Identification of Xanthomicrol as a Major Metabolite of 5-Demethyltangeretin in Mouse Gastrointestinal Tract and Its Inhibitory Effects on Colon Cancer Cells.
5-Demethyltangeretin (5DT) is a unique polymethoxyflavone mainly found in the peel of citrus, and has shown potent suppressive effects on multiple human cancer cells. Biotransformation plays a critical role in the biological activities of dietary bioactive components because their metabolites may exert significant bioactivities. In the present study, the metabolic fate of 5DT in mouse gastrointestinal (GI) tract after long-term oral intake and the anti-cancer effects of its major metabolite were determined. It was found that 5DT underwent extensive biotransformation after oral ingestion in mice. A major demethylated metabolite was produced via phase I metabolism, while conjugates (glucuronide and sulfate) were generated via phase II metabolism. Specifically, 4'-position on the B ring of 5DT was the major site for demethylation reaction, which led to the production of xanthomicrol (XAN) as a major metabolite. More importantly, the level of XAN in the colon was significantly higher than that of 5DT in 5DT-fed mice. Thus, we further determined the suppressive effects of XAN on human colon cancer HCT116 cells. We found that XAN effectively inhibited the proliferation of HCT116 cells by arresting cell cycle and inducing cellular apoptosis, which was further evidenced by upregulated p53 and p21 and downregulated cyclin D and CDK4/6 level. In conclusion, this study identified XAN as a major metabolite of 5DT in mouse GI tract, and demonstrated its suppressive effects on HCT116 colon cancer cells.


Urinary estrogen metabolites and breast cancer: differential pattern of risk found with pre- versus post-treatment collection.
The products of estrogen metabolism may affect breast carcinogenesis. The 16alpha-hydroxyestrone (16-OHE) metabolite has a higher affinity for the estrogen receptor (ER) than the 2-hydroxyestrone (2-OHE) metabolite, while conjugated 2-OHE metabolite may inhibit angiogenesis. We investigated the association between the relative concentrations of these metabolites in urine (2-OHE/16-OHE) and breast cancer in a case-control study of Chinese women living in Shanghai. Incident breast cancer cases between 25 and 65 years of age (n=110) were identified from hospital or population tumor registries in Shanghai, China. Controls (n=110) were randomly selected from a complete registry of the Shanghai population, and individually matched to cases by menopausal status, age, and pre-treatment or post-treatment urine collection time. Urine samples were collected prior to any breast cancer treatment or surgery among 78 case-control pairs, while urine was collected after surgery, and perhaps other treatments, among 32 case-control pairs. A commercial enzyme-immunoassay kit was used to measure urinary estrogen metabolite concentrations. Conditional logistic regression was used to calculate odds ratios summarizing the 2-OHE/16-OHE and breast cancer association within subjects providing either pre-treatment or post-treatment urine samples. Subjects with a higher urinary 2-OHE/16-OHE ratio were less likely to be diagnosed with breast cancer, but only when urine samples were collected prior to breast cancer treatment (OR(Tertile3(T3)versusTertile1(T1))=0.5, 95% CI (0.2, 1.1)). In contrast, a higher 2-OHE/16-OHE ratio was significantly associated with breast cancer among subjects providing urine specimens after treatment initiation (OR(T3versusT1)=8.7, 95% CI (1.6, 47.1)). This observed cross-over modification occurred within both pre-menopausal and post-menopausal women, and independent of body mass index or recent dietary intake. Cross-study differences in urine collection protocols may explain observed inconsistencies in the 2-OHE/16-OHE and breast cancer association. Our case-control analysis using pre-treatment urine samples suggested that a lower 2-OHE/16-OHE ratio was associated with an increased risk of pre-menopausal and post-menopausal breast cancer diagnosis among Chinese women.


Measuring pressure in equilibrium and nonequilibrium lattice-gas models.
We develop an algorithm based on the method proposed by Dickman for directly measuring pressure in lattice-gas models. The algorithm gives the possibility to access the equation of state with a single run by adding multiple ghost sites to the original system. This feature considerably improves calculations and makes the algorithm particularly efficient for systems with inhomogeneous density profiles, both in equilibrium and nonequilibrium steady states. We illustrate its broad applicability by considering some paradigmatic systems of statistical mechanics such as the lattice gas under gravity, nearest-neighbor exclusion models in finite dimension and on regular random graphs, and the boundary-driven simple symmetric exclusion process.


Hypothalamic neurotensin projections promote reward by enhancing glutamate transmission in the VTA.
The lateral hypothalamus (LH) sends a dense glutamatergic and peptidergic projection to dopamine neurons in the ventral tegmental area (VTA), a cell group known to promote reinforcement and aspects of reward. The role of the LH to VTA projection in reward-seeking behavior can be informed by using optogenetic techniques to dissociate the actions of LH neurons from those of other descending forebrain inputs to the VTA. In the present study, we identify the effect of neurotensin (NT), one of the most abundant peptides in the LH to VTA projection, on excitatory synaptic transmission in the VTA and reward-seeking behavior. Mice displayed robust intracranial self-stimulation of LH to VTA fibers, an operant behavior mediated by NT 1 receptors (Nts1) and NMDA receptors. Whole-cell patch-clamp recordings of VTA dopamine neurons demonstrated that NT (10 nm) potentiated NMDA-mediated EPSCs via Nts1. Results suggest that NT release from the LH into the VTA activates Nts1, thereby potentiating NMDA-mediated EPSCs and promoting reward. The striking behavioral and electrophysiological effects of NT and glutamate highlight the LH to VTA pathway as an important component of reward.


Safety and efficacy of Isola instrumentation and arthrodesis for adolescent idiopathic scoliosis: two- to 12-year follow-up.
Retrospective case series including patient outcome assessment. To study the safety and efficacy of Isola instrumentation in comparison with similar series. Both the technique and technology used in the surgical treatment of adolescent idiopathic scoliosis continue to evolve, the common theme since the 1980s being provision of instrumentation stable and strong enough to eliminate the need for postoperative immobilization. The purpose of this study is to determine the safety and efficacy of a system deliberately integrating hook, wire, and screw anchors to deliver torsional and countertorsional corrective loads. A total of 185 consecutive patients, index patient included, were treated by posterior instrumentation and arthrodesis from January 1989 through December 2000. Safety was studied by complications, and reoperation type and occurrence. Effectiveness was studied by deformity correction and health-related quality of life questionnaire response. Variables affecting effectiveness were sought. A total of 179 patients (97%) had outcome assessment at an average of 6 years postoperative, and 176 had radiographic evaluation at an average of 5 years postoperative. There were no deaths, spinal cord or nerve root problems, or acute posterior wound infections. Proven pseudarthrosis occurred in 4 patients (2.2%) and delayed deep wound infection in 2 patients (1.1%). The implant-related reoperation rate was 8% and was necessary more often in the first quarter of the series (17% vs. 4.6%, P = 0.0062). The largest Cobb angle averaged 62 degrees preoperative and 23 degrees at latest follow-up, 63% correction. The largest angle of trunk inclination averaged 16.7 degrees before surgery and 9.9 degrees at latest follow-up, a 39% correction (P < 0.0001). Eighty-eight percent of patients were satisfied or very satisfied with the outcome. The principal problems identified were the need for a stronger transverse connector, stable end-instrumented vertebrae foundations, and convex thoracic anchorage. Isola instrumentation seems to be at least as safe and effective as other instrumentations being used for the surgical treatment of adolescent idiopathic scoliosis.


The WGD-A Dataset of Assembly Line Working Gestures for Ergonomic Analysis and Work-Related Injuries Prevention.
This paper wants to stress the importance of human movement monitoring to prevent musculoskeletal disorders by proposing the WGD-Working Gesture Dataset, a publicly available dataset of assembly line working gestures that aims to be used for worker's kinematic analysis. It contains kinematic data acquired from healthy subjects performing assembly line working activities using an optoelectronic motion capture system. The acquired data were used to extract quantitative indicators to assess how the working tasks were performed and to detect useful information to estimate the exposure to the factors that may contribute to the onset of musculoskeletal disorders. The obtained results demonstrate that the proposed indicators can be exploited to early detect incorrect gestures and postures and, consequently to prevent work-related disorders. The approach is general and independent on the adopted motion analysis system. It wants to provide indications for safely performing working activities. For example, the proposed WGD can also be used to evaluate the kinematics of workers in real working environments thanks to the adoption of unobtrusive measuring systems, such as wearable sensors through the extracted indicators and thresholds.


Cloning and sequence analysis of cDNAs encoding precursors of sarafotoxins. Evidence for an unusual "rosary-type" organization.
Sarafotoxins (SRTXs) are 21-amino acid peptides structurally and functionally similar to endothelins (ETs). To understand how SRTXs are overproduced in venom glands of the snakes Atractaspis engaddensis and hence used as toxins, we cloned cDNAs encoding SRTXs and elucidated their nucleotide sequences. We predict that SRTX precursors are large prepropolypeptide chains with an unusual "rosary-type" structure made of 12 successive similar stretches of 40 residues (39 in the first stretch). Each stretch begins with a "spacer" of 19 invariant residues (18 in the first stretch) immediately followed by the sequence of one SRTX isoform. Six different isoforms are identified within a single precursor molecule. Maturation of the precursor may require endopeptidases that cleave the Leu-Cys bond and the Trp-Arg/Lys bond invariably found at the SRTX N and C termini, respectively.


Aminoguanidine, a selective nitric oxide synthase inhibitor, attenuates cyclophosphamide-induced renal damage by inhibiting protein nitration and poly(ADP-Ribose) polymerase activation.
Cyclophosphamide (CP) is an antineoplastic agent that is used for the treatment of many neoplastic diseases. Renal damage is one of the dose-limiting side effects of CP. Recent studies show that nitrosative stress plays an important role in CP-induced renal damage. The purpose of our study was to investigate whether aminoguanidine (AG), a selective inducible nitric oxide synthase inhibitor, protects against CP-induced nitrosative stress and renal damage. Renal damage was induced in rats by administration of a single injection of CP at a dose of 150 mg/kg body weight intraperitoneally. For the AG pretreatment studies, the rats were injected intraperitoneally with AG at a dose of 200 mg/kg body weight 1 h before administration of CP. The control rats received AG or saline alone. All the rats were killed 16 h after the administration of CP or saline. Pretreatment with AG prevented CP-induced nitration of protein tyrosine and poly(ADP-ribose) polymerase (PARP) activation. Pretreatment with AG attenuated CP-induced renal damage. The present study demonstrates that AG is effective in preventing CP-induced renal damage and also that the protective effect is from its ability to inhibit nitric oxide-induced protein nitration and PARP activation. The present study shows that AG can prevent CP-induced renal damage by inhibiting protein tyrosine nitration and PARP activation. Thus, a more efficient and comfortable therapy can be achieved for patients in need of CP treatment. AG appears to be a promising drug for the prevention of nephrotoxicity of CP.


Calcium Inhibits Phase Shifting of the Circadian Conidiation Rhythm of Neurospora crassa by the Calcium Ionophore A23187.
Effects of the calcium ionophore, A23187, and antimycin A on the circadian conidiation rhythm of Neurospora crassa were examined. A23187 at a concentration of 1 mum in medium not containing divalent cations delayed the phase by 10 hours at CT 10 and advanced it by 5 hours at CT 14 (CT 12 corresponds to the time that discs are transferred from light to dark). This phase shifting was completely inhibited by addition of 0.1 millimolar CaCl(2) but not by MgCl(2) at any concentrations examined.Antimycin A inhibited respiration by 90% at a concentration of 0.2 micrograms per milliliter and lowered the ATP content by 85%. Antimycin A alone caused small phase advances but in combination with A23187 resulted in a large phase delay at CT 10. This phase shifting was not reversed by addition of CaCl(2) lower than 10 millimolar.


Autoantibodies to beta-amyloid and neurotransmitters in patients with Alzheimer's disease and senile dementia of the Alzheimer type.
The content of autoantibodies to beta-amyloid protein Abeta(1-42), its neurotoxic fragment Abeta(25-35), and neurotransmitters were studied in the blood of patients with presenile Alzheimer's disease and senile dementia of the Alzheimer type. Significant differences in the relative content of autoantibodies to Abeta(1-42)and autoantibodies to biogenic amines were demonstrated. These results can be used for the development of a biochemical method for differential diagnosis of Alzheimer dementias.


Treating profound hyponatremia: a strategy for controlled correction.
An alcoholic patient presented with profound hyponatremia (serum sodium concentration, 96 mEq/L) caused by the combined effects of a thiazide diuretic, serotonin reuptake inhibitor, beer potomania, and hypovolemia. A computed tomographic scan of the brain was indistinguishable from one obtained 3 weeks earlier when he was normonatremic. Concurrent administration of 3% saline solution and desmopressin controlled the rate of correction to an average of 6 mEq/L daily and resulted in full neurologic recovery without evidence of osmotic demyelination. This case illustrates the value of controlled correction of profound hyponatremia.


Improving body posture while fueling with a newly designed pump nozzle.
Although petrol pumps are a very common and highly used simple technology, their design consideration for comfort and safety to prevent high-pressure load and musculoskeletal injuries to the body is a neglected area in many countries including Iran. This study attempted to design a new pump nozzle, and to assess the differences in musculoskeletal load related to body posture when a price/volume display is mounted on the pump nozzle. For postural analysis, photographs recording the posture of 100 randomly selected customers while fueling at petrol pumps and the rapid upper limb assessment (RULA) technique were used. The results of this study showed that RULA scores improved significantly after the newly designed pump nozzle was used. The newly designed pump nozzle is useful in reducing load related to body posture while fueling.


[Characteristics of extreme temperature variation in the Loess Plateau and its correlation with average temperature].
In recent decades, extreme climate occurred frequently on the Loess Plateau. It is thus particularly important to study and predict the occurrence of extreme climate. Available researches on extreme climate mainly focus on the changing characteristics of the event itself, but ignore the correlation between average temperature and its changing trend. We used linear trend estimation, Mann-Kendall test, sliding t test and Pearson correlation analy-sis to study the variation trend of extreme temperature and its correlation with average temperature on the Loess Plateau based on the daily maximum temperature, minimum temperature and average temperature data of 79 meteo-rological stations from 1986 to 2019. The results showed that the extreme warmth index in the Loess Plateau region showed a significant upward trend, the extreme cold index showed a significant downward trend, and the frequency of extreme high temperature events increased. Most of the extreme temperature indices had abrupt changes in the middle and late 1990s and in 2012, and the extreme temperature showed a downward trend from 1998 to 2012, which better responded to the phenomenon of global warming hiatus. The increasing trend of mean temperature in the gully region, the rocky mountain region and the valley plain region was more obvious than that in other regions. The stations with large trend of extreme temperature index almost all occurred in the region with large increases of mean temperature. The increases of average temperature in small increments increased the frequency of extreme high temperature event, with the change range of extreme low temperature and its rate being greater than the extreme high temperature. Climate warming on extreme temperature index had a different effect, small changes in the average temperature in the Loess Plateau made the climate transfer towards the direction of more frequent heat.


Infrared optical spectrum of topological crystalline insulator SnTe (001) surface states.
We investigate the effects of varying temperature and chemical potential on the optical absorption spectrum of (001) surface states of topological crystalline insulator SnTe using a four-band effective k ⋅ p Hamiltonian. The spectrum is characterized by a narrow peak at 52 meV and a shoulder feature at 160 meV. Both absorptions have maximal intensity at 0 K or when chemical potential is located at the charge neutrality point. Then, as temperature increases or as chemical potential diverges, they both decrease in intensity. The 52 meV peak originates from transitions between high density of states regions surrounding van Hove singularities and is the spectrum's most prominent feature. Additionally, a third absorption from 110 meV to 150 meV, initially absent at 0 K or chemical potential at charge neutrality point, gradually builds in intensity as temperature increases or as chemical potential diverges. This absorption arises from transitions between low and high energy bands of opposite helicity. Importantly, we find that all distinct spectral features are diminished if the magnitude of chemical potential diverges to values above the van Hove singularity energies. If a given sample's chemical potential is well-controlled, conventional infrared spectroscopy may be used to identify the spectral signatures of SnTe (001) surface states at room temperatures and without use of large magnetic fields.


A Survey of Clinical Isolates of Neisseria Gonorrhoeae For Betalactamase Producers.
Strains of Neisseria gonorrhoeae isolated from smear-positive male patients suffering from acute gonococcal urethritis were studied. Their identity was confirmed on the basis of morphology, oxidase reaction, fermentation of glucose only and failure to grow on nutrient agar. A total of 100 such isolates were, tested for beta-lactamase production by use of nitrocefin. None of these isolates were beta-lactamase producers. A who reference beta lactamase positive strain (No. 5731) was the control.


Relations of Demographic and Clinical Factors With Cardiovascular Autonomic Function in a Population-Based Study: An Assessment By Quantile Regression.
The relationships of many factors with cardiovascular autonomic function (CVAF) outcome parameters may not be uniform across the entire distribution of the outcome. We examined how demographic and clinical factors varied with different subgroups of CVAF parameters. Quantile regression was applied to a cross-sectional analysis of 4,167 adults (56% male; age range, 50-84 years) from 4 ethnic groups (3,419 New Zealand European, 303 Pacific, 227 Maori, and 218 South Asian) and without diagnosed cardiac arrhythmia. Pulse rate variability (root mean square of successive differences (RMSSD) and SD of pulse intervals) and baroreflex sensitivity were response variables. Independent variables were age, sex, ethnicity, brachial and aortic blood pressure (BP) variables, body mass index (BMI), and diabetes. Ordinary linear regression showed that age, sex, Pacific and Maori ethnicity, BP variables, BMI, and diabetes were associated with CVAF parameters. But quantile regression revealed that, across CVAF percentiles, the slopes for these relationships: (i) varied by more than 10-fold in several cases and sometimes changed direction and (ii) noticeably differed in magnitude often (by >3-fold in several cases) compared to ordinary linear regression coefficients. For instance, age was inversely associated with RMSSD at the 10th percentile of this parameter (β = -0.12 ms/year, 95% confidence interval = -0.18 to -0.09 ms/year) but had a positive relationship at the 90th percentile (β = 3.17 ms/year, 95% confidence interval = 2.50 to 4.04 ms/year). The relationships of demographic and clinical factors with CVAF parameters are, in many cases, not uniform. Quantile regression provides an improved assessment of these associations.


Quality of evidence supporting the role of Chinese herbal medicine for the treatment of poststroke depression: A protocol for systematic review and meta-analysis.
Poststroke depression (PSD) is a syndrome that occurs after stroke. The efficacy of Chinese herbal medicine (CHM) for PSD has also received widespread attention, but there is still a lack of clinical evidence because this overview evaluates the published meta-analyses (MAs)/Systematic reviews (SRs). To provide evidence for the clinical application of CHM in the treatment of PSD. Two researchers searched 7 databases for SRs/MAs which are about randomized controlled trials on CHM for PSD. Two investigators use the systematic review assessment tool (AMSTAR-2), the risk of bias in systematic scale, the list of preferred reporting items for systematic reviews and meta-analysis, and the classification of recommended assessments for evaluation, development and evaluation system to assess the included SRs/MAs. Our findings will be published in peer-reviewed journals. This study provides evidence-based medical evidence for the impact of CHM on PSD. INPLASY202210001.


A comparison of radiographic degeneration features of older Chinese women and older Italian Caucasian women with a focus on thoracic spine.
Compared with Caucasians, East Asians have a lower incident of back pain, lower prevalence and severity of osteoporotic vertebral fracture and lumbar spine degeneration. This study compares radiographic spine degeneration features of older Chinese women (as an example of East Asians) and older Italian women (as an example of Caucasians) with a focus on the thoracic spine. From two population-based epidemiological studies conducted in Hong Kong, China and Rome, Italy, 297 pairs (mean age: 73.6 years) age-matched older community women's lateral spine radiographs were sampled. Existence (or absence) of seven degeneration features were assessed including: (1) hyper-kyphosis, (2) disc space narrowing (T3/T4 ~ T11/T12), (3) osteoarthritic (OA) wedging (T4 ~ T12), (4) generalised osteophyte formation (T4 ~ T12); (5) acquired short vertebrae (T4 ~ T12), (6) Schmorl node (T4 - L5), (7) disc calcification (T4-L5). Italian women were more likely to have hyper-kyphosis (53.4% vs 25.6%), disc space narrowing (34.4% vs. 17.2%), OA wedging (6.4% vs. 0.67%), Schmorl node (19.5% vs. 4.4%, all P < 0.001). However, there was no statistically significant difference in osteophyte formation (7.7% vs. 9.4%, P > 0.1) and acquired short vertebrae (8.0% vs. 10.4%, P > 0.1). Disc calcification was uncommon among both Chinese and Italians. For the first time, this study documented a lower prevalence of a number of thoracic spine degeneration features among Chinese. This study further affirms the concept of a generally healthier spine in older Chinese relative to older Caucasians. The observed differences may reflect a foundational background influence of genetic predisposition that represents an important line of future research.


On the Design and Implementation of a Highly Accurate Pulse Predictor for Exercise Equipment.
This study aims to develop highly accurate heart rate monitoring from the hand-held contact signal within a noisy environment during exercise. The periodic pattern and uncertainties of a physiological signal are modeled by a Laplacian random process. Based on this statistical model, a highly accurate pulse predictor (HAPPEE) is derived and implemented in real-time on a Cypress PSoC 5LP development board. A real-time experiment is designed to compare HAPPEE with a commercial heart rate monitor from POLAR. The percentage of credible estimates and the mean square error (MSE) of credible estimates are reported for experiments with seven healthy subjects. The overall percentage of credible estimates is 99.2% for HAPPEE and 93.6% for POLAR. The overall MSE of credible estimates is 3.1 for HAPPEE and 7.7 for POLAR. These results show that HAPPEE is more accurate than POLAR. HAPPEE is able to accurately monitor heart rate within a noisy environment during exercise. Unlike existing heart rate estimation methods, HAPPEE does not require pulse detection or tuning parameters. It can be easily implemented in real-time on a low power and low cost development board for exercise equipment and outperforms a commercial heart rate monitor.


Reliability of ultrasonography compared to radiography in revealing intestinal sand accumulations in horses.
In order to investigate the reliability of ultrasonography in revealing intestinal sand accumulations, the cranioventral parts of the abdomen of 32 horses (age 3-20 years) with signs suggestive of gastrointestinal sand were examined radiographically and ultrasonographically. The amount of sand and its localisation, relative to the ventral abdominal wall, were evaluated radiographically and graded (scale 0-4). Motility of the intestine and whether it was lying against the ventral abdominal wall were evaluated ultrasonographically and the findings summed and graded (scale 0-3) to express the likelihood of the presence of sand. The ultrasonographic and radiographic grades were significantly associated. Out of 14 horses with a moderate or large ventral sand accumulation radiographically, 13 (92.9%) had a positive sand finding ultrasonographically. Out of 8 horses with no signs of sand radiographically, 7 were considered negative for sand ultrasonographically. The specificity of ultrasonography in detecting sand accumulations was 87.5% (7/8) and the sensitivity 87.5% (21/24). Small and more dorsally located accumulations were more difficult to detect ultrasonographically. Of the horses with a small or moderate amount of sand relatively ventrally or only a small part of sand close to the ventral abdominal wall, 70% (7/10) were considered suggestive or positive for sand ultrasonographically. The ventral aspect of sand accumulations was hyperechoic, causing varying acoustic shadowing and the intestine had decreased or absent motility. Ultrasonography revealed the length of the accumulations but gave very limited information of their height. Ultrasonography is a practical and reliable method for detecting sand accumulations but it cannot replace radiography.


Respiratory syncytial virus infection in anti-mu-treated mice.
BALB/c mice were depleted of B cells by anti-mu treatment to investigate the pathogenesis of respiratory syncytial virus (RSV) infection in the absence of antibody. Termination of RSV replication after primary infection occurred with the same kinetics in anti-mu-treated mice as in phosphate-buffered saline (PBS)-treated controls. Yet, when rechallenged, anti-mu-treated mice were more permissive to RSV replication than PBS-treated controls. Anti-mu-treated mice also experienced greater illness than PBS-treated controls during both primary infection and rechallenge. Passive transfer of RSV-specific immune serum to anti-mu-treated mice before rechallenge reconstituted complete protection from RSV replication and diminished illness. Thus, RSV-specific antibody is not required to terminate RSV replication in primary infection, but without antibody, only partial immunity against rechallenge is induced. While it is unknown whether the mechanism is a direct effect on RSV titer or modulation of the illness-causing cellular immune response, the presence of RSV-specific antibody reduces illness in both primary RSV infection and rechallenge of mice.


Further Evidence that Cytoplasmic Acidosis Is a Determinant of Flooding Intolerance in Plants.
We present two pieces of evidence that regulation of cytoplasmic pH near neutrality is a prerequisite for survival of root tips during hypoxia. First, blackeye peas and navy beans show earlier cytoplasmic acidosis under hypoxia than soybeans or pumpkin or maize, and die earlier. Second, when cytoplasmic acidosis in maize root tips is greatly retarded by treatment with 25 millimolar Ca(NO(3))(2), they remain viable under hypoxia for a much longer period of time than untreated hypoxic root tips. We also show that viability of maize root tips is unaffected by the supply of exogenous sugar (and so on the rate of ethanolic fermentation) for at least 16 hours of hypoxia.


The efficacy and safety of traditional Chinese medicine Guilu Erxian Jiao in the treatment of knee osteoarthritis: A systematic review and meta-analysis.
A systematic review was conducted to investigate the efficacy of Guilu Erxian Jiao (GEJ) in the treatment of knee osteoarthritis (OA). We searched PubMed, MEDLINE, EMBASE, the Cochrane Central Register of Controlled Trials, Chinese Electronic Periodical Services, and ClinicalTrials.gov to identify relevant randomized controlled trials or controlled clinical trials, from the inception of each source to April 20, 2021. Primary outcome included overall efficacy, pain score, and Lequesne index score; secondary outcome included adverse events. Methodological quality was assessed using the Cochrane risk of bias tool (RoB 1.0). The meta-analysis was performed based on a random-effects model due to anticipated clinical heterogeneity. The grading of overall evidence was assessed using the GRADE system. The study protocol was registered on PROSPERO (CRD42021233573). Eight studies were included. Compared to controls, GEJ exhibited superior overall efficacy for treating OA (risk ratio (RR) = 1.20; 95% confidence interval (CI) = 1.06-1.35). Regarding pain score, there was no statistical difference between GEJ and controls (standardized mean difference (SMD) = 0.27; 95% CI = -0.91 - 1.46). No significant difference was found in Lequesne score between GEJ and controls (MD = -0.25; 95% CI = -0.52 - 0.01). No statistical difference in adverse reactions was observed between GEJ and controls (risk difference (RD) = -0.01; 95% CI = -0.05-0.03). Our findings suggest that GEJ may have positive effects on overall efficacy in treating OA. However, there is insufficient evidence regarding pain score, Lequesne score, and knee joint function score.


Single molecule optical measurements of orientation and rotations of biological macromolecules.
Subdomains of macromolecules often undergo large orientation changes during their catalytic cycles that are essential for their activity. Tracking these rearrangements in real time opens a powerful window into the link between protein structure and functional output. Site-specific labeling of individual molecules with polarized optical probes and measurement of their spatial orientation can give insight into the crucial conformational changes, dynamics, and fluctuations of macromolecules. Here we describe the range of single molecule optical technologies that can extract orientation information from these probes, review the relevant types of probes and labeling techniques, and highlight the advantages and disadvantages of these technologies for addressing specific inquiries.


Mechanism of alpha-amino acids decomposition in the gas phase. experimental and theoretical study of the elimination kinetics of N-benzyl glycine ethyl ester.
The gas-phase elimination kinetics of N-benzylglycine ethyl ester was examined in a static system, seasoned with allyl bromide, and in the presence of the free chain radical suppressor toluene. The working temperature and pressure range were 386.4-426.7 degrees C and 16.7-40.0 torr, respectively. The reaction showed to be homogeneous, unimolecular, and obeys a first-order rate law. The elimination products are benzylglycine and ethylene. However, the intermediate benzylglycine is unstable under the reaction conditions decomposing into benzyl methylamine and CO(2) gas. The variation of the rate coefficients with temperature is expressed by the following Arrhenius equation: log k(1) (s(-1)) = (11.83 +/- 0.52) - (190.3 +/- 6.9) kJ mol(-1) (2.303RT)(-1). The theoretical calculation of the kinetic parameters and mechanism of elimination of this ester were performed at B3LYP/6-31G*, B3LYP/6-31+G**, MPW1PW91/6-31G*, and MPW1PW91/6-31+G** levels of theory. The calculation results suggest a molecular mechanism of a concerted nonsynchronous six-membered cyclic transition state process. The analysis of bond order and natural bond orbital charges implies that the bond polarization of C(=O)O-C, in the sense of C(=O)O(delta-)...C(delta+), is rate determining. The experimental and theoretical parameters have been found to be in reasonable agreement.


Low-Dimensional Organic Crystals: From Precise Synthesis to Advanced Applications.
Low-dimensional organic crystals (LOCs) have attracted increasing attention recently for their potential applications in miniaturized optoelectronics and integrated photonics. Such applications are possible owing to their tunable physicochemical properties and excellent charge/photon transport features. As a result, the precise synthesis of LOCs has been examined in terms of morphology modulation, large-area pattern arrays, and complex architectures, and this has led to a series of appealing structure-dependent properties for future optoelectronic applications. This review summarizes the recent advances in the precise synthesis of LOCs in addition to discussing their structure-property relationships in the context of optoelectronic applications. It also presents the current challenges related to organic crystals with specific structures and desired performances, and the outlook regarding their use in next-generation integrated optoelectronic applications.


[Elucidation of the mechanism of fertilization and clinical application of assisted reproductive technology].
Fertilization is the process including many events such as maturation of egg and sperm, attachment, binding, acrosomal reaction, penetration, fusion, cortical reaction, zona reaction and nuclear fusion of both gamete, whereby individual gametes from the female and male unite to create offspring. Although the reason for mechanism of fertilization is still not clearly understood, this process may accelerate the rate adaptation in evolution. In this special lecture, I would like to present our experimental and clinical results especially concerning with morphological, physiological, biochemical and molecular approach on the mechanism of fertilization. 1. Development and maturation of follicles and oocytes. It is well known that pituitary FSH, LH control the ovarian function. Follicular development and ovum maturation are also controlled by both pituitary gonadotropins and local factors such as autocrine and paracrine agents. When hMG is injected during 1-6 day of menstrual cycle, several dominant follicles are developed. If hMG is injected after selection of dominant follicles, only one dominant follicle develop in the ovary. When PMS-treated immature rats were injected with immature or mature follicle fluids, rats injected with mature follicular fluid showed strongly suppress in the ovarian weights and numbers of ovulated follicles. Also mature follicle suppress aromatization from and androstenedione to estradiol. These findings mean that mature follicular fluid contains inhibitory factors. Apoptosis of granulosa cells and follicular steroids are related to fertilization. 2. Intracellular calcium of oocyte. Intracellular calcium concentration is known to start to increase in a periodic manner after fertilization in oocytes of mammalians. In 65% of tested mouse oocytes, fertilization occurred during 4 hours observation after sperm insemination in vitro. An initial long lasting intracellular calcium concentration was observed and followed by periodic manner. This calcium oscillation is inhibited by calcium blockers such as verpamil and nifedipine, but increased by high concentration of extracellular calcium concentration in the medium. Role of increase of intracellular calcium are understood to prevent polysperm and activate metabolism of oocytes. 3. Glucose metabolism of oocytes. Mouse embryo utilizes pyruvate as an essential nutrient until the 8-cell stage, and glucose thereafter. We have devised non-radiometrie and enzymatic microassay method to measure glucose, deoxyglucose, deoxyglucose 6-phosphate incorporated into individual mouse oocytes and preimplantation embryo. In parallel, the activities of several enzymes of glycolytic pathway were also determined. In this study, glucose metabolism is necessary to develop in fertilized ova with changing activity of enzymes. 4. Molecular bases of ovarian fluid. The zona pellucida ZP is involved in a number of events in fertilization, all these fertilization events occur in the oviduct. Oviductal glycoprotein 200-240 KD has been identified from oviductal zona pellucida. Monoclonal antibody of oviductal glycoprotein reacted with ZP of oviductal egg but not with the ovarian egg. Anti-ZPO antibody inhibit to bind sperm to ZP. Sequences in mouse and hamster oviduct specific glycoprotein are estimated, this glycoprotein mRNA was observed in only oviduct by northern blotting method. These molecular gene expression was observed by in situ hybridization in the oviduct of estrous cycle of hamster. 5. Microinsemination of sperm. Microinsemination of sperm into oocyte is widely used in clinical medicine. Sperm penetration assay (hamster test) is useful method to estimate fertilization capacity of sperm. But immotile sperm cannot estimate it. So modified micro sperm penetration assay was established to estimate fertilization capacity of sperm by using micro-manipulator. Subzonal sperm injection (SUZI) and intracytoplasmic sperm injection (ICSI) promotes fertilization and cleavage rate in immotile


Comparing SF-36 Scores Collected Through Web-Based Questionnaire Self-completions and Telephone Interviews: An Ancillary Study of the SENTIPAT Multicenter Randomized Controlled Trial.
The 36-Item Short Form Health Survey (SF-36) is a popular questionnaire for measuring the self-perception of quality of life in a given population of interest. Processing the answers of a participant comprises the calculation of 10 scores corresponding to 8 scales measuring several aspects of perceived health and 2 summary components (physical and mental). Surprisingly, no study has compared score values issued from a telephone interview versus those from an internet-based questionnaire self-completion. This study aims to compare the SF-36 score values issued from a telephone interview versus those from an internet-based questionnaire self-completion. Patients with an internet connection and returning home after hospital discharge were enrolled in the SENTIPAT multicenter randomized trial on the day of discharge. They were randomized to either self-completing a set of questionnaires using a dedicated website (internet group) or providing answers to the same questionnaires administered during a telephone interview (telephone group). This ancillary study of the trial compared SF-36 data related to the posthospitalization period in these 2 groups. To anticipate the potential unbalanced characteristics of the responders in the 2 groups, the impact of the mode of administration of the questionnaire on score differences was investigated using a matched sample of individuals originating from the internet and telephone groups (1:1 ratio), in which the matching procedure was based on a propensity score approach. SF-36 scores observed in the internet and telephone groups were compared using the Wilcoxon-Mann-Whitney test, and the score differences between the 2 groups were also examined according to Cohen effect size. Overall, 29.2% (245/840) and 75% (630/840) of SF-36 questionnaires were completed in the internet and telephone groups, respectively (P<.001). Globally, the score differences between groups before matching were similar to those observed in the matched sample. Mean scores observed in the telephone group were all above the corresponding values observed in the internet group. After matching, score differences in 6 out of the 8 SF-36 scales were statistically significant, with a mean difference greater than 5 for 4 scales and an associated mild effect size ranging from 0.22 to 0.29, and with a mean difference near this threshold for 2 other scales (4.57 and 4.56) and a low corresponding effect size (0.18 and 0.16, respectively). The telephone mode of administration of SF-36 involved an interviewer effect, increasing SF-36 scores. Questionnaire self-completion via the internet should be preferred, and surveys combining various administration methods should be avoided. ClinicalTrials.gov NCT01769261; https://www.clinicaltrials.gov/ct2/show/record/NCT01769261.


Discriminant analyzing system for wood wastes using a visible-near-infrared chemometric imaging technique.
A new optical system was developed and applied to automated separation of wood wastes, using a combined technique of visible-near-infrared (Vis-NIR) imaging analysis and chemometrics. Three kinds of typical wood wastes were used, i.e., non-treated, impregnated, and plastic-film overlaid wood. The classification model based on soft independent modeling of class analogy (SIMCA) was examined using the difference luminance brightness of a sample. Our newly developed system showed a good/promising performance in separation of wood wastes, with an average rate of correct separation of 89%. Hence, it is concluded that the system is efficiently feasible for online monitoring and separation of wood wastes in recycling mills.


Old challenges and new areas for international co-operation in population.
Taking stock of accomplishments in the field of population reveals that significant progress has been made since the late 1960s but that much remains to be done. Important challenges in population for 1990 and beyond include the implementation of more effective family planning programs, greater accessibility to better family planning services at the local level, a wider range of choices in contraceptive methods, and better training and supervision of family planning delivery personnel. Another major challenge is to give attention to the various aspects of the role of women--beyond mere acknowledgement and to the actual implementation of programs. Further, policies need to be formulated and implemented across several sectors to deal with the complex interaction between population, resources, and the environment. To devise such policies, knowledge of the interrelationships needs to be clarified and refined. Finally, still greater emphasis will have to be placed on improving the integration of population and development. Accomplishing that will require wider awareness, enhanced coordination and adequate resources--an increase of at least $100 million per year from now to the end of the century over the annual current level of some $550 million for all external assistance for population.


Assessment of retinal hemodynamics with the Canon laser blood flowmeter after a single dose of 2% dorzolamide hydrochloride eyedrops.
Dorzolamide hydrochloride is a carbonic anhydrase inhibitor that reduces intraocular pressure (IOP) by decreasing the production of aqueous humour in the ciliary body. Theoretically, topical use of this agent has the potential to directly affect retinal vasculature through local induced acidosis. We performed a study to determine whether there are changes in retinal arteriole hemodynamics, as assessed with the Canon laser blood flowmeter, in healthy subjects following topical administration of dorzolamide. We recruited 17 healthy volunteers, nine men and eight women aged 25 to 55 years (mean 31.4 [standard deviation (SD) 9.88] years). The inclusion criteria were Snellen visual acuity of 20/30 or better, normal anterior eye examination, IOP of 21 mm Hg or less, and a normal fundus appearance. One eye of each subject was randomly assigned to receive a drop of 2% dorzolamide. The contralateral eye of 10 of the subjects received a placebo drop (artificial tears). Before and 1 hour after drop administration, we obtained blood flow measurements from an inferotemporal arteriole approximately 1 disc diameter from the optic nerve head rim using the Canon laser blood flowmeter, model 100. The IOP was measured by means of Goldmann applanation tonometry before and 1 hour after drop administration. The mean IOP was significantly reduced in the dorzolamide-treated eyes, from 14.4 mm Hg (SD 2.94 mm Hg) to 11.7 mm Hg (SD 2.50 mm Hg) (p < 0.001). The IOP was also reduced in the placebo group (15.6 mm Hg [SD 3.41 mm Hg] vs. 14.6 mm Hg [SD 3.28 mm Hg]), but the difference was not significant. There was no significant difference in mean arteriole diameter, mean blood velocity or mean blood flow after drug administration in the dorzolamide-treated eyes. Our results indicate that a single topical application of dorzolamide in healthy subjects has no effect on retinal arteriole diameter, blood velocity or blood flow, as measured with the Canon laser blood flowmeter. Longer-term studies of retinal hemodynamics in patients with glaucoma are warranted as evolving treatments aim to improve ocular blood flow as well as reduce IOP.


The gene distribution of the maize genome.
Previous investigations from our laboratory showed that the genomes of plants, like those of vertebrates, are mosaics of isochores, i.e., of very long DNA segments that are compositionally homogeneous and that can be subdivided into a small number of families characterized by different GC levels (GC is the mole fraction of guanine+cytosine). Compositional DNA fractions corresponding to different isochore families were used to investigate, by hybridization with appropriate probes, the gene distribution in vertebrate genomes. Here we report such a study on the genome of a plant, maize. The gene distribution that we found is most striking, in that almost all genes are present in isochores covering an extremely narrow (1-2%) GC range and only representing 10-20% of the genome. This gene distribution, which seems to characterize other Gramineae as well, is remarkably different from the gene distribution previously found in vertebrate genomes.


Respiratory arrest in an obstetric patient using remifentanil patient-controlled analgesia.
Remifentanil patient-controlled analgesia is well established in many centres and provides satisfactory pain relief for many women in labour. We describe a patient using remifentanil patient-controlled analgesia who suffered a respiratory arrest requiring a brief period of ventilation. In our institution, remifentanil patient-controlled analgesia has been offered to women in labour since 2009. Up to this point, we had not observed any critical incidents in over 130 patients using this mode of analgesia in our labour suite.


Gender, education, and fertility: a cross-national analysis of Sub-Saharan African nations.
According to the demographic transition theory and the wealth flows model, it is expected that fertility will decline with socioeconomic development, manifested in part through increasingly greater proportions of the population with formal education. Since their independence in the 1960s, most sub-Saharan African nations have experienced rapid changes in educational levels. However, recent estimates indicate that high levels of fertility are being maintained as reflected in the high rates of population increase of approximately 3%/year. Controlling for socioeconomic development as measured by per capita energy consumption and percentage of labor force in agriculture, this article examines the relationship between education and fertility for men and women in 37 sub-Saharan nations. Results indicate that primary school enrollment in 1960 and 1980 for both males and females had a weak negative and nonsignificant relationship with the total fertility rate 15-30 years later. Secondary school enrollment in 1960 for both males and females had weak relationships with the total fertility rate. However, secondary school enrollment for males in 1980 had a significant negative effect on the total fertility rate 10-25 years later. Implications are discussed.


Prevalence of and Theoretical Explanation for Type 2 Benign Paroxysmal Positional Vertigo.
A variant of benign paroxysmal positional vertigo (BPPV) involves the subjective report of vertigo without the coinciding nystagmus. This presentation includes truncal retropulsion when sitting up from the ipsilesional provocative test (ie, Dix-Hallpike), which we term type 2 BPPV. The primary objective of this study is to prospectively determine the prevalence and describe the clinical course of type 2 BPPV. We offer a theoretical explanation for the absence of nystagmus. Prospective, observational study carried out in 2 tertiary hospitals. One hundred eighty patients (134 women, 46 men) met the inclusion criteria and were included between January 10, 2018, and October 30, 2019. Efficacy of physical therapy maneuvers was determined at 1-week follow-up. Three-dimensional reconstructions of the planes of the semicircular canal cupula from histological preparations are offered as evidence for the theoretical explanation. One-third of the patients met the criteria for type 2 BPPV; the remainder had typical posterior or horizontal semicircular canal involvement. Symptoms from type 2 BPPV were longer in duration yet responded favorably to physical therapy maneuvers. Upon repeat testing, 19 patients treated for posterior canalithiasis developed a slight, persistent positional downbeat nystagmus in the Dix-Hallpike position that we propose as evidence the otoconia entered the short arm of the posterior semicircular canal. Our data and 3-dimensional rendering suggest the report of vertigo, yet absent nystagmus in type 2 BPPV is from otoconia aligning with the gravitoinertial vector during provocative testing that precludes cupular stimulation. Type 2 BPPV appears to be a common and treatable form of vertigo.Video Abstract available for more insights from the authors (see Video, Supplemental Digital Content 1 available at: http://links.lww.com/JNPT/A372).


Intralesional (incision) biopsy for melanoma diagnosis: the rules and the exception.
Intralesional (incision) biopsy for melanoma diagnosis can be warranted for large lesions or for those lesions whose in-toto excision leads to cosmetic and/or functional impairment. However, this diagnostic approach carries a risk of underdiagnosis, if a clinicopathologic diagnostic approach is not implemented. As a rule, in large pigmented lesions from special body areas (scalp and acral skin), clinicodermoscopic differential diagnosis of melanoma includes non-melanocytic skin lesions, traumatic skin changes, and nevi. The unique indication to incision biopsy for the differential diagnosis between nevus and melanoma is a relatively small nodular proliferation developing within a medium-large congenital nevus.


The E8E2C protein, a negative regulator of viral transcription and replication, is required for extrachromosomal maintenance of human papillomavirus type 31 in keratinocytes.
The viral E2 protein is a major regulator of papillomavirus DNA replication. An important way to influence viral replication is through modulation of the activity of the E2 protein. This could occur through the action of truncated E2 proteins, called E2 repressors, whose role in the replication cycle of human papillomaviruses (HPVs) has not been determined. In this study, using cell lines that contain episomal copies of the "high-risk" HPV type 31 (HPV31), we have identified viral transcripts with a splice from nucleotide (nt) 1296 to 3295. These transcripts are similar to RNAs from other animal and human papillomaviruses and have the potential to fuse a small open reading frame (E8) to the C terminus of E2, resulting in an E8E2C fusion protein. E8E2C transcripts were present throughout the complete replication cycle of HPV31. A genetic analysis of E8E2C in the context of the HPV31 genome revealed that mutation of the single ATG of the E8 gene, introduction of a stop codon downstream of the ATG, or disruption of the splice donor site at nt 1296 led to a dramatic 30- to 40-fold increase in the transient DNA replication levels in both normal and immortalized human keratinocytes. High-level expression of E8E2C from heterologous vectors was found to inhibit E1-E2-dependent DNA replication of an HPV31 origin of replication construct as well as to interfere with E2's ability to transactivate reporter gene constructs. In addition, HPV31 E8E2C strongly repressed the basal activity of the major viral early promoter P97 independent of E2. E8E2C may therefore exert its negative effect on viral DNA replication through modulating E2's ability to enhance E1-dependent DNA replication as well as by regulating viral gene expression. Surprisingly, HPV31 genomes that were unable to express E8E2C could not be maintained extrachromosomally in human keratinocytes in long-term assays despite high transient DNA replication levels. This suggests that the E8E2C protein may play a role in copy number control as well as in the stable maintenance of HPV episomes.


Experimental tooth clenching. A model for studying mechanisms of muscle pain.
The overall goal of this thesis was to broaden knowledge of pain mechanisms in myofascial temporomandibular disorders (M-TMD). The specific aims were to: Develop a quality assessment tool for experimental bruxism studies (study I). Investigate proprioceptive allodynia after experimental tooth clenching exercises (study II). Evaluate the release of serotonin (5-HT), glutamate, pyruvate, and lactate in healthy subjects (study III) and in patients with M-TMD (study IV), after experimental tooth clenching exercises. In (I), tool development comprised 5 steps: (i) preliminary decisions, (ii) item generation, (iii) face-validity assessment, (iv) reliability and discriminative validity testing, and (v) instrument refinement. After preliminary decisions and a literature review, a list of 52 items to be considered for inclusion in the tool was generated. Eleven experts were invited to participate on the Delphi panel, of which 10 agreed. After four Delphi rounds, 8 items remained and were included in the Quality Assessment Tool for Experimental Bruxism Studies (Qu-ATEBS). Inter-observer reliability was acceptable (k = 0.77), and discriminative validity high (phi coefficient 0.79; P < 0.01). During refinement, 1 item was removed; the final tool comprised 7 items. In (II), 16 healthy females participated in three 60-min sessions, each with 24- and 48-h follow-ups. Participants were randomly assigned to a repetitive experimental tooth clenching task with a clenching level of 10%, 20%, or 40% of maximal voluntary clenching force (MVCF). Pain intensity, fatigue, perceived intensity of vibration (PIV), perceived discomfort (PD), and pressure pain threshold (PPT) were measured throughout. A significant increase in pain intensity and fatigue but not in PD was observed over time. A significant increase in PIV was only observed at 40 min, and PPT decreased significantly over time at 50 and 60 min compared to baseline. In (III), 30 healthy subjects (16 females, and 14 males) participated in two sessions at a minimum interval of 1 wk. Microdialysis was done to collect 5-HT, glutamate, pyruvate, and lactate and to measure masseter muscle blood flow. Two hours after the start of microdialysis, participants were randomized to a 20-min repetitive experimental tooth clenching task (50% of MVCF) or a control session (no clenching). Pain intensity was measured throughout the experiment. Substance levels and blood flow were unaltered at all time points between sessions, and between genders in each session. Pain intensity was significantly higher after clenching in the clenching session compared to the same time point in the control session. In (IV), 15 patients with M-TMD and 15 healthy controls participated in one session and the methodology described above was used. M-TMD patients had significantly higher levels of 5-HT and significantly lower blood flows than healthy controls. No significant differences for any substance at any time point were observed between groups. Time and group had significant main effects on pain intensity. Qu-ATEBS, the 7-item evidence-based quality assessment tool, is reliable, exhibits face-validity, and has excellent discriminative validity. Tooth clenching was associated with pain, fatigue, and short-lasting mechanical hyperalgesia, but not with proprioceptive allodynia. It seems that tooth clenching is not directly related to delayed onset muscle soreness. In healthy subjects and in patients with M-TMD, levels of 5-HT, glutamate, pyruvate, and lactate were unaltered after tooth clenching. But 5-HT levels were significantly higher and blood flows significantly lower in M-TMD patients than in healthy controls at all time points. These two factors may facilitate the release, and enhance the effects, of other algesic substances that may cause pain.


Protective effect of early low-dose hydrocortisone on ventilator-associated pneumonia in the cancer patients: a propensity score analysis.
Ventilator-associated pneumonia (VAP) is a care-related event that could be promoted by immune suppression caused by critical diseases, malignancies and cancer treatments. Low dose of hydrocortisone was proposed for modulation of immune response in the critically ill population. In this monocentric observational study, all cancer patients mechanically ventilated for more than 48 h were included. Effect of low-dose hydrocortisone administered during the first 48 h of mechanical ventilation was evaluated applying inverse probability weighting analysis after propensity score assessment. VAP impact on 1-year mortality, ICU length of stay and mechanical ventilation duration was secondarily determined. Within this cohort, 190 cancer patients were followed. VAP was confirmed in 22.1% of cases in the early hydrocortisone group and confirmed in 42.6% of cases in the no or late hydrocortisone group. Early hydrocortisone exhibited a protective effect on the risk of VAP (OR 0.23; 95% CI 0.12-0.44; P < 0.0001). VAP was associated with 1-year mortality (HR 1.60; 95% CI 1.10-2.34; P = 0.017) and increased ICU length of stay (mean extra length of stay: 4.2 days; 95% CI 0.6-7.8). Immune modulation with low-dose hydrocortisone administered in the first days of mechanical ventilation could protect from VAP occurrence in cancer patients.


The mechanism of white cell reduction by synthetic fiber cell filters.
The mechanism of white cell (WBC) retention by synthetic fiber-based WBC filters was studied. Filters were made of nonwoven fleece prepared from polyester, surface-modified polyester, or polypropylene fibers. Human platelet concentrates were filtered through experimental filters consisting of 8 to 54 layers of nonwoven fleece with mean pore sizes from 7.3 to 14.2 microns. Filters made of fleece of smaller pore size removed WBCs less effectively than filters with larger-pore fleece. Retention of lymphocytes and granulocytes gradually dropped to 0 percent as increasing loads were applied to the filters. The maximal retention capacity for these cell types (i.e., the number of cells retained when "saturating" numbers of WBCs were applied) was proportional to the number of layers of filter material used. Platelet retention did not correlate with WBC retention. Depth filtration, rather than mechanical sieving, seems to be the principal means of WBC removal by nonwoven fiber filters. A low initial number of WBCs in the component to be filtered is important for successful WBC filtration.


Associations Between Genetic Polymorphisms in the VEGFA, ACE, and SOD2 Genes and Susceptibility to Diabetic Nephropathy in the Han Chinese.
Objective: To investigate the association between polymorphisms in the rs2010963 and rs69947 loci of the vascular endothelial growth factor A (VEGFA) gene; the rs4646994 locus of the angiotensin I converting enzyme (ACE) gene; and the rs4880 locus of the superoxide dismutase 2 (SOD2) gene with genetic susceptibility to type 2 diabetic nephropathy (T2DN) in the Chinese Han population. Methods: A total of 650 Chinese Han patients with T2DN and 580 non-nephropathy patients with type 2 diabetes were enrolled in this study. Sanger sequencing was used to detect the genotypes of the rs2010963 and rs69947 loci within VEGFA, the rs4646994 locus of for the ACE gene, and the rs4880 locus of the SOD2 gene in all subjects. Enzyme-linked immunosorbent assays were used to detect VEGFA, ACE, and SOD2 levels in serum. Results: The risk of T2DN was significantly increased (odds ratio [OR] = 1.15, confidence interval [95% CI]: 1.03-1.30) in patients with the GC/CC genotypes compared to those with the GG genotype at the rs2010963 locus of the VEGFA gene under the dominant model of inheritance. Similarly, the risk of T2DN was significantly increased (OR = 1.17, 95% CI: 1.05-1.31) in patients with the CA/AA genotypes compared to those with the CC genotype at the rs69947 locus of the VEGFA gene under the dominant model of inheritance. In addition, the risk of T2DN was increased (OR = 1.57, 95% CI: 1.37-1.74) in patients with the AA genotype compared to the CC/CA genotypes under the recessive model of inheritance. The risk of T2DN was also significantly increased (OR = 1.54, 95% CI: 1.24-1.97) in patients with the ID/DD genotypes compared to those with the II genotype at the rs4646994 locus of the ACE gene under the dominant model; and (OR = 1.41, 95% CI: 1.26-1.57) for DD genotype compared to the II/ID genotypes under the recessive model. We also found the risk of T2DN was significantly increased (OR = 1.25, 95% CI: 1.11-1.39) under the dominant model in patients with the TC/CC genotypes compared to the TT genotype, and for the CC genotype (compared to the TT/TC genotype) (OR = 1.45, 95% CI: 1.18-1.66) under the recessive model at the rs4880 locus of the SOD2 gene rs4880. The haplotypes GAC (OR = 1.17, 95% CI: 1.04-1.29), CAT (OR = 1.12, 95% CI: 1.03-1.60), and CAC (OR = 1.13, 95% CI: 1.01-1.24) constructed from VEGFA gene rs2010963 and rs69947 loci, and the SOD2 gene rs4880 locus were associated with a higher risk for T2DN. Finally, we found that VEGFA protein levels from subjects with the rs2010963 GG genotype and the rs69947 CC genotype were higher in both case groups and the control group than in subjects with rs2010963 GA/AA genotypes and rs69947 CA/AA genotypes, respectively (p < 0.05). The ACE protein levels for variants at the rs4646994 locus showed that the case group and control group subjects with the DD genotype had the highest levels, followed by the ID genotype and the II genotype (p < 0.05). Conclusion: Genetic variation in the VEGFA gene at the rs2010963 and rs69947 loci, the ACE gene at the rs4646994 locus, and the SOD2 gene at the rs4880 locus may increase the risk of developing T2DN.


Intrathecal gadolinium-enhanced MR cisternography: a comprehensive review.
CE-MRC has been in use for the past 15 years and was reported to be a useful method in the evaluation of CSF disorders and hydrocephalus. The use of CE-MRC in conjunction with other MR imaging techniques has been shown to be effective in selected cases for the evaluation of several disorders of cerebrospinal system. CE-MRC has certain advantages over other cisternographic studies with fewer side effects if performed properly. Although intrathecal Gd administration is not widely accepted yet, several recent studies have reported the safety of small-dose intrathecal gadolinium injection. In this review, we describe CE-MRC and review recent applications in several clinical conditions.


Changes of Proteases, Antiproteases, and Pathogens in Cystic Fibrosis Patients' Upper and Lower Airways after IV-Antibiotic Therapy.
In cystic fibrosis (CF) the upper (UAW) and lower airways (LAW) are reservoirs for pathogens like Pseudomonas aeruginosa. The consecutive hosts' release of proteolytic enzymes contributes to inflammation and progressive pulmonary destruction. Objectives were to assess dynamics of protease : antiprotease ratios and pathogens in CF-UAW and LAW sampled by nasal lavage (NL) and sputum before and after intravenous- (IV-) antibiotic therapy. From 19 IV-antibiotic courses of 17 CF patients NL (10 mL/nostril) and sputum were collected before and after treatment. Microbiological colonization and concentrations of NE/SLPI/CTSS (ELISA) and MMP-9/TIMP-1 (multiplex bead array) were determined. Additionally, changes of sinonasal symptoms were assessed (SNOT-20). IV-antibiotic treatment had more pronounced effects on inflammatory markers in LAW, whereas trends to decrease were also found in UAW. Ratios of MMP-9/TIMP-1 were higher in sputum, and ratios of NE/SLPI were higher in NL. Remarkably, NE/SLPI ratio was 10-fold higher in NL compared to healthy controls. SNOT-20 scores decreased significantly during therapy (P = 0.001). For the first time, changes in microbiological patterns in UAW and LAW after IV-antibiotic treatments were assessed, together with changes of protease/antiprotease imbalances. Delayed responses of proteases and antiproteases to IV-antibiotic therapy were found in UAW compared to LAW.


X-band cavity for ESR experiments.
We describe an x-band cavity for ESR experiments built to produce the modulation of the magnetic field without using any coil. The modulation is obtained with a current circulating in the cavity walls. The mechanical construction of this device is simple and inexpensive and allow the reduction of the magnetic air gap of the spectrometer.


Down-regulation of NF-kappa B protein levels in activated human lymphocytes by 1,25-dihydroxyvitamin D3.
The effect of 1,25-dihydroxyvitamin D3 [1,25(OH)2)D3], a steroid hormone with immunomodulating properties, on nuclear factor kappa B (NF-kappa B) proteins was examined in in vitro activated normal human lymphocytes by Western blot analysis. Over a 72-hr period of activation, the expression of the 50-kDa NF-kappa B, p50, and its precursor, p105, was increased progressively. When cells were activated in the presence of 1,25(OH)2D3, the levels of the mature protein as well as its precursor were decreased. The effect of the hormone on the levels of p50 was demonstrable in the cytosolic and nuclear compartments; it required between 4 and 8 hr and was specific, as 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 were ineffective. Besides p50, 1,25(OH)2D3 decreased the levels of another NF-kappa B protein, namely c-rel. In addition, 1,25(OH)2D3 decreased the abundance of a specific DNA-protein complex formed upon incubation of nuclear extracts from activated lymphocytes with a labeled NF-kappa B DNA binding motif. Further, 1,25(OH)2D3 inhibited the transcriptional activity of NF-kappa B in Jurkat cells transiently transfected with a construct containing four tandem repeats of the NF-kappa B binding sequence of the immunoglobulin kappa light chain gene linked to the chloramphenicol acetyltransferase reporter gene. These observations demonstrate directly that there is de novo synthesis of NF-kappa B during human lymphocyte activation and suggest that this process is hormonally regulated.


Patterns of innervation of sympathetic vascular neurons by peptide-containing primary sensory fibers.
The purpose of this study was to determine whether there is a specific organization of the primary sensory innervation on to identified vascular neurons in the inferior mesenteric ganglion (IMG) in guinea-pig. Retrograde tracers were placed intraluminally in inferior mesenteric artery (IMA) or inferior mesenteric vein (IMV) in vitro to identify ganglionic neurons as arterial, venous or unlabeled neurons. The distribution of primary sensory nerve fibers containing calcitonin gene-related peptide (CGRP), neuronal nitric oxide synthase (NOS) and substance P immunoreactivity (SP-IR) was compared before and after treatment with capsaicin. In control animals the density of immunoreactivity varied both with the transmitter and the type of neuron innervated. The density of immunoreactivity for all the three substances was reduced by capsaicin treatment. The degree of reduction of immunoreactivity in the fibers varied with the transmitter and the type of neuron. The density of CGRP and SP immunoreactive fibers was greatest around unlabeled neurons; 78% of the CGRP fibers were of primary sensory origin and all of the SP fibers were primary sensory. Around arterial neurons 44% of the CGRP fibers were of primary sensory origin and around venous 68% were primary sensory. NOS positive innervation around venous neurons was denser than around arterial neurons and all of it was completely (97%) eliminated by capsaicin, indicating that it was solely of primary sensory origin. We conclude that the primary sensory fibers innervating the IMG are differentially distributed to arterial and venous neurons and that the pattern of distribution is characteristic for each sensory neurotransmitter.


Regulatory response to washout of amniotic fluid in sheep.
To test the hypothesis that a substance present in the amniotic fluid could serve as a regulator of amniotic fluid volume, we drained and discarded amniotic fluid while replacing it with lactated Ringer solution that was isotonic to amniotic fluid. Seven ewes with singleton fetuses at 119 +/- 1 days of gestation (mean +/- SE) were instrumented with multiple indwelling catheters in the pedal artery, pedal vein, and amniotic cavity. During the exchange periods, an average of 3,019 +/- 171 ml/day of lactated Ringer solution was infused into the amniotic cavity while an equal amount of amniotic fluid was pumped out and discarded. During the control period, amniotic fluid composition and volume were not altered. Exchange and control periods started with the same amniotic fluid volume, lasted 3 or 4 days, and were randomized with regard to order. Amniotic fluid volume measured by vacuum drainage was 556 +/- 98 ml at the end of the control period and 986 +/- 209 ml (P = 0.03) at the end of the exchange period. Fetal arterial blood gases, hemodynamic parameters and the osmolality gradient between fetal plasma and amniotic fluid were not altered by the exchange process. A linear relationship between the control amniotic fluid volume and the volume at the end of the exchange period (P = 0.003) suggests that the animals with larger control volumes responded to isovolumic dilution with a larger volume increase. We conclude that amniotic fluid may contain a substance that regulates amniotic volume.


Correlation of the percentage of activated, CD3 + DR + lymphocytes to serum neopterin level in HIV-seropositive haemophiliacs.
The percentage of activated, CD3+ DR+ and CD8+ Leu7+ lymphocytes and the serum neopterin concentration were determined in 17 HIV-seropositive and 10 HIV-seronegative haemophiliacs and in 11 healthy control subjects. All three parameters tested were found to be significantly higher in the seropositive patients than in the seronegative controls. In the seropositive group, a significant positive correlation was found between the neopterin levels and the percentage of CD3+ DR+ cells. By contrast, no significant negative or positive correlation was observed between the neopterin levels and the percentage of the CD8+ Leu7+ subset. These data suggest that in the HIV-infected patients the activated T cells responsible for the stimulation of macrophages to produce neopterin are those that do not carry CD8.


Intraocular pressure in free-ranging anuran species in Oklahoma.
Ocular disease appears to be a common issue in anurans. Intraocular pressures were measured for six species of free-ranging anurans in central Oklahoma. No significant differences were identified between left or right eyes. There was a significant negative relationship between the weight of the anuran and intraocular pressure. The intraocular pressure range for the six species was 3-10 mm Hg. Tonometry values in anurans are, to the authors' knowledge, previously unreported and this study provides initial information on intraocular pressure measurement in anurans.


[Enzyme immunoassay of substances related to allergy].
The substances related to allergy that we can measure by EIA were described. Allergic reactions begin from the reaction of an antigen and IgE on the membrane surface of basophils and mast cells. The chemical mediators are released from the cells activated by the reaction of antigen and IgE. The chemical mediators proceed to the reactions of focal or total body inflammation. Measurements of serum IgE, chemical mediators, and cytokines are important parameters for the diagnosis and therapy of allergic disease. Antigen analyses are possible by application of EIA. It is also possible to apply EIA in the prevention of allergic diseases.


Canadian Cardiovascular Society 2023 Guidelines on the Fitness to Drive.
Cardiovascular conditions are among the most frequent causes of impairment to drive, because they might induce unpredictable mental state alterations via diverse mechanisms like myocardial ischemia, cardiac arrhythmias, and vascular dysfunction. Accordingly, health professionals are often asked to assess patients' fitness to drive (FTD). The Canadian Cardiovascular Society previously published FTD guidelines in 2003-2004; herein, we present updated FTD guidelines. Because there are no randomized trials on FTD, observational studies were used to estimate the risk of driving impairment in each situation, and recommendations made on the basis of Canadian Cardiovascular Society Risk of Harm formula. More restrictive recommendations were made for commercial drivers, who spend longer average times behind the wheel, use larger vehicles, and might transport a larger number of passengers. We provide guidance for individuals with: (1) active coronary artery disease; (2) various forms of valvular heart disease; (3) heart failure, heart transplant, and left ventricular assist device situations; (4) arrhythmia syndromes; (5) implantable devices; (6) syncope history; and (7) congenital heart disease. We suggest appropriate waiting times after cardiac interventions or acute illnesses before driving resumption. When short-term driving cessation is recommended, recommendations are on the basis of expert consensus rather than the Risk of Harm formula because risk elevation is expected to be transient. These recommendations, although not a substitute for clinical judgement or governmental regulations, provide specialists, primary care providers, and allied health professionals with a comprehensive list of a wide range of cardiac conditions, with guidance provided on the basis of the level of risk of impairment, along with recommendations about ability to drive and the suggested duration of restrictions.


Trends in Frailty and Its Association With Mortality: Results From the Longitudinal Aging Study Amsterdam, 1995-2016.
The aim of this study was to investigate trends in frailty and its relationship with mortality among older adults aged 64-84 years across a period of 21 years. We used data from 1995 to 2016 from the Longitudinal Aging Study Amsterdam. A total of 7,742 observations of 2,874 respondents in the same age range (64-84 years) across 6 measurement waves were included. Frailty was measured with a 32-item frailty index, with a cutpoint of ≥0.25 to indicate frailty. The outcome measure was 4-year mortality. Generalized estimating equation analyses showed that among older adults aged 64-84 years the 4-year mortality rate declined between 1995 and 2016, while the prevalence of frailty increased. Across all measurement waves, frailty was associated with 4-year mortality (odds ratio = 2.79, 95% confidence interval: 2.39, 3.26). There was no statistically significant interaction effect between frailty and time on 4-year mortality, indicating a stable association between frailty and mortality. In more recent generations of older adults, frailty prevalence rates were higher, while excess mortality rates of frailty remained the same. This is important information for health policy-makers and clinical practitioners, showing that continued efforts are needed to reduce frailty and its negative health consequences.


Contribution of PET imaging to the initial staging and prognosis of patients with Hodgkin's disease.
Positron emission tomographic (PET) scanning utilizing [18F]fluorodeoxyglucose (FDG) is a new method of tumor imaging based on the increased glucose metabolic activity of malignant tumors. In Hodgkin's disease (HD), PET has proven value for the evaluation of residual masses following treatment and for the early diagnosis of relapse. In the initial staging of HD, PET frequently shows a higher stage than conventional methods (upstaging by PET). In the present study, we evaluated the frequency of stage changes by PET in a multicenter setting and determined its prognostic relevance. A total of 73 patients with newly diagnosed HD were staged with both conventional methods and whole-body PET scanning. All histological types and stages were represented. The median time of follow-up after the initial diagnosis was 25 months (range 1 month to 5 years). The response to treatment was determined by standard clinical and diagnostic criteria. For the purpose of this analysis, data from a PET center associated with a university medical center and a PET center associated with a group oncology practice were combined. A total of 21 patients (28.8%) were upstaged by PET compared with conventional methods. In two cases (2.7%), a lower stage was suggested by PET scanning. With one possible exception, the upstaging had no obvious clinical or biological correlate. Among 12 patients in stage I (A + B) by conventional methods, seven were upstaged by PET (58.3%), four to stage II, one to stage III and two to stage IV. Among 42 patients in stage II, eight were upstaged by PET (19.0%), six to stage III and two to stage IV. Among 12 patients in stage III, six (50%) were upstaged to stage IV by PET. If only early-stage patients and major changes are considered (stages IA-IIB to III or IV), among 49, 10 were upstaged to III or IV, whereas in 39 staging was unchanged following PET. In the former group, three relapsed or were refractory compared with none in the latter group (P<0.006). In advanced stage patients (IIIA or IIIB) a trend toward treatment failure was apparent in patients who were upstaged by PET. PET scanning is an interesting new modality for the accurate staging of patients with HD and frequently shows a higher stage than conventional methods. PET should be performed at initial diagnosis and should be included in prospective studies of patients with HD. Upstaging by PET may represent a risk factor for a more advanced stage or a biologically more aggressive tumor. Patients with early-stage disease as identified by conventional methods have a significant risk of treatment failure if a more advanced stage is indicated by PET. At present, major stage changes suggested by PET imaging should be confirmed by an independent diagnostic method.


Lack of sexual activity from erectile dysfunction is associated with a reversible reduction in serum testosterone.
The role of androgenic hormones in human sexuality, in the mechanism of erection and in the pathogenesis of impotence is under debate. While the use of testosterone is common in the clinical therapy of male erectile dysfunction, hypogonadism is a rare cause of impotence. We evaluated serum testosterone levels in men with erectile dysfunction resulting either from organic or non-organic causes before and after non-hormonal impotence therapy. Eighty-three consecutive cases of impotence (70% organic, 30% non-organic, vascular aetiology being the most frequent) were subjected to hormonal screening before and after various psychological, medical (prostaglandin E1, yohimbine) or mechanical therapies (vascular surgery, penile prostheses, vacuum devices). Thirty age-matched healthy men served as a control group. Compared to controls, patients with impotence resulting from both organic and non-organic causes showed reduced serum levels of both total testosterone (11.1 +/- 2.4 vs. 17.7 +/- 5.5 nmol/L) and free testosterone (56.2 +/- 22.9 vs. 79.4 +/- 27.0 pmol/L) (both p < 0.001). Irrespective of the different aetiologies and of the various impotence therapies, a dramatic increase in serum total and free testosterone levels (15.6 +/- 4.2 nmol/L and 73.8 +/- 22.5 pmol/L, respectively) was observed in patients who achieved normal sexual activity 3 months after commencing therapy (p < 0.001). On the contrary, serum testosterone levels did not change in patients in whom therapies were ineffective. Since the pre-therapy low testosterone levels were independent of the aetiology of impotence, we hypothesize that this hormonal pattern is related to the loss of sexual activity, as demonstrated by its normalization with the resumption of coital activity after different therapies. The corollary is that sexual activity may feed itself throughout the increase in testosterone levels.


Loci of invariant-azimuth and invariant-ellipticity polarization States of an optical system.
The loci of polarization states for which either the ellipticity alone or the azimuth alone remains invariant upon passing through an optical system are introduced. The cartesian equations of these two loci are derived in the complex plane in which the polarization states are represented. The equations are quartic and are conveniently expressed in terms of the elements of the Jones. matrix of the optical system. As an exple the loci are determined for a system composed of a pi/4 rotator followed by a quarter-wave retarder.


Post-Synthesis Modification of the Aurivillius Phase Bi2SrTa2O9 via In Situ Microwave-Assisted "Click Reaction".
A new strategy for the functionalization of layered perovskites is presented, based on the in situ post-synthesis modification of a prefunctionalized phase by copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC). The microwave-assisted protonation and grafting of an alkyne alcohol provides the alkyne-functionalized precursor within a few hours, starting from Bi2SrTa2O9. The subsequent microwave-assisted in situ "click reaction" allows the post-synthesis modification of the precursor within ∼2 h, providing a layered perovskite functionalized by an alcohol-grafted 1,4-disubstituted-1H-1,2,3-triazole. Two compounds are described here, bearing an aliphatic and an aromatic substituent, which illustrates the general application of the method. This work opens new perspectives for the functionalization of layered perovskites, going beyond mere insertion/grafting reactions, and thus broadens the design possibilities and the range of applications of these hybrid systems.


Utility of orcein stain and comparison with Masson trichrome stain in chronic intestinal pseudo-obstruction.
Chronic intestinal pseudo-obstruction (CIPO) is caused by a plethora of primary and secondary causes, dominantly involving the neuromuscular tissue, interstitial cells of Cajal, or the connective tissue framework. The lack of the connective tissue framework, known as desmosis, is evaluated by Masson's trichrome (MT) or picrosirius red stains, both of which are recommended in the London classification. We evaluated the orcein stain in detecting desmosis in comparison to the MT stain. We performed both orcein and MT stains in six previously published cases of complete or partial desmosis along with six age-matched controls. Our results showed comparable results of the orcein stain as compared to the MT stain. Additional advantages of lower cost and a clearer background in orcein stain were noteworthy, whereas MT stain can be used for the detection of additional pathology. We believe that orcein stain can be used as a cheap alternative in resource-limited settings.


Impact of changes in CLSI and EUCAST breakpoints for susceptibility in bloodstream infections due to extended-spectrum β-lactamase-producing Escherichia coli.
The impact of recent changes in and discrepancies between the breakpoints for cephalosporins and other antimicrobials, as determined by CLSI and European Committee on Antimicrobial Susceptibility Testing (EUCAST), was analysed in patients with bloodstream infections caused by extended-spectrum β-lactamase (ESBL) producing Escherichia coli in Spain, was analysed. We studied a cohort of 191 episodes of bloodstream infection caused by ESBL-producing E. coli in 13 Spanish hospitals; the susceptibility of isolates to different antimicrobials was investigated by microdilution and interpreted according to recommendations established in 2009 and 2010 by CLSI, and in 2011 by EUCAST. Overall, 58.6% and 14.7% of isolates were susceptible to ceftazidime, and 35.1% and 14.7% to cefepime using the CLSI-2010 and EUCAST-2009/2011 recommendations, respectively (all isolates would have been considered resistant using the previous guidelines). Discrepancies between the CLSI-2010 and the EUCAST-2011 recommendations were statistically significant for other antimicrobials only in the case of amikacin (98.4% versus 75.9% of susceptible isolates; p <0.01). The results varied depending on the ESBL produced. No significant differences were found in the percentage of patients classified as receiving appropriate therapy, following the different recommendations. Four out of 11 patients treated with active cephalosporins according to CLSI-2010 guidelines died (all had severe sepsis or shock); these cases would have been considered resistant according to EUCAST-2011. In conclusion, by using current breakpoints, extended-spectrum cephalosporins would be regarded as active agents for treating a significant proportion of patients with bloodstream infections caused by ESBL-producing E. coli.


Uncoupling Meat From Animal Slaughter and Its Impacts on Human-Animal Relationships.
Slaughter sets the debate about what is acceptable to do to animals at an extremely low bar. Recently, there has been considerable investment in developing cell-based meat, an alternative meat production process that does not require the raising and slaughtering of animals, instead using muscle cells cultivated in a bioreactor. We discuss the animal ethics impacts of cell-based and plant-based meat on human-animal interactions from animal welfare and rights perspectives, focusing on industrial meat production scenarios. Our hypothesis is that the insertion of cell-based meat in the global meat market may alleviate farm animal suffering and potentially restore resources for wild fauna. We employed a conservative estimation of the cell-based meat contribution to the global meat market in the year 2040 to analyze the consequences for human-animal relationships for both wild animals and farmed domesticated animals. We discuss possible effects of an animal cell domestication process, previously described as the second domestication, on human-animal relationships. We consider its potential to reduce the impact of human demographic changes and land use on animal life, in particular whether there would be increased biomass availability and free land for wild animals. We anticipate a major reduction in animal suffering due to the decrease in the number of individual animals involved in food production, which justifies the adoption of cell-based meat from a utilitarian perspective. For the conventional animal food production that remains, further consideration is needed to understand which systems, either high or low welfare, will be retained and the impact of the innovation on the average farm animal welfare. Additionally, it seems likely that there will be less acceptance of the necessity of animal suffering in farming systems when meat production is uncoupled from animal raising and slaughter, supported by a deontological perspective of animal ethics. Consequent to this is anticipated the mitigation of relevant barriers to animal protection and to the recognition of animals as subjects by legislation. Thus, the development of the alternative meats may be related to a significant change in our relationship with non-human animals, with greater benefits than the prima facie effects on farm animals.


Antimicrobial peptides from the skin secretions of the South-East Asian frog Hylarana erythraea (Ranidae).
Peptidomic analysis of norepinephrine-stimulated skin secretions of the South-East Asian frog Hylarana erythraea (formerly Rana erythraea partim) has led to the identification of multiple peptides with antimicrobial activity. Structural characterization of the peptides demonstrated that they belong to the brevinin-1 (3), brevinin-2 (2), esculentin-2 (4), and temporin (1) families. The values in parentheses indicate the number of paralogs. In addition, a peptide (GVIKSVLKGVAKTVALG ML.NH(2)) was isolated that shows some structural similarity to the brevinin-2-related peptides (B2RP) previously isolated from North American frogs of the genus Lithobates. A synthetic replicate of the species B2RP showed broad-spectrum growth inhibitory activity against reference strains of Escherichia coli (MIC=12.5 microM), Staphylococcus aureus (MIC=12.5 microM) and Candida albicans (MIC=50 microM) and was active against multidrug-resistant clinical isolates of Acetinobacter baumannii (MIC in the range 6-12.5 microM). The hemolytic activity of the peptide was relatively low (LC(50)=280 microM). Phylogenetic analysis based upon the amino acid sequences of 47 brevinin-2 peptides from 17 Asian species belonging to the family Ranidae provides support for the placement of H. erythraea in the genus Hylarana.


Effect of incubation duration, growth temperature, and abiotic surface type on cell surface properties, adhesion and pathogenicity of biofilm-detached Staphylococcus aureus cells.
The goal of this study was to investigate the effect of growth conditions such as the temperature (20, 30 and 37 °C), incubation duration (24 and 48 h) and surface type (stainless steel and polycarbonate) on the cell surface physicochemical properties and adhesion to abiotic surfaces of biofilm-detached and planktonic Staphylococcus aureus cells. This study tested also the hypothesis that S. aureus planktonic cells exhibit distinct pathogenic properties compared with their sessile counterparts. The results showed that the changes of the growth conditions promoted changes in the zeta potential, hydrophobicity, electron donor/acceptor character of the studied cell populations. Biofilm-detached cells showed a greater adhesion to stainless steel and polycarbonate compared with planktonic cells. Compared with planktonic cells, sessile ones showed higher cytotoxic effect against HeLa cells, DNase activity, and siderophore levels. The higher cytotoxic effect and production of DNase and siderophore increased with the increase of temperature and duration of incubations. Based on the obtained data, the S. aureus biofilm-detached cells were found to be distinct in many physiological properties compared with their planktonic counterparts.


MRCK activates mouse oocyte myosin II for spindle rotation and male pronucleus centration.
Asymmetric meiotic divisions in oocytes rely on spindle positioning in close vicinity to the cortex. In metaphase II mouse oocytes, eccentric spindle positioning triggers cortical polarization, including the build-up of an actin cap surrounded by a ring of activated myosin II. While the role of the actin cap in promoting polar body formation is established, ring myosin II activation mechanisms and functions have remained elusive. Here, we show that ring myosin II activation requires myotonic dystrophy kinase-related Cdc42-binding kinase (MRCK), downstream of polarized Cdc42. MRCK inhibition resulted in spindle rotation defects during anaphase II, precluding polar body extrusion. Remarkably, disengagement of segregated chromatids from the anaphase spindle could rescue rotation. We further show that the MRCK/myosin II pathway is activated in the fertilization cone and is required for male pronucleus migration toward the center of the zygote. These findings provide novel insights into the mechanism of myosin II activation in oocytes and its role in orchestrating asymmetric division and pronucleus centration.


International Physical Activity Questionnaire: Validity against fitness.
This study was conducted to criterion-validate the short format of International Physical Activity Questionnaire (IPAQ) against health-related fitness. Participants included 951 men, aged 21-43 yr. VO2max by ergometer was used to measure cardiorespiratory fitness. Muscular fitness tests included the number of sit-ups, push-ups, and squats performed during 60 s. Physical activity was assessed from IPAQ by categories for health (health-enhancing physical activity (HEPA)) and by calculating total and vigorous MET-hours per week. The participants were classified in five groups of almost equal size by the IPAQ outcomes. Moreover, the frequency of vigorous activity was assessed by one precoded question (single-item question on leisure-time vigorous physical activity (SIVAQ)). The mean fitness improved from the first through the fourth IPAQ group. VO2max in the most active IPAQ group, however, was lower than in the fourth group (P < 0.05), both with HEPA categories and vigorous METs as the outcome. In contrast, the weekly frequency of vigorous physical activity showed a positive association with fitness through all six categories. The 65 sedentary (according to SIVAQ) and unfit individuals who belonged to the highest 20% (by IPAQ total METs) were older (30.6 +/- 5.1 vs 28.9 +/- 3.8, P = 0.02), more obese (waist 93 +/- 12 vs 89 +/- 10 cm), more often current smokers (59.4 vs 42.0%; P = 0.03), and less educated (<12 yr of education 70.8 vs 52.0%; P = 0.02), compared with the others in the highest IPAQ group. Almost 10% of young men had poor fitness and apparently low physical activity, but they reported very high physical activity by IPAQ. An evident need therefore exists to develop IPAQ further to solve the apparent overreporting by a considerable proportion of sedentary individuals.


IL-6 receptor blockade for allograft dysfunction after lung transplantation in a patient with COPA syndrome.
COPA syndrome is a genetic disorder of retrograde cis-Golgi vesicle transport that leads to upregulation of pro-inflammatory cytokines (mainly IL-1β and IL-6) and the development of interstitial lung disease (ILD). The impact of COPA syndrome on post-lung transplant (LTx) outcome is unknown but potentially detrimental. In this case report, we describe progressive allograft dysfunction following LTx for COPA-ILD. Following the failure of standard immunosuppressive approaches, detailed cytokine analysis was performed with the intention of personalising therapy. Multiplexed cytokine analysis was performed on serum and bronchoalveolar lavage (BAL) fluid obtained pre- and post-LTx. Peripheral blood mononuclear cells (PMBCs) obtained pre- and post-LTx were stimulated with PMA, LPS and anti-CD3/CD28 antibodies. Post-LTx endobronchial biopsies underwent microarray-based gene expression analysis. Results were compared to non-COPA LTx recipients and non-LTx healthy controls. Multiplexed cytokine analysis showed rising type I/II IFNs, and IL-6 in BAL post-LTx that decreased following treatment of acute rejection but rebounded with further clinical deterioration. In vitro stimulation of PMBCs suggested that myeloid cells were driving deterioration, through IL-6 signalling pathways. Tocilizumab (IL-6 receptor antibody) administration for 3 months (4 mg kg-1, monthly) effectively suppressed IL-6 levels in BAL. Mucosal gene expression profile following tocilizumab suggested greater similarity to normal. Clinical effectiveness of IL-6 receptor blockade was not observed. However, we identified IL-6 upregulation associated with graft injury, effective IL-6 suppression with tocilizumab and evidence of beneficial effect on molecular transcripts. This mechanistic analysis suggests a role for IL-6 blockade in post-LTx care that should be investigated further.


Protective effect of Fructus Lycii polysaccharides against time and hyperthermia-induced damage in cultured seminiferous epithelium.
Lycium barbarum L. (Solanaceae) is a Chinese medicinal plant whose fruits (Fructus Lycii) are used by Chinese physicians for treatment of infertility. However, the active ingredients and the mechanism of action underlying Lycium's fertility-facilitating effects remain unknown. Here we report that Fructus Lycii polysaccharides (FLPS) inhibit time- and hyperthermia-induced structural damage in murine seminiferous epithelium, in vitro. Moreover, we found that FLPS delayed apoptosis in this system, both at normothermic and hyperthermic culture conditions. Oxidative stress was reported to be a major cause of structural degradation and apoptosis in hyperthermic testes, and thus the protective effect of FLPS could implicate an antioxidant mechanism of action. To test this hypothesis we assayed the effect of FLPS on ultraviolet light-induced lipid peroxidation, and cytochrome c reduction by free radicals. We found that FLPS is a potent inhibitor of both of these reactions. Together, these results demonstrate the protective effect of FLPS on time- and hyperthermia-induced testicular degeneration in vitro, indicate the potential mechanism of action for this protective effect, and provide a scientific basis for the traditional use of this plant.