Lactase decline in weaning rats is regulated at the transcriptional level and not caused by termination of milk ingestion.
Lactase activity declines during postnatal development in rats, but little is known about the underlying molecular mechanism of this phenomenon. We attempted to clarify whether the regulation was at the transcriptional or post-transcriptional level and to examine the effects of dietary factors on that regulation. Newborn rats were divided into two groups, prolonged nursing and weaning, at d 21. The prolonged nursing rats were nursed for a further 6 d, whereas weaning rats were separated from their dams and fed nonpurified diet for the same period. The patterns of declining lactase protein and mRNA concentrations during weaning were determined by Western blot and Northern blot analyses, respectively, and compared with lactase activity. There were significant (P < 0.001) correlations between them: r = 0.97 for specific activity vs. protein, r = 0.99 for specific activity vs. mRNA and r = 0.96 for protein vs. mRNA. The lactase activity per milligram DNA showed a pattern similar to that of the specific activity. This result argues against the decline in lactase activity being due to the dilution caused by newly synthesized materials during the weaning period and suggests transcriptional regulation. Furthermore, the prolonged nursing rats showed the same results as weanlings for lactase protein, mRNA, specific activity and activity per milligram DNA. These observations indicate that the regulation of lactase expression is at the transcriptional level and that it is not affected by the termination of milk ingestion.


Location-specific responses to nociceptive input support the purposeful nature of motor adaptation to pain.
Movement is changed in pain, but the mechanisms remain unclear. Key questions are unresolved such as whether activation can be inhomogeneously distributed within a muscle in a manner that is specific to the location of noxious input. This study addressed this question using high-density electromyography (EMG) to study regional redistribution of muscle activation within the vasti muscles and changes in knee extension force direction in response to noxious stimulation applied to muscular and nonmuscular tissues around the knee. Fourteen participants performed a low-force knee extension contraction at baseline, during, and after pain induced in 4 locations (infrapatellar fat pad, vastus lateralis, distal vastus medialis, or proximal vastus medialis). The knee extension force direction was estimated from a 3-dimensional load cell positioned just above the ankle. Regional muscle activation was estimated from amplitude of high-density surface EMG signals from vastus medialis and lateralis. Pain-induced spatial variations of activation were identified as the position of the 5 channels that showed the largest decrease (or smallest increase) in amplitude from baseline to pain or after pain. Knee extension force was produced more medially during pain after infrapatellar pad injection only (P = 0.01). Preferential reduction of activation of the distal region of vastus medialis was observed when distal vastus medialis (P < 0.001) or vastus lateralis (P < 0.05) was injected. Both adaptations persisted after pain resolution. These results support the hypothesis that specific adaptation depends on the location of a noxious stimulus and imply that recovery of pain is not necessarily concomitant with return of the EMG to prepain patterns.


Control of IgG glycosylation by in situ and real-time estimation of specific growth rate of CHO cells cultured in bioreactor.
The cell-specific growth rate (µ) is a critical process parameter for antibody production processes performed by animal cell cultures, as it describes the cell growth and reflects the cell physiological state. When there are changes in these parameters, which are indicated by variations of µ, the synthesis and the quality of antibodies are often affected. Therefore, it is essential to monitor and control the variations of µto assure the antibody production and achieve high product quality. In this study, a novel approach for on-line estimation of µ was developed based on the process analytical technology initiative by using an in situ dielectric spectroscopy. Critical moments, such as significant µ decreases, were successfully detected by this method, in association with changes in cell physiology as well as with an accumulation of nonglycosylated antibodies. Thus, this method was used to perform medium renewals at the appropriate time points, maintaining the values of µ close to its maximum. Using this method, we demonstrated that the physiological state of cells remained stable, the quantity and the glycosylation quality of antibodies were assured at the same time, leading to better process performances compared with the reference feed-harvest cell cultures carried out by using off-line nutrient measurements.


[A new tool to measure orthorexia nervosa - applying the Eating Habits Questionnaire (Revised) in Hungary].
Orthorexia nervosa is a new type of disordered eating, characterized by an unhealthy fixation on healthy food. Those who suffer from it are showing compulsive behaviors and increasing dietary restrictions, that can lead to anxiety and physical health problems. Measuring tools for estimating the seve rity of its symptoms are available in Hungary only to a limited extent. The aim of this study was to adapt the Eating Habits Questionnaire's revised version (EHQ-R) to Hungarian, and to perform a psychometric analysis, moreover, to explore the demographic features of orthorexic tendencies. The cross-sectional online questionnaire included adults who perform physical exercize regularly (n = 175, 29% male; mean age 35.4 [SD = 7.59] years). sociodemographic data, anthropometric data (body - weight, height), EHQ-R, ORTO-11-Hu questionnaires. Confirmatory factor analysis supported the EHQ-R's five-factor theoretical structure (c2 (395) = 856.2; p < 0.001; CFI = 0.914; TLI = 0.906; RMSEA = 0.082 [CI 90: 0.074-0.089]). Both the internal reliability of the scales (Cronbach-a = 0.75-0.87) and its consistency over time (r / r = 0.67-0.86) were adequate. The construct validity of the questionnaire is supported by the close correlation of the scales with the ORTO-11-Hu questionnaire (r / r = | 0.63-0.68 |). Regarding gender differences, women scored higher than men on the EHQ-R's Rigidity subscale (t [173] = -1.807; p = 0.073). Age showed a significant, weak, negative relationship with the Negative Emotionality (r = -0.18; p = 0.017) and Time Impairment (r = -0.21; p = 0.005) subscales. The EHQ-R has proven to be a reliable and valid measurement tool, thus, we recommend utilizing it as it can provide a quick and valid assessment about the severity of orthorexia symptoms, enabling appropriate interventions.


Managing hydroclimatic risks in federal rivers: a diagnostic assessment.
Hydroclimatic risks and adaptive capacity are not distributed evenly in large river basins of federal countries, where authority is divided across national and territorial governments. Transboundary river basins are a major test of federal systems of governance because key management roles exist at all levels. This paper examines the evolution and design of interstate water allocation institutions in semi-arid federal rivers prone to drought extremes, climatic variability and intensified competition for scarce water. We conceptualize, categorize and compare federal rivers as social-ecological systems to analyse the relationship between governance arrangements and hydroclimatic risks. A diagnostic approach is used to map over 300 federal rivers and classify the hydroclimatic risks of three semi-arid federal rivers with a long history of interstate allocation tensions: the Colorado River (USA/Mexico), Ebro River (Spain) and Murray-Darling River (Australia). Case studies review the evolution and design of water allocation institutions. Three institutional design trends have emerged: adoption of proportional interstate allocation rules; emergence of multi-layered river basin governance arrangements for planning, conflict resolution and joint monitoring; and new flexibility to adjust historic allocation patterns. Proportional allocation rules apportion water between states based on a share of available water, not a fixed volume or priority. Interstate allocation reform efforts in the Colorado and Murray-Darling rivers indicate that proportional allocation rules are prevalent for upstream states, while downstream states seek reliable deliveries of fixed volumes to increase water security. River basin governance arrangements establish new venues for multilayered planning, monitoring and conflict resolution to balance self governance by users and states with basin-wide coordination. Flexibility to adjust historic allocation agreements, without risk of defection or costly court action, also provides adaptive capacity to manage climatic variability and shifting values. Future research should develop evidence about pathways to adaptive capacity in different classes of federal rivers, while acknowledging limits to transferability and the need for context-sensitive design.


Influence of menopausal status on sleep and hot flashes throughout breast cancer adjuvant chemotherapy.
To examine the patterns and differences among sleep and hot flashes based on menopausal status (pre, peri, or post) in breast cancer adjuvant chemotherapy patients before, during, and after treatment. Secondary analysis from a randomized clinical trial. Outpatient clinics and participants' homes. Two hundred nineteen (219) women; mean age of 52 years (29-79); postoperative, Stages I to IIIA breast cancer. Participants were randomized to a behavioral sleep intervention or a healthy eating control group. Variables measured were menopausal status (self-report), subjective sleep (Pittsburgh Sleep Quality Index), objective sleep (actigraphy), perceived interruption of sleep by hot flashes (diary), and demographic/medical characteristics. Data reported were collected 48 hours before initial chemotherapy, at treatment 4, and 30 days after the last treatment. Analysis included descriptives and repeated measures mixed model analysis of variance. Objective sleep and the perceived interruption of sleep by hot flashes changed significantly over time. Sleep and hot flashes differed over time by menopausal status. Only hot flashes were significantly different by study group and by menopausal status over time. Women's menopausal status when starting chemotherapy was associated with differing patterns of sleep and hot flashes over time.


Personality traits and cognitive function in old-adults with type-2 diabetes.
Objectives: Personality may constitute an important domain of influence on cognitive function in old-adults. We assessed the relationship of personality traits and cognitive performance in individuals with Type-2 Diabetes (T2D), and explored possible mediators. Method: The sample includes 377 dementia-free subjects with T2D participating in the Israel Diabetes and Cognitive Decline study who underwent assessment of cognition and personality (mean age 72 ± 4y; 42% females). We assessed the relationships of personality traits with episodic memory, semantic categorization, attention/working memory, executive function and overall cognition using linear regression models adjusting for age, education, sex, BMI, T2D duration, Hemoglobin A1C (HbA1C), hypertension, c-reactive protein, total- to HDL-cholesterol ratio and ApoEɛ4 genotype. A post-hoc mediation analysis was conducted with HbA1C, proportion of days covered (PDC) by T2D prescription claims and depressive symptoms. Results: After adjustment for multiple covariates, high neuroticism levels were associated with poorer performance overall (β= -0.16 ± 0.05; p = 0.001) and with poorer episodic memory, attention/working memory, and semantic categorization (β= -0.14 ± 0.05; p = 0.007, β= -0.12 ± 0.05; p = 0.017 and β= -0.12 ± 0.05; p = 0.018, respectively). High scores on openness to experience were associated with better global cognition (β = 0.11 ± 0.05; p = 0.026), executive functions (β = 0.13 ± 0.05; p = 0.013) and semantic categorization (β = 0.17 ± 0.05; p = 0.001, respectively). Depressive symptoms mediated the association of neuroticism with executive function, and the association of openness with executive function and overall cognition. Conclusion: Personality may play an important role in cognitive health among elderly subjects with T2D. Future studies should address the mechanisms underlying these relationships and specifically the potential role of depressive symptoms which may be in the causal pathway between personality traits and cognitive outcomes.


Synthesis of phenolics and flavonoids in ginger (Zingiber officinale Roscoe) and their effects on photosynthesis rate.
The relationship between phenolics and flavonoids synthesis/accumulation and photosynthesis rate was investigated for two Malaysian ginger (Zingiber officinale) varieties grown under four levels of glasshouse light intensity, namely 310, 460, 630 and 790 μmol m(-2)s(-1). High performance liquid chromatography (HPLC) was employed to identify and quantify the polyphenolic components. The results of HPLC analysis indicated that synthesis and partitioning of quercetin, rutin, catechin, epicatechin and naringenin were high in plants grown under 310 μmol m(-2)s(-1). The average value of flavonoids synthesis in leaves for both varieties increased (Halia Bentong 26.1%; Halia Bara 19.5%) when light intensity decreased. Photosynthetic rate and plant biomass increased in both varieties with increasing light intensity. More specifically, a high photosynthesis rate (12.25 μmol CO(2) m(-2)s(-1) in Halia Bara) and plant biomass (79.47 g in Halia Bentong) were observed at 790 μmol m(-2)s(-1). Furthermore, plants with the lowest rate of photosynthesis had highest flavonoids content. Previous studies have shown that quercetin inhibits and salicylic acid induces the electron transport rate in photosynthesis photosystems. In the current study, quercetin was an abundant flavonoid in both ginger varieties. Moreover, higher concentration of quercetin (1.12 mg/g dry weight) was found in Halia Bara leaves grown under 310 μmol m(-2)s(-1) with a low photosynthesis rate. Furthermore, a high content of salicylic acid (0.673 mg/g dry weight) was detected in Halia Bara leaves exposed under 790 μmol m(-2)s(-1) with a high photosynthesis rate. No salicylic acid was detected in gingers grown under 310 μmol m(-2)s(-1). Ginger is a semi-shade loving plant that does not require high light intensity for photosynthesis. Different photosynthesis rates at different light intensities may be related to the absence or presence of some flavonoid and phenolic compounds.


Do the immunosuppressive drugs used as treatment for graft-versus-host disease directly inhibit lymphoid tumor cell growth?
Hematopoietic stem cell transplantation (HSCT) is a curative treatment for certain leukemias and lymphomas that prove resistant to standard chemotherapy. The immunosuppressive drugs prednisone and cyclosporin A (CsA) are routinely used during HSCT to prevent or treat graft-versus-host disease (GVHD) or to inhibit antibody-mediated inflammation. However, little is known about the direct impact of prednisone and CsA on the growth of malignant lymphoid cells in the setting of HSCT. To address this issue, we measured tumor cell growth in vitro and in vivo after treatment with prednisone and CsA, used separately, together, and in combination with irradiation. Our results showed that: (i) combinations of CsA and prednisone inhibited the growth of a variety of lymphoid tumors in vitro, particularly in combination with irradiation, and (ii) tumor size was significantly reduced in a mouse B-lymphoid tumor model following CsA or CsA plus prednisone treatments, with or without HSCT.


Site of the abnormal stimulus of gastric secretion after small bowel exclusion.
In dogs with either vagally innervated or denervated pouches, the acid responses to a standard meal, a meal which bypassed the greater part of the small bowel, and a meal introduced directly into the small intestine were measured before and after exclusion of 75% of the small bowel.After intestinal exclusion an increase in acid secretion was found in the latter part of the acid response to the standard meal and the intestinal meal. No increase in acid secretion resulted from the meals which bypassed the small bowel. These findings suggest that an increase in stimulation from the remaining small intestine is responsible for the elevation in acid secretion following small bowel exclusion.


Imaging of microglia in patients with neurodegenerative disorders.
Microglia constitute the main immune defense in the central nervous system. In response to neuronal injury, microglia become activated, acquire phagocytic properties, and release a wide range of pro-inflammatory mediators that are essential for the annihilation of the neuronal insult. Although the role of microglial activation in acute neuronal damage is well defined, the pathophysiological processes underlying destructive or protective role to neurons following chronic exposure to microglial activation is still a subject of debate. It is likely that chronic exposure induces detrimental effects by promoting neuronal death through the release of neurotoxic factors. Positron emission tomography (PET) imaging with the use of translocator protein (TSPO) radioligands provides an in vivo tool for tracking the progression and severity of neuroinflammation in neurodegenerative disease. TSPO expression is correlated to the extent of microglial activation and the measurement of TSPO uptake in vivo with PET is a useful indicator of active disease. Although understanding of the interaction between radioligands and TSPO is not completely clear, there is a wide interest in application of TSPO imaging in neurodegenerative disease. In this article, we aim to review the applications of in vivo microglia imaging in neurodegenerative disorders such as Parkinson's disease, Huntington's disease, Dementias, and Multiple Sclerosis.


Clonal analysis of NK cell development from bone marrow progenitors in vitro: orderly acquisition of receptor gene expression.
In the mouse, two families of MHC class I-specific receptors, namely Ly49 and CD94/NKG2, have been identified on NK cells. Individual NK cells can express several Ly49 molecules as well as members of the CD94/NKG2 family. The expression of multiple receptors with different specificities for MHC class I is thus thought to generate NK cells with diverse recognition patterns. To delineate the mechanism by which NK cells begin to express different patterns of Ly49 and CD94/NKG2 molecules, we developed a clonal assay in which NK1.1(-), IL-2/ IL-15 receptor beta+ NK precursors generated by culture of multipotential Lin(-), c-kit+ progenitors in IL-7, stem cell factor and flt3 ligand are induced to differentiate into NK1.1+ , Ly49+ NK cells. Examination of the clonal populations thus generated revealed heterogeneity in the pattern of Ly49 and CD94/NKG2 gene expression. In addition, a distinct kinetic pattern of expression was observed. CD94, NKG2A, NKG2C and Ly49B were expressed first followed by Ly49G, then Ly49C and I and finally, Ly49A, D, E and F. The data suggest a stochastic but ordered acquisition of class I receptors on NK cells in which developing NK cells become capable of expressing distinct receptors at different times but show no absolute prerequisite to express the receptors that are acquired early in NK development for the expression of those that are acquired later.


The microbiotest battery as an important component in the assessment of snowmelt toxicity in urban watercourses--preliminary studies.
The aim of the study was to use a battery of biotests composed of producers (Selenastrum capricornutum, Sorghum saccharatum, Lepidium sativum, and Sinapis alba), consumers (Thamnocephalus platyurus), and decomposers (Tetrahymena thermophila) to evaluate the toxicity of snowmelt and winter storm water samples. The toxicity of the samples collected in the winter period December to February (2010-2011), in one of the largest agglomerations in Poland, the city of Lodz, was compared to that of storm water samples taken under similar conditions in June. The most toxic snowmelt samples were found to be high acute hazard (class IV), while the remaining samples were rated as slight acute hazard (class II). L. sativum (in the Phytotox test) was the most sensitive test organism, giving 27 % of all toxic responses, followed by S. capricornutum with 23 % of all responses. T. thermophila was the least sensitive, with only 2 % of all toxic responses. The greatest range of toxicity was demonstrated by samples from the single family house catchment: no acute hazard (class I) to high acute hazard (class IV).


Matrix metalloproteinase 9/neutrophil gelatinase associated lipocalin/tissue inhibitor of metalloproteinasess type 1 complexes are localized within cardiomyocytes and serve as a reservoir of active metalloproteinase in porcine female myocardium.
Matrix metalloproteinase 9 (MMP-9) is crucial for physiological tissue repair and pathophysiological myocardial remodeling. The regulation of its functioning has been shown to be mediated by formation of complexes with tissue inhibitor of metalloproteinases 1 (TIMP-1) and neutrophil gelatinase associated lipocalin (NGAL). We investigated the mRNA and protein expression of MMP-9, TIMP-1 and NGAL, the formation of complexes, their gelatinolytic activity and cellular localization in left ventricle (LV) from 10 female pigs with induced systolic heart failure (HF), 5 control pigs, and a woman with severe HF. The MMP-9, TIMP-1 and NGAL mRNA in LV did not differ between diseased and healthy pigs. In all pigs MMP-9, TIMP-1 and NGAL proteins were present in LV as high molecular weight (HMW) complexes (115, 130, 170 and 220 kDa), and no monomers were found. A 80 and 115 kDa gelatinolytically active bands were present in all LV homogenates. A 130-kDa active band was seen only in LV from pigs with severe HF. Similar results were found in the explanted heart of a female patient with severe HF. The incubation of the homogenates of porcine LV at 37°C resulted in appearance of 88 kDa active band, which was accompanied by a decreased intensity of HMW bands. The incubation of the homogenates of porcine LV (depleted of active MMP-9) with trypsin generated 80 and 115 kDa active bands. Immunohistochemistry revealed the presence of MMP-9 in the cytoplasm of porcine cardiomyocytes, but not in cardiofibroblasts. Our data suggest that MMP-9 originates from cardiomyocytes, forms the gelatinolytically inactive complexes with TIMP-1 and NGAL, present in normal and failing myocardium, likely serving as a reservoir of active MMP-9. Further studies are needed to elucidate the role of these HMW complexes in the extracellular matrix remodeling during the progression of HF, which presence should be considered when developing efficient strategies inhibiting myocardial matrix metalloproteinases.


Downregulation of ALDH1A1 expression in non-small cell lung carcinomas--its clinicopathologic and biological significance.
ALDH1A1 metabolizes a variety of endogenous and exogenous aldehyde, and also oxidizes retinol to synthesize retinoic acid and modulate cell differentiation. Moreover, ALDH1A1 is also suggested to participate in the maintenance of cancer stem cells. To investigate the potential role of ALDH1A1 in carcinogenesis of the lung, the present study examined two hundred and sixty eight cases of non-small cell lung carcinoma (NSCLC) for its immunohistochemical expression and analyzed associations between ALDH1A1 levels and a series of clinicopathologic parameters. Also, the biological significance of the aberrant expression of ALDH1A1 was investigated in vitro. ALDH1A1 expression was markedly reduced in 39.9% (107/268) of NSCLCs. The incidence of this reduction was significantly higher in adenocarcinomas (ADC: 41.6%, 85/207) and large cell carcinomas (61.1%, 11/18) than squamous cell carcinomas (25.5%, 11/43). Among ADCs, the downregulation tended to be more remarkable in high grade, poorly differentiated tumors, and tumors with stronger proliferating activity. It also occurred with a significantly higher incidence in smokers than non-smokers. Forced expression of ALDH1A1 in NSCLC cell lines, which had lost ALDH1A1 expression, markedly attenuated their growth. Taken together, loss of ALDH1A1 expression is suggested to promote carcinogenesis especially in the smoking-related ADCs.


Endogenous leptin contributes to baroreflex suppression within the solitary tract nucleus of aged rats.
The decline in cardiovagal baroreflex function that occurs with aging is accompanied by an increase in circulating leptin levels. Our previous studies showed that exogenous leptin impairs the baroreflex sensitivity for control of heart rate in younger rats, but the contribution of this hormone to baroreflex dysfunction during aging is unknown. Thus we assessed the effect of bilateral leptin microinjection (500 fmol/60 nl) within the solitary tract nucleus (NTS) on the baroreflex sensitivity in older (66 ± 2 wk of age) urethane/chloralose anesthetized Sprague-Dawley rats with elevated circulating leptin levels. In contrast to the 63% reduction observed in younger rats, leptin did not alter the baroreflex sensitivity for bradycardia evoked by phenylephrine in older rats (0.76 ± 0.19 baseline vs. 0.71 ± 0.15 ms/mmHg after leptin; P = 0.806). We hypothesized that this loss of sensitivity reflected endogenous suppression of the baroreflex by elevated leptin, rather than cardiovascular resistance to the peptide. Indeed, NTS administration of a leptin receptor antagonist (75 pmol/120 nl) improved the baroreflex sensitivity for bradycardia in older rats (0.73 ± 0.13 baseline vs. 1.19 ± 0.26 at 10 min vs. 1.87 ± 0.32 at 60 min vs. 1.22 ± 0.54 ms/mmHg at 120 min; P = 0.002), with no effect in younger rats. There was no effect of the leptin antagonist on the baroreflex sensitivity for tachycardia, responses to cardiac vagal chemosensitive fiber activation, or resting hemodynamics in older rats. These findings suggest that the actions of endogenous leptin within the NTS, either produced locally or derived from the circulation, contribute to baroreflex suppression during aging.


Search for CP violation in the decays D(s)+ --> KS0pi+ and D(s)+ --> KS0K+.
We have searched for CP violation in the charmed meson decays D((s))(+) --> K(S)(0)pi(+) and D((s))(+) --> K(S)(0)K(+) using 673 fb(-1) of data collected with the Belle detector at the KEKB asymmetric-energy e(+)e(-) collider. No evidence for CP violation is observed. We report the most sensitive CP asymmetry measurements to date for these decays: A(CP)(D(+)-->K(S)(0)pi(+)) = (-0.71 +/- 0.19 +/- 0.20)%, A(CP)(D(s)(+) --> K(S)(0)pi(+)) = (+5.45 +/- 2.50 +/- 0.33)%, A(CP)(D(+) --> K(S)(0)K(+)) = (-0.16 +/- 0.58 +/- 0.25)%, and A(CP)(D(s)(+) --> K(S)(0)K(+)) = (+0.12 +/- 0.36 +/- 0.22)%, where the first uncertainties are statistical and the second are systematic.


Activation of protein kinase C-α/heme oxygenase-1 signaling pathway improves mitochondrial dynamics in lipopolysaccharide-activated NR8383 cells.
Mitochondrial function and morphology are dynamically regulated by fusion and fission. Heme oxygenase-1 (HO-1), which may be upregulated by protein kinase C-α (PKC-α), improves mitochondrial dynamics by controlling the balance between fusion and fission in vivo and in vitro. However, whether the PKC-α/HO-1 signaling pathway is one of the underlying mechanisms in adjusting mitochondrial dynamics in lipopolysaccharide (LPS)-activated macrophages has remained elusive. To explore this, NR8383 cells were pre-treated with PKC-α inhibitor Go6976 or PKC-α activator phorbol-12-myristate-13-acetate for 30 min and then stimulated with LPS for 24 h. Next, the expression of PKC-α, HO-1, mitofusin 1 (Mfn1) and mitofusin 2 (Mfn2), optic atrophy 1 (OPA1), dynamin-related protein 1 (Drp1) and fission 1 (Fis1) was detected to evaluate the possible implication of the PKC-α/HO-1 signaling pathway in the LPS-induced NR8383 cells. The results indicated that activation of the PKC-α/HO-1 signaling pathway increased superoxide dismutase activities and the respiratory control ratio (RCR), decreased the levels of malondialdehyde, reactive oxygen species (ROS), Drp1 and Fis1, and simultaneously enhanced the levels of Mfn1, Mfn2 and OPA1. In contrast, the PKC-α inhibitor decreased the expression of RCR, Mfn1, Mfn2 and OPA1, and increased the expression of MDA and ROS in NR8383 cells. The results suggest that activation of the PKC-α/HO-1 signaling pathway is necessary for the balance of mitochondrial dynamics and oxidative stress in macrophages, which provides clues for probing novel strategies against the detrimental effects of sepsis and other disease states.


Reliability of Triaxial Accelerometry for Measuring Load in Men's Collegiate Ice Hockey.
Van Iterson, EH, Fitzgerald, JS, Dietz, CC, Snyder, EM, and Peterson, BJ. Reliability of triaxial accelerometry for measuring load in men's collegiate ice hockey. J Strength Cond Res 31(5): 1305-1312, 2017-Wearable microsensor technology incorporating triaxial accelerometry is used to quantify an index of mechanical stress associated with sport-specific movements termed PlayerLoad. The test-retest reliability of PlayerLoad in the environmental setting of ice hockey is unknown. The primary aim of this study was to quantify the test-retest reliability of PlayerLoad in ice hockey players during performance of tasks simulating game conditions. Division I collegiate male ice hockey players (N = 8) wore Catapult Optimeye S5 monitors during repeat performance of 9 ice hockey tasks simulating game conditions. Ordered ice hockey tasks during repeated bouts included acceleration (forward or backward), 60% top-speed, top-speed (forward or backward), repeated shift circuit, ice coasting, slap shot, and bench sitting. Coefficient of variation (CV), intraclass correlation coefficient (ICC), and minimum difference (MD) were used to assess PlayerLoad reliability. Test-retest CVs and ICCs of PlayerLoad were as follows: 8.6% and 0.54 for forward acceleration, 13.8% and 0.78 for backward acceleration, 2.2% and 0.96 for 60% top-speed, 7.5% and 0.79 for forward top-speed, 2.8% and 0.96 for backward top-speed, 26.6% and 0.95 for repeated shift test, 3.9% and 0.68 for slap shot, 3.7% and 0.98 for coasting, and 4.1% and 0.98 for bench sitting, respectively. Raw differences between bouts were not significant for ice hockey tasks (p > 0.05). For each task, between-bout raw differences were lower vs. MD: 0.06 vs. 0.35 (forward acceleration), 0.07 vs. 0.36 (backward acceleration), 0.00 vs. 0.06 (60% top-speed), 0.03 vs. 0.20 (forward top-speed), 0.02 vs. 0.09 (backward top-speed), 0.18 vs. 0.64 (repeated shift test), 0.02 vs. 0.10 (slap shot), 0.00 vs. 0.10 (coasting), and 0.01 vs. 0.11 (bench sitting), respectively. These data suggest that PlayerLoad demonstrates moderate-to-large test-retest reliability in the environmental setting of male Division I collegiate ice hockey. Without previously testing reliability, these data are important as PlayerLoad is routinely quantified in male collegiate ice hockey to assess on ice physical activity.


Mid-trimester abortion induced by intravaginal administration of prostaglandin E2 suppositories.
Mid-trimester abortion was successfully induced in 70 of 71 patients by administration of vaginal PGE2 suppositories. The one patient who failed to abort with this method was pregnant in the blind horn of a duplex uterus. The mean abortion time for the successful inductions was 11.88 hours. Multiparous patients aborted somewhat faster than nulliparous patients, but the difference was not significant. Among the 70 successful inductions 42 patients aborted in 12 hours or less and only one patient had an abortion time of more than 24 hours. The drug appeared effective throughout the stages of gestation included in this series--from 8 to 27 weeks. Eight patients were monitored throughout the abortion procedure and uterine activity was calculated and analyzed. The development of uterine activity was gradual without the sudden rise in frequency of contractions and intrauterine baseline tonus that characterized prostaglandin administered by other methods. The most frequently encountered side effect of vaginal PGE2 suppositories was a temperature elevation, which returned to normal within a few hours of the last dose of the drug. Gastrointestinal disturbances--vomiting and diarrhea--were also common, despite a low initial dose of PGE2 and premedication with antiemetic and antidiarrheal agents. These side effects were in general well tolerated by the patients and never required termination of therapy. The cardiovascular effects of PGE2 in this series could be considered minimal. In a single patient surgical intervention was required to remove the placenta. In seven patients the placenta was removed by sponge forceps and in five patients the placenta was removed manually. There was an estimated blood loss exceeding 250 ml. in 10 patients, but transfusion was not required. Although white blood cell count rose significantly during the abortion period there were no significant changes in hematocrit or platelet count. Mid-trimester abortion with intravaginal administration of PGE2 suppositories appears to offer a valid alternative to the presently available techniques, with a rapid abortion time, high success rate, and low incidence of complications.


Serologic diagnosis of Lyme borreliosis by using enzyme-linked immunosorbent assays with recombinant antigens.
Class-specific enzyme-linked immunosorbent assays (ELISAs) with purified recombinant antigens of Borrelia burgdorferi sensu stricto and Western blot analyses with whole cells of this spirochete were used to test human sera to determine which antigens were diagnostically important. In analyses for immunoglobulin M (IgM) antibodies, 14 (82%) of 17 serum samples from persons who had erythema migrans reacted positively by an ELISA with one or more recombinant antigens. There was frequent antibody reactivity to protein 41-G (p41-G), outer surface protein C (OspC), and OspF antigens. In an ELISA for IgG antibodies, 13 (87%) of 15 serum samples had antibodies to recombinant antigens; reactivity to p22, p39, p41-G, OspC, and OspF antigens was frequent. By both ELISAs, serum specimens positive for OspB, OspE, and p37 were uncommon. Analyses of sera obtained from persons who were suspected of having human granulocytic ehrlichiosis (HGE) but who lacked antibodies to ehrlichiae revealed IgM antibodies to all recombinant antigens of B. burgdorferi except OspB and IgG antibodies to all antigens except OspE. Immunoblotting of sera from the study group of individuals suspected of having HGE reaffirmed antibody reactivity to multiple antigens of B. burgdorferi. There was minor cross-reactivity when sera from healthy subjects or persons who had syphilis, oral infections, or rheumatoid arthritis were tested by ELISAs with p37, p41-G, OspB, OspC, OspE, and OspF antigens. Although the results of class-specific ELISAs with recombinant antigens were comparable to those recorded for assays with whole-cell antigen and for individuals with confirmed clinical diagnoses of Lyme borreliosis, immunoblotting is still advised as an adjunct procedure, particularly when there are low antibody titers by an ELISA.


Replication of hepatitis C virus (HCV) RNA in mouse embryonic fibroblasts: protein kinase R (PKR)-dependent and PKR-independent mechanisms for controlling HCV RNA replication and mediating interferon activities.
Hepatitis C virus (HCV) infection causes chronic hepatitis and is currently treated with alpha interferon (IFN-alpha)-based therapies. The underlying mechanisms of chronic HCV infection and IFN-based therapies, however, have not been defined. Protein kinase R (PKR) was implicated in the control of HCV replication and mediation of IFN-induced antiviral response. In this report, we demonstrate that a subgenomic RNA replicon of genotype 2a HCV replicated efficiently in mouse embryonic fibroblasts (MEFs), as determined by cell colony formation efficiency and the detection of HCV proteins and both positive- and negative-strand RNAs. Additionally, the subgenomic HCV RNA was found to replicate more efficiently in the PKR knockout (PKR(-/-)) MEF than in the wild-type (PKR(+/+)) MEF. The knockdown expression of PKR by specific small interfering RNAs significantly enhanced the level of HCV RNA replication, suggesting that PKR is involved in the control of HCV RNA replication. The level of ISG56 (p56) was induced by HCV RNA replication, indicating the activation of PKR-independent antiviral pathways. Furthermore, IFN-alpha/beta inhibited HCV RNA replication in PKR(-/-) MEFs as efficiently as in PKR(+/+) MEFs. These findings demonstrate that PKR-independent antiviral pathways play important roles in controlling HCV replication and mediating IFN-induced antiviral effect. Our findings also provide a foundation for the development of transgenic mouse models of HCV replication and set a stage to further define the roles of cellular genes in the establishment of chronic HCV infection and the mediation of intracellular innate antiviral response by using MEFs derived from diverse gene knockout animals.


Examining mechanisms of action in 12-Step community outpatient treatment.
Twelve-step theory hypothesizes that changes in specific cognitions and behaviors, including adoption of disease model beliefs and involvement in self-help programs, lead to symptom reduction. We examined the role of self-help affiliation in treatment outcome using causal chain analysis. Baseline, end-treatment and follow-up self-help affiliation and substance use were assessed in 252 drug and alcohol abusers attending a community program. Participants were randomly assigned to one of three cognitive-behavioral or 12-Step oriented individual counseling conditions. Participants also were assigned, based on problem severity, to one of three treatment settings (partial hospitalization; intensive outpatient; standard outpatient) that varied according to intensity of additional 12-Step programming. Repeated-measures ANOVA indicated that self-help affiliation increased overall during treatment, particularly in the 12-Step counseling condition (versus cognitive-behavioral), and the partial hospitalization setting (versus standard outpatient). Multiple regression indicated that self-help affiliation was negatively associated with substance use 6 months post-treatment, and that this was not moderated by condition. Self-help affiliation predicted outcome most strongly in the partial hospitalization setting and not at all in outpatient; however, this effect was accounted for by participants' initial problem severity: high-severity patients had poorer outcomes if they failed to increase self-help affiliation, relative to those who increased affiliation and to low-severity patients regardless of affiliation. Results indicate that, in this community-based program, self-help affiliation increased as a function of exposure to 12-Step oriented treatment programming, and significantly predicted better outcome among patients with high levels of problem severity. Implications for future treatment process and dissemination research are discussed.


Transient kinetics of subunit-III-depleted cytochrome c oxidase.
Cytochrome c oxidase from ox heart was depleted of subunit III and its transient kinetic properties studied by stopped-flow and flash photolysis. It was found that the overall mechanism of electron transfer is very similar for subunit-III-depleted and native oxidase, although significant differences in some kinetic parameters have been detected. These include the second-order rate constant for cytochrome c oxidation and the rate-limiting step of the overall process. Moreover, at low cytochrome c/oxidase ratios (where the number of reducing equivalents is insufficient), the rate of reoxidation of cytochrome a was found to be very slow, even in air, and in fact for the subunit-III-depleted enzyme is even slower than for the native oxidase. The stability of reduced cytochrome a excludes the likelihood that removal of subunit III leads to a new O2-binding site, and the result may be relevant to the lowered vectorial H+/e- stoichiometry. The subunit-III-depleted oxidase can be pulsed under appropriate conditions and its combination with CO is unchanged, as shown by kinetic experiments and difference spectroscopy.


Burns: where are we standing with propranolol, oxandrolone, recombinant human growth hormone, and the new incretin analogs?
The hypermetabolic response in critically ill patients is characterized by hyperdynamic circulatory, physiologic, catabolic and immune system responses. Failure to satisfy overwhelming energy and protein requirements after, and during critical illness, results in multiorgan dysfunction, increased susceptibility to infection, and death. Attenuation of the hypermetabolic response by various pharmacologic modalities is emerging as an essential component of the management of severe burn patients. This review focuses on the more recent advances in therapeutic strategies to attenuate the hypermetabolic response and its associated insulin resistance postburn. At present, beta-adrenergic blockade with propranolol represents probably the most efficacious anticatabolic therapy in the treatment of burns. Other pharmacological strategies include growth hormone, insulin-like growth factor, oxandrolone and intensive insulin therapy. Novel approaches to the management of critical illness by judicious glucose control and the use of pharmacologic modulators to the hypercatabolic response to critical illness have emerged. Investigation of alternative strategies, including the use of metformin, glucagon-like-peptide-1 and the PPAR-γ agonists are under current investigation.


Effects of reaction time variability and age on brain activity during Stroop task performance.
Variability in reaction time during task performance may reflect fluctuations in attention and cause reduced performance in goal-directed tasks, yet it is unclear whether the mechanisms behind this phenomenon change with age. Using fMRI, we tested young and cognitively healthy older adults with the Stroop task to determine whether aging affects the neural mechanisms underlying intra-individual reaction time variability. We found significant between-group differences in BOLD activity modulated by reaction time. In older adults, longer reaction times were associated with greater activity in frontoparietal attentional areas, while in younger adults longer reaction times were associated with greater activity in default mode network areas. Our results suggest that the neural correlates of reaction time variability change with healthy aging, reinforcing the concept of functional plasticity to maintain high cognitive function throughout the lifespan.


Towards development of a statistical framework to evaluate myotonic dystrophy type 1 mRNA biomarkers in the context of a clinical trial.
Myotonic dystrophy type 1 (DM1) is a rare genetic disorder, characterised by muscular dystrophy, myotonia, and other symptoms. DM1 is caused by the expansion of a CTG repeat in the 3'-untranslated region of DMPK. Longer CTG expansions are associated with greater symptom severity and earlier age at onset. The primary mechanism of pathogenesis is thought to be mediated by a gain of function of the CUG-containing RNA, that leads to trans-dysregulation of RNA metabolism of many other genes. Specifically, the alternative splicing (AS) and alternative polyadenylation (APA) of many genes is known to be disrupted. In the context of clinical trials of emerging DM1 treatments, it is important to be able to objectively quantify treatment efficacy at the level of molecular biomarkers. We show how previously described candidate mRNA biomarkers can be used to model an effective reduction in CTG length, using modern high-dimensional statistics (machine learning), and a blood and muscle mRNA microarray dataset. We show how this model could be used to detect treatment effects in the context of a clinical trial.


Observation of Quantum-Limited Spin Transport in Strongly Interacting Two-Dimensional Fermi Gases.
We measure the transport properties of two-dimensional ultracold Fermi gases during transverse demagnetization in a magnetic field gradient. Using a phase-coherent spin-echo sequence, we are able to distinguish bare spin diffusion from the Leggett-Rice effect, in which demagnetization is slowed by the precession of a spin current around the local magnetization. When the two-dimensional scattering length is tuned to be comparable to the inverse Fermi wave vector k_{F}^{-1}, we find that the bare transverse spin diffusivity reaches a minimum of 1.7(6)ℏ/m, where m is the bare particle mass. The rate of demagnetization is also reflected in the growth rate of the s-wave contact, observed using time-resolved spectroscopy. The contact rises to 0.28(3)k_{F}^{2} per particle, which quantifies how scaling symmetry is broken by near-resonant interactions, unlike in unitary three-dimensional systems. Our observations support the conjecture that, in systems with strong scattering, the local relaxation rate is bounded from above by k_{B}T/ℏ.


Systematic review of TCF2 anomalies in renal cysts and diabetes syndrome/maturity onset diabetes of the young type 5.
There is a paucity of published works that systematically evaluate gene anomalies or clinical features of patients with renal cysts and diabetes syndrome (RCAD)/maturity onset diabetes of the young type 5 (MODY5). The purpose of this review was to systematically assess the detection rate, genetic and phenotypic implications of heterozygous autosomal dominant TCF2 anomalies. MEDLINE database was searched to select articles recorded in English from 1997 to 2008. The focus was monoallelic germline TCF2 gene mutations/deletions. Biallelic inactivation, polymorphisms, DNA modification (hypomethylation and hypermethylation), loci associated with cancer risk, and somatic TCF2 anomalies were all excluded. After searching the literature, 50 articles were selected. The detection rate of TCF2 anomalies was 9.7% and varied considerably among MODY (1.4%), renal structure anomalies (RSA) (21.4%) and RSA with MODY (41.2%) subgroups. Mutations were strikingly located within the DNA binding domain and varied among exons of the DNA binding domain: exons 2 and 4 were the hottest spots, while mutations were sporadically distributed in exon 3. The consistent phenotypes were RSA (89.6%) and diabetes mellitus (DM) (45.0%). However, the concurrence of RSA and DM was relatively low (27.5%), which hinders the optimal performance of genetic testing and obtainment of timely diagnosis. Other organ involvements were complementary and necessary for the early identification of patients with TCF2 anomalies. Analysis of phenotypes of TCF2 point mutations showed significant differences in the detection rates of RSA, impaired renal function (IRF) and DM according to mutation type but not mutation location. These valuable features of TCF2 anomalies that previously did not receive sufficient attention should not be neglected.


Bordetella pertussis isolates studied at the National Health Institute, Wellington, between 1977 and 1981.
Between 1977 and 1982, 199 isolates of Bordetella pertussis were referred to the National Health Institute for confirmation of identity and from mid 1982 for serotyping. The information gained from cultures referred during 1982 has been of assistance to the New Zealand Department of Health in investigating the causes of the present outbreak and has also enabled comparison with the serotypes involved in the present outbreak in the United Kingdom.


Familial aggregation of ankylosing spondylitis in Southern China.
To study the familial aggregation of ankylosing spondylitis (AS) in southern China and to compare the clinical and laboratory characteristics between the probands and their first-degree relatives with AS. On the basis of questionnaires to 473 patients with AS, 402 responded and 36 self-reported having first-degree relatives with symptoms related to spondyloarthropathies. All together, 144 of 150 first-degree relatives of these 36 probands were examined for clinical and radiographic characteristics. HLA typing for HLA-B27 was performed by standard microlymphocytotoxicity method. The disease duration of the 36 probands was 5.03 +/- 3.76 years (0.5-14 yrs). Forty seven first-degree relatives of the 36 probands were diagnosed as having AS. The prevalence of AS among the first-degree relatives of these 36 aggregated families was 31.3%. The overall prevalence of AS among the first-degree relatives in these 402 families was estimated to be 2.8%. The recurrence risk of the first-degree relatives within these aggregated families was 31.3%, suggesting an excess risk to them of 120.4, while it was 10.8 to the general families. The probands more often had peripheral arthritis and enthesopathies (p < 0.001 and p = 0.01, respectively) than the AS patients among the first-degree relatives. HLA-B27 was associated with development of AS in the probands and the patients among the first-degree relatives. Although familial aggregation of AS in southern China is uncommon in general, the recurrence risk of the first-degree relatives of AS probands within the aggregated families is extremely high, according to our study among hospital based patients. As the disease of the first-degree relatives is often mild and atypical, familial background analysis should be encouraged to assist early diagnosis.


A review of accelerometer-derived physical activity in the idiopathic inflammatory myopathies.
The idiopathic inflammatory myopathies (IIMs) are a group of rare conditions characterised by muscle inflammation (myositis). Accurate disease activity assessment is vital in both clinical and research settings, however, current available methods lack ability to quantify associated variation of physical activity, an important consequence of myositis.This study aims to review studies that have collected accelerometer-derived physical activity data in IIM populations, and to investigate if these studies identified associations between physical and myositis disease activity. A narrative review was conducted to identify original articles that have collected accelerometer-derived physical activity data in IIM populations. The following databases were searched from February 2000 until February 2019: Medline via PubMed, Embase via OVID and Scopus. Of the 297 publications screened, eight studies describing accelerometer use in 181 IIM cases were identified. Seven out of the eight studies investigated juvenile dermatomyositis (JDM) populations and only one reported on an adult-onset population. Population sizes, disease duration, accelerometer devices used, body placement sites, and study duration varied between each study.Accelerometer-derived physical activity levels were reduced in IIM cohorts, compared to healthy controls, and studies reported improvement of physical activity levels following exercise programme interventions, thus demonstrating efficacy.Higher levels of accelerometer-derived physical activity measurements were associated with shorter JDM disease duration, current glucocorticoid use and lower serum creatine kinase. However, no clear association between muscle strength and accelerometer-derived physical activity measures was identified. The use of accelerometer-derived physical activity in IIM research is in its infancy. Whilst knowledge is currently limited to small studies, the opportunities are promising and future research in this area has the potential to improve disease activity assessment for clinical and research applications.


Evidence of recent natural selection on the Southeast Asian deletion (--(SEA)) causing α-thalassemia in South China.
The Southeast Asian deletion (--(SEA)) is the most commonly observed mutation among diverse α-thalassemia alleles in Southeast Asia and South China. It is generally argued that mutation --(SEA), like other variants causing hemoglobin disorders, is associated with protection against malaria that is endemic in these regions. However, little evidence has been provided to support this claim. We first examined the genetic imprint of recent positive selection on the --(SEA) allele and flanking sequences in the human α-globin cluster, covering a genomic region spanning ~410 kb, by genotyping 28 SNPs in a Chinese population consisting of 76 --(SEA) heterozygotes and 138 normal individuals. The pattern of linkage disequilibrium (LD) and the long-range haplotype test revealed a signature of positive selection. The network of inferred haplotypes suggested a single origin of the --(SEA) allele. Thus, our data support the hypothesis that the --(SEA) allele has been subjected to recent balancing selection, triggered by malaria.


Growth and Metal Accumulation of an Alyssum murale Nickel Hyperaccumulator Ecotype Co-cropped with Alyssum montanum and Perennial Ryegrass in Serpentine Soil.
The genus Alyssum (Brassicaceae) contains Ni hyperaccumulators (50), many of which can achieve 30 g kg(-1) Ni in dry leaf. Some Alyssum hyperaccumulators are viable candidates for commercial Ni phytoremediation and phytomining technologies. It is not known whether these species secrete organic and/or amino acids into the rhizosphere to solubilize Ni, or can make use of such acids within the soil to facilitate uptake. It has been hypothesized that in fields with mixed plant species, mobilization of metals by phytosiderophores secreted by Graminaceae plants could affect Alyssum Ni, Fe, Cu, and Mn uptake. We co-cropped the Ni hyperaccumulator Alyssum murale, non-hyperaccumulator A. montanum and perennial ryegrass in a natural serpentine soil. All treatments had standard inorganic fertilization required for ryegrass growth and one treatment was compost amended. After 4 months A. murale leaves and stems contained 3600 mg kg(-1) Ni which did not differ significantly with co-cropping. Overall Ni and Mn concentrations were significantly higher in A. murale than in A. montanum or L. perenne. Copper was not accumulated by either Alyssum species, but L. perenne accumulated up to 10 mg kg(-1). A. montanum could not compete with either A. murale or ryegrass, and neither Alyssum species survived in the compost-amended soil. Co-cropping with ryegrass reduced Fe and Mn concentrations in A. murale but not to the extent of either increasing Ni uptake or affecting plant nutrition. The hypothesized Alyssum Ni accumulation in response to phytosiderophores secreted by co-cropped grass did not occur. Our data do not support increased mobilization of Mn by a phytosiderophore mechanism either, but the converse: mobilization of Mn by the Alyssum hyperaccumulator species significantly increased Mn levels in L. perenne. Tilling soil to maximize root penetration, adequate inorganic fertilization and appropriate plant densities are more important for developing efficient phytoremediation and phytomining approaches.


Peptide mimetics of immunoglobulin A (IgA) and FcαRI block IgA-induced human neutrophil activation and migration.
The cross-linking of the IgA Fc receptor (FcαRI) by IgA induces release of the chemoattractant LTB4, thereby recruiting neutrophils in a positive feedback loop. IgA autoantibodies of patients with autoimmune blistering skin diseases therefore induce massive recruitment of neutrophils, resulting in severe tissue damage. To interfere with neutrophil mobilization and reduce disease morbidity, we developed a panel of specific peptides mimicking either IgA or FcαRI sequences. CLIPS technology was used to stabilize three-dimensional structures and to increase peptides' half-life. IgA and FcαRI peptides reduced phagocytosis of IgA-coated beads, as well as IgA-induced ROS production and neutrophil migration in in vitro and ex vivo (human skin) experiments. Since topical application would be the preferential route of administration, Cetomacrogol cream containing an IgA CLIPS peptide was developed. In the presence of a skin permeation enhancer, peptides in this cream were shown to penetrate the skin, while not diffusing systemically. Finally, epitope mapping was used to discover sequences important for binding between IgA and FcαRI. In conclusion, a cream containing IgA or FcαRI peptide mimetics, which block IgA-induced neutrophil activation and migration in the skin may have therapeutic potential for patients with IgA-mediated blistering skin diseases.


An inducible nuclear factor binds to a growth hormone-regulated gene.
Transcription of the serine protease inhibitor (Spi) 2.1 gene, a member of the serine protease inhibitor family, is induced by growth hormone (GH) in rat liver. To further study the mechanism involved in this process, we have isolated and characterized the Spi 2.1 gene from a rat genomic library. Examination of the 5'-flanking region of the Spi 2.1 gene from normal animals revealed the presence of a DNase I hypersensitive site within 500 base pairs of the transcriptional initiation site, which was not detectable in hypophysectomized animals. Portions of the 5'-flanking region of the Spi 2.1 gene were fused to a heterologous promoter and reporter gene and introduced into primary rat hepatocytes by lipofection. Spi 2.1 sequences from -275 to -54 gave a 2-3-fold induction of reporter gene activity in cells grown in the presence of GH, similar to the level of induction of the endogenous Spi 2.1 mRNA in isolated hepatocytes. Further definition of the essential sequences revealed that a segment from -147 to -102 could confer GH responsiveness when linked in tandem copies in front of a heterologous promoter. Using the gel shift assay, a nuclear factor(s) from normal rat liver was identified which could interact with this minimal response fragment. The importance of this activity to GH regulation was suggested by the fact that it was absent in hypophysectomized animals but reappeared by 1 h after treatment of such animals with GH. The appearance of this activity was not blocked by pretreatment of animals with an inhibitor of protein synthesis, suggesting a preexisting factor is modified by GH to yield an activity which interacts with the Spi 2.1 gene.


Predictors of partner numbers in homosexual men: psychosocial factors in four societies.
Six hundred four homosexual men in four countries (Sweden, Finland, Australia, and the Republic of Ireland) were investigated with regard to their number of sexual partners over the past year and a number of psychological variables including masculinity and femininity, number of infections with sexually transmitted agents, relationships with parents, attitudes toward homosexuality, degree of homosexuality, sex-role conservatism, and a number of demographic variables. By use of multiple linear regression on partner numbers for each sample, it was found that between 15% and 28% of the variance of number of partners was accounted for by psychosocial factors, and that the great majority of these were significant in more than one country. Predictors included masculine and feminine personality traits, relationships with mother, degree of homosexuality, perceived societal attitudes toward homosexuality, age, and education. The data strongly suggest that psychosocial factors are significant predictors of numbers of partners of homosexual men, and that psychological approaches to treatment and prevention of sexually transmitted diseases may be warranted.


Candida glabrata PDR1, a transcriptional regulator of a pleiotropic drug resistance network, mediates azole resistance in clinical isolates and petite mutants.
Candida glabrata, a yeast with intrinsically low susceptibility to azoles, frequently develops increased azole resistance during prolonged treatment. Transposon mutagenesis revealed that disruption of CgPDR1 resulted in an 8- to 16-fold increase in fluconazole susceptibility of C. glabrata. CgPDR1 is a homolog of Saccharomyces cerevisiae PDR1, which encodes a transcriptional regulator of multidrug transporters. Northern blot analyses indicated that CgPDR1 regulated both constitutive and drug-induced expression of CgCDR1, a multidrug transporter gene. In agreement with the Northern analysis, the Cgpdr1 mutant had increased rhodamine accumulation, in contrast to the decreased accumulation in the CgPDR1-overexpressing strain. Northern analyses also indicated the importance of CgPDR1 in fluconazole resistance arising during therapy. Two clinically resistant isolates had higher expression of CgPDR1 and CgCDR1 compared to their paired susceptible isolates. Integrative transformation of CgPDR1 from the two resistant isolates converted the Cgpdr1 mutant into azole-resistant strains with upregulated CgPDR1 expression. Two different amino acid substitutions, W297S in one isolate and F575L in the other, accounted for the upregulated CgPDR1 expression and the resistance. Finally, CgPDR1 was shown to be required for the azole resistance due to mitochondrial deficiency. Thus, CgPDR1 encodes a transcriptional regulator of a pleiotropic drug resistance network and contributes to the azole resistance of clinical isolates and petite mutants.


TUBE VERSUS TRABECULECTOMY IN JUVENILE-ONSET OPEN ANGLE GLAUCOMA - TREATMENT OUTCOMES IN TERTIARY HOSPITALS IN MALAYSIA.
To compare the intraocular pressure (IOP) lowering effect and postoperative complications between primary augmented trabeculectomy and glaucoma drainage device (GDD) implantation as primary surgical intervention in patients with juvenile-onset open angle glaucoma (JOAG). A retrospective review study involving 20 eyes that underwent primary augmented trabeculectomy with mitomycin (MMC) and 10 eyes GDD implantation in 3 tertiary centres in Malaysia between 1 January 2013 and 31 December 2019. They were followed up for at least 12 months postsurgical intervention. Intraocular pressure (IOP), number of topical IOP lowering medication and complications were evaluated at 1, 3, 6 and 12 months post-intervention. Based on the IOP, the success was divided into complete and partial success, and failure. IOP and postsurgical complications were compared using the Repetitive Measure Analysis of Variance (RM ANOVA) and the Pearson chi-square test. Both methods were effective in lowering the IOP. Eyes with primary augmented trabeculectomy have significant lower IOP compared to GDD implantation (p = 0.037). There was a higher incidence of postoperative hypotony (30%) in the trabeculectomy group. There was also a significant reduction of mean number of topical pressure-lowering drugs required postoperatively (p = 0.015). Complete success was achieved in 100% of eyes with trabeculectomy and 67% in GDD implantation (p = 0.047). Primary augmented trabeculectomy and GDD implantation are good surgical options for the treatment of JOAG. Both methods provide IOP lowering at 1 year. However, trabeculectomy provides better pressure lowering, compared to GDD implantation in patients with JOAG.


[A case of heritable pulmonary arterial hypertension treated with long-term oral low-dose imatinib].
Heritable pulmonary arterial hypertension (HPAH) is a rare type of pulmonary arterial hypertension that often presents with progressive exertional dyspnea and for which there is no significant effective drug. A HPAH patient was admitted to our hospital more than three years ago, and the gene mutation was bone morphogenetic protein 2 (BMPR2). For the first 45 months, she was given oral imatinib 100 mg once daily, and her symptoms and hemodynamics improved significantly, with no apparent side effects. It is reported that, in combination with the characteristics of the case and related literatures, it provides clinicians with other feasible treatment options for HPAH.


Advanced diffusion imaging to track progression in Parkinson's disease, multiple system atrophy, and progressive supranuclear palsy.
Advanced diffusion imaging which accounts for complex tissue properties, such as crossing fibers and extracellular fluid, may detect longitudinal changes in widespread pathology in atypical Parkinsonian syndromes. We implemented fixel-based analysis, Neurite Orientation and Density Imaging (NODDI), and free-water imaging in Parkinson's disease (PD), multiple system atrophy (MSAp), progressive supranuclear palsy (PSP), and controls longitudinally over one year. Further, we used these three advanced diffusion imaging techniques to investigate longitudinal progression-related effects in key white matter tracts and gray matter regions in PD and two common atypical Parkinsonian disorders. Fixel-based analysis and free-water imaging revealed longitudinal declines in a greater number of descending sensorimotor tracts in MSAp and PSP compared to PD. In contrast, only the primary motor descending sensorimotor tract had progressive decline over one year, measured by fiber density (FD), in PD compared to that in controls. PSP was characterized by longitudinal impairment in multiple transcallosal tracts (primary motor, dorsal and ventral premotor, pre-supplementary motor, and supplementary motor area) as measured by FD, whereas there were no transcallosal tracts with longitudinal FD impairment in MSAp and PD. In addition, free-water (FW) and FW-corrected fractional anisotropy (FAt) in gray matter regions showed longitudinal changes over one year in regions that have previously shown cross-sectional impairment in MSAp (putamen) and PSP (substantia nigra, putamen, subthalamic nucleus, red nucleus, and pedunculopontine nucleus). NODDI did not detect any longitudinal white matter tract progression effects and there were few effects in gray matter regions across Parkinsonian disorders. All three imaging methods were associated with change in clinical disease severity across all three Parkinsonian syndromes. These results identify novel extra-nigral and extra-striatal longitudinal progression effects in atypical Parkinsonian disorders through the application of multiple diffusion methods that are related to clinical disease progression. Moreover, the findings suggest that fixel-based analysis and free-water imaging are both particularly sensitive to these longitudinal changes in atypical Parkinsonian disorders.


Morphological Changes Induced By Extremely Low-Frequency Electric Fields.
In this paper, morphological effects of electric fields on avian erythrocytes (nucleated red blood cells) have been studied in detail. Morphological changes include rounding and cytoplasm transparency. It has been shown that the effect is non-thermal. Careful imaging and image analyses have been carried out to show that the degree of this effect is frequency-dependent, and has a higher conversion rate at higher temperatures. Furthermore, to better understand the mechanisms behind the morphological changes, we investigated the dedifferentiation hypothesis and performed a series of tests on avian erythrocytes including fluorescence spectroscopy for hemoglobin, and tests on human umbilical cord blood, mesenchymal stem cells, and bone marrow mesenchymal stem cells including flow-cytometry analysis for expression of certain markers and calcium staining. Bioelectromagnetics. 2019;40:375-390. © 2019 Bioelectromagnetics Society.


Early palliative care referral in patients with end stage liver disease is associated with reduced resource utilisation.
Palliative care referral for end stage liver disease (ESLD) is uncommon and usually occurs late. We aimed to review the rate of early palliative care referral (EPCR) in ESLD patients, its associations, and its impacts on resource utilisation and survival. A retrospective review of all patients with ESLD admitted to a single hepatology unit between 2013 and 2016. Inclusion criteria for study entry were at least two admissions for decompensated liver disease within a six month period and not eligible for liver transplantation. The EPCR group was defined as those patients who received palliative care referral at least 30 days prior to mortality. 74 patients were included in the study. EPCR rate was 19%. On multivariate analysis EPCR was associated with hepatocellular carcinoma (OR 4.47, 95% CI 1.02-19.5, p=0.047), and negatively associated with alcoholic liver disease (OR 0.16, 95% CI 0.032-0.88 p=0.035). There was no difference in survival based on EPCR status. Hospitalization costs were lower in the EPCR group (p=0.027). There was also a significantly lower number of endoscopies (p=0.009), and blood transfusions (p=0.001) in the EPCR group. EPCR was also associated with higher rates of outpatient palliative care and advanced care planning. EPCR in ESLD was uncommon and associated with hepatocellular carcinoma and lack of alcoholic liver disease. EPCR was associated with decreased resource utilisation and further high quality studies are required to confirm the benefits of EPCR in ESLD.


Small sample bias in the gamma frailty model for univariate survival.
The gamma frailty model is a natural extension of the Cox proportional hazards model in survival analysis. Because the frailties are unobserved, an E-M approach is often used for estimation. Such an approach is shown to lead to finite sample underestimation of the frailty variance, with the corresponding regression parameters also being underestimated as a result. For the univariate case, we investigate the source of the bias with simulation studies and a complete enumeration. The rank-based E-M approach, we note, only identifies frailty through the order in which failures occur; additional frailty which is evident in the survival times is ignored, and as a result the frailty variance is underestimated. An adaption of the standard E-M approach is suggested, whereby the non-parametric Breslow estimate is replaced by a local likelihood formulation for the baseline hazard which allows the survival times themselves to enter the model. Simulations demonstrate that this approach substantially reduces the bias, even at small sample sizes. The method developed is applied to survival data from the North West Regional Leukaemia Register.


Characteristics of pneumonia with negative chest radiography in cases confirmed by computed tomography.
Background: Chest radiography is commonly used for diagnosing community-acquired pneumonia (CAP). Computed tomography (CT) is not routinely recommended for initial assessment of CAP patients but is more sensitive and more specific than chest radiography. Objectives: To investigate characteristics of pneumonia with negative chest radiography in cases confirmed by CT. Methods: We included patients diagnosed with CAP in the emergency department, and chest radiography and CT were performed and sputum cultures were collected. The CR- group was defined as patients for whom infiltration of pneumonia was detected only on CT. The CR+ group was defined as patients for whom infiltration was detected on both chest radiography and CT. Data were collected retrospectively from medical records. Results: A total of 138 patients were included, with 58 patients in the CR- group and 80 patients in the CR+ group. Mean age was higher in the CR- group than in the CR+ group, and white blood cell counts and C-reactive protein (CRP) levels were lower in the CR- group than in the CR+ group (8.4 × 103/μL vs 12.4 × 103/μL, p = 0.01; 4.7 mg/dL vs 15.6 mg/dL, p < 0.001, respectively). Laterality of the infiltrated lungs differed between groups (right:left:bilateral = 14:30:14 vs 48:20:12, p = 0.006). Multivariate logistic analysis identified leukocytosis, elevated CRP levels (odds ratio (OR) 3.57, p = 0.003), laterality (OR 2.16, p = 0.006) as predictors of pneumonia in the CR- group. Conclusion: In pneumonia with negative chest radiography in cases confirmed by CT, milder inflammation and infiltration in the left lung tended to be seen.


ERGOBOSS: Ergonomic Optimization of Body-Supporting Surfaces.
Humans routinely sit or lean against supporting surfaces and it is important to shape these surfaces to be comfortable and ergonomic. We give a method to design the geometric shape of rigid supporting surfaces to maximize the ergonomics of physically based contact between the surface and a deformable human. We model the soft deformable human using a layer of FEM deformable tissue surrounding a rigid core, with measured realistic elastic material properties, and large-deformation nonlinear analysis. We define a novel cost function to measure the ergonomics of contact between the human and the supporting surface. We give a stable and computationally efficient contact model that is differentiable with respect to the supporting surface shape. This makes it possible to optimize our ergonomic cost function using gradient-based optimizers. Our optimizer produces supporting surfaces superior to prior work on ergonomic shape design. Our examples include furniture, apparel and tools. We also validate our results by scanning a real human subject's foot and optimizing a shoe sole shape to maximize foot contact ergonomics. We 3D-print the optimized shoe sole, measure contact pressure using pressure sensors, and demonstrate that the real unoptimized and optimized pressure distributions qualitatively match those predicted by our simulation.


Pathways to vocational services: factors affecting entry by veterans enrolled in Veterans Health Administration mental health services.
The current study provides naturalistic data documenting the pathways-to-care to vocational services for 155 veterans who were receiving some form of mental health care from the Veterans Health Administration and had a vocational need but were not currently enrolled in vocational services. Of the participants, 94.2% had recognized their vocational need, 80.6% reported that they or someone else had sought help to alleviate the need, and 77.4% had previously received some form of vocational services. The median length of the participants' vocational need was more than 4.2 years. Delays associated with recognition, help-seeking, and treatment entry all contributed to the overall delay in entering appropriate care. Filtering factors associated with quicker recognition, seeking help, and receiving services included diagnosis, level of disability, type of vocational need, and support from primary providers, family, and friends. The results provide information for designing interventions to improve service entry by adults with mental health problems and vocational needs.


A novel BMP expressed in developing mouse limb, spinal cord, and tail bud is a potent mesoderm inducer in Xenopus embryos.
The bone morphogenetic proteins (BMPs) play critical roles in patterning the early embryo and in the development of many organs and tissues. We have identified a new member of this multifunctional gene family, BMP-11, which is most closely related to GDF-8/myostatin. During mouse embryogenesis, BMP-11 is first detected at 9.5 dpc in the tail bud with expression becoming stronger as development proceeds. At 10.0 dpc, BMP-11 is expressed in the distal and posterior region of the limb bud and later localizes to the mesenchyme between the skeletal elements. BMP-11 is also expressed in the developing nervous system, in the dorsal root ganglia, and dorsal lateral region of the spinal cord. To assess the biological activity of BMP-11, we tested the protein in the Xenopus ectodermal explant (animal cap) assay. BMP-11 induced axial mesodermal tissue (muscle and notochord) in a dose-dependent fashion. At higher concentrations, BMP-11 also induced neural tissue. Interestingly, the activin antagonist, follistatin, but not noggin, an antagonist of BMPs 2 and 4, inhibited BMP-11 activity on animal caps. Our data suggest that in Xenopus embryos, BMP-11 acts more like activin, inducing dorsal mesoderm and neural tissue, and less like other family members such as BMPs 2, 4, and 7, which are ventralizing and anti-neuralizing signals. Taken together, these data suggest that during vertebrate embryogenesis, BMP-11 plays a unique role in patterning both mesodermal and neural tissues.


Dietary modulation of phase 1 and phase 2 activities with benzo(a)pyrene and related compounds in the intestine but not the liver of the channel catfish, Ictalurus punctatus.
These studies demonstrated that intestinal mucosa of the channel catfish contained activities comparable with liver for several phase 2 xenobiotic-metabolizing enzymes, and showed that CYP1A-dependent monooxygenase activities were inducible in intestine but not liver by dietary exposure to low concentrations of the Ah agonist, beta-naphthoflavone (BNF). The diets administered were laboratory-prepared, semisynthetic pellets of known composition, commercial chow, or chow supplemented with BNF at 10 or 100 mg BNF/kg chow. Very low intestinal benzo(a)pyrene hydroxylase [aryl hydrocarbon hydroxylase (AHH)] and ethoxyresorufin O-deethylase (EROD) activities were found in catfish fed the semisynthetic diet. Intestinal EROD and AHH activities were elevated by the commercial chow diet and further induced by supplementation with 10, but not 100, mg BNF/kg diet. In vitro studies showed that catfish EROD and AHH activities were sensitive to inhibition by BNF, with mean IC50 values of 0.078 and 2.2 microM, respectively. Thus, residues of BNF retained in intestinal mucosa may have masked monooxygenase induction in catfish fed the 100 mg BNF/kg diet. Microsomal UDP-glucuronosyltransferase and cytosolic PAPS-sulfotransferase activities with 3-hydroxybenzo(a)pyrene as substrate were largely unaffected by the diets studied, and intestinal activities were similar to hepatic activities. Glutathione S-transferase activity was slightly induced in intestinal, but not hepatic cytosol of catfish treated with BNF at the 10 mg/kg diet level relative to chow controls. Epoxide hydrolase activity with styrene oxide as substrate was not affected by diet in intestinal microsomes.


Infected cell protein 0 encoded by bovine herpesvirus 1 can activate caspase 3 when overexpressed in transfected cells.
Infection of cattle or bovine cells with bovine herpesvirus 1 (BHV-1) leads to increased apoptosis. Previous studies indicated that BHV-1 infected cell protein 0 (bICP0), the major transcriptional regulatory protein of BHV-1, is toxic in transiently transfected cells. Point mutations within the zinc RING finger of bICP0 reduced toxicity and eliminated the ability of bICP0 to activate viral gene expression. In mouse neuroblastoma cells (neuro-2A) and bovine turbinate cells, bICP0 activated caspase 3, a key regulatory protein in the apoptotic pathway. A pro-apoptotic gene (Bax), but not bICP0, induced caspase 3 cleavage and activation by 8 h after transfection of neuro-2A cells. Conversely, bICP0 or the N-terminal 356 aa of bICP0 did not induce caspase 3 cleavage in neuro-2A cells until 30 h after transfection, suggesting that bICP0 stimulates caspase 3 cleavage by an indirect mechanism. These studies indicate that the toxic functions of bICP0 correlate with caspase 3 cleavage and activation.


Finding a dominance order most consistent with a linear hierarchy: a new procedure and review.
A procedure for ordering a set of individuals into a linear or near-linear dominance hierarchy is presented. Two criteria are used in a prioritized way in reorganizing the dominance matrix to find an order that is most consistent with a linear hierarchy: first, minimization of the numbers of inconsistencies and, second, minimization of the total strength of the inconsistencies. The linear ordering procedure, which involves an iterative algorithm based on a generalized swapping rule, is feasible for matrices of up to 80 individuals. The procedure can be applied to any dominance matrix, since it does not make any assumptions about the form of the probabilities of winning and losing. The only assumption is the existence of a linear or near-linear hierarchy which can be verified by means of a linearity test. A review of existing ranking methods is presented and these are compared with the proposed method. Copyright 1998 The Association for the Study of Animal Behaviour. Copyright 1998 The Association for the Study of Animal Behaviour.


A component of platypus (Ornithorhynchus anatinus) venom forms slow-kinetic cation channels.
The lipid bilayer technique is used to examine the biophysical properties of anion and cation channels frequently formed by platypus (Ornithorhynchus anatinus) venom (OaV). The OaV-formed anion channel in 250/50 mm KCl cis/trans has a maximum conductance of 857 +/- 23 pS (n = 5) in 250/50 mm KCl cis/trans. The current-voltage relationship of this channel shows strong inward rectification. The channel activity undergoes time-dependent inactivation that can be removed by depolarizing voltage steps more positive than the reversal potential for chloride, E(Cl), (+40 mV). The reversal potential of the OaV-formed slow current activity in 250/50 mm KCl cis/trans is close to the potassium equilibrium potential (E(K)) of -40 mV. The conductance values for the slow channel are 22.5 +/- 2.6 pS and 41.38 +/- 4.2 pS in 250/50 and 750/50 mm cis/trans, respectively. The gating kinetics of the slow ion channels are voltage-dependent. The channel open probability (P(o)) is between 0.1 and 0.8 at potentials between 0 and +140 mV. The channel frequency (F(o)) increases with depolarizing voltages between 0 and +140 mV, whereas mean open time (T(o)) and mean closed time (T(c)) decrease. Ion substitution experiments of the cis solution show that the channel has conductance values of 21.47 +/- 2. 3 and 0.53 +/- 0.1 pS in 250 mm KCl and choline Cl, respectively. The amplitude of the single channel current is dependent on [K(+)](cis) and the current reversal potential (E(rev)) responds to increases in [K(+)](cis) by shifting to more negative voltages. The increase in current amplitude as a function of increasing [K(+)](cis) can be best described by a third order polynomial fit. At +140 mV, the values of the maximal single channel conductance (gamma(max)) and the concentration for half maximal gamma (K(s)) are 38.6 pS and 380 mm and decline to 15.76 pS and 250 mm at 0 mV, respectively. The ion selectivity of the channel to K(+), Na(+), Cs(+) and choline(+) was determined in ion substitution experiments. The permeability values for P(K(+)):P(Na(+)):P(Cs(+)):P(choline(+)) were 1:1:0.63:0.089, respectively. On the other hand, the activity of the slow channel was eliminated (Fig. 7B). The slow channel was reversibly inhibited by [TEA(+)](trans) and the half-maximal inhibitory concentration (K(i)) was approximately 48 mm.


Etiology, pathophysiology, and treatment of atrial fibrillation: part 1.
Atrial fibrillation (AF) is associated with a higher incidence of mortality, stroke, and coronary events than is sinus rhythm. AF with a rapid ventricular rate may cause a tachycardia-related cardiomyopathy. Immediate direct-current cardioversion should be performed in patients with AF and acute myocardial infarction, chest pain due to myocardial ischemia, hypotension, severe heart failure, or syncope. Intravenous beta blockers, verapamil, or diltiazem may be given to immediately slow a very rapid ventricular rate in AF. An oral beta blocker, verapamil, or diltiazem should be used in persons with AF if a fast ventricular rate occurs at rest or during exercise despite digoxin. Amiodarone may be used in selected patients with symptomatic life-threatening AF refractory to other drugs. Digoxin should not be used to treat patients with paroxysmal AF. Nonpharmacologic therapies should be used in patients with symptomatic AF in whom a rapid ventricular rate cannot be slowed by drugs. This is part 1 of a 2-part review of the etiology, pathophysiology, and treatment of atrial fibrillation. The second part will be published in the subsequent issue of Cardiology in Review.


Discretionary time among older adults: how do physical activity promotion interventions affect sedentary and active behaviors?
Investigation goals were to document discretionary time activities among older adults, determine whether time spent in discretionary activities varied by gender, and investigate whether participation in a prescribed physical activity (P) intervention increased the time that older adults spend in discretionary time physical activities that were not specifically prescribed by interventions. Longitudinal data were drawn from 2 published studies of older adults. Study 1 compared 2 PA interventions in healthy older men and women (N = 103, M =70.2 years), and Study 2 compared a PA intervention with a nutrition intervention in healthy older women (N =93, M =63.1 years). Participants in both studies completed similar assessments of their discretionary time activities using the Community Healthy Activities Model Program for Seniors questionnaire. Across both studies, at baseline, over 95% of participants reported talking on the telephone and reading as frequent sedentary discretionary time activities; over 80% reported visiting with friends and watching television or listening to the radio. Women engaged in significantly greater hours of social activities and household maintenance activities than did men (p <.05). From baseline to 12-month posttest, social, recreational, and household activities remained stable by gender and across time after participating in a PA intervention. Despite previously documented 2- to 3-hr increases in physical activities occurring in response to the study interventions, increases did not generalize for most participants to activities not prescribed by the intervention. Older adults are participating in numerous sedentary social and recreational activities that appear to remain stable across time and in the face of PA intervention prescriptions.


Chinese Herbal Medicine in the Treatment of Chronic Heart Failure: Three-Stage Study Protocol for a Randomized Controlled Trial.
Background. Chinese herbal medicine (CHM) has been used in the treatment of chronic heart failure (CHF) for a long time. Treatment based on syndrome differentiation and the main characteristic of TCM is the fundamental principle of TCM practice. In this study protocol, we have designed a trial to assess the efficacy and safety of CHM on CHF based on syndrome differentiation. Methods/Design. This is a three-stage trial of CHM in the treatment of CHF. The first stage is a literature review aiming to explore the common syndromes of CHF. The second is a multicentral, randomized, placebo-controlled trial to evaluate the efficacy and safety of CHM for the treatment of CHF. The third is a multicentral, randomized controlled clinical trial aiming to make cost-effectiveness analysis and evaluate the feasibility, compliance, and universality of CHM on CHF. Discussion. This trial will evaluate the efficacy, safety, feasibility, compliance, and universality of CHM on CHF. The expected outcome is to provide evidence-based recommendations for CHM on CHF and develop a prescription of CHM in the treatment of CHF. This trial is registered with NCT01939236 (Stage Two of the whole trial).


MiR-24 is required for hematopoietic differentiation of mouse embryonic stem cells.
Overexpression of miRNA, miR-24, in mouse hematopoietic progenitors increases monocytic/ granulocytic differentiation and inhibits B cell development. To determine if endogenous miR-24 is required for hematopoiesis, we antagonized miR-24 in mouse embryonic stem cells (ESCs) and performed in vitro differentiations. Suppression of miR-24 resulted in an inability to produce blood and hematopoietic progenitors (HPCs) from ESCs. The phenotype is not a general defect in mesoderm production since we observe production of nascent mesoderm as well as mesoderm derived cardiac muscle and endothelial cells. Results from blast colony forming cell (BL-CFC) assays demonstrate that miR-24 is not required for generation of the hemangioblast, the mesoderm progenitor that gives rise to blood and endothelial cells. However, expression of the transcription factors Runx1 and Scl is greatly reduced, suggesting an impaired ability of the hemangioblast to differentiate. Lastly, we observed that known miR-24 target, Trib3, is upregulated in the miR-24 antagonized embryoid bodies (EBs). Overexpression of Trib3 alone in ESCs was able to decrease HPC production, though not as great as seen with miR-24 knockdown. These results demonstrate an essential role for miR-24 in the hematopoietic differentiation of ESCs. Although many miRNAs have been implicated in regulation of hematopoiesis, this is the first miRNA observed to be required for the specification of mammalian blood progenitors from early mesoderm.


Computation of the mid-sagittal plane in 3-D brain images.
We present a new method to automatically compute, reorient, and recenter the mid-sagittal plane in anatomical and functional three-dimensional (3-D) brain images. This iterative approach is composed of two steps. At first, given an initial guess of the mid-sagittal plane (generally, the central plane of the image grid), the computation of local similarity measures between the two sides of the head allows to identify homologous anatomical structures or functional areas, by way of a block matching procedure. The output is a set of point-to-point correspondences: the centers of homologous blocks. Subsequently, we define the mid-sagittal plane as the one best superposing the points on one side and their counterparts on the other side by reflective symmetry. Practically, the computation of the parameters characterizing the plane is performed by a least trimmed squares estimation. Then, the estimated plane is aligned with the center of the image grid, and the whole process is iterated until convergence. The robust estimation technique we use allows normal or abnormal asymmetrical structures or areas to be treated as outliers, and the plane to be mainly computed from the underlying gross symmetry of the brain. The algorithm is fast and accurate, even for strongly tilted heads, and even in presence of high acquisition noise and bias field, as shown on a large set of synthetic data. The algorithm has also been visually evaluated on a large set of real magnetic resonance (MR) images. We present a few results on isotropic as well as anisotropic anatomical (MR and computed tomography) and functional (single photon emission computed tomography and positron emission tomography) real images, for normal and pathological subjects.


A comparison of intravenous hydrocortisone, methylprednisolone, and dexamethasone in acute bronchial asthma.
The short-term airway response of 14 subjects with severe asthma to equivalent low doses of intravenous hydrocortisone, methylprednisolone, and dexamethasone was examined. Overall the subjects improved without any significant differences between the three groups in subject age, baseline percent predicted FEV1, or percent predicted FEV1 at 3, 6, 9, 24, 48, and 72 hours (P greater than .05).


Atypical age-related cortical thinning in episodic migraine.
Prior studies demonstrate reduced cortical thickness and volume in migraineurs. However, the effect of age on cortical thickness has not been assessed in migraineurs. In this study we investigated whether the process of aging on cortical thickness affects migraineurs differently compared to age-matched healthy controls, i.e. whether aging exacerbates cortical thinning in migraineurs. Cortical thickness was estimated using a general linear model vertex-by-vertex approach for 32 healthy controls (mean age=35.3 years; SD=11.6) and 27 episodic migraine patients (mean age=33.6 years; SD=12.3). Results were modeled using a main effect analysis to estimate the effect of age on cortical thickness for each group separately, and an age-by-group analysis to estimate differences in age-related cortical thinning between migraine patients and normal controls. Although migraineurs and normal controls both have expected age-related thinning in many regions along the cortical mantle, migraineurs have age-related thinning of regions that do not thin in healthy controls, including: bilateral postcentral, right fusiform, and right temporal pole areas. Cortical thinning of these regions is more prominent with advancing age. Results suggest that migraine is associated with atypical cortical aging, suggesting that the migraine disease process interacts with aging to affect cortical integrity.


The effect of intravenous fixed-dose heparin during total hip arthroplasty on the incidence of deep-vein thrombosis. A randomized, double-blind trial in patients operated on with epidural anesthesia and controlled hypotension.
Heparin was given in fixed doses intravenously during unilateral primary total hip-replacement operations in a prospective, double-blind trial to assess the effect on the incidence of deep-vein thrombosis. One hundred and fifty patients were randomly assigned to one of two groups before the operation. Twenty-four patients were excluded from the study, leaving 126 patients. Group I consisted of sixty-six patients who received saline solution intravenously, and Group II comprised sixty patients who received heparin. All patients had epidural anesthesia with controlled hypotension. Fixed doses of heparin were administered five minutes before the operative incision was made and every thirty minutes throughout the operation. Mean arterial pressures were maintained at between fifty and sixty millimeters of mercury in all patients. Ascending venography was done on the seventh day after the operation. The incidence of deep-vein thrombosis was 24 per cent (sixteen of sixty-six patients) in Group I and 8 per cent (five of sixty patients) in Group II; the difference is significant (p = 0.03). The intraoperative loss of blood averaged 220 +/- 79 milliliters in Group I compared with 269 +/- 109 milliliters in Group II. An average of less than one unit of blood was transfused for each patient in each group. Postoperatively, there was no difference between the groups with regard to the amount of drainage that was collected in a Hemovac device or the values for hematocrit.


Activation of the endomitotic spindle assembly checkpoint and thrombocytopenia in Plk1-deficient mice.
Polyploidization in megakaryocytes is achieved by endomitosis, a specialized cell cycle in which DNA replication is followed by aberrant mitosis. Typical mitotic regulators such as Aurora kinases or Cdk1 are dispensable for megakaryocyte maturation, and inhibition of mitotic kinases may in fact promote megakaryocyte maturation. However, we show here that Polo-like kinase 1 (Plk1) is required for endomitosis, and ablation of the Plk1 gene in megakaryocytes results in defective polyploidization accompanied by mitotic arrest and cell death. Lack of Plk1 results in defective centrosome maturation and aberrant spindle pole formation, thus impairing the formation of multiple poles typically found in megakaryocytes. In these conditions, megakaryocytes arrest for a long time in mitosis and frequently die. Mitotic arrest in wild-type megakaryocytes treated with Plk1 inhibitors or Plk1-null cells is triggered by the spindle assembly checkpoint (SAC), and can be rescued in the presence of SAC inhibitors. These data suggest that, despite the dispensability of proper chromosome segregation in megakaryocytes, an endomitotic SAC is activated in these cells upon Plk1 inhibition. SAC activation results in defective maturation of megakaryocytes and cell death, thus raising a note of caution in the use of Plk1 inhibitors in therapeutic strategies based on polyploidization regulators.


Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses.
A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5+/6+ (P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus.


Sensory recovery of innervated and non-innervated radial forearm free flaps: functional implications.
Findings reported in the literature on the sensation provided by intraorally applied innervated vs. non-innervated radial forearm free flaps differ. In an effort to understand these differences in sensory recovery, the authors carried out sensory evaluations in 12 patients who had undergone radial forearm free flaps. Seven patients had innervated flaps for defects of the tongue and floor of mouth; five had non-innervated flaps to various sites. Flap sensitivity to temperature, light touch, dull touch, and sharpness and two-point discrimination was assessed at the donor site and contralaterally, and at the recipient site and contralateral mirror-image oral mucosa. Patients subjectively rated post-reconstruction sensation and provided quality of life (QOOL) data. The innervated flaps demonstrated better sensory recovery than the non-innervated flaps, although the latter did restore reasonable sensation. This paper describes the results, compares the study to other similar studies, and discusses various factors in the sensory recovery of both innervated and non-innervated intraoral radial forearm free flaps. The authors conclude that, although the trend in this study is toward improved function with the innervated flaps, these flaps do not appear to offer major intraoral functional advantage over the non-innervated flaps, which attain reasonably effective sensory recovery from neural ingrowth, if the lingual nerve is intact.