resilient. Objectives: In this study, we examine the differential effects of physiological arousal across three police skill domains: verbal communication, nonverbal communication, and tactical skill. Methods: A sample of Swedish police cadets (N = 17) participated in a critical incident simulation, which was a reenactment of a real-life incident that had resulted in a police officer death; the simulation included multiple calls, dynamic environments, and surprise threats. An expert rater evaluated the cadets across multiple domains of skill, and physiological arousal was monitored by continuous heart rate monitoring and measures of circulating cortisol and antithrombin taken before and after the incident simulation. Results: The simulation increased police officer arousal, as reflected in elevated heart rate, but this alone did not predict differences in performance. Greater increase in antithrombin was associated with better general performance, but a specific deficit in verbal communication as compared to tactical performance and nonverbal communication. Change in cortisol was unrelated to the skill assessments. Conclusions: Police officer performance during a critical incident simulation is affected by physiological arousal. The findings are discussed with implications for police officer decision-making and real-world performance.


Mechanisms for generating cell membrane antigens that are recognized by cytotoxic T lymphocytes.
Studies with many viruses have revealed that viral specific protein synthesis is an obligatory step in generating antigens on target cells for antiviral cytotoxic T lymphocytes. This has been most clearly demonstrated with DI particles, virions that are structurally complete but lack infectious RNA. Adsorption of such particles onto target cell membranes does not render these cells susceptible to lytic attack by antiviral effector cells, unless some viral protein synthesis transpires. However, some viruses, such as Sendai virus, circumvent the requirement for viral protein synthesis via fusion of the viral envelope with the target cell membrane, a process mediated by a specialized fusion protein. Once inserted into the lipid bilayer, it is likely that viral components and self H-2 noncovalently associate so that the complex can be recognized by antiviral cytotoxic T cells. This idea is supported by the demonstration that viral proteins and H-2 containing membrane proteins, incorporated into reconstituted membrane vesicles or liposomes are recognized by cytotoxic T cells. These data further show that native rather than altered viral and H-2 molecules are the moieties recognized. Associations between antigen and H-2 have been detected by a variety of techniques and in some cases are not random but selective; that is, viral antigens perferentially associate with some H-2 alleles and not others. In summary, these findings indicate that although viral antigens are present in the mature virions, these components are not recognized by antiviral killer cells until integrated into the plasma membrane. This may be achieved either through direct fusion of the viral envelope with the target cell or following viral protein synthesis and insertion of viral antigens into the plasma membrane.


Monocyte chemoattractant protein-1 blockade inhibits lung cancer tumor growth by altering macrophage phenotype and activating CD8+ cells.
The role of chemokines in the pathogenesis of lung cancer has been increasingly appreciated. Monocyte chemoattractant protein-1 (MCP-1, also known as CCL2) is secreted from tumor cells and associated tumor stromal cells. The blockade of CCL2, as mediated by neutralizing antibodies, was shown to reduce tumorigenesis in several solid tumors, but the role of CCL2 in lung cancer remains controversial, with evidence of both protumorigenic and antitumorigenic effects. We evaluated the effects and mechanisms of CCL2 blockade in several animal models of non-small-cell lung cancer (NSCLC). Anti-murine-CCL2 monoclonal antibodies were administered in syngeneic flank and orthotopic models of NSCLC. CCL2 blockade significantly slowed the growth of primary tumors in all models studied, and inhibited lung metastases in a model of spontaneous lung metastases of NSCLC. In contrast to expectations, no significant effect of treatment was evident in the number of tumor-associated macrophages recruited into the tumor after CCL2 blockade. However, a change occurred in the polarization of tumor-associated macrophages to a more antitumor phenotype after CCL2 blockade. This was associated with the activation of cytotoxic CD8(+) T lymphocytes (CTLs). The antitumor effects of CCL2 blockade were completely lost in CB-17 severe combined immunodeficient mice or after CD8 T-cell depletion. Our data from NSCLC models show that CCL2 blockade can inhibit the tumor growth of primary and metastatic disease. The mechanisms of CCL2 blockade include an alteration of the tumor macrophage phenotype and the activation of CTLs. Our work supports further evaluation of CCL2 blockade in thoracic malignancies.


Butorphanol dependence increases hippocampal kappa-opioid receptor gene expression.
Butorphanol is a synthetic opioid agonist/antagonist analgesic agent, which exerts its effects mainly via kappa-opioid receptors. Characterizations of the gene expression levels of the mRNA for and protein levels of the kappa-opioid receptor in different brain regions of rats are essential for investigating possible mechanisms in the development of physical dependence on and withdrawal from butorphanol. Animals were rendered dependent by intracerebroventricular (i.c.v.) infusion of butorphanol (26 nmol/microl/hr) via osmotic minipumps for 3 days. Rats were sacrificed immediately (dependent group) or 7 hr after discontinuation of i.c.v. butorphanol infusion (withdrawal group). Expression levels of the mRNA for the kappa-opioid receptor, as detected by reverse transcription-polymerase chain reaction followed by Southern blot analysis, were significantly increased in the cerebral cortex, striatum, and midbrain, including thalamus, hippocampus, and pons, in animals dependent on butorphanol. In both dependent and withdrawal groups, Western blot analysis of kappa-opioid receptor protein levels showed significant increases in the amygdaloid nucleus, paraventricular thalamus, and thalamus. However, in the withdrawal group, there were significant decreases in the hippocampus and cortical regions, including the frontal, parietal, and temporal cortex. Regional changes in the mRNA for and protein levels of the kappa-opioid receptor focus attention on highly special roles for this receptor in the development of physical dependence on and the expression of withdrawal from butorphanol dependence.


Trapping of Brønsted acids with a phosphorus-centered biradicaloid - synthesis of hydrogen pseudohalide addition products.
The trapping of classical hydrogen pseudohalides (HX, X = pseudohalogen = CN, N3, NCO, NCS, and PCO) utilizing a phosphorus-centered cyclic biradicaloid, [P(μ-NTer)]2, is reported. These formal Brønsted acids were generated in situ as gases and passed over the trapping reagent, the biradicaloid [P(μ-NTer)]2, leading to the formation of the addition product [HP(μ-NTer)2PX] (successful for X = CN, N3, and NCO). In addition to this direct addition reaction, a two-step procedure was also applied because we failed in isolating HPCO and HNCS addition products. This two-step process comprises the generation and isolation of the highly reactive [HP(μ-NTer)2PX]+ cation as a [B(C6F5)4]- salt, followed by salt metathesis with salts such as [cat]X (cat = PPh4, n-Bu3NMe), which also gives the desired [HP(μ-NTer)2PX] product, with the exception of the reaction with the PCO- salt. In this case, proton migration was observed, finally affording the formation of a [3.1.1]-hetero-propellane-type cage compound, an OC(H)P isomer of a HPCO adduct. All discussed species were fully characterized.


Availability of Rubisco small subunit up-regulates the transcript levels of large subunit for stoichiometric assembly of its holoenzyme in rice.
Rubisco is composed of eight small subunits coded for by the nuclear RBCS multigene family and eight large subunits coded for by the rbcL gene in the plastome. For synthesis of the Rubisco holoenzyme, both genes need to be expressed coordinately. To investigate this molecular mechanism, the protein synthesis of two subunits of Rubisco was characterized in transgenic rice (Oryza sativa) plants with overexpression or antisense suppression of the RBCS gene. Total RBCS and rbcL messenger RNA (mRNA) levels and RBCS and RbcL synthesis simultaneously increased in RBCS-sense plants, although the increase in total RBCS mRNA level was greater. In RBCS-antisense plants, the levels of these mRNAs and the synthesis of the corresponding proteins declined to a similar extent. The amount of RBCS synthesized was tightly correlated with rbcL mRNA level among genotypes but not associated with changes in mRNA levels of other major chloroplast-encoded photosynthetic genes. The level of rbcL mRNA, in turn, was tightly correlated with the amount of RbcL synthesized, the molar ratio of RBCS synthesis to RbcL synthesis being identical irrespective of genotype. Polysome loading of rbcL mRNA was not changed. These results demonstrate that the availability of RBCS protein up-regulates the gene expression of rbcL primarily at the transcript level in a quantitative manner for stoichiometric assembly of Rubisco holoenzyme.


Estimation of the parameters of a gaussian mixture using the method of moments.
Given a general n-dimensional bimodal Gaussian mixture, this paper shows how unknown parameters may be found by the method of moments. Three cases are considered-equal modal probabilities, known but not necessarily equal probabilities, and all parameters unknown. The solution involves sample moments no higher than fourth order. For Gaussian mixtures where the number of modes is unknown, fourth-order moments can be used to count them, provided all modes have the same covariance matrix, and their multiplicity is not greater than data dimensionality. Examples of mode-counting and the determination of bimodal parameters are included.


[Construction and specificity of porcine bmp15 gene reporter vector].
The aim of this study is to identify the express specificity of bone morphogenetic protein 15 (Bmp15) in porcine. The pBMP15-EGFP reporter vector was constructed from the 2.2 kb fragment of porcine bmp15 promoter to trace the differentiation process of stem cells into oocyte-like cells. We used porcine ovary and Chinese Hamster Ovary cell line (CHO), mouse myoblast cell line (C2C12) and porcine amniotic fluid stem cell (pAFSC) to investigate the expression and regulation of this gene via RT-PCR, immunofluorescence, cell transfection, and microinjection methods. We also used single layer cell differentiation to detect the application potential of bmp15. The results show that bmp15 gene was specifically expressed in the porcine ovary and CHO rather than in C2C12 and pAFSC. In addition, the characteristic of tissue-specific of Bmp15 was detected on CHO instead of other cell lines by transient transfection. We also detected the expression of Bmp15 in oocyte at different development stages by immunofluorescence of fixed paraffin-embedded ovary sections. Furthermore, microinjection results show that bmp15 expressed in oocytes at 18 h of maturation in vitro, and continued up to 4-cell stage embryos. Most importantly, we found that the expression of Bmp15 started at day 12 after inducing pAFSC into oocyte-like cells by transfection; green fluorescent was visible in round cell masses. It indicated that bmp15 has the expression specificity and the pBMP15-EGFP reporter vector can be used to trace Bmp15 action in the differentiation of stem cells into germ cells.


Antigenic and genetic properties of viruses linked to hemorrhagic fever with renal syndrome.
Hemorrhagic fever with renal syndrome (HFRS) comprises a variety of clinically similar diseases of viral etiology that are endemic to and sporadically epidemic throughout the Eurasian continent and Japan. Although HFRS has not been reported in North America, viruses that are antigenically similar to HFRS agents were recently isolated from rodents in the United States. Examination and comparison of eight representative isolates from endemic disease areas and from regions with no known associated HFRS indicate that these viruses represent a new and unique group that constitutes a separate genus in the Bunyaviridae family of animal viruses.