Intrinsic bending of microtubule protofilaments.
The complex polymerization dynamics of the microtubule (MT) plus end are closely linked to the hydrolysis of the GTP nucleotide bound to the β-tubulin. The destabilization is thought to be associated with the conformational change of the tubulin dimers from the straight conformation in the MT lattice to a curved conformation. It remains under debate whether this transformation is directly related to the nucleotide state, or a consequence of the longitudinal or lateral contacts in the MT lattice. Here, we present large-scale atomistic simulations of short tubulin protofilaments with both nucleotide states, starting from both extreme conformations. Our simulations indicate that both interdimer and intradimer contacts in both GDP and GTP-bound tubulin dimers and protofilaments in solution bend. There are no observable differences between the mesoscopic properties of the contacts in GTP and GDP-bound tubulin or the intradime and interdimer interfaces.


Physical Activity and Breast Cancer Survival-Epidemiologic Evidence and Potential Biologic Mechanisms.
To review the biologic mechanisms that may be operative between physical activity and survival after breast cancer. Physical activity decreases risk of mortality from breast cancer by up to 40%. Several biologic mechanisms have been hypothesized to explain this association. Previous cohort studies and randomized trials have examined the primary mechanisms that appear to be operative, which involve a decrease in sex hormone levels, insulin resistance, and inflammation. The evidence is still inconsistent and several limitations in the existing literature exist. Understanding the biologic mechanisms involved in the association of physical activity and breast cancer survival will provide more precision to physical activity guidelines for cancer survival. To achieve this objective, future research should include direct measurements of physical activity, sedentary behaviour, and health-related fitness to provide a more comprehensive assessment of these factors and their association with biomarkers and survival after breast cancer.


How are response properties in the middle temporal area related to inference on visual motion patterns?
Neurons in the primate middle temporal area (MT) respond to moving stimuli, with strong tuning for motion speed and direction. These responses have been characterized in detail, but the functional significance of these details (e.g. shapes and widths of speed tuning curves) is unclear, because they cannot be selectively manipulated. To estimate their functional significance, we used a detailed model of MT population responses as input to convolutional networks that performed sophisticated motion processing tasks (visual odometry and gesture recognition). We manipulated the distributions of speed and direction tuning widths, and studied the effects on task performance. We also studied performance with random linear mixtures of the responses, and with responses that had the same representational dissimilarity as the model populations, but were otherwise randomized. The width of speed and direction tuning both affected task performance, despite the networks having been optimized individually for each tuning variation, but the specific effects were different in each task. Random linear mixing improved performance of the odometry task, but not the gesture recognition task. Randomizing the responses while maintaining representational dissimilarity resulted in poor odometry performance. In summary, despite full optimization of the deep networks in each case, each manipulation of the representation affected performance of sophisticated visual tasks. Representation properties such as tuning width and representational similarity have been studied extensively from other perspectives, but this work provides new insight into their possible roles in sophisticated visual inference.


Poly(ε-caprolactone) scaffolds of highly controlled porosity and interconnectivity derived from co-continuous polymer blends: model bead and cell infiltration behavior.
Porous structures destined for tissue engineering applications should ideally show controlled and narrow pore size distributions with fully interconnected pores. This study focuses on the development of novel poly(ε-caprolactone) (PCL) structures with fully connected pores of 84, 116, 141, and 162 μm average diameter, from melt blending of PCL with poly(ethylene oxide) (PEO) at the co-continuous composition, followed by static annealing and selective extraction of PEO. Our results demonstrate a low onset concentration for PEO continuity and a broad region of phase inversion. A novel in vitro assay was used to compare scaffold infiltration by 10-μm diameter polystyrene beads intended to mimic trypsinized human bone marrow stromal cells (hBMSCs). Beads showed a linear increase in the extent of scaffold infiltration with increasing pore size, whereas BMSCs infiltrated 162 and 141 μm pores, below which the cells aggregated and adhered near the seeding area with low infiltration into the porous device. While providing a baseline for non-aggregated systems, the beads closely mimic trypsinized cells at pore sizes equal to or larger than 141 μm, where optimal retention and distribution of hBMSCs are detected. A cytotoxicity assay using L929 cells showed that these scaffolds were cytocompatible and no cell necrosis was detected. This study shows that a melt blending approach produces porous PCL scaffolds of highly controlled pore size, narrow size distribution and complete interconnectivity, while the bead model system reveals the baseline potential for a homogeneous, non-aggregated distribution of hBMSCs at all penetration depths.


Persistent diffuse pulmonary interstitial emphysema mimicking pulmonary emphysema.
A 69-year-old male non-smoker with a history of atopic asthma presented with symptoms suggestive of chronic obstructive pulmonary disease and this appeared to be corroborated by lung function testing and a chest radiograph. However, a chest CT showed no evidence of pulmonary emphysema and instead demonstrated free air along the bronchovascular sheaths indicative of pulmonary interstistial emphysema, possibly caused by repeated prior exacerbations of asthma. His lung function tests and symptoms improved within months of being treated for his airways disease but the CT findings were unchanged after 2 years.


Shear properties of some dental and other polymers.
The shear modulus and elastic limit in shear were determined for a number of polymers of clinical interest, using a static torsion method. In particular, ultra-high modulus polyethylene was studied as a function of draw ratio, and compared with corresponding Young's modulus data. Materials of high shear moduli of potential clinical value are described.


Protein hydrophobicity and stability support the thermodynamic theory of protein degradation.
Rat liver enzymes were used to study the relationship between their in vivo half-lives and their apparent hydrophobicity or their resistance to inactivation by mechanical shaking. The apparent hydrophobicity of these enzymes, measured as the percent of the protein recovered from an octyl-Sepharose column, is correlated with their known half-lives (r = 0.75, P less than 0.01). The presence of specific ligands which are known to increase compactness by impeding unfolding of proteins decreased the apparent hydrophobicity of fructose-1,6-bisphosphatase, glucose-6-phosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, and pyruvate kinase. Resistance of enzymes to inactivation by mechanical shaking correlated well with their in vivo half-lives (r = 0.90, P less than 0.01). When the shaking experiments were done in the presence of substrates, fructose-1,6-bisphosphatase, glucose-6-phosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase and lactate dehydrogenase were protected from inactivation.


Injury epidemiology in Chinese undergraduate classical dance students: A 14-week prospective study.
The aim of this study was to investigate the injury prevalence, incidence rate, injury proportions, and characteristics of injuries in Chinese undergraduate classical dance students by using the modified Oslo Sports Trauma Research Centre Questionnaire on Health Problems. Prospective study. Online survey. 63 Chinese classical undergraduate students (female: 40, male: 23; median age: 20 years, range: 17-20 years) completed the survey. The injury prevalence (proportions) and incidence rate were calculated. The characteristics (severity, locations, injury type) of injuries were investigated. During the 14-weeks, 84% students reported injury occurrence more than one time. The injury incidence rate was 3.28 injuries per 1000h across 14-weeks. Weekly proportion for all injuries ranged from 38.2% to 61.9% and substantial injuries ranged from 7.5% to 22.7%. The lower back was the most frequently injured body location (38.9%), followed by the knee (17.3%) and the ankle (12.9%). In all reported injuries, overuse injury was the most prevalent type (78.9%, 95% CI: 73.2%-81.1%). Chinese classical dance students are at a relatively high risk for injuries. Injury prevention programs in Chinese classical dance students should focus on the lower back and lower extremity.


The platelet storage capability of different plastic containers.
Platelet concentrates (PC), prepared by platelet apheresis, were stored in four different types of blood bags. One of the bags, manufactured with a thinner PVC film than previously, was tested in three different bag volumes. From 25 donors a total number of 99 PC were prepared. Platelet numbers varied from 20 to 140 X 10(9) platelets per bag. The cell count, pH, pO2, pCO2 and lactate were determined initially and on days 1, 3 and 5 of storage. In a separate test, the oxygen diffusion capacity of the bags was determined by oxidation of sodium sulfite in the presence of cobaltous chloride. The oxygen diffusion capacity found was 16 (PL 732, 300 ml), 13.5 (Teruflexa 800 ml), 11.5 (PL 1240, 400 ml), 10.6 (Teruflexa 600 ml), 9 (Teruflexa 400 ml) and 4 (PL 146, 300 ml) mumol O2/h, respectively. For each bag type, the minimum and maximum platelet number stored with maintained pH levels (6.9-7.4) was defined. The maximum platelet number stored with maintained aerobic metabolism, correlated to the oxygen diffusion capacity of the bag, r = 0.998, p less than 0.001, n = 6; thus the maximum platelet number successfully stored for 5 days in each container can be predicted by determination of the oxygen diffusion capacity. In PC with a low platelet yield, pH values above 7.4 were observed after 1 and 3 days. When the results are compared with platelet yield data from routine blood banking, the optimal bags for platelet storage can be chosen. These conclusions must be further investigated in studies in vivo.


Health status in survivors older than 70 years after hospitalization with COVID-19: observational follow-up study at 3 months.
To analyze factors associated with mortality at 3 months and readmissions, functional and cognitive decline, anorexia and affective disorders in patients aged > 70 years surviving after hospital admission for SARS-CoV-2. Patients aged > 70 years, discharged after hospitalization with COVID-19. mortality, readmissions, functional and cognitive impairment, anorexia and mood disorder. 165 cases at 3 months after hospital discharge, 8.5% died and 20% required at least one hospital readmission. The presence of severe dependence at discharge (BI < 40) was associated at 3 months with a higher risk of mortality (OR 5.08; 95% CI 1.53-16.91) and readmissions (OR 4.53; 95% CI 1.96-10.49). The post-hospitalization functional deterioration was associated with persistence of deterioration at 3 months (OR 24.57; 95% CI 9.24-65.39), cognitive deterioration (OR 2.32; 95% CI 1.03-5.25) and affective (OR 4.40; 95% CI 1.84-10.55) CONCLUSIONS: Loss function in older people after hospitalization by COVID-19 may contribute to identify patients with a higher risk of sequelae in the short term that require closer follow-up.


Report of the results of a double-blind, randomized, single-dose trial of a topical 2% escin gel versus placebo in the acute treatment of experimentally-induced hematoma in volunteers.
The efficiency of topically applied 2% escin gel in reducing tenderness to pressure on experimentally induced hematoma is demonstrated. A double-blind, randomized signal dose trial versus placebo in healthy volunteers was used.


Electrochemical DNA biosensor for the detection of DNA hybridization with the amplification of Au nanoparticles and CdS nanoparticles.
In this article, colloidal gold nanoparticles (Au NPs) and carboxyl group-functionalized CdS Nanoparticles (CdS NPs) were immobilized on the Au electrode surface to fabricate a novel electrochemical DNA biosensor. Both Au NPs and CdS NPs, well known to be good biocompatible and conductive materials, could provide larger surface area and sufficient amount of binding points for DNA immobilization. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) experiments were performed to follow the whole electrode fabrication process. DNA immobilization and hybridization were characterized with differential pulse voltammetry (DPV) by using [Co(phen)(2)(Cl)(H(2)O)]Cl.2H(2)O as an electrochemical hybridization indicator. With this approach, the target DNA could be quantified at a linear range from 2.0x10(-10) to 1.0x10(-8) M, with a detection limit of 2.0x10(-11) M by 3sigma. In addition, the biosensor exhibited a good repeatability and stability for the determination of DNA sequences.


Relevance of the dinoflagellate Gonyaulax fragilis in mucilage formations of the Adriatic Sea.
Oceanographic cruises were carried out monthly from June 1999 to July 2002 to follow the mucilage formation process in the Northern Adriatic Sea. Results show that in correspondence with these events the dinoflagellate Gonyaulax fragilis (Schütt) Kofoid was observed both in the water column and within mucilage aggregates. In the water column, increasing abundances were observed from May until July, with values never exceeding 8500 cells l(-1). Much higher densities were observed within superficial gelatinous aggregates (22800-3400000 cells l(-1)). In mucilage samples, a large number of decomposing cells were present, together with abundant alive cells, enveloped in exudates. G. fragilis isolated from mucilage samples was cultured in three different culture media; it was characterized by a low growth rate but it produced a high amount of polysaccharides. The highest yield both in terms of cell number and carbohydrate production was observed in the medium having the highest nitrogen and phosphorus content and the lowest N/P ratio. The monomeric composition of G. fragilis carbohydrates, compared with that of mucilage samples, showed that in both natural and cultured samples galactose was the most abundant sugar; in addition, an overall good correlation, especially between the monomeric carbohydrate composition of G. fragilis grown in f/2 medium and that of a mucilage sample in which this species was present in high density, was observed.


Crash report data analysis for creating scenario-wise, spatio-temporal attention guidance to support computer vision-based perception of fatal crash risks.
Reducing traffic fatal crashes has been an important mission of transportation. With the rapid development of sensor and Artificial Intelligence (AI) technologies, the computer vision (CV)-based crash anticipation in the near-crash phase is receiving growing attention. The ability to perceive fatal crash risks in an early stage is of paramount importance as well because it can improve the reliability of crash anticipation. Yet this task is challenging because it requires establishing a relationship between the driving scene information that CV can recognize and the fatal crash features that CV will not get until the crash occurrence. Image data with the annotation for directly training a reliable AI model for the early visual perception of fatal crash risks are not abundant. The Fatality Analysis Reporting System (FARS) contains big data on fatal crashes, which is a reliable data source for finding fatal crash clusters and discovering their distribution patterns to tell the association between driving scene characteristics and fatal crash features. To enhance CV's ability to perceive fatal crash risks earlier, this paper develops a data analytics model from fatal crash report data, which is named scenario-wise, spatio-temporal attention guidance. First, the paper identifies five descriptive variables that are sparse and thus allow for decomposing the 5-year (2013-2017) fatal crash dataset to develop scenario-wise attention guidance. Then, an exploratory analysis of location- and time-related descriptive variables suggests dividing fatal crashes into spatially defined groups. A group's temporal distribution pattern is an indicator of the similarity of fatal crashes in the group. Hierarchical clustering and K-means clustering further merge the spatially defined groups into six clusters according to the similarity of their temporal patterns. After that, association rule mining discovers the statistical relationship between the temporal information of driving scenes with fatal crash features, such as the first harmful event and the manner of collisions, for each cluster. The paper illustrates how the developed attention guidance supports the design and implementation of a preliminary CV model that can identify agents of a possibility to involve in fatal crashes from their environmental and context information.


Pain and sleep: from reaction to action.
Sleep disruption by painful stimuli is frequently observed both in clinical and experimental conditions. Nociceptive stimuli produce significantly more arousals (30% of stimuli) than non-nociceptive ones. However, even if they do not interrupt sleep, they can trigger a variety of other reactions. Reflex behaviours in response to nociceptive stimuli can be observed during all sleep stages, and are more likely to occur in association with an arousal than alone. Cardiac activation represents a robust sympathetically driven effect preserved whatever the state of vigilance, even if its magnitude can be modulated by a concomitant cortical arousal. Not withstanding these reactions, incorporation of nociceptive stimuli into dream content remains limited. At cortical level, laser-evoked potential studies demonstrate that the processing of nociceptive stimulations is partly conserved during all sleep stages. Furthermore, when nociceptive stimulations interrupt sleep, the cortical response presents a late component suggesting that the stimulation has to be cognitively processed in order to produce a subsequent arousal. More complex reactions to nociceptive stimulations were occasionally reported. In this context, an epileptic patient with intracerebral electrodes implanted for therapeutic purposes allowed us extending these observations. This patient exhibited finger lifts in response to stimulations delivered during paradoxical (REM) sleep. This motor reaction was previously used during wakefulness to indicate that the stimulation had been perceived. When these finger lifts occurred a systematic re-activation of the anterior cingulate preceded each movement. This observation suggests that during PS, not only the processing of sensory inputs but also the capacity for the sleeper to intentionally indicate his perception could be preserved under particular circumstances.


The "teacup sign": Significance in breast imaging.
Milk of calcium (MOC) is a term used for sedimented calcifications within small cysts (Milk of Calcium - an overview|ScienceDirect Topics, n.d.). MOC in the breast is a benign entity and present in 4-6% of women that undergo diagnostic mammography (Park et al., 2008).2 Calcium particles within cysts produce 'teacup'-shaped calcifications on true lateral views, and smudged calcifications on craniocaudal (CC) views (Veloso Gomes et al., n.d.).3 On the CC projection, the calcifications will have a cloudlike or smudgy appearance like tea leaves in the bottom of a teacup (Milk of Calcium - an overview|ScienceDirect Topics, n.d.). Because this is a characteristic benign finding, there is no need for the patient to undergo a biopsy.


Integrated Behavioral Health for Preschool Children in Pediatric Primary Care.
Behavioral problems in preschool children have led to increased rates of expulsion and suspension from preschool. This case report describes the management of children aged 0-5 with behavioral concerns in a pediatric practice. Identified children were referred to an integrated behavioral health provider who provided accessible evaluation and treatment to parents and children, as well as consultation to childcare providers. The report describes case examples of how these services prevented adverse outcomes at the preschool age for three children.


A defensin antimicrobial peptide from the venoms of Nasonia vitripennis.
Although many antimicrobial components (i.e. antimicrobial peptides) have been found in many social Hymenoptera venoms, no antimicrobial compound is purified and characterized from parasitic Hymenoptera. From the venoms of the ectoparasitic wasp, Nasonia vitripennis, a defensin-like antimicrobial peptide named defensin-NV was purified and characterized. Defensin-NV is composed of 52 amino acid residues including 6 cysteines forming 3 disulfide bridges. Its amino acid sequence is VTCELLMFGGVVGDSACAANCLSMGKAGGSCNGGLCDCRKTTFKELWDKRFG. By BLAST search, defensin-NV showed significant sequence similarity to other insect defensin antimicrobial peptides. Defensin-NV exerted strong antimicrobial activity against tested microorganisms including Gram-positive bacteria, Gram-negative bacteria and fungi. The cDNA encoding defensin-NV was cloned from the venom reservoir cDNA library of N. vitripennis. The current work firstly purified and characterized an antimicrobial peptide from parasitic Hymenoptera.


In situ phytoremediation of arsenic- and metal-polluted pyrite waste with field crops: effects of soil management.
Sunflower, alfalfa, fodder radish and Italian ryegrass were cultivated in severely As-Cd-Co-Cu-Pb-Zn-contaminated pyrite waste discharged in the past and capped with 0.15m of unpolluted soil at Torviscosa (Italy). Plant growth and trace element uptake were compared under ploughing and subsoiling tillages (0.3m depth), the former yielding higher contamination (∼30%) in top soil. Tillage choice was not critical for phytoextraction, but subsoiling enhanced above-ground productivity, whereas ploughing increased trace element concentrations in plants. Fodder radish and sunflower had the greatest aerial biomass, and fodder radish the best trace element uptake, perhaps due to its lower root sensitivity to pollution. Above-ground removals were generally poor (maximum of 33mgm(-2) of various trace elements), with Zn (62%) and Cu (18%) as main harvested contaminants. The most significant finding was of fine roots proliferation in shallow layers that represented a huge sink for trace element phytostabilisation. It is concluded that phytoextraction is generally far from being an efficient management option in pyrite waste. Sustainable remediation requires significant improvements of the vegetation cover to stabilise the site mechanically and chemically, and provide precise quantification of root turnover.


miR-18a-5p shuttled by mesenchymal stem cell-derived extracellular vesicles alleviates early brain injury following subarachnoid hemorrhage through blockade of the ENC1/p62 axis.
Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have therapeutic potential in various diseases due to their capacity to transfer bioactive cargoes such as microRNAs (miRNAs or miRs) to recipient cells. The present study isolated EVs from rat MSCs and aimed to delineate their functions and molecular mechanisms in early brain injury following subarachnoid hemorrhage (SAH). We initially determined the expression of miR-18a-5p and ENC1 in hypoxia/reoxygenation (H/R)-induced brain cortical neurons and rat models of SAH induced by the endovascular perforation method. Accordingly, increased ENC1 and decreased miR-18a-5p were detected in H/R-induced brain cortical neurons and SAH rats. After MSC-EVs were co-cultured with cortical neurons, the effects of miR-18a-5p on neuron damage, inflammatory response, endoplasmic reticulum (ER) stress, and oxidative stress markers were evaluated based on ectopic expression and depletion experiments. miR-18a-5p overexpression in brain cortical neurons co-cultured with MSC-EVs was shown to impede neuron apoptosis, ER stress and oxidative stress while augmenting neuron viability. Mechanistically, miR-18a-5p bound to the 3'UTR of ENC1 and reduced its expression, weakening the interaction between ENC1 and p62. Through this mechanism, transfer of miR-18a-5p by MSC-EVs contributed to the eventual inhibition of early brain injury and neurological impairment following SAH. Overall, miR-18a-5p/ENC1/p62 may be a possible mechanism underlying the cerebral protective effects of MSC-EVs against early brain injury following SAH.


[The demonstration of the vessels of the microcirculatory bed of the dura mater by using ATPase].
In order to reveal the microcirculatory bed vessels, a histochemical method is suggested, lead technique for determination of ATFase localization being taken as its base. Owing to this method, it is possible to reveal constantly and rather fully microvessels long time after death and differentiate various links of the microcirculatory bed in the connective tissue membranes.


Effects of manganese on microglia M1/M2 polarization and SIRT1-mediated transcription of STAT3-dependent genes in mouse.
Overexposure to manganese (Mn) can lead to neurological diseases, characterized by behavioral and motor impairments. Microglia-mediated neuroinflammation is involved in the pathogenesis and progression of neurodegenerative diseases. Microglial M1 and M2 phenotypes play pro-inflammatory and anti-inflammatory roles, respectively, in response to microenvironmental disturbances. Silent information regulator (SIRT1) has been demonstrated to play an important role in the neuroinflammatory response by deacetylating various transcription factors, such as proliferator-activated receptor γ coactivator 1α (PGC-1α), nuclear factor kappa B (NF-κB), and signal transducer and activator of transcription 3 (STAT3). In addition, PGC-1α and STAT3 have been implicated in microglial polarization and inflammatory response. In this study, Mn exposure (50, 100, 200 μmol/kg) induced neuroinflammatory injury and interfered with microglial M1/M2 polarization in mice, as indicated by the upregulated expression of M1 polarization marker mRNA (IL-1β, IL-6, TNF-α, and iNOS), whereas changes in M2 polarization markers (IL-4, TGF-β, and Arg1) varied, with some increasing and some decreasing, in response to increasing Mn doses, which was consistent with the flow cytometry results used to detect the percentages of each microglial type. We found that Mn could downregulate SIRT1 expression and activate NF-κB signaling. Following mice in the 200 μmol/kg Mn treatment, STAT3 and PGC-1α levels in the nuclear fraction both significantly decreased, and the interaction between the proteins decreased, affecting the transcription of STAT3-mediated genes. These findings provide new insights regarding the role played by Mn neurotoxicity in the suppression of neuroinflammation through the regulation of microglial polarization.


Systematic detection and repair of subclinical inguinal hernias at radical retropubic prostatectomy.
Inguinal hernias occur in 15% of men within 1 to 2 years after radical retropubic prostatectomy (RRP). This suggests that many of these hernias may have been present at surgery, but not diagnosed. Beginning in September 2001, all patients undergoing RRP were carefully examined intraoperatively for the presence of a hernia or a dilated internal inguinal ring. In this study we define the frequency of inguinal hernias detected at RRP and the durability of repair. Between September 2001 and December 2004, 430 patients underwent RRP by one surgeon. Hernias, detected in 142 men (33%), were repaired preperitoneally: direct hernias were repaired with mesh and indirect hernias with 2-0 Prolene suture. The patients were followed up for an average of 19 months (range 9 to 36). Information on postoperative hernia status was available for 315 men (73.3%). Of the 430 patients, 142 (33%) had hernias repaired at RRP. Of these hernias, 32% were right sided, 28% left sided, and 40% bilateral; 34% were direct hernias and 66% indirect. Of the 142 patients who underwent intraoperative hernia repair, 95 (67%) had follow-up data, and 5 (5.3%) of that group developed a hernia postoperatively. None of the hernias repaired at RRP recurred. Recurrent hernias on the side of the repair occurred in 3.1%. Incidental inguinal hernias were commonly found (33%) at RRP. Two thirds of the defects detected were indirect. Clinical recurrence after repair was rare.


The value of hysteroscopic management of cesarean scar pregnancy: a report of 44 cases.
With the incidence of cesarean scar pregnancy (CSP) rising, the reports of serious adverse outcomes of it have increased gradually. The management of CSP remains an inadequately explored clinical field, and there is no consensus on it presently. The present study was performed to investigate the efficacy and safety of operative hysteroscopy in the diagnosis and treatment of CSP. Forty-four patients with CSP underwent operative hysteroscopy for removal of scar ectopic pregnancy in our institution. Among them, hysteroscopy was combined with laparoscopy in two patients, three cases with massive hemorrhage were pretreated with bilateral uterine artery embolization before hysteroscopic surgery, and four patients were pretreated with mifepristone (200 mg for 3 days) and methotrexate (25 mg for 2 days). Clinical data, serum β-human chorionic gonadotropin, myometrial thickness, residual conceptus, cesarean scar defect, operation time, blood loss, and hospital stay were recorded. All of the ectopic gestations were removed entirely by operative hysteroscopy. Mean operation time was 34.8±16.5 minutes (range 20-120 minutes), and mean blood loss was 35.3±24.4 mL (range 5-100 mL). The mean hospital stay was 5.0±3.01 days (range 1-19 days). Cesarean scar defect could be diagnosed in 70% (31/44) of patients, while in 20/32 cases (63%), a conceptus remained after uterine curettage only was performed. Operative hysteroscopy might be recommended as a first-line treatment modality for patients with a cesarean scar ectopic pregnancy, especially when myometrium thickness between bladder and gestational sac is more than 3 mm.


Development of a triplex TaqMan real-time RT-PCR assay for differential detection of wild-type and HCLV vaccine strains of classical swine fever virus and bovine viral diarrhea virus 1.
In this study, genomic sequences of pestiviruses available in GenBank were aligned to design three primer pairs and TaqMan probes: two targeting the NS5A region of the viral genome of classical swine fever virus (CSFV) for the differentiation of wild-type CSFV and hog cholera lapinized vaccine (HCLV) vaccine, and one targeting the 5'-untranslated region of bovine viral diarrhea virus 1 (BVDV-1). With these primers and probes, a triplex TaqMan real-time RT-PCR assay was developed for differentiating wild-type CSFV, the HCLV strain, and BVDV-1. The detection limit of the assay was 4.5 TCID(50) for wild-type CSFV, 10 TCID(50) for HCLV-strain CSFV, and 3.2 TCID(50) for BVDV-1. The triplex real-time RT-PCR had at least 98% (248 samples) agreement with other RT-PCR methods. The assay provides a sensitive tool for simultaneous detection and differentiation of wild-type CSFV and HCLV from BVDV-1.


A comparative study of haloperidol and diazepam in the treatment of anxiety.
In a single-blind study in general practice, 60 patients with anxiety neuroses were randomly allocated to receive either 0.5 mg haloperidol twice daily or 2 mg diazepam 3-times daily for 6 weeks. Eighteen patients (6 on haloperidol and 12 on diazepam) were excluded from the analysis of efficacy. On the Hamilton Rating Scale both haloperidol and diazepam reduced the anxiety and depression scores. The investigator's assessment of 14 anxiety symptoms and signs indicated that haloperidol produced significantly (p=0.05) greater symptomatic improvement than diazepam after 4 and 6 weeks of treatment, and the patients' overall response was also significantly (p less than 0.005) greater with haloperidol. After 6 weeks, 93% of patients felt 'better' or 'much better' on haloperidol, compared with 83% on diazepam. A few, minor side-effects were reported, slightly fewer on haloperidol than on diazepam. In the parameters tested in this study, haloperidol has been shown to be more effective than diazepam in the treatment of anxiety neuroses and appears to provide significantly better overall symptomatic relief and to be more acceptable to patients than diazepam.


Latexin expression is downregulated in human gastric carcinomas and exhibits tumor suppressor potential.
Latexin, also known as endogenous carboxypeptidase inhibitor (CPI), has been found to inhibit mouse stem cell populations and lymphoma cell proliferation, demonstrating its potential role as a tumor suppressor. Our previous study also suggested a correlation between latexin expression and malignant transformation of immortalized human gastric epithelial cells. Here, we examined latexin expression in human gastric carcinomas and investigated the effect of differential latexin expression on proliferation of gastric cancer cells in vitro and in vivo. Monoclonal antibody against human latexin was prepared and immunohistochemical analysis was performed to detect latexin expression in 41 paired gastric carcinomas and adjacent normal control tissues. Human gastric cancer cells MGC803 (latexin negative) stably transfected with LXN gene and BGC823 cells (latexin positive) stably transfected with antisense LXN gene were established for anchorage-dependent colony formation assay and tumorigenesis assay in nude mice. Differentially expressed genes in response to exogeneous latexin expression were screened using microarray analysis and identified by RT-PCR. Bisulfite sequencing was performed to analyze the correlation of the methylation status of LXN promoter with latexin expression in cell lines. Immunohistochemical analysis showed significantly reduced latexin expression in gastric carcinomas (6/41, 14.6%) compared to control tissues (31/41, 75.6%) (P < 0.05). Overexpression of LXN gene in MGC803 cells inhibited colony formation and tumor growth in nude mice. Conversely, BGC823 cells transfected with antisense LXN gene exhibited enhanced tumor growth and colony formation. Additionally, several tumor related genes, including Maspin, WFDC1, SLPI, S100P, and PDGFRB, were shown to be differentially expressed in MGC803 cells in response to latexin expression. Differential expression of Maspin and S100P was also identified in BGC823 cells while latexin expression was downregulated. Further bisulfite sequencing of the LXN gene promoter indicated CpG hypermethylation was correlated with silencing of latexin expression in human cells. Latexin expression was reduced in human gastric cancers compared with their normal control tissues. The cellular and molecular evidences demonstrated the inhibitory effect of latexin in human gastric cancer cell growth and tumorigenicity. These results strongly suggest the possible involvement of latexin expression in tumor suppression.


Parkinson disease with mild cognitive impairment: Domain-specific cognitive complaints predict dementia.
The presence of subjective cognitive complaints (SCC) as a predictor of cognitive impairment in Parkinson´s disease (PD) has shown conflicting results. Most previous studies only assessed complaints in the memory domain. We investigate the association of SCCs across cognitive domains with development of mild cognitive impairment (PD-MCI) and dementia (PDD) in PD and to assess agreement between SCCs and objective cognitive impairments in this population. This is a retrospective analysis of a prospective cohort study. Participants were enrolled at six North-American movement disorders centers. They underwent neuropsychological and non-cognitive clinical evaluations, including the modified Neurobehavioral Inventory to elicit SCC (rated by each patient and independently by their close contact (CC)). Associations between SCCs and development of future cognitive impairment were assessed. Agreement between SCCs and objective impairment within the same domain was also calculated. Of 138 included PD patients, 42% fulfilled criteria for PD-MCI. None of the NBI items predicted development of cognitive impairment after one and two years in PD with normal cognition. In PD-MCI patients, SCCs related to attention predicted dementia at year one. CC ratings of SCCs related to memory and language problems predicted PDD in PD-MCI patients. According to CC reported patients' complaints, there was a significant agreement between SCCs and objective cognitive test scores on attention. Eliciting SCCs including cognitive domains other than memory is crucial for a complete evaluation, including both patient and CC report. Memory, language, and especially attention SCCs in PD-MCI may predict progression to dementia.


Intricacies of cardiac damage in coxsackievirus B3 infection: implications for therapy.
Heart disease is the leading cause of death in humans, and myocarditis is one predominant cause of heart failure in young adults. Patients affected with myocarditis can develop dilated cardiomyopathy (DCM), a common reason for heart transplantation, which to date is the only viable option for combatting DCM. Myocarditis/DCM patients show antibodies to coxsackievirus B (CVB)3 and cardiac antigens, suggesting a role for CVB-mediated autoimmunity in the disease pathogenesis; however, a direct causal link remains to be determined clinically. Experimentally, myocarditis can be induced in susceptible strains of mice using the human isolates of CVB3, and the disease pathogenesis of postinfectious myocarditis resembles that of human disease, making the observations made in animals relevant to humans. In this review, we discuss the complex nature of CVB3-induced myocarditis as it relates to the damage caused by both the virus and the host's response to infection. Based on recent data we obtained in the mouse model of CVB3 infection, we provide evidence to suggest that CVB3 infection accompanies the generation of cardiac myosin-specific CD4 T cells that can transfer the disease to naïve recipients. The therapeutic implications of these observations are also discussed.


A procedure for measuring gastric bleeding caused by drugs.
Seven subjects were given one day's treatment with full doses of either aspirin, ibuprofen, or placebo, and then their stomachs were washed out three times at intervals of 10 min, for five successive days. The blood in the recoveries was estimated chemically. After compensating for gastric emptying, the rate of bleeding was expressed as ml blood per day. Relative to placebo, one day's treatment with aspirin increased the rate of blood loss significantly at day 1, median 2.7 ml/day, and not thereafter. A corresponding dose of ibuprofen did not produce any significant bleeding. The procedure gives specific information about gastric bleeding. It can measure rates of bleeding down to 0.01 ml/day.


Surface Attachment of Ammonia-Oxidizing Bacteria in Soil.
A BSTRACTIndigenous ammonia-oxidizing bacteria (AOB) in a clay loam soil were extremely difficult to release from soil particles compared to most heterotrophic bacteria; less than 1% of indigenous AOB (estimated as potential ammonia oxidation rate) were extractable by the dispersion-density-gradient centrifugation technique. This is at least 10-fold less than the extractability of heterotrophic bacteria. Urea applications to the same soil induced a 5-fold increase in the potential ammonia oxidation rate, and this resulted in a much higher percentage (8%) extractability of AOB. Thus, the newly grown AOB in the urea-treated soil were less strongly attached to the soil particles. The contrast suggests that the strong attachment of indigenous AOB is a gradual process taking place due to a long residence time (infrequent/slow cell division) compared to heterotrophic organisms. However, the contrast could also reflect differences in species composition of the original AOB community and those growing in response to urea inputs. Specific detection of AOB in extinction dilution cultures was done by PCR and sequencing of the products. Considerable diversity was found within the genus Nitrosospira, but severe problems with the specificity of the primers were observed. Two allegedly AOB specific PCR primers pairs were used: one specific for Nitrosospira (SPIRA) and one which should encompass all AOB within the beta- Proteobacteria (GAOB). Only 33% of the cultures that gave PCR products with GAOB also gave products with the SPIRA primer pair, suggesting the presence of AOB other than Nitrosospira. However, the phylogeny based on the sequencing placed all the cultures in various clusters of the Nitrosospira clade, suggesting that the SPIRA primers do not match all members of the Nitrosospira genus. The cultures obtained from the urea-treated soil were different from the others in giving PCR products only with the SPIRA primers and not with the GAOB. Since sequencing also here confirmed the presence of Nitrosospira, these observations suggest that the GAOB primers do not match all AOB species.


Plaques en prairie fauchée.
We report a clinical picture of a mowed meadow pattern of the tongue in a 29-year-old sexually active man who has sex with men. The painful lesions also called "Plaques en prairie fauchée", occur in the context of the discovery of a postive HIV serology. The diagnosis of secondary syphilis was confirmed by a positive syphilis serology (nontreponemal RPR test title of 64) and a positive Treponema pallidum PCR, amplifying the tpp47 gene, on a tongue swab. The lesions completely resolve with a penicillin treatment.


Hybrid neurofibroma/schwannoma of the orbit.
Hybrid neurofibroma/schwannoma is a rare variant of hybrid peripheral nerve sheath tumours (HPNST). A Medline search up to December 2021 identified only six cases of this tumour in the orbit. We report the case of a 78-year-old man who presented with left exophthalmos. Computed tomography showed a left intraconal orbital mass. The clinico-radiological diagnosis was consistent with an intraconal cavernous angioma. Orbitotomy was performed, obtaining an 18×16×11mm mass. Two different morphologies were seen microscopically, diagnostic of hybrid neurofibroma/schwannoma. HPNSTs of the orbit are uncommon and most reported cases showed a hybrid neurofibroma/schwannoma morphology. Hybrid neurofibroma/schwannomas have been associated with neurofibromatosis and schwannomatosis. Local recurrences have been reported. The correct identification of these tumours is important due to their potential use as a syndromic marker.


A study of the effects of chromium exposure on the growth of Pseudokirchneriella subcapitata (Korshikov) hindak evaluated by Central Composite Design and Response Surface Methodology.
The aim of this study was to evaluate the effects of chromium exposure on the growth of P. subcapitata using the Central Composite Design (CCD) and Response Surface Methodology (RSM). The highest values for algal density and biomass were obtained in the longest exposure times and for the lowest chromium concentrations. The CCD used for the analysis of treatment combinations showed that a second order polynomial regression model was in good agreement with experimental results, with R (2) = 81.50 and 89.90; for algal density and biomass (p < 0.05), respectively. Only the exposure time was significant for algal density. For chlorophyll, in contrast, the exposure time, chromium concentration and their interaction significantly affected the growth of P. subcapitata. The findings confirmed the sensitivity of P. subcapitata to chromium (VI), which makes it a suitable bioindicator of environmental contamination for this metal.


Rheumatologic manifestations of pregnancy.
Pregnancy, whether normal or complicated, induces change in nearly every system of the body. Because most rheumatologic disorders are multisystemic and often affect young women, it may be difficult to differentiate pregnancy-related change from new onset or exacerbation of rheumatic disease. Familiarity with common manifestations of pregnancy is important in evaluating young women of childbearing age,whether or not they have known rheumatologic disease. Presentation of new connective tissue disease during pregnancy is often associated with poorer prognosis, so it is especially important to distinguish between pregnancy-induced change and true autoimmune inflammation requiring prompt and aggressive therapy.


A Prenatal Ultrasound Scanning Approach: One-Touch Technique in Second and Third Trimesters.
This study was aimed at evaluating the performance of the innovative technique Smart Fetus (SF) developed to recognize the planes and obtain the basic biometric measurements of fetuses automatically. This prospective study included 1005 uncomplicated singleton pregnancies undergoing routine examinations. For every pregnancy, planes, including the transverse section of the thalami, transverse section of the abdomen and longitudinal section of the femur, were acquired, and standard biometric measurements, including biparietal diameter, head circumference, abdominal circumference and femur length, were obtained using SF and traditional ultrasound technique (TUT). The accuracy, reproducibility and time required for the analysis of SF were compared with those of TUT. In 998 of 1005 cases (99.30%), SF successfully acquired the sections and made all measurements. The agreement between the techniques was high for all measurements. The time to obtain sections and measure biometric parameters or solely measure biometric parameters was significantly shorter with SF than with TUT. No significant differences were found in SF repeated measurements obtained by two independent observers. The SF technique helped in the acquisition of reliable standard sections and biometric measurements and saved time. It might serve as a novel ultrasound scanning approach and improve workflow efficiency.


Influence of a bleaching agent on surface and mechanical properties of orthodontic thermoplastic retainer materials : An in vitro study.
To investigate the effect of a tooth whitening agent on surface roughness, surface hardness, and force delivery properties of polyethylene terephthalate glycol (PETG) thermoplastic retainer materials of two different thicknesses. PETG sheets (1 mm and 1.5 mm) were thermoformed over a 30 × 60 × 10 mm3 rectangular stone model. Surface hardness, surface roughness, and results of a three-point bending test were evaluated before and after treatment (5 h daily for 14 days) with a 15% carbamide peroxide home bleaching agent. Data were analyzed using an independent sample t‑test. Exposure of PETG specimens to the bleaching agent for 14 days significantly reduced surface hardness and increased surface roughness of samples of both thicknesses. The magnitude of force at different deflection points decreased significantly in the 1.5 mm thickness specimens treated with the bleaching agent, but no significant change was observed in the specimens with 1 mm thickness. Use of a home bleaching agent could have adverse effects on surface roughness and hardness of PETG materials leading to reduced survival time of thermoplastic orthodontic retainers.


Olfactory dysfunction in LRRK2 G2019S mutation carriers.
Olfactory dysfunction is an established nonmotor feature of idiopathic Parkinson disease (PD), which may precede disease onset. Olfaction is likely disturbed in patients with PD with leucine-rich repeat kinase (LRRK2) G2019S mutations, although the degree of impairment is debated. It is also unclear whether mutation carriers who have not yet manifested with PD have olfactory disturbances. Thirty-one subjects with LRRK2 G2019S mutation-related PD (PD-manifesting carriers [PD-MC]), 30 subjects with PD without mutations (PD noncarriers [PD-NC]), 28 mutation carrier family members (nonmanifesting carriers [NMC]), and 46 controls completed the University of Pennsylvania Smell Identification Test (UPSIT). Generalized estimating equations were applied to determine whether olfactory score was associated with PD and LRRK2 mutation status. As expected, having PD was associated with impaired olfaction regardless of LRRK2 mutation status. More importantly, however, impaired olfaction was increased overall in LRRK2 carriers both with and without PD, though the impairment was only present in a subset of NMCs. Compared to controls, the mean score was lower among NMC (difference = -3.518, p = 0.006), MC (difference = -7.677, p < 0.0001), and idiopathic PD (PD-NC) (difference = -13.810, p < 0.0001). Olfaction was better among MC (PD-MC) than non-LRRK2 PD (PD-NC) (difference = 6.13, p = 0.0012). Group differences from the continuous analysis were maintained in dichotomous analysis stratifying at 15th percentile for age and gender. Olfaction is impaired in LRRK2 G2019S-mutation related PD, although less overall than other PD. Further, olfaction is impaired in a subset of LRRK2 NMC, suggesting that olfaction may be a marker for development of PD in this group, and that longitudinal studies are warranted.


Individual and household-level predictors of health related quality of life among middle-aged people in rural Mid-east China: a cross-sectional study.
China has an enormous and growing middle-aged population. Little is known about health-related quality of life (HRQOL) for this group, especially in rural areas. We examined HRQOL and its individual and household predictors among middle-aged people in rural Mid-east China. HRQOL questionnaires and information about individual and household characteristics were collected from 428 subjects aged 45 to 65 in 12 villages in Mid-east China. We examined the eight dimensions of the SF-36 instrument, along with the Physical Component Summary (PCS) and Mental Component Summary (MCS) using a reference sample in Hong Kong for standardization. Individual and household predictors of PCS and MCS were examined by one-way ANOVA and binary logistic regression analysis. Self-reported HRQOL was similar to that seen in middle-aged populations elsewhere. Based on univariate analyses, PCS differed by age, education, occupation, household per capita income, drinking water supply, and frequency of household members caring about each other; MCS differed by education, household per capita income, drinking water supply, and frequency of caring about each other. Individual and household-level factors accounted for 12.5% and 8.2% of the variance in PCS, respectively, and for 3.1% and 10.7% of the variance in MCS. HRQOL among middle-aged people in rural China appears similar to that observed elsewhere, and varies by income, education, and other factors. Household factors, particularly the extent to which household members care about each other, are significant predictors of physical and mental health. In addition to improving general socioeconomic conditions, efforts to improve HRQOL for middle-aged people in rural China need to focus on the family environment.


Two-column simulated moving-bed process for binary separation.
A two-column simulated moving-bed system has been developed for binary separation. The system combines a flexible node design, robust pump configuration, and cyclic flow-rate modulation to exploit the benefits of simulated counter-current operation. The feasibility of the proposed two-column system is demonstrated on the linear separation of two nucleosides by reversed phase. Emphasis is given to the potentialities of the process compared to single-column batch chromatography with recycling for the same amount of stationary phase. The performance of the proposed two-column process is verified with laboratory-scale experiments and detailed simulations for different difficulties in separation and desorbent-to-feed ratios.


The role of basking in the development of endothermy and torpor in a marsupial.
Marsupials have a slow rate of development and this allows a detailed examination of thermoregulatory developmental changes and stages. We quantified the cooling rates of marsupial dunnarts (Sminthopsis crassicaudata) at 40-56 days (d) old, and torpor and basking behaviour in animals given the option to bask in four age groups from 60 to 150 d. The development of thermoregulation was a continuum, but was characterised by three major thermoregulatory stages: (1) at 40 d, animals were unable to maintain a constant high body temperature during short-term cold exposure; (2) at 60 d, animals could maintain a high T b for the first part of the night at an ambient temperature of 15.0 ± 0.7 °C; later in the night, they entered an apparent torpor bout but could only rewarm passively when basking under a heat lamp; (3) from ~90 d, they expressed prolonged torpor bouts and were able to rewarm endogenously. Young newly weaned 60 d animals were able to avoid hypothermia by basking. In this case, basking was not an optional behavioural method of reducing the cost of rewarming from torpor, but was essential for thermoregulation independent of the nest temperature. Results from our study suggest that basking is a crucial behavioural trait that permits young marsupials and perhaps other juvenile altricial mammals to overcome the developmental stage between poikilothermy early in development and full endothermy later in life.


COVID-19 vaccination program at a student-run free clinic: A descriptive study.
People historically excluded from receiving medical care in the United States, in addition to being at greater risk for SARS-CoV-2 infection, have had slower vaccine uptake due to structural barriers to availability. We present one student-run free clinic's SARS-CoV-2 vaccination program from January 15 to August 1, 2021, in Nashville, Tennessee. We tracked SARS-CoV-2 vaccine primary series completion among 273 free clinic patients with the help of medical student volunteers, who scheduled appointments and answered vaccine-related questions. We worked with our academic medical center partner to host a single-dose vaccination event at our clinic. We compared vaccine series completion in our clinic to adult vaccine completion in Davidson County, Tennessee on August 1, 2021. Of the 273 free clinic participants, 144 identified as Spanish-speaking (52.7%) and 172 (63%) had at least one qualifying comorbidity per the December 30, 2020, Tennessee COVID-19 Vaccination Plan. As such, 183 (67%) were characterized as vaccine eligible in Phase 1a2, 1b, or 1c. On August 1, 2021, 63.1% of free clinic patients had completed their primary SARS-CoV-2 vaccination series compared with 58.9% of adults in Davidson County, Tennessee (RD 4.2%, 95% CI: -1.5% to 9.9%). Spanish-speaking free clinic patients were most likely to have completed their vaccination series. We describe a framework for a patient-centered vaccination effort to reach individuals traditionally missed by large vaccination campaigns. We highlight structural hurdles experienced by vulnerable populations, including language barriers, lack of technology or reliable internet access, inflexible working schedules, lack of transportation, and vaccine misinformation.


The relationship between expression of PD-L1 and HIF-1α in glioma cells under hypoxia.
Hypoxia inducible factor-1α (HIF-1α) up-regulates the expression of programmed death ligand-1 (PD-L1) in some extracranial malignancies. However, whether it could increase PD-L1 expression in intracranial tumor is still unknown. Here, we explored the relationship between HIF-1α and PD-L1 expression in glioma, and investigated their clinical significance. In glioma patients, HIF-1α and PD-L1 were overexpressed in high grade glioma tissues and were significantly associated with poor survival. In glioma cells, PD-L1 expression was induced under hypoxia condition, and the enhanced PD-L1 expression was abrogated by either HIF-1α knock-down or HIF-1α inhibitor treatment. Furthermore, ChIP-qPCR analysis showed the direct binding of HIF-1α to PD-L1 proximal promoter region, providing evidence that HIF-1α up-regulates PD-L1 in glioma. In glioma murine model, the combination treatment with HIF-1α inhibitor and anti-PD-L1 antibody caused a more pronounced suppressive effect on tumor growth compared to either monotherapy. Immunologically, the combination treatment improved both dendritic cell (DC) and CD8+ T cell activation. Overall, our results demonstrated that positive correlation between PD-L1 and HIF-1α in glioma, and provide an alternative strategy, inhibiting HIF-1α, as combination therapies with immunotherapies to advance glioma treatment.


Paper-based mediatorless enzymatic microfluidic biofuel cells.
In this study, eco-friendly and disposable paper-based membraneless microfluidic enzymatic fuel cells (EFCs) were developed without any mediators to reduce the toxicity and cost of EFCs. Glucose oxidase and laccase were immobilized on multi-walled carbon nanotube electrodes to catalyze the redox reaction of glucose and oxygen. Micromachining techniques well-suited for mass production were used to precisely fabricate micro-scale Y-shaped and cross-shaped EFCs. Experimental measurements showed that the concentration of glucose in the fuel solution affects the cell performance, which occurs because the flow speed of the fuel stream decreases as the concentration of glucose increases. The highest performance of power density (104.2 ± 3.35 μW cm-2) and current density (615.6 ± 3.14 μA cm-2) were obtained with the Y-shaped channel configuration at a glucose concentration of 100 mM. This performance is the best of all paper-based single EFCs reported to date. The new paper-based co-laminar flow mediatorless EFC exhibits strong potential to power miniaturized and portable on-site diagnostic devices.


Pharmacokinetics and tissue distribution of schizonepetin in rats.
Schizonepetin, a natural monoterpene from Herba Schizonepetae, is a potential antiviral agent. In this paper, a simple, rapid and sensitive HPLC-UV method was first developed and validated for the determination of schizonepetin in rat plasma and tissue homogenates after oral and intravenous administration. The results showed that schizonepetin was absorbed and eliminated rapidly, and its oral absolute bioavailability in rats achieved about 75%. The drug distributed widely in various tissues of rats, and had no long-term accumulation in vivo. The research provides reliable scientific data for designing drug treatment regimens of schizonepetin.


Adaptive management as a foundational framework for developing collaborative estuary management programs.
Collaborative nonregulatory programs can benefit the long-term sustainability of environmental resources. Such programs benefit from extensive planning and assessment relative to ecological systems as well as public participation. While many programs use adaptive management as a guiding programmatic framework, few programs successfully integrate social and human context into their adaptive management frameworks. While this adaptive governance framework can be successful, many potential challenges arise when incorporating public stakeholders into the adaptive management framework. To reduce those challenges, programs need participation from diverse stakeholder groups that represent multiple communities of interest, place, and identity. The participatory process benefits from a diverse group of stakeholders and can result in successful management of environmental resources. We highlight the participatory co-management process of three newly developing nonregulatory programs that are modeled after the United States EPA's National Estuary Program in the Perdido and Pensacola Bay systems, Choctawhatchee Bay, and the St. Andrew and St. Joseph Bay systems (Florida USA). This case study illustrates how collaborative nonregulatory programs can be implemented not only in the United States, but also in other regions of the world.


Digital financial inclusion and environmental entrepreneurship: evolution of state legal environmental responsibility in China.
This investigation attempts to look at how digital finance and environmental taxes affect environmental entrepreneurship. We have used the QARDL model to evaluate the short- and long-run estimations. The main findings of the QARDL model confirm that digital finance is vital in promoting the environmental entrepreneurship model in the long run. In contrast, in the short run, digital finance does not significantly impact environmental entrepreneurship. Moreover, environment-related taxes, environmental innovations, and economic development promote environmental entrepreneurship in the long run across most quantiles. However, in the short run, environment-related taxes and economic development do not exert any significant impact on environmental entrepreneurship in almost all quantiles, while environmental innovation promotes environmental entrepreneurship in most quantiles. Further, asymmetric effects of digital finance (ATM and debit card), environment-related taxes, environmental innovations, and economic development are confirmed in the long run; in the short run, asymmetric effects are observed in the case of environmental innovation in both the models. Following the results, we suggest that the public and private sectors should do more financing to promote ecopreneurship in society. Moreover, more taxes should be imposed on entrepreneurs who do not adopt green practices.


The Mealtime Audit Tool (MAT) - Inter-Rater Reliability Testing of a Novel Tool for the Monitoring and Assessment of Food Intake Barriers in Acute Care Hospital Patients.
Barriers to food intake (FI) exist in hospital that could exacerbate insufficient FI and malnutrition. The Mealtime Audit Tool (MAT) is a staff-administered clinical assessment tool to identify FI barriers for individual patients. Two studies were completed. The objectives of the first study were to test a draft version of the tool and characterize barriers to food intake in older adults in four diverse hospitals, while the second study aimed to demonstrate the inter-rater reliability of the revised MAT. Multi-site, cross sectional. Four acute care hospitals in Canada. Study 1: 120 older (65+ years, adequate cognition) medical or surgical patients. Study 2: 90 medical or surgical patients. In study 1, participants had barriers experienced at one mealtime assessed with MAT. Descriptive analyses characterized the prevalence of barriers across the hospitals. Revisions were made to the MAT based on recommendations from sites. A revised version was tested for inter-rater reliability in study 2. Intraclass correlation coefficient (ICC) was calculated for total MAT scores from 90 patient meals assessed by two raters. Kappa statistics were calculated for each of the 18 MAT items. Mean (+/- standard deviation) number of barriers experienced in Study 1 was 2.93 +/- 1.58, and in Study 2 was 2.51 +/- 1.19. The revised MAT was reliable with an ICC of 0.68 (95%CI: 0.52-0.79). Ten of 16 items in which kappa could be calculated had at least fair agreement. MAT is sufficiently reliable when used by auditors with minimal training. Routinely auditing mealtimes with MAT could be useful in identifying and removing barriers to food intake for older hospitalized patients.


Breastfeeding and insulin requirements in women with Type 1 diabetes mellitus in the first year postpartum.
To explore whether breastfeeding affects postpartum insulin requirements, HbA1c levels, and pregnancy weight retention in women with Type 1 Diabetes Mellitus (T1DM). This prospective study included 66 women with T1DM. The women were divided into two groups based on whether they were breastfeeding (BF) at 6 months postpartum (BFyes, n = 32) or not (BFno, n = 34). Mean daily insulin requirement (MDIR), HbA1c levels, and pregnancy weight retention at 5 time-points from discharge to 12 months postpartum were compared. MDIR increased by 35% from 35.7 IU at discharge to 48.1 IU at 12 months postpartum (p < 0.001). MDIR in BFyes and BFno were comparable, however in BFyes, MDIR were continuously lower compared to BFno. Postpartum HbA1c increased rapidly from 6.8% at 1 month to 7.4% at 3 months postpartum and settled at 7.5% at 12 months postpartum. The increase in HbA1c during the first 3 months postpartum was most pronounced in BFno (p < 0.001). Although neither were statistically significant, from 3 months postpartum HbA1c levels were highest in the BFno and BFno had a higher pregnancy weight retention compared to BFyes (p = 0.31). In women with T1DM, breastfeeding did not significantly affect postpartum insulin requirements, HbA1c levels or pregnancy weight retention in the first year after delivery.


Monocytes direct T lymphocyte stimulation of human peripheral blood granulocyte-macrophage colony formation.
The committed granulocyte-macrophage progenitor cell (or colony-forming unit, CFU-GM) differentiates in vitro under the influence of the soluble glycoprotein factor(s) termed colony stimulating activity (GM-CSA), which can be derived from several cellular sources, including normal human peripheral blood mononuclear cells (PBMNC). We have investigated the respective roles of the monocyte and T lymphocyte components of PBMNC in generating GM-CSA using as an assay CFU-GM colony formation by purified peripheral blood null cells. Highly purified unstimulated T-lymphocytes did not themselves stimulate null cell CFU-GM colony formation, but when monocytes were also included, large numbers of colonies were observed, far greater than those stimulated by monocytes alone. This synergism was independent of HLA phenotype, occurring with T-lymphocyte and monocyte fractions from unrelated donors. Media conditioned by unstimulated purified monocytes promoted CFU-GM colonies from null cells only in the presence of T-lymphocytes, suggesting that monocytes produce a factor directing the synthesis and/or release of GM-CSA by T cells.


Informed Consent of the Chronic Total Occlusion Patient.
This article focuses on the general principles of informed consent, then highlights the particular risks associated with chronic total occlusion interventions. The goal is to provide a basic framework for the interventional cardiologist to use when having consent discussions with his or her patients.


Phytohormones: Key players in the modulation of heavy metal stress tolerance in plants.
Heavy metal (HM) stress in plants has received considerable global attention as it threatens sustainable growth in agriculture worldwide. Hence, desperate efforts have been undertaken for combating the effects of this stress in plants. Interestingly, the use of phytohormones in reducing the impact of HM toxicity has gained much momentum in the recent past. Phytohormones act as chemical messengers that improve the HM stress resistance in plants, thus allowing them to retain their growth and developmental plasticity. Their exogenous application as well as manipulation of endogenous levels through precise targeting of their biosynthesis/signaling components is a promising approach for providing a protective shield against HM stress in plants. However, for the successful use of phytohormones for field plants exposed to HM toxicity, in-depth knowledge of the key pathways regulated by them is of prime importance. Hence, the present review mainly summarizes the key conceptual developments on the involvement of phytohormones in the mitigation of HM stress in plants. The role of various genes, proteins, and signaling components involved in phytohormones associated HM stress tolerance and their modulation has also been discussed. Thus, this update will pave the way for improving HM stress tolerance in plants with the advent of phytohormones for sustainable agriculture growth in the future.


Warming efficacy of Ranger™ and FT2800 fluid warmer under different room temperatures and flow rates.
The comparison of the heating capabilities with different warming system between 3M™ Ranger™ warmer (3M) and FT2800 fluid warmer (FT) under different room temperatures and infusion rates, has been rarely reported previously. The study was then aimed to compare the warming efficacies of dry heat technology (3M) and coaxial warming system (FT) under different room temperatures and infusion rates, the advantages and disadvantages of both infusion systems would be compared to provide reference for clinical infusion practice. In the study, both target warming temperatures of 3M and FT warmer were set at 41 °C, fluid was administrated under 20, 22 and 24 °C room temperatures and drip rates of 60, 80, 100, 120, 140, 160, 200, 350 drops min-1. The fluid temperature at the outlet of the infusion tube (Toutlet) was measured and compared. The Toutlet of FT was higher than that of 3 M (P < 0.001) under different room temperatures. The Toutlet of FT increased with the room temperature raised (P < 0.05). As for 3M, Toutlet was lowest at 20 °C (P < 0.001) and no statistical difference of Toutlet was found between 22 and 24 °C (P = 0.667). Linear regression showed that the Toutlet of 3M increased with the speed up of drip rate, while the Toutlet of FT was decreased. The relationship between Toutlet & room temperature & drip rate for both 3M and FT warmers was calculated by a formula. 3M Ranger™ and FT2800 show different heating capabilities under different room temperatures and drip rates. 3M is more efficient at high flow rate while FT is more efficient at low flow rate. There is a formula relationship between Toutlet & room temperature & drip rate for both 3M and FT warmers.


PODO: Trial Design: Phase 2 Study of PF-06730512 in Focal Segmental Glomerulosclerosis.
Focal segmental glomerulosclerosis (FSGS) is characterized by proteinuria and a histologic pattern of glomerular lesions of diverse etiology that share features including glomerular scarring and podocyte foot process effacement. Roundabout guidance receptor 2 (ROBO2)/slit guidance ligand 2 (SLIT2) signaling destabilizes the slit diaphragm and reduces podocyte adhesion to the glomerular basement membrane (GBM). Preclinical studies suggest that inhibition of glomerular ROBO2/SLIT2 signaling can stabilize podocyte adhesion and reduce proteinuria. This clinical trial evaluates the preliminary efficacy and safety of ROBO2/SLIT2 inhibition with the ROBO2 fusion protein PF-06730512 in patients with FSGS. The Study to Evaluate PF-06730512 in Adults With FSGS (PODO; ClinicalTrials.gov identifier NCT03448692), an open-label, phase 2a, multicenter trial in adults with FSGS, will enroll patients into 2 cohorts (n = 22 per cohort) to receive either high- or low-dose PF-06730512 (intravenous) every 2 weeks for 12 weeks. Key inclusion criteria include a confirmed biopsy diagnosis of FSGS, an estimated glomerular filtration rate (eGFR) ≥45 ml/min/1.73 m2 based on the Chronic Kidney Disease Epidemiology Collaboration formula (30-45 with a recent biopsy), and urinary protein-to-creatinine ratio (UPCR) >1.5 g/g. Key exclusion criteria include collapsing FSGS, serious/active infection, ≥50% tubulointerstitial fibrosis on biopsy, and organ transplantation. The primary endpoint is change from baseline to week 13 in UPCR; secondary endpoints include safety, changes in eGFR, and PF-06730512 serum concentration. This ongoing trial will report the efficacy, safety, pharmacokinetics, and biomarker results of PF-06730512 for patients with FSGS. Findings from this proof-of-concept study may support further development and evaluation of PF-06730512 to treat FSGS and warrant assessment in phase 3 clinical trials.


Identification of factors that predict outpatient utilisation of a Plastic Dressing Clinic. A retrospective review of 287 paediatric burn cases.
This study investigated aetiology of burn cases presenting to the Royal Hospital for Sick Children in Edinburgh to identify factors that influence the number of outpatient visits patients make to the Plastic Dressing Clinic (PDC) following discharge. A retrospective review was performed of all paediatric burn cases presenting between January 2009 and July 2012. Information gathered included patient demographics, burn aetiology, details of inpatient admission and outpatient PDC attendance. 287 cases were identified, mean age 2.79 years (range 0-15), gender ratio 1.52:1 (M:F). Scald was the most common aetiology, n=172 (59.9%). Most burns were superficial thickness, n=173 (60.3%). One-way ANOVA showed that full thickness burn, skin graft and pressure garment therapy significantly increased the number of PDC appointments (p<0.05). Pearson correlation coefficient found that length of stay in hospital and time spent in theatre were positively correlated to the number of PDC appointments (p<0.01). Outpatient utilisation of the PDC can be predicted from burn characteristics. Full thickness burn, skin graft and pressure garment therapy are identified to significantly increase the number of PDC appointments following paediatric burn.


Photo- and thermo-activated electron transfer system based on a luminescent europium organic framework with spectral response from UV to visible range.
A dual stimuli-responsive system was constructed by introducing methyl viologen into a luminescent europium-organic framework with a compact packing mode. In addition to absorption and emission properties that are sensitive to both light and heat involving electron/energy transfer processes, the photoactive MOF has a spectral response from UV to the visible region.


Involvement of a surface concanavalin A-binding glycoprotein in the adhesion of Trichomonas vaginalis to substrates.
Treatment of Trichomonas vaginalis with EDTA removes their ability to adhere to glass surfaces and changes their affinity to Concanavalin A (ConA) by a different distribution of their surface structures. Filtrates of the EDTA-treated Trichomonas passed through affinity chromatography columns (ConA bound to Sepharose 4B) separate into 2 fractions, one fraction was bound to the ConA-Sepharose beads, the other was not. The Con A - bound fraction appears to be a glycoprotein which restores in a specific way the ability of the EDTA-treated protozoa to adhere to glass.


Comparison of root canal preparation using reciprocating Safesiders stainless steel and Vortex nickel-titanium instruments.
The aim of the present study was to assess several parameters related to the clinical usage of 2 root canal preparation instruments: Vortex .06 rotary nickel-titanium instruments, and Safesiders reciprocating stainless steel instruments. Fifty extracted mandibular molars with mesial root canal curvatures between 20° and 50° were divided into 2 groups and embedded in acrylic resin inside a modified Bramante muffle system. All root canals were prepared to ISO size 40 using either Vortex .06 rotary nickel-titanium-instruments in a low-torque motor or Safesiders stainless steel instruments in a proprietary reciprocating handpiece. The following parameters were evaluated: straightening of curved root canals, working safety issues (perforations, instrument breakages, canal blockages, loss of working length), postpreparation root canal cross-section, and working time. The Vortex .06 instruments maintained canal curvatures well, with the mean degree of straightening recorded as 0.72°. Safesiders instruments demonstrated significantly more canal straightening, with the mean degree of straightening recorded as 15.5°. More than 90% of the root canals prepared with the Vortex .06 instruments resulted in a round or oval cross-section, whereas the Safesiders instruments produced round or oval cross-sections 60% of the time. Neither of the 2 instruments could effectively prepare 100% of the root canal circumference. The area of dentin removed and the remaining dentin thicknesses from each region were similar for the 2 groups. Six procedural incidents were recorded for the Vortex .06 group, compared with 19 for the Safesiders group. There were no instrument fractures recorded in either group. Mean working time was significantly shorter for Vortex .06 (279 s) than for Safesiders (324 s). Vortex .06 maintained the original root canal curvatures well, whereas Safesiders instruments demonstrated significant straightening and irregular preparation shapes when used in sizes larger than ISO 20. Preparation of the complete circumference of the root canal was not possible with either system. Fewer procedural errors occurred with the Vortex instruments.


Interplay between splicing and transcriptional pausing exerts genome-wide control over alternative polyadenylation.
Recent studies have identified multiple polyadenylation sites in nearly all mammalian genes. Although these are interpreted as evidence for alternative polyadenylation, our knowledge of the underlying mechanisms is still limited. Most studies only consider the immediate surroundings of gene ends, even though in vitro experiments have uncovered the involvement of external factors such as splicing. Whereas in vivo splicing manipulation was impracticable until recently, we now used mutants in the Death Inducer Obliterator (DIDO) gene to study their impact on 3' end processing. We observe multiple rounds of readthrough and gene fusions, suggesting that no arbitration between polyadenylation sites occurs. Instead, a window of opportunity seems to control end processing. Through the identification of T-rich sequence motifs, our data indicate that splicing and transcriptional pausing interact to regulate alternative polyadenylation. We propose that 3' splice site activation comprises a variable timer, which determines how long transcription proceeds before polyadenylation signals are recognized. Thus, the role of core polyadenylation signals could be more passive than commonly believed. Our results provide new insights into the mechanisms of alternative polyadenylation and expand the catalog of related aberrations.Abbreviations APA: alternative polyadenylation; bp: basepair; MEF: mouse embryonic fibroblasts; PA: polyadenylation; PAS: polyadenylation site; Pol II: (RNA) polymerase II ; RT-PCR:reverse-transcriptase PCR; SF:splicing factor; SFPQ:splicing factor rich in proline and glutamine; SS:splice site; TRSM:Thymidine rich sequence motif; UTR:untranslated terminal region.


Herpes simplex virus infection limited to the brainstem.
Focal meningoencephalitis is commonly caused by Herpes simplex virus infection, which typically affects temporal or frontal lobes, and carries a mortality rate of 70% if untreated. On rare occasions, however, the infection is restricted to the brain stem. Polymerase chain reaction analysis of cerebrospinal fluid is the gold standard for the diagnosis of herpes simplex encephalitis. A 46-year-old male was admitted to the hospital with a three-day history of headache and fever up to 39 degrees C. Cerebrospinal fluid findings were in accordance with aseptic meningitis. On the third hospital day, the patient presented with double vision followed by confusion, and gaze paresis developed. The condition rapidly progressed from stupor to coma. A second examination of cerebrospinal fluid revealed a low glucose level (1.2 mmol/l) and cefotaxime with ampicillin were started empirically. All cerebrospinal fluid specimens were negative for bacteria and fungi. Serum IgG antibodies for herpes simplex virus type 1 were found with no intrathecal specific antibody synthesis. A polymerase chain reaction analysis of cerebrospinal fluid sample performed on the seventh day of his illness was negative for herpes simplex virus 1 and 2. A computer tomography scan of the brain did not show any abnormality. Despite antimicrobial and supportive intensive care, the condition of the patient progressively deteriorated and he died on the 11th day after admission. An autopsy revealed hemorrhagic and necrotic brainstem meningoencephalitis, and herpes simplex virus type 1 infection was confirmed by hybridization in situ. Herpes simplex virus encephalitis carries a mortality rate of 70% if untreated. The atypical location of the infection, as well as an atypical clinical manifestation with negative radiological and microbiological tests, could be the reasons for false diagnoses and mistreatment. Many authors advocate the use of empiric acyclovir in any patients with unexplained encephalopathy, since delay in treatment may greatly affect outcome. We describe a patient who died due to a herpes simplex virus 1 encephalitis affecting the brainstem, where nucleic acids were found post mortem by in situ hybridization. On rare occasions, the herpes simplex viral infection, as well as clinical manifestations and pathological changes, is restricted solely to the brainstem.


Impact of the European Union on access to medicines in low- and middle-income countries: A scoping review.
This Scoping Review synthesises evidence of the impacts of European Union (EU) law, regulation, and policy on access to medicines in in non-EU low- and middle-income countries (LMICs), and the mechanisms and nature of those impacts. We searched eight scholarly databases and grey literature published between 1995-2021 in four languages. The EU exerts global influence on pharmaceuticals in LMICs in three ways: explicit agreements between EU-LMICs (ex. accession, trade, and economic agreements); LMICs' reliance on EU internal regulation, standards, or methods (ex. market authorisation); 'soft' forms of EU influence (ex. research funding, capacity building). This study illustrates that EU policy makers adopt measures with the potential to influence medicines in LMICs despite limited evidence of their positive and/or negative impact(s). The EU's fragmented internal and external actions in fields related to pharmaceuticals reveal the need for principles for global equitable access to medicines to guide EU policy.


Ultra-sensitive determination of Formoterol in human serum by high performance liquid chromatography and electrospray tandem mass spectrometry.
An analytical method was developed and validated to determine Formoterol in human serum in the range from 0.40 to 100.24 pg/mL by high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) due to the lack of efficient methods to determine very low levels of Formoterol in serum and plasma. Serum was diluted by water and mixed with the internal standard (d6-Formoterol). Formoterol and internal standard were extracted using a cation-exchange solid phase column (SCX-3). After eliminating endogenous serum constituents through washing steps with water and methanol, elution took place using methanol/ammonia. After evaporation of the elution liquid the residue was redissolved and analyzed by HPLC-MS/MS with electrospray ionisation (ESI) in positive mode. A gradient between 10 mM ammonium formate and acetonitrile was used. The inter-batch precision of the calibration standards ranged from 1.55% to 9.01%. The inter-batch accuracy of the calibration standards ranged from 93.37% to 107.30%. The lower limit of quantitation (LLOQ, 0.40 pg/mL) had a precision of 19.67% and an accuracy of 96.78%. Comparable results were obtained for quality control samples. Stability in human serum was given over three freeze/thaw cycles and 2h at room temperature. Formoterol in human serum was stable for at least 6 months below -20 degrees C. This method has been used widely for quantifying Formoterol after inhalation of 9-36 microg of the drug by volunteers. A cross validation with human plasma versus serum was performed after this method was successfully validated in human serum.


Use of orthopedic manual physical therapy to manage chronic orofacial pain and tension-type headache in an adolescent.
Temporomandibular dysfunction (TMD) is frequently associated with multiple headache types. While the efficacy of orthopedic manual physical therapy (OMPT) intervention for TMD with/without headache symptoms has been investigated, it has received less attention than other musculoskeletal conditions. This case describes the OMPT treatment and outcome of a 14 year-old female with a 2-year history of facial pain surrounding the right temporomandibular joint (TMJ), and bilateral occipital and temporal tension-type headaches (TTHs). Intervention included manual mobilization of the TMJ and surrounding cervical/facial soft tissue structures, postural re-education, and patient/family education on prevention, self-treatment, and postural adaptations. Outcomes included pain free and maximal mouth opening (MMO), pressure pain threshold (PPT) over the masseter muscle belly, fear avoidance beliefs questionnaire-physical activity subscale (FABQ-PA), jaw pain and function questionnaire (JPFQ), visual analog pain scale (VAS) for facial pain, and headache frequency, duration, and intensity on a VAS. Results included increased pain free and MMO by 22 mm, improved PPT bilaterally by 403.05 kPa on the affected, and 360.88 kPa on the unaffected side, and decreased reported headache frequency, duration, and intensity. Score decreases of 5 points on VAS with MMO, 8 points on FABQ-PA, and 5 points on JPFQ were also noted. Further research is warranted to investigate the effects of OMPT on pain processing and functional outcomes in patients with TMD and headache.


[Transfer of skills: implementing post-transplant follow-up care status for transplant nurses: a report by the SFGM-TC].
The number of Hematopoietic Stem Cell Transplantations has globally taken off in the past decade. However, this increase in transplantation activity has put in the spotlight the need to create a special transplantation-skilled population of nurses. This type of specialisation allocated solely to this activity has not existed within the French nursing community until now. In the attempt to harmonize clinical practices between different French transplantation centers, the French Society of Bone Marrow Transplantation and Cell Therapy (SFGM-TC) sets up its forth annual series of workshops which brought together practitioners from all member centers and took place in September 2013 in Lille. Here we report our results and recommendations regarding the implementation of a transplant nurse status for post-transplant follow-up care.


Enhanced Echo Intensity of Skeletal Muscle Is Associated With Exercise Intolerance in Patients With Heart Failure.
Skeletal muscle is quantitatively and qualitatively impaired in patients with heart failure (HF), which is closely linked to lowered exercise capacity. Ultrasonography (US) for skeletal muscle has emerged as a useful, noninvasive tool to evaluate muscle quality and quantity. Here we investigated whether muscle quality based on US-derived echo intensity (EI) is associated with exercise capacity in patients with HF. Fifty-eight patients with HF (61 ± 12 years) and 28 control subjects (58 ± 14 years) were studied. The quadriceps femoris echo intensity (QEI) was significantly higher and the quadriceps femoris muscle thickness (QMT) was significantly lower in the patients with HF than the controls (88.3 ± 13.4 vs 81.1 ± 7.5, P= .010; 5.21 ± 1.10 vs 6.54 ±1.34 cm, P< .001, respectively). By univariate analysis, QEI was significantly correlated with age, peak oxygen uptake (VO2), and New York Heart Association class in the HF group. A multivariable analysis revealed that the QEI was independently associated with peak VO2 after adjustment for age, gender, body mass index, and QMT: β-coefficient = -11.80, 95%CI (-20.73, -2.86), P= .011. Enhanced EI in skeletal muscle was independently associated with lowered exercise capacity in HF. The measurement of EI is low-cost, easily accessible, and suitable for assessment of HF-related alterations in skeletal muscle quality.


Designing effective amiRNA and multimeric amiRNA against plant viruses.
RNA-mediated virus resistance is increasingly becoming a method of choice for antiviral defense in plants when effective natural resistance is unavailable. In this chapter we discuss the design principles of artificial micro RNA (amiRNA), in which a natural miRNA precursor gene is modified to target a different species of RNA, in particular viral RNA. In addition, we explore the advantages and effectiveness of multiple amiRNAs within one polycistronic amiRNA precursor against a virus, as illustrated with Wheat streak mosaic virus, WSMV. The judicious selection of amiRNAs, which are sequences of short length as compared to other related methodologies of RNA interference, greatly assists in avoiding unintended off-targets in the host plant. The viral sequences targeted can be genomic or replicative and should be derived from conserved regions of the published WSMV genome. In short, using published folding and miRNA selection rules and algorithms, candidate miRNA sequences are selected from conserved regions between a number of WSMV genomes, and are BLASTed against wheat TIGR ESTs. Five miRNAs are selected that are least likely to interfere with the expression of transcripts from the wheat host. Then, the natural miRNA in each of the five arms of the polycistronic rice miR395 is replaced in silico with the chosen artificial miRNAs. This artificial precursor is transformed into wheat behind a ubiquitin promoter, and its integration into transformed wheat plants is confirmed by PCR and Southern blot analysis. We have demonstrated the effectiveness of this methodology using an amiRNA precursor that we have termed Fanguard. The processing of amiRNAs in transgenic leaves is verified through splinted ligation assay, and the functionality of the transgene in preventing viral replication is verified by virus bioassay. Resistance is confirmed using mechanical virus inoculation over two subsequent generations. This example demonstrates the potential of polycistronic amiRNA to achieve stable immunity to economically important viruses.


Estimating Myocardial Motion by 4D Image Warping.
A method for spatio-temporally smooth and consistent estimation of cardiac motion from MR cine sequences is proposed. Myocardial motion is estimated within a 4-dimensional (4D) registration framework, in which all 3D images obtained at different cardiac phases are simultaneously registered. This facilitates spatio-temporally consistent estimation of motion as opposed to other registration-based algorithms which estimate the motion by sequentially registering one frame to another. To facilitate image matching, an attribute vector (AV) is constructed for each point in the image, and is intended to serve as a "morphological signature" of that point. The AV includes intensity, boundary, and geometric moment invariants (GMIs). Hierarchical registration of two image sequences is achieved by using the most distinctive points for initial registration of two sequences and gradually adding less-distinctive points to refine the registration. Experimental results on real data demonstrate good performance of the proposed method for cardiac image registration and motion estimation. The motion estimation is validated via comparisons with motion estimates obtained from MR images with myocardial tagging.


Distractor-evoked deviation in saccade direction suggests an asymmetric representation of the upper and lower visual fields on oculomotor maps.
The programming of rapid eye movements or "saccades" involves a large collection of neural substrates. The subcortical oculomotor center - the superior colliculus (SC) - contains a topographical motor map that encodes saccade vectors. Using a visual distractor task, the present study examined a classic model of the SC motor map, which assumes a symmetrical representation of the upper visual field (UVF) and lower visual field (LVF). Visual distractors are known to attract or repel the saccade trajectory, depending on their angular distance from the target. In the present study, the distractor (if presented) was placed at a location that mirrored the target in the opposite visual field (upper or lower). The symmetrical SC model predicts equivalent directional deviations for saccades into the UVF and LVF. The results, however, showed that the directional deviations evoked by visual distractors were much stronger for saccades directed to the LVF. We argue that this observation is consistent with the recent neurophysiological finding that the LVF is relatively under-represented, as compared to the UVF, in the SC and possibly in other oculomotor centers. We conclude the paper with a suggested revision to the SC model.


Randomized, placebo-controlled trial showing that finasteride reduces prostatic vascularity rapidly within 2 weeks.
To measure expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in the prostates of men after transurethral resection of the prostate (TURP) following 2 weeks of treatment with finasteride. Sixty-four men scheduled to undergo TURP were randomized to receive 5 mg of finasteride or placebo daily for 2 weeks before surgery. Sections of prostatic urothelium were stained for VEGF expression and for CD31 to assess MVD. Ten consecutive, non-overlapping high-power fields were analysed in a blinded fashion. In all, 31 men received finasteride and 33 placebo; the groups were similar in patient age, resected prostate weight, preoperative catheterization, prostate-specific antigen level, aspirin use, spinal anaesthesia and postoperative diagnosis of prostate cancer. The mean (95% confidence interval) MVD was significantly lower in the finasteride group (60, 55-65) than in the placebo group (71, 64-78; P < 0.01). Similarly, the mean expression of VEGF was significantly lower in the finasteride group (47, 43-52 vs 61, 54-67; P < 0.001) Finasteride inhibits angiogenic growth factors leading to reduced vascularity, and this is the basis of its action in reducing haematuria of prostatic origin. The present study shows that finasteride influences the prostatic microvasculature after only 2 weeks exposure.


Automated, High-Throughput Protoplast Transfection for Gene Editing and Transgene Expression Studies.
In an era of cost-efficient gene synthesis and high-throughput construct assembly, the onus of scientific experimentation is on the rate of in vivo testing for the identification of top performing candidates or designs. Assay platforms that are relevant to the species of interest and in the tissue of choice are highly desirable. A protoplast isolation and transfection method that is compatible with a large repertoire of species and tissues would be the platform of choice. A necessary aspect of this high-throughput screening approach is the need to handle many delicate protoplast samples at the same time, which is a bottleneck for manual operation. Such bottlenecks can be mitigated with the use of automated liquid handlers for the execution of protoplast transfection steps. The method described within this chapter utilizes a 96-well head for simultaneous, high-throughput initiation of transfection. While initially developed and optimized for use with etiolated maize leaf protoplasts, the automated protocol has also been demonstrated to be compatible with other established protoplast systems, such as soybean immature embryo derived protoplast, similarly described within. This chapter also includes instructions for a sample randomization design to reduce the impact of edge effects, which might be present when microplates are used for fluorescence readout following transfection. We also describe a streamlined, expedient, and cost-effective protocol for determining gene editing efficiencies using the T7E1 endonuclease cleavage assay with a publicly available image analysis tool.


PI3K/AKT signaling activates HIF1α to modulate the biological effects of invasive breast cancer with microcalcification.
Microcalcification (MC) is a valuable diagnostic indicator of breast cancer, and it is reported to be associated with increased tumor aggressiveness and poor prognosis. Nevertheless, the exact potential molecular mechanism is not completely understood. Here, we find that the mineralized invasive breast cancer (IBC) cells not only increased their proliferation and migration, but also showed the characteristic of doxorubicin resistance. The PI3K/AKT signaling pathway is associated with the generation of calcification in IBC, and it activates the transcription and translation of its downstream hypoxia-inducible factor 1α (HIF1α). Knockdown of HIF1α protein significantly downregulated cell proliferation and migration while calcification persists. Meanwhile, calcified breast cancer cells restored sensitivity to doxorubicin because of suppressed HIF1α expression. In addition, we provide initial data on the underlying value of HIF1α as a biomarker of doxorubicin resistance. These findings provide a new direction for exploring microcalcifications in IBC.


CT colonography in a Korean population with a high residue diet: comparison between wet and dry preparations.
To compare wet and dry preparation methods for computed tomography colonography (CTC) in terms of preparation quality, interpretation time, and diagnostic performance for polyp detection in a population with a high residue diet. Eighty-six patients were divided into two groups. Group 1 (n=24) received a wet preparation of 4l polyethylene glycol (PEG) solution, and group 2 (n=62) received a dry preparation of phosphor-soda. Abnormal findings, including polyps, and the time required to interpret the CTC images in both groups were documented by a radiologist. CTC findings were compared to those of colonoscopy as a reference standard. Two radiologists evaluated the quality of CTC with regard to residual fluid, faeces, and colonic distension using a four-point scale in consensus. Statistical differences for residual fluid, faeces, distensibility on CTC, and interpretation time between the two groups were analysed. The diagnostic performance of CTC in both groups was also compared. One-hundred and ninety polyps in 70 patients were identified using colonoscopy. Regarding the quality of images produced the wet preparation was significantly better than the dry preparation (p<0.05). The average interpretation time was significantly shorter for the wet group (11.7 min) than the dry group (16.4 min) (p<0.05). For per-patient analysis, the positive predictive value (PPV) was significantly better for the wet (100%) than the dry group (79.6%; p=0.025). Sensitivities and PPV for >or=10 mm polyps were comparable between two groups (p>0.05). In a population with a high-residue diet, CTC with wet preparation can be interpreted in a time-efficient manner and is comparable with CTC with dry preparation.