Bone geometry in older adults with subclinical hypothyroidism upon levothyroxine therapy: A nested study within a randomized placebo controlled trial.
The effect of levothyroxine (LT4) therapy for subclinical hypothyroidism (SHypo) on appendicular bone geometry and volumetric density has so far not been studied. In a nested study within the randomized, placebo-controlled Thyroid Hormone Replacement for Subclinical Hypothyroidism (TRUST) trial, we assessed the effect of LT4 therapy on bone geometry as measured by peripheral quantitative computed tomography (pQCT). In the TRUST trial, community-dwelling adults aged ≥65 years with SHypo were randomized to LT4 with dose titration vs. placebo with mock titration. We analyzed data from participants enrolled at the TRUST site in Bern, Switzerland who had bone pQCT measured at baseline and at 1 to 2 years follow-up. The primary outcomes were the annual percentage changes of radius and tibia epi- and diaphysis bone geometry (total and cortical cross-sectional area (CSA) and cortical thickness), and of volumetric bone mineral density (bone mineral content (BMC) and total, trabecular and cortical volumetric bone mineral density (vBMD)). We performed linear regression of the annual percentage changes adjusted for sex, LT4 dose at randomization and muscle cross-sectional area. The 98 included participants had a mean age of 73.9 (±SD 5.4) years, 45.9% were women, and 12% had osteoporosis. They were randomized to placebo (n = 48) or LT4 (n = 50). Annual changes in BMC and vBMD were similar between placebo and LT4-treated groups, without significant difference in bone geometry or volumetric bone mineral density changes, neither at the diaphysis, nor at the epiphysis. For example, in the placebo group, epiphyseal BMC (radius) decreased by a mean 0.2% per year, with a similar decrease of 0.5% per year in the LT4 group (between-group difference in %ΔBMC 0.3, 95% CI -0.70 to 1.21, p = 0.91). Compared to placebo, LT4 therapy for an average 14 months had no significant effect on bone mass, bone geometry and volumetric density in older adults with subclinical hypothyroidism. TRIAL REGISTRATION: The trial was registered on ClinicalTrials.gov numbers NCT01660126 (TRUST Thyroid trial) and NCT02491008 (Skeletal outcomes).


Interactions of bupranolol with the polymorphic debrisoquine/sparteine monooxygenase (CYP2D6).
The beta-adrenoceptor blocker bupranolol turned out to be a competitive inhibitor of the polymorphic cytochrome P450 CYP2D6 of which sparteine is a substrate. There was stereo-selectivity of bupranolol involved: (-)-bupranolol was the weakest inhibitor with an apparent Ki value of 1.32 microM, (+)-bupranolol was the most potent with an apparent Ki value of 0.55 microM, while the therapeutically used racemic bupranolol had an intermediate value of 0.88 microM. A 10 min pre-incubation of 5 microM bupranolol with the enzyme preparation prior to the addition of substrate, reduced the inhibition of sparteine metabolism from 52 to about 25%. This suggests that--during these inhibition studies--bupranolol was much more rapidly metabolized than was sparteine, so that the measured Ki values must represent overestimates. The enzyme catalysing bupranolol metabolism was CYP2D6: microsomes from a liver with the genetic enzyme deficiency did not metabolize bupranolol; in microsomes from livers containing the enzyme and 10 microM bupranolol, 5 microM quinidine caused a 72% inhibition of bupranolol metabolism. Although our methods were not sufficiently sensitive to measure the Km of bupranolol directly, it is undoubtedly the beta-adrenoceptor blocker with the highest-known apparent affinity for CYP2D6. High affinity and rapid metabolism are infrequent combinations in enzymology.


Activation of Src-dependent Smad3 signaling mediates the neutrophilic inflammation and oxidative stress in hyperoxia-augmented ventilator-induced lung injury.
Mechanical ventilation and concomitant administration of hyperoxia in patients with acute respiratory distress syndrome can damage the alveolar epithelial and capillary endothelial barrier by producing inflammatory cytokines and reactive oxygen species. The Src tyrosine kinase and Smad3 are crucial inflammatory regulators used for ventilator-induced lung injury (VILI). The mechanisms regulating interactions between high-tidal-volume mechanical ventilation, hyperoxia, and acute lung injury (ALI) are unclear. We hypothesized that high-tidal-volume mechanical stretches and hyperoxia augment lung inflammation through upregulation of the Src and Smad3 pathways. Wild-type or Src-deficient C57BL/6 mice, aged between 6 and 8 weeks, were exposed to high-tidal-volume (30 mL/kg) ventilation with room air or hyperoxia for 1-4 h after 2-mg/kg Smad3 inhibitor (SIS3) administration. Nonventilated mice were used as control subjects. We observed that the addition of hyperoxia to high-tidal-volume mechanical ventilation further induced microvascular permeability, neutrophil infiltration, macrophage inflammatory protein-2 and matrix metalloproteinase-9 (MMP-9) production, malondialdehyde, nicotinamide adenine dinucleotide phosphate oxidase activity, MMP-9 mRNA expression, hypoxemia, and Src and Smad3 activation (P < 0.05). Hyperoxia-induced augmentation of VILI was attenuated in Src-deficient mice and mice with pharmacological inhibition of Smad3 activity by SIS3 (P < 0.05). Mechanical ventilation of Src-deficient mice with hyperoxia further reduced the activation of Smad3. Our data suggest that hyperoxia-increased high-tidal-volume ventilation-induced ALI partially depends on the Src and Smad3 pathways.


The development of an estimation model for energy expenditure during water walking by acceleration and walking speed.
The aim of this study was to develop an estimation equation for energy expenditure during water walking based on the acceleration and walking speed. Cross-validation study. Fifty participants, males (n=29, age: 27-73) and females (n=21, age: 33-70) volunteered for this study. Based on their physical condition water walking was conducted at three self-selected walking speeds from a range of: 20, 25, 30, 35 and 40 m/min. Energy expenditure during each trial was calculated. During water walking, an accelerometer was attached to the occipital region and recorded three-dimensional accelerations at 100 Hz. A stopwatch was used for timing the participant's walking speed. The estimation model for energy expenditure included three components; (i) resting metabolic rate, (ii) internal energy expenditure for moving participants' body, and (iii) external energy expenditure due to water drag force. When comparing the measured and estimated energy expenditure with the acceleration data being the third component of the estimation model, high correlation coefficients were found in both male (r=0.73) and female (r=0.77) groups. When walking speeds were applied to the third component of the model, higher correlation coefficients were found (r=0.82 in male and r=0.88 in female). Good agreements of the developed estimation model were found in both methods, regardless of gender. This study developed a valid estimation model for energy expenditure during water walking by using head acceleration and walking speed.


Spatiotemporal patterns of the astroglial proliferation in rat brain injured at the postmitotic stage of postnatal development: a combined immunocytochemical and autoradiographic study.
Proliferative response of astrocytes to unilateral injury of the cerebral hemisphere was investigated in 30-day-old rats using a combination of [3H]thymidine autoradiography and immunocytochemical staining for glial fibrillary acidic protein. At different intervals following injury the animals were injected with [3H]thymidine and the distribution of double-labeled cells was recorded 4 h (short-term experiment) or 40-days (long-term experiment) after each injection. Within the region of the lesion a strong reactive proliferation of astrocytes began as early as 2 h after injury, although, at that time all dividing neuroglial cells were located at a relatively long distance from the lesion margin. Forty days later, however, autoradiographically labeled astrocytes were observed in the center of the lesion. This was regarded as evidence for the ability of astrocytes to migrate towards the site of injury. Maximal reactive proliferation of astroglial cells was observed at day 2 after injury and then reached the postmitotic plateau at about day 8. The cells, while dividing, were mostly GFAP-negative. Therefore, the number of astrocytes produced at particular days after injury, and their contribution to post-traumatic scar formation could be assessed only in the long-term experiment, after the newly formed cells became GFAP-positive.


In silico method for predicting infectious strains of influenza A virus from its genome and protein sequences.
Influenza A is a contagious viral disease responsible for four pandemics in the past and a major public health concern. Being zoonotic in nature, the virus can cross the species barrier and transmit from wild aquatic bird reservoirs to humans via intermediate hosts. In this study, we have developed a computational method for the prediction of human-associated and non-human-associated influenza A virus sequences. The models were trained and validated on proteins and genome sequences of influenza A virus. Firstly, we have developed prediction models for 15 types of influenza A proteins using composition-based and one-hot-encoding features. We have achieved a highest AUC of 0.98 for HA protein on a validation dataset using dipeptide composition-based features. Of note, we obtained a maximum AUC of 0.99 using one-hot-encoding features for protein-based models on a validation dataset. Secondly, we built models using whole genome sequences which achieved an AUC of 0.98 on a validation dataset. In addition, we showed that our method outperforms a similarity-based approach (i.e., blast) on the same validation dataset. Finally, we integrated our best models into a user-friendly web server 'FluSPred' (https://webs.iiitd.edu.in/raghava/fluspred/index.html) and a standalone version (https://github.com/raghavagps/FluSPred) for the prediction of human-associated/non-human-associated influenza A virus strains.


Surgery or EUS-guided choledochoduodenostomy for malignant distal biliary obstruction after ERCP failure.
Endoscopic retrograde cholangiopancreatography (ERCP) is the method of choice for drainage in patients with distal malignant biliary obstruction, but it fails in up to 10% of cases. Percutaneous transhepatic cholangiography (PTC) and surgical bypass are the traditional drainage alternatives. This study aimed to compare technical and clinical success, quality of life, and survival of surgical biliary bypass or hepaticojejunostomy (HJT) and endoscopic ultrasound (EUS)-guided choledochoduodenostomy (CDT) in patients with distal malignant bile duct obstruction and failed ERCP. A prospective, randomized trial was conducted. From March 2011 to September 2013, 32 patients with malignant distal biliary obstruction and failed ERCP were studied. The HJT group consisted of 15 patients and the CDT group consisted of 14 patients. Technical and clinical success, quality of life, and survival were assessed prospectively. Technical success was 94% (15/16) in the HJT group and 88% (14/16) in the CDT group (P = 0.598). Clinical success occurred in 14 (93%) patients in the HJT group and in 10 (71%) patients in the CDT group (P = 0.169). During follow-up, a statistically significant difference was seen in mean functional capacity scores, physical health, pain, social functioning, and emotional and mental health aspects in both techniques (P < 0.05). The median survival time in both groups was the same (82 days). Data relating to technical and clinical success, quality of life, and survival were similar in patients who underwent HJT and CDT drainage after failed ERCP for malignant distal biliary obstruction.


Men and women show distinct brain activations during imagery of sexual and emotional infidelity.
Jealousy-related behaviors such as intimate partner violence and morbid jealousy are more common in males. Principal questionnaire studies suggest that men and women have different modules to process cues of sexual and emotional infidelity. We aimed to elucidate the neural response to sentences depicting sexual and emotional infidelity in men and women using functional magnetic resonance imaging. Although there was no sex difference in the self-rating score of jealousy for sexual and emotional infidelity, men and women showed different brain activation patterns in response to the two types of infidelity. During jealous conditions, men demonstrated greater activation than women in the brain regions involved in sexual/aggressive behaviors such as the amygdala and hypothalamus. In contrast, women demonstrated greater activation in the posterior superior temporal sulcus. Our fMRI results are in favor of the notion that men and women have different neuropsychological modules to process sexual and emotional infidelity. Our findings might contribute to a better understanding of the neural basis of the jealousy-related behaviors predominantly observed in males.


Functional status of patients with end-stage renal disease.
A longitudinal study of 979 patients with end-stage renal disease from 27 dialysis centers in the Upper Midwest was conducted to measure the patients' functional status with use of the Karnofsky Activity Scale. At the initiation of dialysis, 50% of all patients were rehabilitated or caring for themselves, and the three variables that most influenced the initial rehabilitation score were age, diabetic status, and sex. Initial functional status was also analyzed for three cohorts of dialysis patients, grouped according to outcome (renal transplantation, continued dialysis, and death). Patients who received a renal transplant had initial rehabilitation scores that were higher than those who underwent dialysis for 2 years or those who died. In the group of patients who underwent dialysis for 2 years, a statistically significant improvement in rehabilitation scores was noted at 2 years in comparison with the scores obtained at the initiation of dialysis. Initial rehabilitation scores were good predictors of the 2-year scores. Of the patients in the 2-year dialysis cohort, 78% maintained or had improvement in their functional status.


Determination of material models for arterial walls from uniaxial extension tests and histological structure.
An approach is proposed that allows the determination of material models from uniaxial tests and histostructural data including fiber orientation of the tissue. A combination of neo-Hookean and Fung-type strain-energy functions is utilized, and inequality constraints imposed on the constitutive parameters are derived providing strict local convexity and preferred fiber orientations. It is shown how the Fung-type model gets a pseudo-structural aspect inherent in the phenomenological model; a correlation between the fiber structure and the parameters of the Fung-type model is explicitly provided. In order to apply the proposed approach, quasi-static uniaxial extension tests of preconditioned prepared strips from the intima, media and adventitia of a human aorta with non-atherosclerotic intimal thickening are acquired in axial and circumferential directions; structural information from histological analyses for each aortic tissue are documented. Data reveal a remarkable thickness, load-bearing capacity and stiffness of the intimal samples in comparison with the media and adventitia. Constitutive parameters for each aortic tissue layer are determined by solving the constrained problem using a penalty function method; a new approach for the estimation of appropriate start values is proposed. Finally, the predictivity and efficacy of the material models is shown by comparing model data with data from the uniaxial extension tests and histological image analyses.


[Clinical and histopathological aspects of 113 necrotizing malignant melanomas of the choroid. 2. Immunogenetic characterization of T-cell receptor-positive tumor-infiltrating lymphocytes and survival of patients with necrotizing melanomas of the choroid].
T-cell receptor (TCR) gene analysis of tumor infiltrating lymphocytes (TIL) has been recognized to play a pivotal role in the immunosurveillance of solid neoplasms. Therefore, 113 necrotizing choroidal melanomas (NMM) were compared to 100 non-necrotizing melanomas (MM) histologically and immunohistochemically. NMM showed TIL more frequently (76.11% vs. 21%, p < 0.001). V alpha/V beta- (70.3% of NMM) and V gamma 1- (39.2% of NMM) and V delta 1- (52.7% of NMM) TCR+ cells were distributed focal or diffuse, and correlated with tumor volume, diameter, and scleral invasion. 74.33% of NMM patients had died after 240 months (5 year survival: 58.1%). Mortality was not significantly associated with TIL (p < 0.33) or V alpha/V beta-TCR+ cells (p < 0.2), however, survival was significantly increased with evidence of V gamma 1- and V delta 1-TCR+ cells (p < 0.026). V alpha/V beta-, V gamma 1- and V delta 1-TCR+ tumor infiltrating lymphocytes can be demonstrated in choroidal melanomas. This is a basis for functional studies providing a rationale for the evaluation of immunotherapeutic modalities.


An update on antimicrobial chemotherapy: 1. The mechanisms of action of antibiotics.
This three-part series will provide an update on the modes of action of currently available antibiotics, review the mechanisms of antibiotic resistance and discuss the impact of antibiotic resistance on prescribing for oral infections.


The "Trinity" smart hospital construction policy promotes the development of hospitals and health management in China.
Recently, in order to comprehensively promote the development of medical institutions and solve the nationwide problems in the healthcare fields, the government of China developed an innovative national policy of "Trinity" smart hospital construction, which includes "smart medicine," "smart services," and "smart management". The prototype of the evaluation system has been established, and a large number of construction achievements have emerged in many hospitals. In this article, the summary of this field was performed to provide a reference for medical workers, managers of hospitals, and policymakers.


Dupilumab significantly improves sleep in adults with atopic dermatitis: results from the 12-week placebo-controlled period of the 24-week phase IV randomized double-blinded placebo-controlled DUPISTAD study.
Sleep disturbance is a prominent symptom of atopic dermatitis (AD) and can result in insomnia, daytime fatigue, drowsiness, reduced productivity and impaired quality of life (QoL). The Dupilumab Effect on Sleep in AD Patients (DUPISTAD) phase IV randomized double-blinded placebo-controlled study evaluated the impact of dupilumab treatment on sleep and other patient- and physician-reported outcomes. Adults with moderate-to-severe AD were randomized 2 : 1 to dupilumab 300 mg once every 2 weeks (q2w) or placebo for 12 weeks; concomitant topical corticosteroids were permitted. Patients subsequently entered an open-label phase and received dupilumab 300 mg q2w for a further 12 weeks. The primary endpoint was the percentage change in sleep quality from baseline to week 12, assessed using a novel numeric rating scale (NRS). Secondary and exploratory endpoints included percentage change in peak pruritus NRS (PP NRS), change in SCORing Atopic Dermatitis (SCORAD), SCORAD sleep visual analogue scale (VAS), Eczema Area and Severity Index, Patient-Reported Outcomes Measurement Information System (PROMIS) sleep-related impairment T-score and the Epworth Sleepiness Scale. Sleep diary and wrist actigraphy measurements were recorded throughout the study. In total, 127 patients received dupilumab and 61 patients received placebo. Demographic and baseline disease characteristics were balanced between groups. Sleep quality NRS significantly improved in patients treated with dupilumab by week 12 vs. placebo [least squares mean of the difference (LSMD) -15.5%, P < 0.001]. PP NRS (LSMD -27.9%, P < 0.001), SCORAD (LSMD -15.1, P < 0.001), SCORAD sleep VAS (LSMD -2.1, P < 0.001) and PROMIS T-score (LSMD -3.6, P < 0.001) were also significantly improved at week 12 with dupilumab vs. placebo. The overall percentage of patients reporting treatment-emergent adverse events was lower in the dupilumab group (56.7%) than in the placebo group (67.2%). Dupilumab significantly improved sleep quality and perception of sleep continuity, itch, metrics of AD severity and QoL in adults with moderate-to-severe AD, with an acceptable safety profile compared with placebo.


Nuclear export signal in CDC25B.
CDC25B is a dual-specificity phosphatase that activates CDK1/cyclin B. The nuclear exclusion of CDC25B is controlled by the binding of 14-3-3 to the nuclear export signal (NES) of CDC25B, which was reported to be amino acids H28 to L40 in the N-terminal region of CDC25B. In studying the subcellular localization of CDC25B, we found a functional NES at V52 to L65, the sequence of which is VTTLTQTMHDLAGL, where bold letters are leucine or hydrophobic amino acids frequently seen in an NES. The deletion of this NES sequence caused the mutant protein to locate exclusively in nuclei, while NES-fused GFP was detected in the cytoplasm. Moreover, the introduction of point mutations at some of the critical amino acids impaired cytoplasmic localization. Treatment with leptomycin B, a potent inhibitor of CRM1/exportin1, disrupted the cytoplasmic localization of both Flag-tagged CDC25B and NES-fused GFP. From these results, we concluded that the sequence we found is a bona fide NES of CDC25B.


Recommendations to eliminate subjective olfactory methods from compendial identification tests.
Substitution of IR and UV absorption spectroscopy and use of the various test papers listed in the compendia provide satisfactory means of identifying drug and other substances that presently require subjective odor identification.


Early and ongoing stable glycaemic control is associated with a reduction in major adverse cardiovascular events in people with type 2 diabetes: A primary care cohort study.
To determine whether achieving early glycaemic control, and any subsequent glycaemic variability, was associated with any change in the risk of major adverse cardiovascular events (MACE). A retrospective cohort analysis from the Oxford-Royal College of General Practitioners Research and Surveillance Centre database-a large, English primary care network-was conducted. We followed newly diagnosed patients with type 2 diabetes, on or after 1 January 2005, aged 25 years or older at diagnosis, with HbA1c measurements at both diagnosis and after 1 year, plus five or more measurements of HbA1c thereafter. Three glycaemic bands were created: groups A (HbA1c < 58 mmol/mol [<7.5%]), B (HbA1c ≥ 58 to 75 mmol/mol [7.5%-9.0%]) and C (HbA1c ≥ 75 mmol/mol [≥9.0%]). Movement between bands was determined from diagnosis to 1 year. Additionally, for data after the first 12 months, a glycaemic variability score was calculated from the number of successive HbA1c readings differing by 0.5% or higher (≥5.5 mmol/mol). Risk of MACE from 1 year postdiagnosis was assessed using time-varying Cox proportional hazards models, which included the first-year transition and the glycaemic variability score. From 26 180 patients, there were 2300 MACE. Compared with group A->A transition over 1 year, those with C->A transition had a reduced risk of MACE (HR 0.75; 95% CI 0.60-0.94; P = .014), whereas group C->C had HR 1.21 (0.81-1.81; P = .34). Compared with the lowest glycaemic variability score, the greatest variability increased the risk of MACE (HR 1.51; 1.11-2.06; P = .0096). Early control of HbA1c improved cardiovascular outcomes in type 2 diabetes, although subsequent glycaemic variability had a negative effect on an individual's risk.


Activity and structural changes of mushroom tyrosinase induced by n-alkyl sulfates.
Catecholase activity and structural changes of mushroom tyrosinase (MT) were studied in the presence of some n-alkyl sulfate derivatives. Experiments showed that MT reached its optimal activity in the presence of 1.5, 0.6, and 0.2 mM of sodium n-octyl sulfate (SOS), sodium n-dodecyl sulfate (SDS) and sodium n-tetradecyl sulfate (STS), respectively. Native and incubated MT with the n-alkyl sulfates were also investigated from structural point of view by far-UV circular dichroism (CD) and intrinsic fluorescence spectroscopy. At the above mentioned concentrations of SOS, SDS, and STS no change in the secondary structure of MT was observed. However, changes in the tertiary structure of the enzyme due to the presence of n-alkyl sulfates were obvious. Results of this research indicate that n-alkyl sulfate with longer chain induces greater conformational changes in MT, hence, can activate it at lower concentrations.


Potential of Nanotechnology-based Formulations in Combating Pulmonary Infectious Diseases: A Current Scenario.
Pulmonary microbial infection is mainly caused by microbes like atypical bacteria, viruses, and fungi, on both the upper and lower respiratory tracts. One of the demands of the present is the use of nanotechnology-based treatments to fight various lung infections. The main aim of the study is to explore all pulmonary infectious diseases and to compare the advanced and novel treatment approaches with the conventional methods which are available to treat infections. This work sheds light on pulmonary infectious diseases with their conventional and present treatment approaches along with a focus on the advantageous roles of nano-based formulations. In the literature, it has been reported that the respiratory system is the key target of various infectious diseases which gives rise to various challenges in the treatment of pulmonary infections. The present review article describes the global situation of pulmonary infections and the different strategies which are available for their management, along with their limitations. The article also highlights the advantages and different examples of nanoformulations currently combating the limitations of conventional therapies. The content of the present article further reflects on the summary of recently published research and review works on pulmonary infections, conventional methods of treatment with their limitations, and the role of nano-based approaches to combat the existing infectious diseases which will jointly help the researchers to produce effective drug formulations with desired pharmacological activities.


PIAS3/SOCS1-STAT3 axis responses to oxidative stress in hepatocellular cancer cells.
The participation of STAT3 and its upstream inhibitors, PIAS3 and SOCS1, in the oxidative response of hepatocellular carcinoma (HCC) cells was uncertain. Here, the expression of PIAS3 and SOCS1 in HCC tissues and cell lines was explored, and we sought to determine whether oxidative stress epigenetically regulated PIAS3 and SOCS1 expression and STAT3 activation in HCC cells. The expression of PIAS3 and SOCS1 was markedly decreased in HCC cell lines and tissues compared to normal hepatic cells and tissues. In HCC patients, low PIAS3 and SOCS1 expression were associated with poor survival. Oxidative stress induced by H2O2 in HepG2 cells was indicated by low antioxidant levels and high protein carbonyl content. Moreover, oxidative stress in HepG2 cells contributed to reduced proliferation but increased apoptosis, migration, and invasion capacity, which might be counteracted by antioxidants, such as tocopheryl acetate (TA). PIAS3 and SOCS1 expression was markedly decreased, while STAT3 was activated in HepG2 cells in response to H2O2 exposure. Co-treatment with antioxidant TA effectively increased the expression of PIAS3 and SOCS1, but it dephosphorylated STAT3 in H2O2-treated cells. PIAS1 or SOCS1 overexpression in HepG2 cells after H2O2 treatment restored cell viability and anti-oxidative responses and decreased apoptosis, migration, and invasion ability, and dephosphorylated STAT3 levels. Co-administration of the STAT3 activator, colivelin, partially abolished the effect of PIAS3 and SOCS1 overexpression in these processes. Therefore, oxidative stress in HCC cells may improve their migration and reduce proliferation through STAT3 activation through the repression of PIAS3 and SOCS1 expression.


Re-purposing is needed for beneficial bugs, not for the drugs.
Between 150 and 200 species of plants, insects, birds or mammals go extinct every day. We do not have any idea what the global extinction rate for microorganisms is. What is clear is that we have already lost a maximum number of the microbes that used to live in and on our skins. Many of our microbial partners are facing extinction as we apply selection pressures that are unprecedented in our long-standing relationships. Recent estimates are that we have lost at least one third of the diversity of our skin microbiome. Every day, most of us bath or shower in water that contains chlorine or fluorine; these additives do a great job of killing pathogenic microbes, but they are probably not helping our skin microbiome. Most of the people apply cosmetic products every day, as these products contain preservatives that prevent microbial growth on the shelf. These same chemicals may well kill microbes on the skin. The daily use of high-pH soaps probably will not help microbial life that is adapted to living on the skin's natural pH of 5. The rise in the rate of C-section births from around 5% in 1970 to more than 30% today is likely to be a contributing factor. Vaginal microbes seed our skins at birth and C-sections disrupt this process. The overuse of broad-spectrum antibiotics has contributed to the loss of our microbial partners in all body sites and the skin is no exception. It is now clear that skin is an ecosystem that is dependent on commensal microbes for optimal health. In general, a diverse ecosystem is a healthy ecosystem that is robust in the face of change. Low-diversity ecosystems are more fragile and susceptible to dysbiosis. Eczema and acne rates have increased rapidly over the last 50 years. These diseases are almost unknown in hunter-gatherer communities. Now, we face two exciting challenges: finding out which species matter and how to get them back.


Self-reported and the actual involvement of community pharmacy professionals in the management of acute childhood diarrhea in Jazan Province, Saudi Arabia: Simulated patient visits and survey study.
Acute childhood diarrhea is one of the most common causes of dehydration, and if severe, can potentially lead to death as well. The present study was aimed at evaluating the knowledge and attitudes of community pharmacy professionals towards the management of acute childhood diarrhea and comparing them with their actual practices in Jazan Province, Kingdom of Saudi Arabia (K.S.A). Simulated patient visits and a cross-sectional survey making use of a 27-item self-report questionnaire were conducted amongst a sample of 303 community pharmacy professionals (51.2 % male and 48.8 % female) with an age range of 25-56 years, between August 1 and December 1, 2021, in Jazan Province, Saudi Arabia. Convenience sampling technique was used for the recruitment of the participants. Significant positive correlations were seen between knowledge score (self-report survey) and practice score (simulated patient visit) regarding patients' history taking (r = 0.65; p < 0.01), drug recommendations (r = 0.71; p < 0.01) and providing information regarding food and fluid intake (r = 0.44; p < 0.01). The alpha coefficients for all the items of the survey instrument were noted to be greater than (or) equal to the 0.70 threshold for almost all sections, hence indicating good reliability and internal consistency of the developed scale. In the present study, even though we observed improved participants' performance during the self report survey, their performance level greatly dropped in actual practice. This warrants for a need for educational programs to improve their actual dispensing practices. The present study has also shown simulated patient visits to be a reliable, simple and a robust method of assessing the actual dispensing practices of community pharmacy professionals.


The morphological relationship in electron microscopy between NOR-silver proteins and intranucleolar chromatin.
The relative distribution of NOR proteins and chromatin fibers in the nucleoli was visualized in human cell line. The chromatin was revealed by a Feulgen-like procedure using osmium-ammine as DNA tracer. This selective staining was combined with NOR-silver staining. We provide morphological evidence for constant overlapping of the silver deposit sites with dispersed intranucleolar chromatin fibers. Silver stained proteins were sometimes observed in contact with the chromatin fibers, suggesting that at least some of the Ag-NOR proteins might be closely connected with the dispersed nucleolar DNA.


Effect of metabolic factors on apoptosis in thymocytes during tumor growth.
Intensive apoptotic death of thymocytes is a possible mechanism of thymus involution during tumor growth. We studied the role of hypercholesterolemia and lactate acidosis in the induction of increased sensitivity of thymocytes to apoptosis during growth of transplanted hepatoma 22a in mice. Spontaneous apoptosis in thymocytes during tumor growth in mice was studied in vitro by acridine orange/ethidium bromide staining and diphenylamine test. Plasma levels of lactate, total cholesterol, alpha-cholesterol, and triglycerides were measured. A positive correlation was found between intensification of apoptosis (diphenylamine test) and increased concentration of total plasma cholesterol on days 21 and 28 after inoculation of tumor cells. Plasma lactate content did not increase at this term. We hypothesize that hypercholesterolemia accompanying tumor growth acts as a factor increasing thymocyte sensitivity to apoptosis.


Comparison of hypertension management between cancer survivors and the general public.
Proper management of hypertension is important for better survival and quality of life of cancer survivors who have hypertension. We aimed to compare hypertension management between cancer survivors and the general population. A nationwide, multicenter, cross-sectional survey was administered to adult cancer patients, currently receiving treatment or follow-up, who had been diagnosed with hypertension. Comparison group was selected from among participants in the health behavior survey of the third Korean National Health and Nutrition Examination Survey. Self-reported hypertension management was surveyed, including antihypertensive medication adherence, frequency of blood pressure (BP) monitoring and perceived BP control. Multivariate logistic regression analysis was used to evaluate the relationship between cancer survivorship and each outcome measure. Compared with the general population, cancer survivors were more likely to report full adherence (92.7% vs. 73.0%; adjusted odds ratio (aOR)=3.45; 95% confidence interval (CI), 2.08-5.73), more frequent BP measurement (≥ 24 per year: 50.1% vs. 24.7%; aOR=2.51; 95% CI, 1.83-3.46), and very good perceived BP control (60.8% vs. 26.2%; aOR=4.34; 95% CI, 3.13-6.02). Cancer survivors appear to be better with antihypertensive medication adherence and BP monitoring than those without cancer, and as a result, they appear to be under better BP control. However, several methodological limitations of our study prompt further research on this issue.


Uncommon defibrinogenation and coagulopathy caused by Trimeresurus stejnegeri stejnegeri envenomation in a patient with swelling above the ankle.
In Taiwan, Trimeresurus stejnegeri stejnegeri (Stejneger's Bamboo pitviper) is responsible for more than half of all venomous snakebites annually. This species often causes local envenoming characterized by tissue swelling and pain, occasional local ecchymosis, bullae and blister formation, and lymphangitis and lymphadenitis. The pathophysiology and treatment of potentially life-threatening coagulopathy and defibrinogenation induced by T. s. stejnegeri systemic envenoming have not been specifically addressed. Here, we describe the case of a man who was bitten by T. s. stejnegeri on his right first toe, which later developed into swelling above the ankle. It was found that there was severe hypofibrinogenemia, prolonged prothrombin time, and reduced activities of factors V and XI, plasminogen, and α2-antiplasmin. Even though a favorable outcome was achieved after repeatedly administering specific antivenom, fresh frozen plasma, and cryoprecipitate, probably low effectiveness of antivenom against the coagulopathy and prodigious amounts of replacement products were observed. To control coagulopathy early and avoid the needless replacement of coagulation factor, which are associated with inherent adverse reactions, more frequent serial blood assessment (e.g., every 6 h) and higher initial antivenom doses may be helpful. Knowledge of the specific coagulation factor deficiencies may improve our understanding of the relationship between hemotoxins and the resulting envenoming syndromes in this snakebite.


Clinicopathologic significance of immunostaining of α-thalassemia/mental retardation syndrome X-linked protein and death domain-associated protein in neuroendocrine tumors.
α-Thalassemia/mental retardation syndrome X-linked protein (ATRX) and death domain-associated protein (DAXX) genes are tumor suppressors whose mutations have been identified in sporadic pancreatic neuroendocrine tumors as well as in patients with MEN1. However, it is unknown whether ATRX and DAXX alterations are specific for pancreatic neuroendocrine tumor. In addition, the association of ATRX/DAXX protein loss with tumor cell proliferation has not been examined. We, therefore, immunostained ATRX and DAXX in 10 gastric, 15 duodenal, 20 rectal, 70 pancreatic, and 22 pulmonary neuroendocrine tumors with 15 nonneoplastic pancreases and 27 pancreatic adenocarcinomas to elucidate the site-specific roles of ATRX/DAXX abnormalities. At least 1 loss of ATRX and DAXX immunoreactivity was detected in all neuroendocrine tumor cases but not in any of nonneoplastic pancreatic tissues or pancreatic adenocarcinomas. The loss of DAXX protein was correlated with the Ki-67 index (ATRX, P = .904; DAXX, P = .044). The status of DAXX immunoreactivity correlated positively with World Health Organization histologic grade (P = .026). These results suggest that the status of ATRX or DAXX protein loss in neuroendocrine tumor differed among the organs in which these tumors arose, and these proteins may play site-specific roles in the development of these tumors.


Prevalence of valproate prescriptions in women of childbearing age in certain regions of Russia.
The goal of this retrospective study was to analyze the proportion of women with epilepsy who had received valproate (VPA) prescriptions in certain regions of Russia in 2018. This retrospective cross-sectional study was based on the IQVIA Russia longitudinal prescriptions (LRx) database and included all individuals with a documented epilepsy code (International Classification of Diseases, Tenth Revision [ICD-10]: G.40) from 13 regions in Russia who had received at least one prescription of an antiepileptic drug (AED). The prevalence of VPA prescriptions in female patients with epilepsy aged 16-45 years was analyzed by age group and epilepsy diagnosis code. A multivariate logistic regression model was used to study the association between predefined variables and the probability of having received a VPA prescription. We found a total of 15,412 patients with epilepsy aged 16-45 who had received AED prescriptions in 2018 in the LRx database; 4488 (29.1%) of those patients were women. Of those, 64% had received at least one VPA prescription in 2018. The highest prevalence of VPA prescriptions was found in the age group 16-20 years (69%). This prevalence decreased with age. When compared with women aged 41-45 years, the 16-20-year-old age group was associated with a 1.6-fold increased probability of having receiving a VPA prescription (odds ratio [OR]: 1.60; p < 0.001), followed by the 21-25-year-old age group (OR: 1.46; p < 0.001). Nevertheless, the majority of women received VPA in low dosages (below 700 MG per day). The prevalence of VPA prescriptions in women of childbearing age was quite high in Russia. The therapeutic doses were in line with international guidelines and had low teratogenic potential. Further research is needed to gain a better understanding of the reasons for prescribing VPA to women with epilepsy who are of childbearing age.


Theoretical study on the mechanism of the side reaction of 1-butyl-3-methylimidazolium cation with D-glucose.
To understand the non-inert nature of ionic liquids in cellulose chemistry, density functional theory calculations were carried out to investigate the reaction of 1-butyl-3-methylimidazolium cation ([bmim](+)) with model compounds α- and β-glucoses (the basic units of cellulose and its degradation products) in the absence and presence of base, triethylamine (TEA).The calculated energy barriers for the reactions of [bmim](+) with α- and β-glucoses without TEA were 67.97 and 66.19 kcal mol(-1), respectively. With the assistance of TEA, the barriers were reduced to 48.17 and 46.96 kcal mol(-1), due to the enhanced electrophilic abilities of H6 and nucleophilic abilities of C2 induced through deprotonation of the C2 atom, respectively. The present results rationalize the experimental finding well and provide a clear explanation on why imidazolium-based ionic liquids are considered as non-inert solvents in cellulose chemistry.


Improvements in the wallpaper industry during the second half of the 19th century: micro-Raman spectroscopy analysis of pigmented wallpapers.
Scientific studies of the pigments used in the manufacturing process of some pigmented wallpapers are presented in this work. Non-destructive micro-Raman spectroscopy was selected for this purpose, and provides important information about how the 19th century wallpaper industry incorporated new materials in their works and designs. At the same time, analysis can help to date the samples of uncatalogued wallpapers. Chrome yellow, burnt Sienna, Prussian blue, ultramarine blue, red lead, carbon black, calcium carbonate, red iron oxide and a red organic pigment were identified. According to the palette used, as well as to the manufacturing process, the wallpapers in this study can be dated to the second half of the 19th century.


Ca(2+)-dependent inhibition of Na+/H+ exchanger 3 (NHE3) requires an NHE3-E3KARP-alpha-actinin-4 complex for oligomerization and endocytosis.
Two PDZ domain-containing proteins, NHERF and E3KARP are necessary for cAMP-dependent inhibition of Na(+)/H(+) exchanger 3 (NHE3). In this study, we demonstrate a specific role of E3KARP, which is not duplicated by NHERF, in Ca(2+)-dependent inhibition of NHE3 activity. NHE3 activity is inhibited by elevation of intracellular Ca(2+) ([Ca(2+)](i)) in PS120 fibroblasts stably expressing E3KARP but not those expressing NHERF. In addition, this Ca(2+)-dependent inhibition requires Ca(2+)-dependent association between alpha-actinin-4 and E3KARP. NHE3 is indirectly connected to alpha-actinin-4 in a protein complex through Ca(2+)-dependent interaction between alpha-actinin-4 and E3KARP, which occurs through the actin-binding domain plus spectrin repeat domain of alpha-actinin-4. Elevation of [Ca(2+)](i) results in oligomerization and endocytosis of NHE3 as well as in inhibition of NHE3 activity. Overexpression of alpha-actinin-4 potentiates the inhibitory effect of ionomycin on NHE3 activity by accelerating the oligomerization and endocytosis of NHE3. In contrast, overexpression of the actin-binding domain plus spectrin repeat domain acts as a dominant-negative mutant and prevents the inhibitory effect of ionomycin on NHE3 activity as well as the oligomerization and internalization of NHE3. From these results, we propose that elevated Ca(2+) inhibits NHE3 activity through oligomerization and endocytosis of NHE3, which occurs via formation of an NHE3-E3KARP-alpha-actinin-4 complex.


Probabilistic tractography of the extracranial branches of the trigeminal nerve using diffusion tensor imaging.
The peripheral course of the trigeminal nerves is complex and spans multiple bony foramen and tissue compartments throughout the face. Diffusion tensor imaging of these nerves is difficult due to the complex tissue interfaces and relatively low MR signal. The purpose of this work is to develop a method for reliable diffusion tensor imaging-based fiber tracking of the peripheral branches of the trigeminal nerve. We prospectively acquired imaging data from six healthy adult participants with a 3.0-Tesla system, including T2-weighted short tau inversion recovery with variable flip angle (T2-STIR-SPACE) and readout segmented echo planar diffusion weighted imaging sequences. Probabilistic tractography of the ophthalmic, infraorbital, lingual, and inferior alveolar nerves was performed manually and assessed by two observers who determined whether the fiber tracts reached defined anatomical landmarks using the T2-STIR-SPACE volume. All nerves in all subjects were tracked beyond the trigeminal ganglion. Tracts in the inferior alveolar and ophthalmic nerve exhibited the strongest signal and most consistently reached the most distal landmark (58% and 67%, respectively). All tracts of the inferior alveolar and ophthalmic nerve extended beyond their respective third benchmarks. Tracts of the infraorbital nerve and lingual nerve were comparably lower-signal and did not consistently reach the furthest benchmarks (9% and 17%, respectively). This work demonstrates a method for consistently identifying and tracking the major nerve branches of the trigeminal nerve with diffusion tensor imaging.


A root-knot nematode effector manipulates the rhizosphere microbiome for establishing parasitism relationship with hosts.
Root-knot nematode (RKN; Meloidogyne spp.) is one of the most infamous soilborne plant diseases, causing severe crop losses every year. Effector proteins secreted by RKNs play crucial roles during plant-nematode interaction. However, less is known about whether RKN effector proteins can impact the rhizosphere microbial environment. In this study, we investigated the rhizosphere microbiome community of MiMIF-2 (a plant immunity-modulating effector) transgenic Arabidopsis thaliana with or without nematode infection using the Illumina high-throughput sequencing analysis. The results showed that the bacterial species richness index increased, while the fungi species richness index decreased in M. incognita-infected MiMIF-2 transgenic A. thaliana plants. The relative abundance of genera such as Clitopilus, Komagataeibacter, Lactobacillus, Prevotella, Moritella, Vibrio, Escherichia-Shigella, and Pseudomonas was reduced in MiMIF-2 transgenic A. thaliana plants compared to wild type, but was significantly increased after inoculation with M. incognita. The Cluster of Orthologous Genes (COG) function classification analysis revealed a decrease in the relative abundance of defense mechanisms, secondary metabolite biosynthesis, transport, and nematode infection catabolism-related functions in MiMIF-2 lines compared to the wild type. These differences may be the reason for the increased susceptibility of MiMIF-2 transgenic A. thaliana to nematode infection. Our results provide a new insight into RKN effector proteins and their association with the microbial community, host, and plant pathogens, which will lead to the exploration of new innovative ideas for future biological control of RKNs.


Stereoselective synthesis of novel antiproliferative steroidal (E, E) dienamides through a cascade aldol/cyclization process.
The stereoselective and metal-free protocol involving a cascade aldol/cyclization process for the synthesis of steroidal (E, E) dienamides from steroidal α, α-dicyanoalkene was reported. This protocol efficiently achieved the construction of C=C bond and selective conversion of cyano group into carboxamide in one-pot procedure under mild condition. Further biological evaluation showed that some of these compounds had moderate to excellent cytotoxic activities against all the tested cancer cell lines and were more potent than well-known drug 5-fluorouracil. Particularly, compound 3c represented excellent inhibitory effect against MCF-7 (IC50=0.76 μM), which was about 10-fold more potent than 5-fluorouracil.


Phylogenetic analysis of fungal heterotrimeric G protein-encoding genes and their expression during dimorphism in Mucor circinelloides.
In fungi, heterotrimeric G proteins are key regulators of biological processes such as mating, virulence, morphology, among others. Mucor circinelloides is a model organism for many biological processes, and its genome contains the largest known repertoire of genes that encode putative heterotrimeric G protein subunits in the fungal kingdom: twelve Gα (McGpa1-12), three Gβ (McGpb1-3), and three Gγ (McGpg1-3). Phylogenetic analysis of fungal Gα showed that they are divided into four distinct groups as reported previously. Fungal Gβ and Gγ are also divided into four phylogenetic groups, and to our understanding this is the first report of a phylogenetic classification for fungal Gβ and Gγ subunits. Almost all genes that encode putative heterotrimeric G subunits in M. circinelloides are differentially expressed during dimorphic growth, except for McGpg1 (Gγ) that showed very low mRNA levels at all developmental stages. Moreover, several of the subunits are expressed in a similar pattern and at the same level, suggesting that they constitute discrete complexes. For example, McGpb3 (Gβ), and McGpg2 (Gγ), are co-expressed during mycelium growth, and McGpa1, McGpb2, and McGpg2, are co-expressed during yeast development. These findings provide the conceptual framework to study the biological role of these genes during M. circinelloides morphogenesis.


Histopathological changes in the ovary following acute and chronic low-level tritium exposure to mice in vivo.
Swiss albino mice were exposed to various doses of tritiated water, either through intraperitoneal injection (acute) or orally (chronic) for various lengths of time. The ovaries were examined for qualitative histopathological and quantitative changes in the number of oocytes. All experimental groups displayed varying degrees of dose dependent radiolesions. It was found that chronic irradiation was more damaging than multiple weekly injection, which in turn was more damaging than a single injection. Also, radiosensitivity varied with the age of the mice.


A reference data base for multilead electrocardiographic computer measurement programs.
In an effort to standardize and evaluate the performance of electrocardiographic computer measurement programs, a 15 lead reference library has been developed based on simultaneously recorded standard 12 lead and orthogonal XYZ lead data. A set of 250 electrocardiograms (ECGs) with selected abnormalities was analyzed by a group of five referee cardiologists and 11 different 12 lead and 6 XYZ computer programs. Attention was focused on the exact determination of the onsets and offsets of P, QRS and T waves. The referees performed their task on highly amplified, selected complexes from the library in a two round process. Median results of the referees coincided best with the median derived from all programs. An analysis of stability proved that the combined program median was a robust reference. However, some individual program results were widely divergent. Paired t tests demonstrated earlier onset for P and QRS (p less than 0.001), as well as later offset for P and T waves in the median 12 lead than in the XYZ results. Significant differences also existed among results obtained by programs analyzing all standard ECG leads at one time, the so-called multilead programs, and those obtained by the conventional standard three lead analysis programs. As a consequence, the derived P, PR, QRS and QT interval measurements varied quite widely among the various programs. Significant differences were also observed among measurements of Q, R and S duration. Some programs showed Q waves that were on the average 6 ms (p less than 0.001) longer than those of others. This may significantly influence diagnostic performance.(ABSTRACT TRUNCATED AT 250 WORDS)


Cooperative binding of transcription factors in the human genome.
Transcription factors (TFs) cooperatively bind to specific DNA sequences to control chromatin and gene transcription in eukaryotes. Here, we searched canonical binding, co-binding and tethered binding regions of a TF within ChIP-seq peaks, and investigated the effect of TF-TF cooperation in human GM12878 and K562 cells. We found that TFs except for CTCF and SPI1 showed a large proportion of co-binding and tethered binding regions, and TFs frequently co-binding with other TFs would also frequently tether other TFs to their binding positions. We further observed lower in vivo nucleosome occupancy, higher in vitro nucleosome occupancy and higher levels of H2A.Z, H3K27ac, H3K9ac, H3K4me1, H3K4me2 and H3K4me3 within distal co-binding regions where other TFs were recruited. In addition, target genes for proximal co-binding regions where other TFs were recruited showed significantly higher expression levels. These results indicated that TF-TF cooperation directly associates with the chromatin structure and gene transcription.


Habitat-dependent trophic transfer of legacy and emerging halogenated flame retardants in estuarine and coastal food webs near a source region.
With the phase-out of legacy halogenated flame retardants (HFRs), such as decabromodiphenyl ether (BDE-209), emerging ones, such as decabromodiphenyl ethane (DBDPE), are being widely produced. We conducted field campaigns to assess the trophic transfer of legacy and emerging HFRs in estuarine and coastal food webs of Laizhou Bay, which are located near the largest HFR manufacturing base in China. Seawater, sediment, plankton, invertebrates, and fish were collected from both sites. BDE-209 was the predominant compound in the estuary, whereas DBDPE was the main contributor to HFRs in the bay, followed by BDE-209. Invertebrates, especially bivalves and sea cucumbers, showed higher levels of BDE-209 and DBDPE than fish. The HFR levels in the organisms of the two coastal zones were comparable to each other, although their concentrations in the estuarine water were one order of magnitude higher than those in the bay. The HFR profiles in benthic organisms were similar to those in the sediments, indicating that the bioaccumulation of HFRs in coastal food webs depended on the habitat. The ΣHFR concentrations followed the order filter-feeding > carnivorous for invertebrates, and demersal non-migratory fish showed higher HFR levels than oceanodromous fish. The trophic magnification factors estimated for BDE-209, dechlorane plus, and DBDPE were lower than 1, suggesting biodilution potential in both food webs, whereas several PBDE congeners exhibited biomagnification capacity. Feeding habits, habitats, hydrophobicity, bioavailability, and metabolism may be the main factors impacting the bioaccumulation of HFRs in organisms in estuarine-coastal ecosystems of northern China.


Update on histone deacetylase inhibitors in peripheral T-cell lymphoma (PTCL).
Peripheral T-cell lymphomas (PTCLs) are a group of highly aggressive malignancies with generally poor prognoses, and the first-line chemotherapy of PTCL has limited efficacy. Currently, several novel targeted agents, including histone deacetylase inhibitors (HDACis), have been investigated to improve the therapeutic outcome of PTCLs. Several HDACis, such as romidepsin, belinostat, and chidamide, have demonstrated favorable clinical efficacy and safety in PTCLs. More novel HDACis and new combination therapies are undergoing preclinical or clinical trials. Mutation analysis based on next-generation sequencing may advance our understanding of the correlation between epigenetic mutation profiles and relevant targeted therapies. Multitargeted HDACis and HDACi-based prodrugs hold promising futures and offer further directions for drug design.


Bionanotechnology approach for FAD-dependent enzymes with nanomaterials sensor.
We have reported the modification of biomolecule with nanomaterials. In this paper, the electrochemical response of different FAD-dependent enzymes at carbon nanomaterials modified electrode. The modified electrode also exhibits a promising enhanced electrocatalytic activity toward the oxidation of substrate. Different methods were used for fabrication of modified electrode. The presence of nanomaterials enhances the enzyme loading and stability. Cyclic voltammograms (CVs) were used for the determination of substrate and the apparent coefficient values for these compounds at different electrodes. Finally, we have studied the surface morphology of the modified electrode using scanning electron microscopy (SEM), which revealed that enzyme is coated on nanomaterials.


Isolation and identification of the new baicalin target protein to develop flavonoid structure-based therapeutic agents.
Proteins are vital constituents of all living organisms. As many therapeutic agents alter the activity of functional proteins, identifying functional target proteins of small bioactive molecules isessential for the rational design of stronger medicines. Flavonoids with antioxidant, anti-allergy, and anti-inflammatory effects are expected to have preventive effects for several diseases closely related to oxidation and inflammation, including heart disease, cancer, neurodegenerative disorders, and eye diseases. Therefore, identifying the proteins involved in the pharmacological actions of flavonoids, and designing a flavonoid structure-based medicine that strongly and specifically inhibits flavonoid target proteins, could aid the development of more effective medicines for treating heart disease, cancer, neurodegenerative disorders, and ocular diseases with few side effects. To isolate the flavonoid target protein, we conducted a novel affinity chromatography in a column wherein baicalin, a representative flavonoid, was attached to Affi-Gel 102. Through affinity chromatography and nano LC-MS/MS, we identified GAPDH as a flavonoid target protein. Then, we performed fluorescence quenching and an enzyme inhibition assay to experimentally confirmbaicalin's binding affinity for, and inhibition of, GAPDH. We also conducted in silico docking simulations to visualize the binding modes of baicalin and the newly identified flavonoid target protein, GAPDH. From the results of this study, it was considered that one of the reasons why baicalin exhibits the effects on cancer and neurodegenerative diseases is that it inhibits the activity of GAPDH. In summary, we showed that Affi-Gel102 could quickly and accurately isolate the target protein for bioactive small molecules, without the need for isotopic labeling or a fluorescent probe. By using the method presented here, it was possible to easily isolate the target protein of a medicine containing a carboxylic acid.


Destabilization of CDC6 upon DNA damage is dependent on neddylation but independent of Cullin E3 ligases.
CDC6 is an important component of the pre-replication complex and plays an essential role in the regulation of DNA replication in eukaryotic cells. Deregulation of CDC6 protein levels results in rereplication and genomic instability. CDC6 expression is tightly regulated during the cell cycle. One major mechanism of cell cycle dependent regulation of CDC6 is APC(Cdh1) mediated protein ubiquitination and degradation during G1 phase. In addition to APC(Cdh1) dependent degradation, alternative, Cullin RING E3 ubiquitin ligase dependent degradation pathways have been characterized in yeast. Here we studied whether Cullin RING E3 ligases also play a role in the turnover of CDC6 protein in mammalian cells. To this end, we used the Nedd8 E1 inhibitor MLN4924, which blocks the activity of all Cullin E3 ligases. We observed that treatment with MLN4924 increased CDC6 protein expression. However, this effect was due to a delay in cell cycle progression from G1 to S phase, resulting in accumulation of cells with high CDC6 protein levels. Therefore, our results indicate that Cullin E3 ligases are not involved in the basal turnover of CDC6 in mammalian cells. Interestingly, we also found that the DNA cross-linker mitomycin C induces marked CDC6 protein degradation. Mitomycin C induced CDC6 degradation is not mediated by APC(Cdh1), Cullin or HUWE1 E3 ubiquitin ligases. Notably, mitomycin C mediated CDC6 degradation requires the neddylation pathway. Our results provide evidence for a novel, cullin independent mechanism of CDC6 posttranslational regulation upon DNA damage that involves the neddylation pathway.


An in vitro assessment of bioaccessibility of arsenicals in rice and the use of this estimate within a probabilistic exposure model.
In this study, an in vitro synthetic gastrointestinal extraction protocol was used to estimate bioaccessibility of different arsenicals present in 17 rice samples of various grain types that were collected across the United States. The across matrix average for total arsenic was 209 ng/g±153 (\[xmacr]±2σ). The bioaccessibility estimate produced an across matrix average of 61%±19 (\[xmacr]±2σ). The across matrix average concentrations of inorganic arsenic (iAs) and dimethylarsinic acid (DMA) were 81 ng/g±67.7 and 41 ng/g±58.1 (\[xmacr]±2σ), respectively. This distribution of iAs concentrations in rice was combined with the distribution of consumption patterns (from WWEIA) in a Stochastic Human Exposure and Dose Simulator model to estimate population-based exposures. The mean consumption rate for the population as a whole was 15.7 g per day resulting in a 0.98 μg iAs per day exposure. The mean consumption rate for children 1-2 years old was 7 g per day resulting in a 0.48 μg iAs per day exposure. Presystemic biotransformation of DMA in rice was examined using an in vitro assay containing the anaerobic microbiota of mouse cecum. This assay indicated that DMA extracted from the rice was converted to dimethylthioarsinic acid, although a second oxygen-sulfur exchange to produce DMDTA was not observed.


Association between the diagnostic classification of newly diagnosed coronary artery disease and future heart failure development.
The first clinical manifestation of coronary artery disease (CAD) varies widely from unheralded myocardial infarction (MI) to mild, incidentally detected disease. The primary objective of this study was to quantify the association between different initial CAD diagnostic classifications and future heart failure. This retrospective study incorporated the electronic health record of a single integrated health care system. Newly diagnosed CAD was classified into a mutually exclusive hierarchy as MI, CAD with coronary artery bypass graft (CABG), CAD with percutaneous coronary intervention, CAD only, unstable angina, and stable angina. An acute CAD presentation was defined when the diagnosis was associated with a hospital admission. New heart failure was identified after the CAD diagnosis. Among 28 693 newly diagnosed CAD patients, initial presentation was acute in 47% and manifested as MI in 26%. Within 30 days of CAD diagnosis, MI [hazard ratio (HR) = 5.1; 95% confidence interval: 4.1-6.5] and unstable angina (3.2; 2.4-4.4) classifications were associated with the highest heart failure risk (compared to stable angina), as was acute presentation (2.9; 2.7-3.2). Among stable, heart failure-free CAD patients followed on average 7.4 years, initial MI (adjusted HR = 1.6; 1.4-1.7) and CAD with CABG (1.5; 1.2-1.8) were associated with higher long-term heart failure risk, but an initial acute presentation was not (1.0; 0.9-1.0). Nearly 50% of initial CAD diagnoses are associated with hospitalization, and these patients are at high risk of early heart failure. Among stable CAD patients, MI remained the diagnostic classification associated with the highest long-term heart failure risk, however, having an initial acute CAD presentation was not associated with long-term heart failure.


Neutrophil phagocytosis during endotoxin-induced lung injury.
Depressed neutrophil (PMN) phagocytosis in patients with ARDS may contribute to the known increased incidence of pulmonary sepsis. To evaluate changes in phagocytosis, circulating PMNs from normal rats were compared to circulating and alveolar PMNs (obtained by bronchoalveolar lavage, BAL) from rats after 72 hr of endotoxin infusion (LPS-Rx)-induced acute lung injury. Since phagocytosis correlates with adherence, PMN adherence to coverslips and to a standard nylon wool column was also measured. PMN adherence to nylon wool was 65% for control, 77% for circulating LPS-Rx, and 20% for BAL PMNs. As a measure of phagocytosis the PMNs were incubated for 30 min with opsonized fluorescent (FITC) tagged yeast. Total PMN with yeast were 95.4 +/- 2.1% for control; 96.4 +/- 1.8% for circulating LPS-Rx; and 78.7 +/- 7.8% (P less than 0.05 compared to control) for BAL PMNs. Total numbers of yeast particles per 100 PMN are 270 +/- 64 for control, 300 +/- 42 for circulating LPS-Rx, and 170 +/- 45 (P less than 0.05 compared to control) for BAL PMN. (1) Intraalveolar (BAL) PMNs have decreased adherence; (2) nonadherent PMNs have decreased uptake of yeast; (3) BAL PMNs, overall, have a significantly decreased uptake of yeast; (4) this depression in BAL PMN phagocytosis may partially explain the known decreased rate of bacterial clearance in injured lungs and the increased risk of pulmonary sepsis with adult respiratory distress syndrome.


Conformational properties of streptokinase.
The conformational properties of streptokinase (SK) have been assessed by the techniques of differential scanning calorimetry, circular dichroism (CD), and through a combinational approach employing several algorithms which are predictive of secondary structural characteristics. In low ionic strength buffers, SK undergoes a reversible two-state thermal transition with a temperature of maximum heat capacity (Tm) of 46.1 +/- 0.9, a delta Hcal of 98 +/- 11 kcal/mol and a delta Hcal/delta HvH of approximately 1. In high ionic strength buffers, similar calorimetric properties were obtained with the exception that the delta Hcal/delta HvH values were considerably less than 1, indicating the existence of an additional irreversible thermally induced alteration in the molecule, most likely resulting in its aggregation. The effect of pH on the thermal unfolding properties of SK was determined. The results demonstrated that single two-state thermal transitions were obtained, with progressively decreasing Tm values, as the pH was reduced from 6.4 to 3.4, indicating a destabilization of the entire molecule at reduced pH. In the alkaline region, between pH 8.4 and 9.4, stabilization of a separate region of the molecule was obtained, as evidenced by an increase in the delta Hcal/delta HvH to values approximating 2. CD analysis was performed in order to estimate secondary structural characteristics of SK. The best fit of secondary structural parameters to the experimental CD spectrum provided estimates of 17% helices, 28% beta-sheet, 21% beta-turns, and 34% disordered structures. Both the intensity of the spectral band at 208 nm and the level of antiparallel beta-sheet strongly suggest that SK is an alpha + beta protein.


Directed evolution of biosynthetic pathways. Recruitment of cysteine thioethers for constructing the cell wall of Escherichia coli.
We report that expansion of thioether biosynthesis in Escherichia coli generates sulfur-containing amino acids that can replace meso-diaminopimelate, the essential amino acid used for cross-linking the cell wall. This was accomplished by jointly overexpressing the metB gene coding for L-cystathionine gamma-synthase and disrupting the metC gene, whose product, L-cystathionine beta-lyase, is responsible for the destruction of L-cystathionine and other L-cysteine thioethers. As a result, meso-lanthionine and L-allo-cystathionine were produced endogenously and incorporated in the peptidoglycan, thereby enabling E. coli strains auxotrophic for diaminopimelate to grow in its absence. Thus, current techniques of metabolic engineering can be applied to evolving the chemical constitution of living cells beyond its present state.


A meta-analysis of randomized controlled trials: Efficacy of selenium treatment for sepsis.
To understand the clinical outcomes of selenium therapy in patients with sepsis syndrome, we conducted a meta-analysis of randomized controlled trials (RCT). A total of 13 RCTs comparing selenium and placebo for patients with sepsis were reviewed systematically. However, we could not detect the association of selenium treatment with a decreased mortality at different time course (relative risk [RR] [95% confidence interval, CI]: 0.94 [0.82-1.06] at day 28; 0.73 [0.36-1.47] at day 90; 1.16 [0.78-1.71] at 6 months; respectively). Selenium supplementation did not show favorable efficacy in the incidence of renal failure, secondary infection or duration of mechanical ventilation (RR [95% CI]: 0.65 [0.41-1.03]; 0.96 [0.87-1.06]; standard mean difference [SMD] [95% CI]: 0.17 [-0.30-0.63]; respectively). Interestingly, we found that selenium therapy was benefit for sepsis patients with reduced duration of vasopressor therapy, staying time in intensive care unit and hospital, and incidence of ventilator-associated pneumonia (SMD [95% CI]: -0.75 [-1.37 to -0.13]; -0.15 [CI: -0.25 to -0.04]; -1.22 [-2.44 to -0.01]; RR [95% CI]: 0.61 [0.42-0.89]; respectively). Based on our findings, intravenous selenium supplementation could not be suggested for routine use.


Soluble epoxide hydrolase inhibition exerts beneficial anti-remodeling actions post-myocardial infarction.
A contributory role for soluble epoxide hydrolase (sEH) in cardiac remodeling post-myocardial infarction (MI) has been suggested; however effects of sEH inhibition following MI have not been evaluated. In this study, we examined in vivo post-MI anti-remodeling effects of a novel sEH inhibitor (GSK2188931B) in the rat, and evaluated its direct in vitro effects on hypertrophy, fibrosis and inflammation. Post-MI administered GSK2188931B (80 mg/kg/d in chow) for 5 weeks improved left ventricular (LV) ejection fraction compared to vehicle-treated (Veh) rats (P<0.01; Sham 65 ± 2%, MI+Veh 30 ± 2%, MI+GSK 43 ± 2%) without affecting systolic blood pressure. Percentage area of LV tissue sections stained positive for picrosirius red (PS) and collagen I (CI) were elevated in LV non-infarct zone (P<0.05; NIZ; PS: Sham 1.46 ± 0.13%, MI+Veh 2.14 ± 0.22%, MI+GSK 1.28 ± 0.14%; CI: Sham 2.57 ± 0.17%, MI+Veh 5.06 ± 0.58%, MI+GSK 2.97 ± 0.34%) and peri-infarct zone (P<0.001; PIZ; PS: Sham 1.46 ± 0.13%, MI+Veh 9.06 ± 0.48%, MI+GSK 6.31 ± 0.63%; CI: Sham 2.57±0.17%, MI+Veh 10.51 ± 0.64%, MI+GSK 7.77 ± 0.57%); GSK2188931B attenuated this increase (P<0.05). GSK2188931B reduced macrophage infiltration into the PIZ (P<0.05). GSK2188931B reduced AngII- and TNFα-stimulated myocyte hypertrophy, AngII- and TGFβ-stimulated cardiac fibroblast collagen synthesis, including markers of gene expression ANP, β-MHC, CTGF and CI (P<0.05). GSK2188931B reduced TNFα gene expression in lipopolysaccharide (LPS)-stimulated monocytes (P<0.05). sEH inhibition exerts beneficial effects on cardiac function and ventricular remodeling post-MI, and direct effects on fibrosis and hypertrophy in cardiac cells. These findings suggest that sEH is an important contributor to the pathological remodeling following MI, and may be a useful target for therapeutic blockade in this setting.


Rapid detection of numerical aberrations of chromosomes 13, 18 and 21 in chorionic mesenchymal cells.
We have devised and evaluated a rapid screening method for the detection of numerical aberrations of chromosomes 13, 18 and 21 in chorionic villus cells. We used non-radioactive in situ hybridization (ISH) with three chromosome-specific probes on overnight-attached mesenchymal cells from chorionic villi. A blind study was performed of 47 karyotypically normal samples, one triploid sample, two samples trisomic for chromosome 21, and two samples from a fetus with putative mosaicism (46/47, +21). All samples were hybridized with the chromosome 18- and 21-specific probes. Thirty samples were additionally hybridized with the chromosome 13-specific probe. The test could be completed within 3-4 days of sampling. In samples disomic with respect to the probed chromosomes, an average of 2 per cent (range 0-9 per cent) had three hybridization signals. By contrast, in the samples trisomic for the probed chromosome(s), 57 per cent (chromosome 13), 51 per cent (chromosome 18), and an average of 74 per cent (55-86 per cent) (chromosome 21) of the nuclei exhibited three signals. In the putative mosaic samples, the number of nuclei with three chromosome 21-specific signals ranged from 41 to 69 per cent. We conclude that this technique rapidly and clearly distinguishes between normal and trisomic/triploid samples, and consequently may be of use in future prenatal diagnosis.


EEG improvements with antiepileptic drug treatment can show a high correlation with behavioral recovery in children with ADHD.
We investigated the relationship between neuropsychological disturbance, assessed using the global assessment of functioning (GAF) and the ADHD-rating scale (ADHD-RS), paroxysmal EEG abnormalities, and treatment with valproate sodium (VPA) in children with both attention deficit hyperactivity disorder (ADHD) and paroxysmal abnormality (PA). Participants with ADHD but without obvious epilepsy were recruited between April 1, 2003 and March 31, 2008. Paroxysmal abnormality was scored by measuring the spike frequency. Of 46 children, 16 showed PA; 3 of the 16 were excluded because no follow-up EEG was available. The EEG improved with VPA treatment in 5 of 8 patients with frontal PA and 3 of 5 patients with rolandic PA. While 83.3% of the patients with improvements in both assessments had frontal PA, only 16.7% had rolandic PA. The patients with frontal PA showed a significantly higher correlation between PA frequency and improvement in ADHD-RS compared with those with rolandic PA. In this study of children with ADHD, EEG improvement with antiepileptic drug treatment showed a high correlation with behavioral improvements as shown by ADHD-RS and GAF scores. However, this was not a population-based study, and the relative importance of detecting and treating PA in ADHD has yet to be determined.


DNA methyltransferase 1 rs16999593 genetic polymorphism decreases risk in patients with transposition of great arteries.
Complete transposition of the great arteries (TGA) is the most frequent cyanotic heart defect diagnosed in neonates. However, the exact etiology of TGA is unknown. The aim of the present study was to assess the association of TGA pathogenesis with single nucleotide polymorphisms (SNPs) in DNA methyltransferases (DNMTs)-1 and 3a- in Chinese children. We genotyped 5 SNPs (rs16999593, rs16999358, and rs2228611 in DNMT1; and rs2276599 and rs2276598 in DNMT3A) in 206 patients with complete TGA and 252 healthy children. Statistical analysis was performed to explore the association of the 5 SNPs with complete TGA susceptibility. Compared with the T/T and C/C genotypes, the heterozygous genotype C/T of rs16999593 correlated with a decreased risk for complete TGA under codominant (OR=0.46; 95% CI=0.29-0.72), dominant (OR=0.58; 95% CI=0.38-0.88), and overdominant (OR=0.44; 95% CI=0.28-0.68) models. In contrast, the genotype C/C of rs16999593 correlated with a higher risk for TGA under a recessive model (OR=3.15; 95% CI=1.14-8.68) compared with the T/T and C/T genotypes. Furthermore, the TGC, TGT, CGC, and CGT haplotypes of DNMT1 did not differ significantly between the two groups, whereas the frequency of the TAC haplotype was lower in the case group (OR<1; P=0.002). No significant differences in the frequencies of the TC, CC, TT, and CT haplotypes of DNMT3A were found between the two groups. Furthermore, logistic regression showed that sex and the rs16999358 SNP were two independent risk factors for complete TGA. Overall, the C/T genotype of the rs16999593 SNP in DNMT1 might decrease the risk of complete TGA pathogenesis in the Southern Chinese population.


Insulin-like material from the digestive tract of the tunicate Pyura pachydermatina (sea tulip).
Gut tissue from the tunicate Pyura pachydermatina (sea tulip) was found to contain a compound or compounds which react with anti-porcine insulin antibodies, but not anti-hag-fish insulin antibodies, and which also stimulate lipogenesis in isolated rat fat cells. The insulin-like material is present in two immunologically active forms, a species of Mr 6000 apparently similar to mammalian insulin, and a high Mr form which expresses biological activity only after further purification by reverse-phase HPLC. The bioactivity of both species is suppressed in the presence of anti-porcine insulin antibodies.


Outbreak response strategies with type 2-containing oral poliovirus vaccines.
Despite exhaustive and fully-financed plans to manage the risks of globally coordinated cessation of oral poliovirus vaccine (OPV) containing type 2 (OPV2) prior to 2016, as of 2022, extensive, continued transmission of circulating vaccine-derived polioviruses (cVDPVs) type 2 (cVDPV2) remains. Notably, cumulative cases caused by cVDPV2 since 2016 now exceed 2,500. Earlier analyses explored the implications of using different vaccine formulations to respond to cVDPV2 outbreaks and demonstrated how different properties of novel OPV2 (nOPV2) might affect its performance compared to Sabin monovalent OPV2 (mOPV2). These prior analyses used fixed assumptions for how outbreak response would occur, but outbreak response implementation can change. We update an existing global poliovirus transmission model to explore different options for responding with different vaccines and assumptions about scope, delays, immunization intensity, target age groups, and number of rounds. Our findings suggest that in order to successfully stop all cVDPV2 transmission globally, countries and the Global Polio Eradication Initiative need to address the deficiencies in emergency outbreak response policy and implementation. The polio program must urgently act to substantially reduce response time, target larger populations - particularly in high transmission areas - and achieve high coverage with improved access to under-vaccinated subpopulations. Given the limited supplies of nOPV2 at the present, using mOPV2 intensively immediately, followed by nOPV2 intensively if needed and when sufficient quantities become available, substantially increases the probability of ending cVDPV2 transmission globally.


The immunomodulatory effect of leflunomide in rat cardiac allotransplantation.
Leflunomide, a novel immunosuppressant, has been the subject of recent preclinical studies using solid organ allo- and xenotransplantation models. The objectives of this study were to evaluate the efficacy and toxicity of leflunomide using a rat cardiac allotransplant model in two different strain combinations (DA x PVG and DA x Lew). Leflunomide, at doses ranging between 5 and 30 mg/kg, prolonged graft survival in both strain combinations as effectively as CsA and FK506 1 mg/kg (P < 0.05). A dose-dependent effect was seen only after a longer treatment course. When ongoing rejection was intercepted early (postoperative day 2), 5 mg/kg was as effective as 1 mg/kg FK506 (P > 0.05) but was inferior to CsA in the DA x PVG combination (P < 0.05). However, in the DA x Lew combination, leflunomide was equally as efficacious as 15 mg/kg CsA and 1 mg/kg FK506 (P > 0.05). If ongoing rejection was treated at postoperative day 4, 10 mg/kg leflunomide was not only as effective as 15 mg/kg CsA and 1 mg/kg FK506, but demonstrated a dose-dependent increase in graft survival in both strain combinations. The toxicity of leflunomide at doses of especially 5-20 mg/kg was minimal in comparison to therapeutic doses of CsA and FK506 using body weight and biochemical parameters of renal and liver function. These in vivo observations convincingly show leflunomide to be equally as potent an immunosuppressant as CsA and FK506 in transplant rejection. It is also well tolerated on long-term administration and, by virtue of this fact, is a potentially suitable candidate for clinical transplantation.


Effect of irbesartan monotherapy compared with ACE inhibitors and calcium-channel blockers on patient compliance in essential hypertension patients: a multicenter, open-labeled, three-armed study.
This multicenter, three-armed, open-labeled study investigated patient compliance of patients receiving irbesartan, angiotensin-converting enzyme (ACE) inhibitors or calcium-channel blockers (CCB) for essential hypertension for a 6-month period. Patients were either newly diagnosed or switched from existing antihypertensive medication due to lack of efficacy or side-effects. Patients were started monotherapy with irbesartan (n=377), ACE inhibitors (n=298) or CCB (n=308) and were reevaluated on 1st, 3rd, and 6th months of the treatment. The primary endpoint was patient compliance, assessed by proportion of patients who had taken their study medication every day. Efficacy was recorded as mean reductions in blood pressure and the proportion of patients whose blood pressure normalized. Tolerability was assessed by reported adverse events. Significantly more patients receiving irbesartan had complied with study medication after 3 and 6 months of treatment than ACE inhibitors or CCB. Significantly fewer patients receiving irbesartan needed to change their antihypertensive medication. All three study treatments exhibited similar efficacy profiles, but irbesartan had significantly less adverse events. This study demonstrated that patient compliance to irbesartan was significantly superior to other study treatments. Irbesartan is therefore a suitable first-line therapy for essential hypertension in everyday clinical practice.


Prognostic Significance of Cyclin D1 Expression in Renal Cell Carcinoma: a Systematic Review and Meta-analysis.
Previous studies indicated that cyclin D1 shown the potential as a tumor biomarker. However, the prognostic value of cyclin D1 in renal cell carcinoma (RCC) remains controversial. This study investigated the correlation of cyclin D1 expression with the prognostic and clinicopathological features in RCC patients. We systematically searched the database of PubMed, Embase, Cochrane, and Web of Science updated on November 26, 2017. Eighteen studies with 2282 patients satisfied the inclusion criteria. Results demonstrated that cyclin D1 overexpression in RCC showed significant favorable prognostic impact on disease-free survival (DFS) (HR 0.57, 95% CI: 0.43-0.74) and disease-specific survival (DSS) (HR 0.59, 95% CI 0.41-0.85) without significant heterogeneity. In subgroup of clear cell RCC, the prognostic effect on DFS was robust and the pooled HR was 0.39 (95% CI: 0.27-0.57). However, no association between overall survival (OS) and cyclin D1 expression was observed. Stratified analysis in DFS studies by sample size, staining patterns race and metastasis status showed similar results. Otherwise, cyclin D1 overexpression predicted a reduced prevalence of high TNM stage (T3 + T4) (OR 0.63, 95% CI: 0.40-0.99), high-grade tumor (G3 + G4) (OR 0.51, 95% CI: 0.31-0.81) and large tumor size (OR 0.35, 95% CI: 0.19-0.62). Our meta-analysis indicated that cyclin D1 overexpression could predict the favorable prognosis in patients with RCC.


Dose-Response Analysis of the Effect of Carbidopa-Levodopa Extended-Release Capsules (IPX066) in Levodopa-Naive Patients With Parkinson Disease.
Parkinson disease is an age-related disorder of the central nervous system principally due to loss of dopamine-producing cells in the midbrain. Levodopa, in combination with carbidopa, is widely regarded as an effective treatment for the symptoms of Parkinson disease. A dose-response relationship is established for carbidopa-levodopa extended-release capsules (IPX066) in levodopa-naive Parkinson disease patients using a disease progression model. Unified Parkinson Disease Rating Scale (UPDRS) part II plus part III scores from 171 North American patients treated with placebo or IPX066 for approximately 30 weeks from a double-blind, parallel-group, dose-ranging study were used to develop the pharmacodynamic model. The model comprised 3 components: a linear function describing disease progression, a component describing placebo (or nonlevodopa) effects, and a component to describe the effect of levodopa. Natural disease progression in early Parkinson disease as measured by UPDRS was 11.6 units/year and faster in patients with more severe disease (Hoehn-Yahr stage 3). Maximum placebo/nonlevodopa response was 23.0% of baseline UPDRS. Maximum levodopa effect from IPX066 was 76.7% of baseline UPDRS, and the ED50 was 450 mg levodopa. Equilibration half-life for the effect compartment was 62.8 days. Increasing age increased and being female decreased equilibration half-life. The quantitative model allowed description of the entire time course of response to clinical trial intervention.


Nucleotide-binding sites can enhance N-acylation of nearby protein lysine residues.
Acyl-CoAs are reactive metabolites that can non-enzymatically S-acylate and N-acylate protein cysteine and lysine residues, respectively. N-acylation is irreversible and enhanced if a nearby cysteine residue undergoes an initial reversible S-acylation, as proximity leads to rapid S → N-transfer of the acyl moiety. We reasoned that protein-bound acyl-CoA could also facilitate S → N-transfer of acyl groups to proximal lysine residues. Furthermore, as CoA contains an ADP backbone this may extend beyond CoA-binding sites and include abundant Rossmann-fold motifs that bind the ADP moiety of NADH, NADPH, FADH and ATP. Here, we show that excess nucleotides decrease protein lysine N-acetylation in vitro. Furthermore, by generating modelled structures of proteins N-acetylated in mouse liver, we show that proximity to a nucleotide-binding site increases the risk of N-acetylation and identify where nucleotide binding could enhance N-acylation in vivo. Finally, using glutamate dehydrogenase as a case study, we observe increased in vitro lysine N-malonylation by malonyl-CoA near nucleotide-binding sites which overlaps with in vivo N-acetylation and N-succinylation. Furthermore, excess NADPH, GTP and ADP greatly diminish N-malonylation near their nucleotide-binding sites, but not at distant lysine residues. Thus, lysine N-acylation by acyl-CoAs is enhanced by nucleotide-binding sites and may contribute to higher stoichiometry protein N-acylation in vivo.


Ecophysiological characteristics and biogas production of cadmium-contaminated crops.
The present study proposes a novel strategy to get a rational production of biogas of the biomass residues from phytoremediation. This study investigates physiological responses, cadmium (Cd) accumulation and biogas production from canola, oat and wheat in pot and batch experiments. The results indicate that (1) aerial biomasses for canola, oat and wheat were enhanced by 5 mg Cd/kg soil by 19.41%, 8.78% and 3.38%, and the upper limit of Cd concentration that canola, oat and wheat can tolerate for aerial biomass production were 50, 10 and 10 mg Cd/kg soil; (2) canola accumulates more Cd than oat and wheat in its aerial parts; (3) cumulative biogas yields were 159.37%, 179.23% and 111.34% of the control when Cd in the shoot were 2.00±0.44, 39.80±1.25 and 6.37±0.15 mg Cd/kg biomass for canola, oat and wheat. Phytoremediation in cooperation with bioenergy production provide new insights for both soil remediation and energy research.


Complement C5a (desArg) generation in serum exposed to damaged aortic endothelium.
Complement activation by injured endothelial cells was investigated using ex vivo rabbit thoracic aortas as a source of endothelium and a neutrophil aggregation (NA) bioassay to detect the complement cleavage product C5a (desArg). Endothelium, oxygen-starved by incubating aortas 30 min with Tyrode's buffer, activated complement as demonstrated by a 57% increase in the NA response induced by serum from buffer-treated aortas as compared to serum from untreated control aortas. Incubation of MgEGTA serum in injured aortas resulted in a 27% (P less than 0.025) weaker NA response than normal serum, indicating participation by both classical and alternative pathways of complement activation. Serum from aortas incubated 30 min with 100 micrograms/ml cholestane-3 beta, 5 alpha, 6 beta-triol, a cytotoxic cholesterol oxidation derivative, induced a NA response comparable to that from serum from aortas treated 30 min with Tyrode's buffer. Heat inactivation of serum prior to aortic incubation abolished NA activity and serum incubated in deendothelialized aortas lacked NA activity. Fractionation of serum samples from these experiments on Sephadex G-100 revealed a single peak of NA activity corresponding to the molecular weight of C5a (desArg). Endothelial cell injury was demonstrated by the inability to exclude Trypan blue dye and by scanning electron microscopy. These data demonstrate that damaged arterial endothelium can effectively activate the complement system, resulting in the production of an anaphylatoxic inflammatory mediator.


Coagulation Disorders, Testing, and Treatment in Exotic Animal Critical Care.
Despite poor recognition in the literature, exotic companion animals are affected by many diseases that can result in disordered coagulation and fibrinolysis. This article outlines current knowledge of hemostasis, common diagnostic tests and reviews reported diseases associated with coagulopathy in small mammals, bird and reptiles. A range of conditions affect platelets and thrombocytes, endothelium and blood vessels, and plasma clotting factors. Improved recognition and monitoring of hemostatic disorders will enable targeted therapy and improved case outcomes.


Modeling residual force enhancement with generic cross-bridge models.
The interaction of actin and myosin through cross-bridges explains much of muscle behavior. However, some properties of muscle, such as residual force enhancement, cannot be explained by current cross-bridge models. There is ongoing debate whether conceptual cross-bridge models, as pioneered by Huxley (A.F. Huxley, Muscle structure and theories of contraction, Prog. Biophys. Biophys. Chem. 7 (1957) 255) could, if suitably modified, fit experimental data showing residual force enhancement. Here we prove that there are only two ways to explain residual force enhancement with these 'traditional' cross-bridge models: the first requires cross-bridges to become stuck on actin (the stuck cross-bridge model) while the second requires that cross-bridges that are pulled off beyond a critical strain enter a 'new' unbound state that leads to a new force-producing cycle (the multi-cycle model). Stuck cross-bridge models cannot fit the velocity and stretch amplitude dependence of residual force enhancement, while the multi-cycle models can. The results of this theoretical analysis demonstrate that current kinetic models of cross-bridge action cannot explain the experimentally observed residual force enhancement. Either cross-bridges in the force-enhanced state follow a different kinetic cycle than cross-bridges in a 'normal' force state, or the assumptions underlying traditional cross-bridge models must be violated during experiments that show residual force enhancement.


The effect of temporal parameters on subjective sensations evoked by electrical tooth stimulation.
The effect of stimulus duration and frequency on subjective sensations evoked by electrical tooth stimulation was studied in 12 subjects. The sensory responses were classified using 5 equi-sensation categories (perception threshold, prepain, pain threshold, moderate pain, intense pain). Both continuously increasing and randomised stimuli were applied. A comparison was made with the activation thresholds of intradental A- and C-fibres in the cat. The mean threshold of intradental A-fibres was lower than the perception threshold at all pulse durations. Perception threshold decreased with increasing stimulus frequency. Current intensities which evoked prepain at a stimulus frequency of 1 Hz were rated as pain at 20 Hz. At supraliminal pain levels the effects of summation were more marked. High-frequency stimulation produced intense pain sensations at intensities well below the activation thresholds of pulpal C-fibres in the cat. We conclude that both perception and pain thresholds and supraliminal pain are modified by temporal summation, and that activation of different pulpal fibre populations is not responsible for production of prepain and pain sensations.


Mycobacterium thermoresistibile infection following knee-replacement surgery.
We report a case of Mycobacterium thermoresistibile as a cause of infection following total knee replacement. This infection was masked by the prior isolation of a vancomycin-resistant enterococcus. The infection was resolved with long-term therapy using moxifloxacin and doxycycline.


A unified analytic solution approach to static bending and free vibration problems of rectangular thin plates.
A unified analytic solution approach to both static bending and free vibration problems of rectangular thin plates is demonstrated in this paper, with focus on the application to corner-supported plates. The solution procedure is based on a novel symplectic superposition method, which transforms the problems into the Hamiltonian system and yields accurate enough results via step-by-step rigorous derivation. The main advantage of the developed approach is its wide applicability since no trial solutions are needed in the analysis, which is completely different from the other methods. Numerical examples for both static bending and free vibration plates are presented to validate the developed analytic solutions and to offer new numerical results. The approach is expected to serve as a benchmark analytic approach due to its effectiveness and accuracy.


Relationship of the lung microbiome with PD-L1 expression and immunotherapy response in lung cancer.
Lung cancer is the primary cause of cancer-related deaths worldwide. The human lung serves as a niche to a unique and dynamic bacterial community that is related to the development of multiple diseases. Here, we investigated the differences in the lung microbiomes of patients with lung cancer. 16S rRNA sequencing was performed to evaluate the respiratory tract microbiome present in the bronchoalveolar lavage fluid. Patients were stratified based on programmed death-ligand 1 (PD-L1) expression levels and immunotherapy responses. In total, 84 patients were prospectively analyzed, of which 59 showed low (< 10%), and 25 showed high (≥ 10%) PD-L1 expression levels. The alpha and beta diversities did not significantly differ between the two groups. Veillonella dispar was dominant in the high-PD-L1 group; the population of Neisseria was significantly higher in the low-PD-L1 group than in the high-PD-L1 group. In the immunotherapy responder group, V. dispar was dominant, while Haemophilus influenzae and Neisseria perflava were dominant in the non-responder group. The abundances of Neisseria and V. dispar differed significantly in relation to PD-L1 expression levels and immunotherapy responses.


Cystic ovarian enlargement resulting from autoimmune oophoritis.
Autoimmune oophoritis is a rare cause of premature ovarian failure. Previous studies of autoimmune oophoritis have detailed the clinical, immunologic, and histologic findings. The gross pathologic features of the involved ovaries have detailed the clinical, immunologic, and histologic findings. The gross pathologic features of the involved ovaries have not been emphasized. Herein, we describe a 35-year-old woman with autoimmune oophoritis who presented with symptoms referable to a right cystic adnexal mass. She underwent a diagnostic laparotomy and right salpingo-oophorectomy for removal of an 8.0-cm-diameter, twisted multicystic ovary. Microscopically, mononuclear inflammatory cells infiltrated multiple follicular and luteal cysts. We have seen two additional patients with histologically verified autoimmune oophoritis who underwent diagnostic laparotomy and oophorectomy because of cystic enlargement of the ovaries. Stimulation of ovarian follicles by elevated levels of pituitary gonadotropins probably causes the cystic changes. Recognition of this aspect of autoimmune oophoritis may help to prevent oophorectomy in patients whose functioning ovarian tissue is already compromised by autoimmune destruction.


Exclusive double outlet right ventricle with atrioventricular concordance and pulmonary stenosis. Results of reconstructive surgery.
Double outlet right ventricle associated with atrioventricular concordance, pulmonary stenosis and situs solitus of the atria is a subset of double outlet right ventricle related through the surgical treatment. From 1974 to 1985, 14 patients, 5 males, 9 females (mean age 8.9 years, range 13 months-22 years) were operated upon. All patients had infundibular stenosis and normal or large pulmonary arteries. The apex of the heart was to the right in 2 patients, the right and left ventricles were superior and inferior in 2 patients and 1 patient had both anomalies. The ventricular septal defect was subaortic in 11 patients (aorto-mitral discontinuity in 5) and non-committed in 3 patients. Three patients had 2 ventricular septal defects. The aorta was anterior in 3 patients and to the right of the pulmonary artery in 11 patients. All patients, through a transventricular and transatrial approach, had a reconstructive surgery. In 3 patients, an aortic homograft valved conduit was used. One patient had the ventricular septal defect enlarged. There was one early death (7.1%) from high residual right ventricle pressure and no late death. One patient had a transient atrioventricular block. One patient was reoperated upon for a residual ventricular septal defect. All survivors had a good clinical result. Re-evaluation in 8 patients confirmed excellent haemodynamics: the right ventricle to pulmonary artery pressure gradient decreased from 80 mm Hg (range 60-95) preoperatively to 24 mm Hg (range 3-32) postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS)