Excess length of stay, charges, and mortality attributable to medical injuries during hospitalization.
Although medical injuries are recognized as a major hazard in the health care system, little is known about their impact. To assess excess length of stay, charges, and deaths attributable to medical injuries during hospitalization. The Agency for Healthcare Research and Quality (AHRQ) Patient Safety Indicators (PSIs) were used to identify medical injuries in 7.45 million hospital discharge abstracts from 994 acute-care hospitals across 28 states in 2000 in the AHRQ Healthcare Cost and Utilization Project Nationwide Inpatient Sample database. Length of stay, charges, and mortality that were recorded in hospital discharge abstracts and were attributable to medical injuries according to 18 PSIs. Excess length of stay attributable to medical injuries ranged from 0 days for injury to a neonate to 10.89 days for postoperative sepsis, excess charges ranged from 0 dollar for obstetric trauma (without vaginal instrumentation) to 57 727 dollars for postoperative sepsis, and excess mortality ranged from 0% for obstetric trauma to 21.96% for postoperative sepsis (P<.001). Following postoperative sepsis, the second most serious event was postoperative wound dehiscence, with 9.42 extra days in the hospital, 40 323 dollars in excess charges, and 9.63% attributable mortality. Infection due to medical care was associated with 9.58 extra days, 38 656 dollars in excess charges, and 4.31% attributable mortality. Some injuries incurred during hospitalization pose a significant threat to patients and costs to society, but the impact of such injury is highly variable.


A novel RNPC3 gene variant expands the phenotype in patients with congenital hypopituitarism and neuropathy.
Pathogenic biallelic RNPC3 variants cause congenital hypopituitarism (CH) with congenital cataracts, neuropathy, developmental delay/intellectual disability, primary ovarian insufficiency, and pituitary hypoplasia. Here, we aimed to evaluate the clinical and molecular characteristics of two patients with CH and neuropathy. Proband was evaluated by clinical, laboratory, and radiological exams followed by exome sequencing (ES). Clinical investigation of an affected sibling and variant segregation in the family was performed by Sanger sequencing. A three-dimensional protein model study was conducted to predict the effect of the variant on the function of the RNPC3 peptide. Proband was a 16-month-old girl who was referred for the evaluation of failure to thrive. Her height, weight, and head circumference were 55.8 cm (-7.6 SDS), 6.5kg (-3.6 SDS), and 41.8 cm (-3.82), respectively. She had a developmental delay and intellectual disability. Central hypothyroidism, growth hormone, and prolactin deficiencies were identified, and MRI revealed pituitary hypoplasia. Electroneuromyography performed for the gait abnormality revealed peripheral neuropathy. A homozygous novel variant c.484C>T/p.(Pro162Ser) in the RNPC3 was detected in the ES. Her brother had the same genotype, and he similarly had pituitary hormone deficiencies with polyneuropathy. Expanding our knowledge of the spectrum of RNPC3 variants, and apprehending clinical and molecular data of additional cases, is decisive for accurate diagnosis and genetic counseling.


How to assess the feeding activity in ecotoxicological laboratory tests using enchytraeids?
The risk assessment of pesticides on soil fauna is an issue to protect agroecosystem sustainability. Enchytraeids are recognized as relevant soil bioindicators of chemical stress in agroecosystems. In laboratory, the reproduction test was found to be sensitive to reveal chemical impacts on enchytraeids. However, it does not allow to assess the impacts on ecological functions in which enchytraeids are involved. The objectives of this study were (i) to explore the feasibility of the bait-lamina test with enchytraeids under laboratory conditions and (ii) to compare its sensitivity with the Enchytraeid Reproduction Test. We exposed individuals of Enchytraeus albidus to two commercial formulations of fungicides widely used in Europe. The EC50 reproduction for the Swing® Gold (50 g L-1 epoxiconazole and 133 g L-1 dimoxystrobin) and the Cuprafor micro® (50% copper oxychloride) were respectively estimated at 1.66 ± 0.93 times the recommended dose and > 496 mg kg-1 of copper. However, no impact was found on the feeding activity of enchytraeids. The bait-lamina test thus appeared less sensitive than the Enchytraeid Reproduction Test to the tested fungicides. Despite that, this test which is achievable under laboratory conditions and allows assessing indirect effects of pesticides is quick, cheap, and easy to perform. It would deserve to be used to explore longer-exposure effects through the repeated addition of bait-lamina sticks.


Biodegradable PCL/fibroin/hydroxyapatite porous scaffolds prepared by supercritical foaming for bone regeneration.
Regenerative medicine seeks advanced solutions for bone repair in the form of bioactive synthetic scaffolds by using simple and reproducible processing techniques. In this work, poly-ε-caprolactone (PCL)-based porous scaffolds with improved osteoconductive and osteoinductive properties were processed by supercritical foaming through a careful tuning of components and processing conditions. Composite scaffolds were prepared from various combinations of PCL, silk fibroin and nano-hydroxyapatite (nHA). The green and cost-effective supercritical CO2 foaming method applied rendered solid scaffolds with 67-70% porosity. The incorporation of fibroin and nHA in the scaffolds increased the compressive modulus, cellular adhesion and calcium deposition. The composite scaffolds were tested in vivo in a large-scale calvarial defect model, and bone regeneration was evaluated for up to 14 weeks after implantation. Histomorphometric results showed that all implanted constructs gave rise to the endochondral bone formation and unveiled the synergistic effect of silk fibroin and nHA on the bone repair extent. The information gathered may shed light on the design and processing criteria of bioactive bone scaffolds.


Comparative study of the effect of Shugan Shuru Granule on pathology and p53 gene expression in patients with hyperplastic disease of breast.
To study the effect of Shugan Shuru Granule (SSG) on the p53 gene expression in patients with hyperplastic disease of breast (HDB) to indirectly explore the mechanism of SSG's effect on HDB on the molecular pathological level. Sixty-six patients with HDB were allocated in the treated group and the control group, with the former treated with SSG and the latter not. All patients underwent breast operation and their diseased mammary tissues were cut out, sectioned, and observed under microscopy with HE staining and immunohistochemical staining, with ABC method adopted to estimate the degree of hyperplasia and p53 gene expression. The severity of HDB was classified into normal, mild, moderate and severe grades (marked as 0 to III), according to the degree of hyperplasia in the mammary gland. Hyperplasia in the control group mostly belonged to grade I-III before treatment, showing overgrowth of gland and proliferation of glandular epithelial cells, which were high columnar shaped, more stratified, with papillary or substantive dysplasia. While in the treated group, most belonged to grade 0-I after SSG treatment, with proliferated gland and dysplasia recovered to normal or disappeared. The positive rate of p53 gene expression in the treated group was 9.09%, and in the control group 39.39%, comparison between the two groups showing significant difference (P < 0.01), the intensity in the former was significantly weaker than that in the latter. SSG could not only inhibit the proliferation of mammary duct epithelia and lobuli, but also inhibit the over-expression of P53. Therefore, it could be regarded as an effective remedy for treatment of HDB and prevention of mammary cancer genesis.


Cytokine-induced killer cells: NK-like T cells with cytotolytic specificity against leukemia.
Cytokine-induced killer (CIK) cells are a unique population of cytotoxic T lymphocytes (CTL) with the characteristic CD3+CD56+ phenotype. These cells have demonstrated higher proliferative and cytolytic activities in comparison to the reported CD3-CD56+ lymphokine activated killer (LAK) cells that are essentially activated natural killer (NK) cells. CIK cells are non-MHC-restricted in target cell recognition and killing. We have shown the feasibility of generating CIK cells from a series of marrow samples of patients with acute myeloid leukemia (AML) collected at diagnosis. At maturity, the CIK cells exhibit potent cytotoxicity against autologous AML targets as well as allogeneic myeloid leukemia cells, regardless of the HLA types of these targets. This observed cytotoxicity is not entirely due to NK cells as prior pre-absorption of the NK cells cytolytic activities does not abolish the subsequent cytotolytic activities against leukemic targets. It has also been reported by others that CIK cells are cytolytic against chronic myeloid leukemia (CML) cells, both in vitro and in the SCID mouse tumor model. In a mouse transplant model across MHC barrier, the CIK cells generated from the donor do not induce graft vs. host disease as observed for unfractionated donor splenocytes. In comparison to untreated control mice, the infusion of CIK cells results in the prolonged survival of murine leukemia-bearing mice. CIK cells also express CD94, part of the NK receptor comprising of CD94-NKG2 heterodimer. However, only low level of the killer immunoglobulin-like receptors are expressed by the CIK cells. In addition, as reported for the classical CTL, CIK cells could interact with dendritic cells (DC) to result in the enhancement of cytotolytic activities against tumor cells. The characteristic biological properties of the CIK cells would, therefore, enable them to be exploited for anti-leukemic therapy.


Effects of Qigong Exercise on Non-Motor Symptoms and Inflammatory Status in Parkinson's Disease: A Protocol for a Randomized Controlled Trial.
Background: Non-motor symptoms such as sleep disturbance, cognitive decline, fatigue, anxiety, and depression in Parkinson's disease (PD) impact quality of life. Increased levels of pro-inflammatory cytokines in individuals with PD have been reported, which may contribute to non-motor symptoms. A mind-body exercise, Qigong, has demonstrated benefits across different medical conditions. However, a lack of evidence causes clinicians and patients to be uncertain about the effects of Qigong in individuals with PD. This study will examine the effects of Qigong on non-motor symptoms and inflammatory status in individuals with PD. Methods: Sixty individuals with PD will be recruited. Qigong and sham Qigong group (n = 30 for each) will receive a 12-week intervention. Participants will practice their assigned exercise at home (2×/day) and attend routinely group exercise meetings. Results: Clinical questionnaires and neuropsychological tests will measure non-motor symptoms including sleep quality (primary outcome). Biomarker assays will measure inflammatory status. A two-way mixed-design analysis of variance (ANOVA) will be utilized. Conclusions: This study may generate evidence for the benefits of Qigong on non-motor symptoms of PD and the effect on inflammatory status. Findings may lead to the development of a novel, safe, and cost-effective rehabilitation approach for individuals with PD.


Role of solvent and dendritic architecture on the redox core encapsulation.
Dendrimers with redox cores can accept, donate, and/or store electrons and are used in nanoscale devices like artificial receptors, magnetic resonance imaging, sensors, light harvesting antennae, and electrical switches. However, the dendrimer molecular architectures can significantly alter the encapsulation of the redox core and charge transfer pathways, thereby changing the electron transfer rates. In this study, we used molecular dynamics simulations to investigate the role of solvent and peripheral groups on molecular structure and core encapsulation of iron-sulfur G2-benzyl ether dendrimers in polar and nonpolar solvent. We found that the dendrimer branches collapse in water and swell in chloroform. The presence of the long hydrophobic alkyl groups at the periphery deters the encapsulation of the core in water which may cause an increase in electron transfer rate. However, in chloroform, the dendrimer branches remain in the extended form, which leads to an increased radius of gyration. Our results suggest that peripheral alkyl chains in dendrimers cause steric hindrance, which prevents branches from back folding in chloroform solvent, but in water it reverses the trend. Overall, the presence of a hydrophobic interior and hydrophilic periphery in a dendrimer improves core encapsulation in water while hindering encapsulation in chloroform.


Varicella zoster immunoglobulin after postnatal exposure to varicella: survey of experts.
A questionnaire based on a hypothetical 6-day-old infant exposed to a sibling with varicella was mailed to 316 Pediatric Infectious Diseases Society members to determine their recommendations for administration of varicella zoster immunoglobulin (VZIG). Of the 137 faculty members who responded 20% would recommend VZIG for the infant described (13% were undecided). When further information including a negative maternal history for varicella, prematurity or maternal varicella infection occurring 1 week after delivery was considered, the numbers who recommended VZIG (43, 46 and 52%, respectively) rose significantly (P less than 0.01). Twenty-two physicians had knowledge of a severe case of varicella following postnatal exposure. Only 33% of faculty members surveyed feel there are no indications for VZIG in infants exposed after 48 hours of age.


Cross-Resistance between Cry1 Proteins in Fall Armyworm (Spodoptera frugiperda) May Affect the Durability of Current Pyramided Bt Maize Hybrids in Brazil.
Genetically modified plants expressing insecticidal proteins from Bacillus thuringiensis (Bt) offer valuable options for managing insect pests with considerable environmental and economic benefits. Despite the benefits provided by Bt crops, the continuous expression of these insecticidal proteins imposes strong selection for resistance in target pest populations. Bt maize (Zea mays) hybrids have been successful in controlling fall armyworm (Spodoptera frugiperda), the main maize pest in Brazil since 2008; however, field-evolved resistance to the protein Cry1F has recently been reported. Therefore it is important to assess the possibility of cross-resistance between Cry1F and other Cry proteins expressed in Bt maize hybrids. In this study, an F2 screen followed by subsequent selection on MON 89034 maize was used to select an S. frugiperda strain (RR) able to survive on the Bt maize event MON 89034, which expresses the Cry1A.105 and Cry2Ab2 proteins. Field-collected insects from maize expressing the Cry1F protein (event TC1507) represented most of the positive (resistance allele-containing) (iso)families found. The RR strain showed high levels of resistance to Cry1F, which apparently also conferred high levels of cross resistance to Cry1A.105 and Cry1Ab, but had only low-level (10-fold) resistance to Cry2Ab2. Life history studies to investigate fitness costs associated with the resistance in RR strain revealed only small reductions in reproductive rate when compared to susceptible and heterozygous strains, but the RR strain produced 32.2% and 28.4% fewer females from each female relative to the SS and RS (pooled) strains, respectively. Consistent with the lack of significant resistance to Cry2Ab2, MON 89034 maize in combination with appropriate management practices continues to provide effective control of S. frugiperda in Brazil. Nevertheless, the occurrence of Cry1F resistance in S. frugiperda across Brazil, and the cross-resistance to Cry1Ab and Cry1A.105, indicates that current Cry1-based maize hybrids face a challenge in managing S. frugiperda in Brazil and highlights the importance of effective insect resistance management for these technologies.


Change in health risks and work productivity over time.
We sought to examine the relationship between changes in health risks and changes in work productivity. Pre- and postanalysis was conducted on 500 subjects who participated in a wellness program at a large national employer. Change in health risks was analyzed using McNemar chi-square tests, and change in mean productivity was analyzed using paired t tests. A repeated measures regression model examined whether a change in productivity was associated with a change in health risks, controlling for age and gender. Individuals who reduced one health risk improved their presenteeism by 9% and reduced absenteeism by 2%, controlling for baseline risk level, age, gender, and interaction of baseline risk and risk change. In conclusion, reductions in health risks are associated with positive changes in work productivity. Self-reported work productivity may have utility in the evaluation of health promotion programs.


Phase II trial of continuous infusion carboplatin, 5-fluorouracil, and radiotherapy for localized cancer of the esophagus.
The aim of this study was to determine the toxicity, response rate, failure-free survival, and overall survival in a treatment program comprising continuous infusion carboplatin, short in-fusion 5-fluorouracil (5-FU) and radiotherapy for localized carcinoma of the thoracic esophagus. To be eligible, patients were required to have Karnofsky performance status greater than or equal to 60, adequate organ function, and have received no prior therapy. Planned radiation dose was 50 Gy in 25 fractions over 5 weeks. 5-FU was to be administered commencing days 1 and 29 of radiotherapy, and given at a dose of 1 g/m2/d for 4 days as a continuous infusion. Carboplatin was to commence on day 1 of radiotherapy and be given throughout the period of radiation as a continuous infusion. The starting dose of carboplatin was 28 mg/m2/d. The protocol specified a 25% dose reduction of carboplatin if more than two of the first six patients experienced dose-limiting toxicity (DLT). DLT was defined as grade IV neutropenia lasting more than 7 days, grade IV thrombocytopenia, or any grade IV nonhematologic toxicity. All 23 patients in the study received protocol radio-therapy, except one who was given an extra 10 Gy. Seven patients received carboplatin at 28 mg/m2/d and 16 received 21 mg/m2/d. Hematologic DLTs were experienced by all of the seven patients receiving the higher dose. No patients in the low-dose group experienced hematologic DLTs, and only 2 of 16 ceased chemotherapy early because of myelosuppression. Three patients in the low-dose group experienced grade IV esophagitis but were able to complete protocol radiotherapy. Apart from esophagitis, nonhematologic toxicity was generally moderate or mild. Six patients had thrombosis complicating the central venous catheters. Endoscopy was performed in 21 patients (91%), with an overall complete response rate of 65% (CI: 43-84%) for the whole group or 71% (CI: 48-89%) for the endoscopically evaluated group. Estimated median failure-free survival time was 8.9 months (CI: 7.1-12.9), and estimated median overall survival time was 21.4 months (CI: 9.6 -35.4). Carboplatin at 21 mg/m2/d as a continuous infusion may be given safely in combination with short infusional 5-FU and radiotherapy for localized carcinoma of the esophagus. This combination has resulted in response data comparable to that of larger studies of cisplatin-containing regimens and warrants further study, ideally in a phase III randomized controlled trial.


Active form of vitamin D ameliorates non-alcoholic fatty liver disease by alleviating oxidative stress in a high-fat diet rat model.
The purpose of this study was to determine whether treatment using the active form of vitamin D (1,25(OH)2D3) could protect against high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) in rats and ameliorate oxidative stress. Male Sprague-Dawley rats were divided into three groups and treated with standard chow, HFD, or HFD plus intraperitoneal injection of 1,25(OH)2D3 (5 μg/kg body weight, twice per week), respectively, for 16 weeks. Serum lipid profiles, hepatic function, intrahepatic lipid, and calcium levels were determined. Hepatic histology was examined using hematoxylin/eosin, Masson's trichrome, and Oil Red O staining. Oxidative stress was assessed by measuring hepatic malondialdehyde (MDA) and F2α-isoprostane content. Expression of nuclear factor-erythroid-2-related factor 2 (Nrf2) and downstream target genes was analyzed using quantitative RT-PCR. 1,25(OH)2D3 treatment improved the serum lipid profile, reduced intrahepatic lipid levels, and attenuated hepatic steatosis and inflammation in HFD rats. Furthermore, MDA and F2α-isoprostane levels in liver tissue were reduced by 1,25(OH)2D3 administration. Although 1,25(OH)2D3 did not regulate the expression of Nrf2 mRNA, it did induce Nrf2 nuclear translocation. The expression of Nrf2 target genes, including Gclc, Nqo1, Sod2, and Cat, was up-regulated by 1,25(OH)2D3. We conclude that 1,25(OH)2D3 protects against HFD-induced NAFLD by attenuating oxidative stress, inducing NRF2 nuclear translocation, and up-regulating the expression of genes encoding antioxidant enzymes.


Encoding of High Frequencies Improves with Maturation of Action Potential Generation in Cultured Neocortical Neurons.
The ability of neocortical neurons to detect and encode rapid changes at their inputs is crucial for basic neuronal computations, such as coincidence detection, precise synchronization of activity and spike-timing dependent plasticity. Indeed, populations of cortical neurons can respond to subtle changes of the input very fast, on a millisecond time scale. Theoretical studies and model simulations linked the encoding abilities of neuronal populations to the fast onset dynamics of action potentials (APs). Experimental results support this idea, however mechanisms of fast onset of APs in cortical neurons remain elusive. Studies in neuronal cultures, that are allowing for accurate control over conditions of growth and microenvironment during the development of neurons and provide better access to the spike initiation zone, may help to shed light on mechanisms of AP generation and encoding. Here we characterize properties of AP encoding in neocortical neurons grown for 11-25 days in culture. We show that encoding of high frequencies improves upon culture maturation, which is accompanied by the development of passive electrophysiological properties and AP generation. The onset of APs becomes faster with culture maturation. Statistical analysis using correlations and linear model approaches identified the onset dynamics of APs as a major predictor of age-dependent changes of encoding. Encoding of high frequencies strongly correlated also with the input resistance of neurons. Finally, we show that maturation of encoding properties of neurons in cultures is similar to the maturation of encoding in neurons studied in slices. These results show that maturation of AP generators and encoding is, to a large extent, determined genetically and takes place even without normal micro-environment and activity of the whole brain in vivo. This establishes neuronal cultures as a valid experimental model for studying mechanisms of AP generation and encoding, and their maturation.


Controlled study of excimer and pulsed dye lasers in the treatment of psoriasis.
The excimer laser delivers high energy monochromatic ultraviolet (UV) B at 308 nm. Advantages over conventional UV sources include targeting of lesional skin, reducing cumulative dose and inducing faster clearance. Studies of the pulsed dye laser (PDL) in psoriasis report between 57% and 82% response rates; remission may extend to 15 months. To our knowledge, this is the first study assessing both excimer and PDL in psoriasis. We conducted a within-patient controlled prospective trial of treatment of localized plaque psoriasis. Twenty-two adult patients, mean Psoriasis Area and Severity Index 7.1, were recruited. Fifteen patients completed the full treatment, of which 13 were followed up to 1 year. Two selected plaques were treated with excimer twice weekly and V Beam PDL, pretreated with salicylic acid (SA), every 4 weeks, respectively. Two additional plaques, treated with SA alone or untreated, served as controls. The primary outcome measures were: (i) changes in plaque-modified Psoriasis Activity and Severity Index (PSI) scores from baseline to end of treatment; (ii) clinical response to treatment (CR(T)), assessed by serial photographs; (iii) percentage of plaques clear at the end of treatment; and (iv) percentage of plaques clear at 1-year follow-up. The secondary outcome measures were: (i) number of laser treatments to clearance; (ii) time to relapse; (iii) frequency of side-effects; and (iv) qualitative observations with SIAscope. The mean improvement in PSI was 4.7 (SD 2.1) with excimer and 2.7 (SD 2.4) with PDL. PSI improvement was significantly greater in excimer than PDL (P = 0.003) or both control plaques (P < 0.001). CR(T) indicated 13 patients responded best with excimer, two patients best with PDL, and in seven patients there was no difference between the two lasers. CR(T) was significantly greater for excimer than PDL (P = 0.003) or both controls (P < 0.001). CR(T) was also significantly greater for PDL than SA alone (P = 0.004) or untreated control (P =0.002). Nine (41%) patients cleared with excimer, after mean 8.7, median 10 weeks treatment. Seven of these nine patients were followed up to 1 year; four remained clear, two relapsed at 1 month, and one at 6 months. Six (27%) patients cleared with PDL, after mean 3.3, median four treatments. All six patients were followed up to 1 year; four remained clear, one relapsed at 4 months and one at 9 months. Despite common side-effects including blistering and hyperpigmentation, patient satisfaction was high. Serial images obtained with the SIAscope during treatment indicated different mechanisms of action of the two lasers. Excimer and V Beam PDL are useful treatments for plaque psoriasis. Although the excimer appears to be on average more efficacious, a subset of patients may respond better to PDL. Long-term remission is achievable with both lasers.


Treatment of Gustilo grade III leg fractures by external fixation associated with limited internal fixation.
To explore the clinical effects of external fixation associated with limited internal fixation on treatment of Gustilo grade III leg fractures. From July 2006 to December 2008, 40 cases of Gustilo grade III leg fractures were emergently treated in our unit with external fixation frames. Soft tissue injuries were grouped according to the Gustilo classification as IIIA in 17 cases, IIIB in 13 cases, and IIIC in 10 cases. All the patients were debrided within 8 hours, and then fracture reposition was preformed to reestablish the leg alignment. Limited internal fixation with plates and screws were performed on all the Gustilo IIIA cases and 10 Gustilo IIIB cases at the first operation. But all the Gustilo IIIC cases and 3 Gustilo IIIB cases who had severe soft tissue injuries and bone loss only received Vacuum-sealing drainage (VSD). Broad-spectrum antibiotics were regularly used and VSD must be especially maintained easy and smooth for one week or more after operation. Limited internal fixation and transplanted free skin flaps or adjacent musculocutaneous flaps were not used to close wounds until the conditions of the wounds had been improved. The first operations were completed within 90-210 minutes (170 minutes on average). The blood transfusions were from 400 ml to 1500 ml (those used for anti-shock preoperatively not included). All the 40 patients in this study were followed up for 6-28 months, 20.5 months on average. The lower limb function was evaluated according to the comprehensive evaluation standards of leg function one year after operation and the results of 28 cases were excellent, 9 were good and 3 were poor. External fixation associated with limited internal fixation to treat Gustilo grade III leg fractures can get satisfactory early clinical therapeutic effects.


The mop1 (mediator of paramutation1) mutant progressively reactivates one of the two genes encoded by the MuDR transposon in maize.
Transposons make up a sizable portion of most genomes, and most organisms have evolved mechanisms to silence them. In maize, silencing of the Mutator family of transposons is associated with methylation of the terminal inverted repeats (TIRs) surrounding the autonomous element and loss of mudrA expression (the transposase) as well as mudrB (a gene involved in insertional activity). We have previously reported that a mutation that suppresses paramutation in maize, mop1, also hypomethylates Mu1 elements and restores somatic activity to silenced MuDR elements. Here, we describe the progressive reactivation of silenced mudrA after several generations in a mop1 background. In mop1 mutants, the TIRA becomes hypomethylated immediately, but mudrA expression and significant somatic reactivation is not observed until silenced MuDR has been exposed to mop1 for several generations. In subsequent generations, individuals that are heterozygous or wild type for the Mop1 allele continue to exhibit hypomethylation at Mu1 and mudrA TIRs as well as somatic activity and high levels of mudrA expression. Thus, mudrA silencing can be progressively and heritably reversed. Conversely, mudrB expression is never restored, its TIR remains methylated, and new insertions of Mu elements are not observed. These data suggest that mudrA and mudrB silencing may be maintained via distinct mechanisms.


Disseminated Cutaneous Rhinosporidiosis.
A 41-year-old man from the state of Chhattisgarh, India, had a previous history of nasopharyngeal rhinosporidosis for 27 years, for which he had surgical intervention seven times, most recently endoscopically 2 months earlier. For the last 8 months, he had noticed skin lesions on the shoulders and forehead. Of interest was a history of bathing in ponds as a child. Examination revealed a soft, skin-colored, strawberry-shaped papule on the forehead (Figure 1A), a papule with a hemorrhagic crust on the left shoulder (Figure 1B), and a crusted friable plaque on the right shoulder (Figure 1C). Endoscopy revealed a 1 cm × 1 cm red polypoidal mass on the lateral wall of the nasopharynx. A diagnosis of disseminated cutaneous rhinosporidiosis with nasal-nasopharyngeal rhinosporidiosis was considered.


Urinary L-valyl proline in patients with aortic aneurysms.
Recent evidence indicates that metabolism of elastin may be altered in patients with different types of infrarenal aortic disease and that the phenotypic expression of aortic disease may be dependent on the balance between aortic elastase and antiprotease activity. The dipeptide L-valyl proline (LVP) is a specific amino acid sequence for elastin and can be quantitated by high performance liquid chromatography analysis of the urine. This study was done to determine if alterations in systemic elastin metabolism could be detected in patients with different types of infrarenal aortic disease by quantitating urinary LVP. Patients were divided into one of five groups and had urine analyzed for LVP. These are control, no known aortic disease (n = 12); occlusive aortic disease (n = 10); elective abdominal aortic aneurysms (AAA) (n = 26); ruptured AAA (n = 5), and multiple aneurysms (n = 4). Urine values were correlated with aortic elastase and aortic antiprotease activity. Urinary LVP was significantly higher in patients with multiple aneurysms (1,209 micrograms per milliliter of urine) as compared with all of the other groups. Patients with elective AAA had significantly higher urinary LVP (40.5 micrograms per milliliter of urine) than patients with occlusive disease (9.1 micrograms per milliliter of urine) and those in the control group (4.2 micrograms per milliliter of urine). Patients with ruptured AAA did not have significantly elevated urinary LVP compared with other groups (18.6 micrograms per milliliter of urine). Urinary LVP increased significantly as aortic elastase and aortic elastase and antiprotease activity increased. These data suggest that elastin metabolism, as reflected by urinary LVP, is altered in patients with aortic aneurysmal disease and provide further evidence to support the concept that systemic elastin metabolism is altered in patients with different types of infrarenal aortic pathologic findings.


Behavioral responses and mortality in German cockroaches (Blattodea: Blattellidae) after exposure to dishwashing liquid.
Behavioral responses and mortality of the German cockroach, Blattella germanica (L.), after exposure to solutions of common household dishwashing liquid diluted in tap water without other known insecticidal active ingredients are described. Soap solutions of 0.05, 0.1, 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, and 5.0% were tested on first and second instars, fifth and sixth instars, and adults. Cockroaches were treated individually or in groups. Behavioral observations after treatment included an immediate knockdown period, an awakening and a struggle period, an unresponsive period, and either death or recovery between 30 min and 18 h after treatment. Probit analysis gave an excellent fit of unresponsive rates 18 h after treatment with the various soap concentrations, and LD50 and LD99 values were calculated to be 0.4 and 3.0% soap, respectively. Adult females were the most difficult to kill, but at soap concentrations of 1% or higher, 95% or greater unresponsiveness and death occurred. Soap concentrations of 3% or greater resulted in 100% unresponsiveness after 3 min, and eventually resulted in 100% mortality within 72 h for all adults and nymphs treated. Observations on the mode and site of action for soap are consistent with the blockage of spiracles and/or tracheae that results in asphyxiation and death. Future investigations into the efficacy of soaps against other arthropod groups are warranted and may be beneficial in certain situations.


Interventional endoscopy in the pancreatobiliary tree.
Since the advent of endoscopic retrograde cholangiopancreatography (ERCP) in 1970 and endoscopic sphincterotomy (EST) in 1974, there has been rapid proliferation of endoscopic techniques for the diagnosis and therapy of benign and malignant biliary and pancreatic disorders. The ability to biopsy under direct vision, reliably obtain a pancreatogram, exclude other upper gastrointestinal disorders, and reexamine with ease, combined with the lack of hepatic puncture, has given the endoscopic route an undeniable advantage over the percutaneous transhepatic route for these interventions. Although some have become routine procedures (EST for postcholecystectomy choledocholithiasis; transpapillary stent insertion for inoperable biliary stenoses), the exact place of many of these interventions remains unclear, and the subject of prospective controlled studies where possible. This review attempts to give an overview of present and developing ERCP- and EST-based diagnostic and interventive techniques and their application to the wide spectrum of pancreatobiliary diseases.


In situ fluorescent visualization of nucleolar organizer region-associated proteins with a thiol reagent.
Nucleolar organizer regions (NORs) are nucleolus-forming rDNA loops associated with argyrophil proteins, the amount of which varies according to the proliferative state of the cell. It has been presumed that the nucleolar protein-related thiol groups may have a role in selective silver staining. We investigated the nuclear thiol distribution with a fluorescent thiol reagent, coumarinyl-phenyl-maleimide (CPM) in human K-562 myeloblast cultures and found that SH group-related fluorescence was brightest in the area of nucleoli, which became highly selective after RNAse digestion. A remarkable co-localization of AgNOR silver reaction and CPM fluorescence was observed, although occupation of the SH groups by CPM did not prevent the silver staining. We applied the stain to dual-parameter flow cytometry in combination with DNA content measurements, which provide further information on nucleolar function and changes in experimental and pathological specimens.


Dietary selenium intake in Belgium for different population groups at risk for deficiency.
An estimation of the dietary selenium intake in different population groups in Belgium has been carried out. 24 h duplicate meals and food consumption statistics combined with concentration levels in different foods, as determined by hydride generation atomic absorption spectrometry, are used. Dietary intake of selenium is low to very low, especially for vegetarians (12.7 +/- 9.1 micrograms) and patients on long-term total parenteral nutrition (1.2 +/- 0.8 microgram). The different intakes are discussed and compared with intakes published for other countries, while the importance of bioavailability of the selenium in food in the risk of deficiency is stressed.


Genome Sequence of Roseovarius mucosus Strain SMR3, Isolated from a Culture of the Diatom Skeletonema marinoi.
We present the genome of Roseovarius mucosus strain SMR3, a marine bacterium isolated from the diatom Skeletonema marinoi strain RO5AC sampled from top layer sediments at 14 m depth. Its 4,381,426 bp genome consists of a circular chromosome and two circular plasmids and contains 4,178 coding sequences (CDSs).


Expressed CpG island sequence tag microarray for dual screening of DNA hypermethylation and gene silencing in cancer cells.
We present a novel concept by using expressed CpG island sequence tags (ECISTs)for dual analysis of DNA methylation and gene expression in cancer cells. ECISTs are present in the genome and are DNA fragments expected to be located in the promoter and first exon region of genes. Their GC-rich segments can be used for screening hypermethylated CpG sites in cancer cells, and their exon-containing portions can be used for measuring levels of the corresponding transcripts. A total of 1162 loci met the criteria of ECISTs from an initial screening of 7776 CpG island tags. This ECIST panel was used to analyze the breast cancer cell line MDA-MB-231, which was treated with a demethylating agent. Microarray profile analysis identified 30 methylation-silenced genes, the expression of which could be directly reactivated by demethylation. An additional group of 96 up-regulated genes was also identified but appeared to be downstream from this epigenetic cascade. Thus, this study shows that the ECIST microarray can be used to differentiate the primary and secondary causes of demethylation and provides an effective tool to elucidate the mechanisms of aberrant DNA methylation in cancer.


Investigation and standardization of prothrombin times in chickens.
This investigation was undertaken to standardize the determination of the one-stage prothrombin time for use with chickens. Homologous thromboplastin was essential and the most active thromboplastin was obtained from chickens four-weeks old or younger. Acetone-dried brain powder could be stored for at least 4 months at -15 degrees C. without loss of activity. Extraction of brain powder with 0.025 M CaCl2 at 42 degrees C. gave better thromboplastic activity than the standard extraction with physiological saline at room temperature. Thromboplastin solutions could be stored in ice water for only 6 hours without loss of activity. Citrate concentration had to be increased from the usual 0.10 M to 0.18 M to prevent premature clotting of plasma. Plasma donor age had no effect on the prothrombin times. Freezing and thawing as well as storage of plasma in the frozen state increased the prothrombin times. Using the best conditions, the mean prothrombin time for 1200 birds determined over a 6-month period was 9.4 sec. with an individual range of 7.18-11.4 sec. This represents a significantly lower prothrombin time with lower variability than that reported in the literature.


Venoarterial extracorporeal membrane oxygenation decannulation using the novel Manta vascular closure device.
Removal of large calibre arterial sheaths, as employed in venoarterial extracorporeal membrane oxygenation, is still associated with a high risk for vascular access site complications. In this study, we investigated the large bore Manta closure device as an option for venoarterial extracorporeal membrane oxygenation decannulation. Sixteen patients weaned from venoarterial extracorporeal membrane oxygenation between January-June 2019 were prospectively included in a registry (eight during extracorporeal cardiopulmonary resuscitation, seven with severe cardiogenic shock and one patient with fulminant pulmonary embolism). All patients had an arterial sheath removed at bedside using the Manta device achieving rapid haemostasis without clinically relevant complications afterwards. A computed tomography angiography was available for 11 patients (68.8%) and reassessed with regards to relevant pathologies at the closure device. Average intraluminal area was 40.4 mm2 (12.2-76.3 mm2) at the closure site. All devices but one were correctly deployed. Three patients (27.3%) had a visible thrombus at the closure device resulting in a >60% stenosis, two of which had a puncture site distal of the femoral artery bifurcation. Decannulation from venoarterial extracorporeal membrane oxygenation using the Manta closure device is a viable approach for rapid haemostasis, but is also associated with a significant risk of thrombotic vessel stenosis. These preliminary findings indicate that puncture in near proximity or distal to the femoral bifurcation might identify patients at risk and further studies are needed.


Gluconeogenesis, non-essential amino acid synthesis and substrate partitioning in chicken embryos during later development.
We aimed to quantify the rate of gluconeogenesis (GNG), non-essential amino-acid (NEAA) synthesis, and substrate partitioning to the Krebs cycle in embryonic (e) day e14 and e19 chicken embryos. An in ovo continuous tracer infusion approach was employed to test the hypotheses that GNG and NEAA synthesis in developing chicken embryo increases from e14 to e19. [13C6]Glucose or [13C3]glycerol was continuously infused (8 h) into the chorio-allantoic compartment of eggs on e14 and e19. Glucose entry rate, Cori cycling, and GNG were higher (P < 0.05) in e19 compared to e14 embryos, presumably to support higher glycogen deposition in liver and muscle. Whereas de novo synthesis of alanine, aspartate, and glutamate via glycolysis and the Krebs cycle was higher (P < 0.01) in e14 embryos, synthesis of these NEAA from glycerol was higher (P < 0.05) in e19 compared to e14 embryos. These patterns of glucose and glycerol utilization suggest a metabolic shift to conserve glucose for glycogen synthesis and an increased utilization of yolk glycerol (from triacylglyceride) after e14. Although the contribution of glycerol to GNG in e19 embryos was higher (P < 0.05) than that in e14 embryos, the contribution of glycerol to GNG (1.3 to 6.0%) was minor. Based on [13C6]glucose tracer kinetics, the activities of both pyruvate carboxylase (PC) and pyruvate dehydrogenase (PDH) in the liver were higher (P < 0.05) in e19 embryos; whereas the higher (P < 0.01) relative activity of liver PC compared to PDH in e14 embryos suggests a greater anaplerotic flux into the Krebs cycle. In summary, the in ovo continuous tracer infusion approach allowed for a measurement of chicken embryo whole body and liver metabolism over a shorter window of development. This study provided quantitative estimates of the developmental shifts in substrate utilization, GNG, and NEAA synthesis by chicken embryos, as well as qualitative estimates of the activities of enzymes central to the Krebs cycle, glucose, and fatty acid metabolism.


Investigations of a novel self-emulsifying osmotic pump tablet containing carvedilol.
Carvedilol has been made into a novel osmotic pump tablet which includes Gelucire 44/14, Lutrol F68, Transcutol P, silicon dioxide, mannitol, citric acid, and sodium hydrogen carbonate. The Self-emulsifying osmotic pump tablet (SEOPT) has two outstanding features. It could improve the bioavailability of carvedilol by self-emulsifying drug delivery system (SEDDS), control the release rate and make the plasma concentrations more stable by elementary osmotic pump tablet. The results of transmission electron microscope (TEM) and particle size assessment showed that the shape of the resultant emulsion was round and regular, the average diameter of the particles was 246 nm. Since the solubility of carvedilol was improved by the emulsion, the elementary SEOPT could guarantee a complete release of carvedilol under the osmotic pressure of mannitol. The cumulative release at 12 hr was 85.18%. Therefore the disadvantage that lipophilic drugs can not be released completely when prepared into elementary osmotic pump tablet was resolved. The results of Differential scanning calorimetry (DSC), Infrared spectroscopy (IR) and X-ray diffraction diffraction (XRD) proved that carvedilol was amorphous in the preparation. The relative bioavailability of carvedilol in beagle dogs was 156.78%. The plasma concentrations were more stable compared with that of commercially available tablet (Luode). And the in vitro and in vivo correlation was good (r = 0.9725). Therefore, the elementary SEOPT developed in this paper might provide a new idea for preparing lipophilic drugs into osmotic pump tablet conveniently.


Optimisation of telephone triage of callers with symptoms suggestive of acute cardiovascular disease in out-of-hours primary care: observational design of the Safety First study.
In the Netherlands, the 'Netherlands Triage Standard' (NTS) is frequently used as digital decision support system for telephone triage at out-of-hours services in primary care (OHS-PC). The aim of the NTS is to guarantee accessible, efficient and safe care. However, there are indications that current triage is inefficient, with overestimation of urgency, notably in suspected acute cardiovascular disease. In addition, in primary care settings the NTS has only been validated against surrogate markers, and diagnostic accuracy with clinical outcomes as the reference is unknown. In the Safety First study, we address this gap in knowledge by describing, understanding and improving the diagnostic process and urgency allocation in callers with symptoms suggestive of acute cardiovascular disease, in order to improve both efficiency and safety of telephone triage in this domain. An observational study in which 3000 telephone triage recordings (period 2014-2016) will be analysed. Information is collected from the recordings including caller and symptom characteristics and urgency allocation. The callers' own general practitioners are contacted for the final diagnosis of each contact. We included recordings of callers with symptoms suggestive of acute coronary syndrome (ACS) or transient ischaemic attack (TIA)/stroke. With univariable and multivariable logistic regression analyses the diagnostic accuracy of caller and symptom characteristics will be analysed in terms of predictive values with urgency level, and ACS and TIA/stroke as outcomes, respectively. To further improve our understanding of the triage process at OHS-PC, we will carry out additional studies applying both quantitative and qualitative methods: (i) case-control study on serious adverse events (SAE), (ii) conversation analysis study and (iii) interview study with triage nurses. The Medical Ethics Committee Utrecht, the Netherlands endorsed this study (National Trial Register identification: NTR7331). Results will be disseminated at scientific conferences, regional educational sessions and publication in peer-reviewed journals.


Block copolymer micelles as nanoreactors for self-assembled morphologies of gold nanoparticles.
Self-assembled gold (Au) nanoparticles (NPs) were synthesized in micelle surface cavities of a L121 block polymer in the presence of zwitterionic (viz. DPS, TPS, and HPS) and sugar surfactants (OG and DDM) in aqueous phase at 70 °C by using the surface cavities of L121 as reducing sites for converting Au(III) into Au(0). All reactions were monitored simultaneously by UV-visible spectroscopy to determine the growth kinetics in gold nucleating centers on the basis of surface plasmon resonance that also helped in tracing the structure micelle transitions over a wide temperature range of 10-70 °C. The surfactant/L121 mole ratio was changed systematically from 0.5 to 2.5 by keeping L121 and HAuCl4 concentrations constant at 10 and 0.25 mM, respectively, to determine the shape and size of the micelles and their relation to the self-assembled behavior of Au NPs. TEM studies were used to have a direct insight into the morphology of micelle templates and their shape and size for self-assembled NPs. L121 along with DPS (C12 carbon chain) produced well-defined micelles loaded with tiny NPs of 3-6 nm in the L121-rich region of the mixture, while large flower-like compound micelles with a clear core-shell morphology were produced in the DPS-rich region. TPS and HPS (C14 and C16 hydrocarbon chains, respectively) with stronger hydrophobicity than DPS also produced almost similar micelles loaded with tiny NPs in the L121-rich region, but they disappear in the surfactant-rich region. Replacement of zwitterionic with ionic surfactants did not yield micelle templates for self-assembled NPs. Results conclude that well-defined micelles of L121 are the fine templates for self-assembled NPs that can only be achieved in the presence of a neutral surfactant with low concentration and low hydrophobicity.


Peak parking determination of the obstruction factor in lauryl acrylate monolithic CEC columns.
The peak parking method was used to determine the obstruction factor of lauryl acrylate porous polymer monoliths. Polymers were prepared in situ in fused-silica capillaries using thermally initiated polymerization. These columns have been used for CEC of neutral analytes. Thiourea, which is unretained, was used as the test analyte for the obstruction factor measurement. The obstruction factor was determined to be 0.72 with a SD of (+/-0.01), which is consistent with the concept that organic porous polymer monoliths are more permeable than traditional LC stationary phases.


The Effects of a Simple Sensor Reorientation Procedure on Peak Tibial Accelerations during Running and Correlations with Ground Reaction Forces.
While some studies have found strong correlations between peak tibial accelerations (TAs) and early stance ground reaction forces (GRFs) during running, others have reported inconsistent results. One potential explanation for this is the lack of a standard orientation for the sensors used to collect TAs. Therefore, our aim was to test the effects of an established sensor reorientation method on peak Tas and their correlations with GRFs. Twenty-eight runners had TA and GRF data collected while they ran at a self-selected speed on an instrumented treadmill. Tibial accelerations were reoriented to a body-fixed frame using a simple calibration trial involving quiet standing and kicking. The results showed significant differences between raw and reoriented peak TAs (p < 0.01) for all directions except for the posterior (p = 0.48). The medial and lateral peaks were higher (+0.9-1.3 g), while the vertical and anterior were lower (-0.5-1.6 g) for reoriented vs. raw accelerations. Correlations with GRF measures were generally higher for reoriented TAs, although these differences were fairly small (Δr2 = 0.04-0.07) except for lateral peaks (Δr2 = 0.18). While contingent on the position of the IMU on the tibia used in our study, our results first showed systematic differences between reoriented and raw peak accelerations. However, we did not find major improvements in correlations with GRF measures for the reorientation method. This method may still hold promise for further investigation and development, given that consistent increases in correlations were found.


Susceptibility of irradiated B6D2F1/J mice to Klebsiella pneumoniae administered intratracheally: a pulmonary infection model in an immunocompromised host.
Bacteria such as Klebsiella pneumoniae can invade and colonize an immunocompromised host and complicate clinical recovery. In the study reported here, an experimental model of induced pneumonia was developed in 60Co gamma-photon-irradiated mice for the purpose of evaluating efficacy of therapeutic agents. The model was characterized by use of probit analysis of bacterial dose, and microbiologic, and histopathologic results. Bacterial colony-forming-unit (CFU) values producing 50% mortality within 30 days (LD50/30) and their 95% confidence intervals were 4.0 x 10(4) [1.7 x 10(4) - 8.9 x 10(4)] for 0-Gray (Gy)-irradiated mice, 1.9 x 10(4) [7.0 x 10(3) - 4.8 x 10(4)] for 5-Gy-irradiated mice, and 1.0 x 10(3) [2.8 x 10(2) - 3.3 x 10(3)] for 7-Gy-irradiated mice. Probit regression line fits calculated by use of an iterative, weighted least-squares fit, were used to assess a dose-modifying factor (DMF). The DMFs for mortality, compared with that for the 0-Gy dose, with their 95% confidence intervals, were 2.2 [0.63 - 7.7] for the 5-Gy and 38.9 [9.6 -165.0] for 7-Gy doses. The 5-Gy probit line did not significantly differ (P = 0.21) from the 0-Gy probit line (dose ratios did not significantly differ from 1), whereas the 7-Gy probit line differed significantly from the 0-Gy probit line (P < 0.001). These results demonstrate that 7-Gy 60Co gamma-photon radiation in combination with intratracheal K. pneumoniae challenge induces a valid pulmonary infection model in immunocompromised female B6D2F1/J mice.


Determination of Human Papillomavirus Type 18 Lineage of E6: A Population Study from Iran.
The epidemiological studies in Iran on HPV18 nucleotide changes are rare. This type of virus is prevalent in the Iranian population. Therefore, in the present study, we aimed to identify the genetic variability in HPV18 in the E6 region to evaluate the prevalence of lineage distribution and sublineages in a sample population in Iran. Overall, 60 HPV18 confirmed cases were investigated between 2019 and 2021. The specimens were collected, and molecular genotyping was done using the Linear Array HPV Genotyping Test. DNA extraction was performed by a viral DNA/RNA kit. The HPV E6 gene was amplified by using type-specific primers designed according to the HPV18 genome prototype sequence. The sequencing of the E6 region was successfully done on 43 samples which were then compared to the reference sequence. The most frequent sublineage of HPV18 in this study was A4 (69.7%), followed by A1 (18.6%) and A3 (11.6%). Neither A2 nor A5 sublineage was not detected in this study. The related nucleotide acid changes according to the main references were as follows: A3: T104C/T232G/T485C/C549A, A4: T104C/T485C/C549A. The predominance of A lineage with the high frequency of A4 sublineage was found in the present research. The importance of sublineages in susceptibility to a progressive form of infection requires to be more investigated among the different population.


Role of food perceptions in food selection of the elderly.
Concerns with the nutritional status and well being of older adults and the related potential social and economic impact have generated much research interest in their food intake patterns. Identifying determinants of food use and analyzing their interactions in the food selection process is critical for making nutritional recommendations and for suggesting modifications to unhealthy food behaviors. This article reviews a conceptual model of food perceptions and its applications for the study of food selection. It also discusses the relevance of specific perceptual dimensions in planning and implementing tailored nutrition interventions for a new generation of senior citizens.


Role of Bag-1 in the survival and proliferation of the cytokine-dependent lymphocyte lines, Ba/F3 and Nb2.
The expression and function of the newly identified Bcl-2- and Raf-1- binding protein, Bag-1, during the cytokine-regulated growth of B and T cell lines was examined. Immunoblot analysis of lysates from the interleukin-3 (IL-3)-dependent B cell line Ba/F3, and the PRL-dependent T cell line Nb2, revealed that variations in Bag-1 levels paralleled alterations in cellular proliferation, viability, and apoptosis induced by the presence or absence of growth factor. To test whether up-regulation of Bag-1 levels altered cellular survival and proliferation, Ba/F3 cells were transfected with a Bag-1 expression construct. The overexpression of Bag-1 in transfected Ba/F3 cells induced an IL-3-independent state. Such transfectants demonstrated sustained viability and proliferation, with minimal apoptosis, in the complete absence of exogenous IL-3. Bag-1 expression was also compared in glucocorticoid-sensitive Nb2 cells and a PRL-independent, glucocorticoid-resistant subline, SFJCD1, during culture of these lines in dexamethasone (Dex). Bag-1 levels were profoundly decreased by the addition of Dex to Nb2 cells, precedent to the onset of apoptotic cell death. In contrast, Dex treatment or PRL withdrawal had no effect on levels of Bag-1 within the SFJCD1 line. These findings establish that the overexpression of Bag-1 in the appropriate cellular context promotes cellular survival and growth, events that may result from the juxtaposition of this protein with mitogenic and antiapoptotic signaling pathways.


Isorhynchophylline, a Potent Plant Alkaloid, Induces Apoptotic and Anti-Metastatic Effects in Human Hepatocellular Carcinoma Cells through the Modulation of Diverse Cell Signaling Cascades.
Isorhynchophylline (Rhy) is an active pharmacological component of Uncaria rhynchophylla that has been reported previously to exert significant antihypertensive and neuroprotective effects. However, very little is known about its potential anti-cancer activities. This study was carried out to evaluate the anticancer effects of Rhy against various human carcinoma cell lines. We found that Rhy exhibited substantial cytotoxic effect against human hepatocellular carcinoma HepG2 cells when compared with other human carcinoma cell lines including those of lung, pancreas, prostate, head and neck, breast, multiple myeloma, brain and renal cell carcinoma. Rhy induced apoptosis as characterized by accumulation of cells in sub G1 phase; positive Annexin V binding; activation of caspase-8, -9, and -3; and cleavage of PARP (poly-ADP ribose polymerase). This effect of Rhy correlated with the down-regulation of various proteins that mediated cell proliferation, cell survival, metastasis, and angiogenesis. Moreover, cell proliferation, migration, and constitutive CXCR4 (C-X-C chemokine receptor type 4), MMP-9 (Matrix metallopeptidase-9), and MMP-2 expression were inhibited upon Rhy treatment. We further investigated the effect of Rhy on the oncogenic cell signaling cascades through phospho-kinase array profiling assay. Rhy was found to abrogate phospho-p38, ERK, JNK, CREB, c-Jun, Akt, and STAT3 signals, but interestingly enhanced phospho-p53 signal. Overall, our results indicate, for the first time, that Rhy could exert anticancer and anti-metastatic effects through regulation of multiple signaling cascades in hepatocellular carcinoma cells.


rIFN-gamma-activated rat neutrophils induce tumor cell apoptosis by nitric oxide.
We have previously shown that 1) neutrophils activated with various cytokines, including rat recombinant interferon gamma (rIFN-gamma), inhibit tumor cell growth and that 2) nitric oxide (NO) is the effector molecule in tumor inhibition by rIFN-gamma-stimulated rat peritoneal exudate neutrophils. In this study, we examined the nature of tumor cell death induced by rat peritoneal neutrophils activated by rIFN-gamma in order to clarify the mechanism of apoptosis in neoplastic tumor cell death. DNA of 3 syngeneic rat tumor cell lines was significantly fragmented within 3 hr of incubation in the presence of rIFN-gamma-activated neutrophils, and this effect was dependent on both the concentration of rIFN-gamma and the number of neutrophils. This DNA fragmentation was inhibited by L-N-(I-iminoethyl)-ornithine (L-NIO), a NO synthase inhibitor, but not by superoxide dismutase (SOD). Tumor cells treated with the activated neutrophils were shown by electron microscopy to be apoptotic, exhibiting necrotic features with a longer incubation. On the other hand, cytolysis of tumor cells, as determined by a [3H]-uridine release assay, was first observed only at 24 hr of incubation with the rIFN- gamma-activated neutrophils. Taken together, our results suggest that tumor cell apoptosis by activated neutrophils is NO-dependent and that apoptotic tumor cells undergo necrosis as a secondary process. We suggest that tumor cell apoptosis induced by activated neutrophils plays an important role in regulation of neoplastic tumor cell growth and death in vivo.


In vitro bone exposure to strontium improves bone material level properties.
In rats treated with strontium ranelate, the ultimate load of intact bone is increased and associated with changes in microstructure and material level properties. Evaluation by micro-computed-tomography-based finite element analysis has shown that these changes independently contribute to the improvement of bone strength induced by strontium ranelate treatment. However, the mechanism by which Sr ion acts on bone material level properties remains unknown. The vertebrae of intact female rats were exposed overnight to 0.5, 1 or 2M chloride salt solutions of Sr, Ca and Ba. The latter two were used to assess the specificity of Sr. Bone material level properties were evaluated by measuring hardness, elastic modulus and working energy in a nanoindentation test. Wavelength dispersive X-ray spectroscopy provided semi-quantitative elemental analysis and mapping. Incubation with Sr rendered bone stiffer, harder and tougher. Among the divalent ions tested, Sr had the greatest effect. Sr affinity was also assessed on in vivo treated bone specimens. After in vitro exposure, the highest improvements were observed in ovariectomized rats. However, anti-osteoporotic treatments did not influence the capacity of Sr to modify bone material level properties. Our findings demonstrated that in vitro incubation with Sr selectively improved bone material level properties, which may contribute to the macroscopic increase of bone properties observed under Sr therapy.


Vascular basement membrane alterations and β-amyloid accumulations in an animal model of cerebral small vessel disease.
Non-amyloid cerebral small vessel disease (CSVD) and cerebral amyloid angiopathy (CAA) may be interrelated through the damaged basement membranes (BMs) and extracellular matrix changes of small vessels, resulting in a failure of β-amyloid (Aβ) transport and degradation. We analyzed BM changes and the pattern of deposition of Aβ in the walls of blood vessels in spontaneously hypertensive stroke-prone rats (SHRSP), a non-transgenic CSVD model. In 45 SHRSP and 38 Wistar rats aged 18 to 32 weeks: (i) the percentage area immunostained for vascular collagen IV and laminin was quantified; (ii) the capillary BM thickness as well as endothelial and pericyte pathological changes were analysed using transmission electron microscopy (TEM); and (iii) the presence of vascular Aβ was assessed. Compared with controls, SHRSP exhibited a significantly higher percentage area immunostained with collagen IV in the striatum and thalamus. SHRSP also revealed an age-dependent increase of the capillary BM thickness and of endothelial vacuoles (caveolae) within subcortical regions. Endogenous Aβ deposits in the walls of small blood vessels were observed in the cortex (with the highest incidence found within fronto-parietal areas), striatum, thalamus and hippocampus. Vascular β-amyloid accumulations were frequently detected at sites of small vessel wall damage. Our data demonstrate changes in the expression of collagen IV and of the ultrastructure of BMs in the small vessels of SHRSP. Alterations are accompanied by vascular deposits of endogenous Aβ. Impaired β-amyloid clearance along perivascular and endothelial pathways and failure of extracellular Aβ degradation may be the key mechanisms connecting non-amyloid CSVD and CAA.


The Impact of Depression on Functioning in Elderly Patients With Low Vision.
The authors examined the interrelationships between low vision, depression, and functioning in older adults. Seventy patients over age 65 attending a low-vision clinic were interviewed by a geriatric nurse-practitioner, using a structured clinical format to diagnose depression according to DSM-III-R criteria. In all, 38.6% of patients met criteria for major depression. There was no association between depression and severity of visual impairment, but a direct relationship was found between depression and functional impairment. Depression was the most powerful variable associated with functional impairment (adjusted R2 = 0.20; P < 0.0001). Depression frequently co-occurs with visual impairment in this population. Appropriate recognition and treatment may reduce functional disability and improve quality of life.


What Are the Costs of Hip Osteoarthritis in the Year Prior to a Total Hip Arthroplasty?
The majority of the cost analysis literature on total hip arthroplasties (THAs) has been focused around the perioperative and postoperative period, with preoperative costs being overlooked. The Humana Administrative Claims database was used to identify Medicare Advantage (MA) and Commercial beneficiaries undergoing elective primary THAs. Preoperative healthcare resource utilization in the year prior to a THA was grouped into the following categories: office visits, X-rays, magnetic resonance imagings, computed tomography scans, intra-articular steroid and hyaluronic acid injections, physical therapy, and pain medications. Total 1-year costs and per-patient average reimbursements for each category have been reported. Total 1-year preoperative costs amounted to $21,022,883 (average = $512/patient) and $4,481,401 (average = $764/patient) for MA and Commercial beneficiaries, respectively. The largest proportion of total 1-year costs was accounted for by office visits (35% in Commercial; 41% in MA) followed by pain medications (28% in Commercial; 35% in MA). Conservative treatments (steroid injections, hyaluronic acid injections, physical therapy, and pain medications) alone accounted for 40%-44% of the total 1-year costs prior to a THA. A high healthcare utilization within the last 3 months prior to surgery was noted for opioids and steroid injections. On average, $500-$800/patient is spent on hip osteoarthritis-related care in the year prior to a THA. Despite their potential risks, opioids and steroid injections are often utilized in the last 3 months prior to surgery.


Waterlogging in late dormancy and the early growth phase affected root and leaf morphology in Betula pendula and Betula pubescens seedlings.
The warmer winters of the future will increase snow-melt frequency and rainfall, thereby increasing the risk of soil waterlogging and its effects on trees in winter and spring at northern latitudes. We studied the morphology of roots and leaves of 1-year-old silver birch (Betula pendula Roth) and pubescent birch (Betula pubescens Ehrh.) seedlings exposed to waterlogging during dormancy or at the beginning of the growing season in a growth-chamber experiment. The experiment included 4-week dormancy (Weeks 1-4), a 4-week early growing season (Weeks 5-8) and a 4-week late growing season (Weeks 9-12). The treatments were: (i) no waterlogging, throughout the experiment ('NW'); (ii) 4-week waterlogging during dormancy (dormancy waterlogging 'DW'); (iii) 4-week waterlogging during the early growing season (growth waterlogging 'GW'); and (iv) 4-week DW followed by 4-week GW during the early growing season ('DWGW'). Dormancy waterlogging affected the roots of silver birch and GW the roots and leaf characteristics of both species. Leaf area was reduced in both species by GW and DWGW. In pubescent birch, temporarily increased formation of thin roots was seen in root systems of GW seedlings, which suggests an adaptive mechanism with respect to excess soil water. Additionally, the high density of non-glandular trichomes and their increase in DWGW leaves were considered possible morphological adaptations to excess water in the soil, as was the constant density of stem lenticels during stem-diameter growth. The higher density in glandular trichomes of DWGW silver birch suggests morphological acclimation in that species. The naturally low density of non-glandular trichomes, low density of stem lenticels in waterlogged seedlings and decrease in root growth seen in DWGW and DW silver birch seedlings explain, at least partly, why silver birch grows more poorly relative to pubescent birch in wet soils.


Instantaneous Wave-Free Ratio Measurement During Intracranial Submaximal Angioplasty: Case Series and 2-Dimensional Operative Video.
Instantaneous wave-free ratios (iFRs) are functional measures of arterial stenosis that have become essential to interventional cardiology procedures. Their use for intracranial submaximal angioplasty (angioplasty with an undersized balloon) has not been studied extensively. To describe the feasibility and technique of iFR measurement for stenosis assessment during intracranial angioplasty. We present a series of consecutive patients treated between January 1, 2017 and June 30, 2018 with submaximal intracranial angioplasty in whom pre- and postprocedure iFR measurements were obtained with a Verrata-Volcano pressure wire (Philips, Amsterdam, The Netherlands). We collected patient data on age, sex, comorbid conditions, presenting complaints, modified Rankin scale (mRS) score at admission, neurological findings, procedure duration, fluoroscopy time, intraprocedural complications, length of hospital stay, and mRS score at last clinical follow-up (favorable outcome, 0-2). Angiographic stenosis severity and iFR values were recorded before and after angioplasty. A total of 12 patients underwent iFR-guided angioplasty during the study period. The median patient age was 69.5 yr (range 48-81 yr). All patients had symptomatic intracranial arterial stenosis (3-basilar, 2-vertebral, 6-middle cerebral, 1-internal carotid). Preangioplasty stenosis ranged from 55% to 90%. The median postangioplasty reduction in stenosis was 17% (range 9%-30%). Preangioplasty values ranged from 0.30 to 0.40 (n = 4). Postangioplasty values ranged from 0.6 to 0.9 (n = 5). iFR values improved considerably in all patients. No procedure-related complications occurred. The median follow-up was 8.9 mo (range 3-25 mo). Follow-up outcomes were favorable in 10 patients. iFR measurement before and after intracranial angioplasty is feasible. It may be used to assess the adequacy of intracranial angioplasty.


Exciton scattering approach for branched conjugated molecules and complexes. III. Applications.
The exciton scattering (ES) approach is an efficient tool to calculate the excited states electronic structure in large branched polymeric molecules. Using the previously extracted parameters, we apply the ES approach to a number of phenylacetylene-based test molecules. Comparison of ES predictions with direct quantum chemistry results for the excitation energies shows an agreement within several meV. The ES framework provides powerful insights into photophysics of macromolecules by revealing the connections between the molecular structure and the properties of the collective electronic states, including spatial localization of excitations controlled by the energy.


Impact of Direction of Unloading Influence on Template Rate of Perceived Exertion.
Greer, BK, Young, PR, Thompson, B, Rickert, BJ, and Moran, MF. Impact of direction of unloading influence on template rate of perceived exertion. J Strength Cond Res 32(12): 3407-3413, 2018-It is suggested that exercisers engage in a process of teleoanticipation and create an exercise template based on previous experience with the exercise task that guides their perceptions of the amount of effort required for task completion. This study examined how altering workload intensity during a positive-pressure treadmill task may impact Rating of Perceived Exertion (RPE). In a counterbalanced design, 15 collegiate cross-country runners (7 men and 8 women) performed 2 25-minute runs at a constant velocity, while body mass (BM) was either increased from 60 to 100% (low-to-high progression trial [INC]) or decreased from 100 to 60% (high-to-low progression trial) in 5-minutes increments. Oxygen consumption (V[Combining Dot Above]O2), heart rate (HR), and respiratory exchange ratio (RER) were collected. RPE was recorded at the end of each stage, and energy expenditure (EE) was calculated with V[Combining Dot Above]O2 and RER data. There were no significant differences between direction of loading conditions for V[Combining Dot Above]O2, EE, HR, and RER (p > 0.05). Between-trial differences in RPE at 100, 90, and 80% BM were statistically significant (p < 0.001), with higher RPEs observed during the INC. Differences in RPE observed between conditions cannot be explained by physiological mechanisms. These findings suggest that RPE is a multifaceted construct that can be impacted by subjectively based anticipatory factors such as exercise intensity.


Single-dose pharmacokinetics and safety pharmacodynamics of formoterol delivered by two different dry powder inhalers.
The present study aimed at assessing the pharmacokinetics (PK) and safety pharmacodynamics (PD) of 24 microg formoterol delivered via a Novolizer and via an Aerolizer in healthy subjects. This was a randomized, open-label, crossover study. Beside PK, serum potassium, and glucose profiles, vital signs, and ECG were recorded. Twenty-nine subjects (15 males) were enrolled. The inhalation maneuver had to be repeated by 19 subjects using the Aerolizer and 1 subject using a Novolizer. While eight (28%) subjects completely failed to inhale correctly via the Aerolizer (four were identified by the investigators immediately after inhalation, another four by bioanalytics later), all did it correctly via the Novolizer. The bioanalytical evaluation indicated two distinct serum peaks. The shapes of serum concentration-time profiles were more homogeneous after inhaling via the Novolizer than via the Aerolizer. After adjusting for the delivered dose the Cmax of formoterol predicting pulmonary absorption was higher after the Novolizer than after the Aerolizer, while the average AUC0-infinity levels indicating total systemic exposure were equivalent. There was no evidence for different pharmacodynamic behavior with respect to serum potassium and glucose profiles, vital signs, and ECG. The Novolizer yields higher pulmonary absorption of formoterol than the Aerolizer and equivalent safety profiles. Considering the lower variability of PK profiles and the higher proportion of correct inhalations, formoterol is more reliably inhaled via Novolizer.


[The efficacy of prevention of postoperative cognitive dysfunction in cardiac surgeries with the use of the cerebrolysin].
To assess the efficacy of postoperative cytoprotection with cerebrolysin in cardiac surgeries without using cardiopulmonary bypass and to analyze the changes in the blood circulation in the postoperative period in groups with- and without cerebrolysin preconditioning. Thirty-eight patients, who underwent coronary and mammaro-coronary bypass grafting without using cardiopulmonary bypass, were included in the study. Fifteen patients received cerebrolysin before surgery. Cerebrolysin improved cognitive test scores. Positive changes on anxiety and depression scales were observed as well. In the group of patients treated with cerebrolysin, quantitative parameters of the cerebral blood flow were in stable condition, with a slight increase on the 10th day after surgery, which may indicate increasing stress resistance of cells of the central nervous system after appropriate pharmacological protection.


Radiation from high-intensity ultrashort-laser-pulse and gas-jet magnetized plasma interaction.
Using a gas-jet flow, via the interaction between an ultrashort high-intensity laser pulse and plasma in the presence of a perpendicular external dc magnetic field, the short pulse radiation from a magnetized plasma wakefield has been observed. Different nozzles are used in order to generate different densities and gas profiles. The neutral density of the gas-jet flow measured with a Mach-Zehnder interferometer is found to be proportional to back pressure of the gas jet in the range of 1 to 8 atm. Strength of the applied dc magnetic field varies from 0 to 8 kG at the interaction region. The frequency of the emitted radiation with the pulse width of 200 ps (detection limit) is in the millimeter wave range. Polarization and spatial distributions of the experimental data are measured to be in good agreement with the theory based on the V(p)xB radiation scheme, where V(p) is the phase velocity of the electron plasma wave and B is the steady magnetic field intensity. Characteristics of the radiation are extensively studied as a function of plasma density and magnetic field strength. These experiments should contribute to the development of a new kind of millimeter wavelength radiation source that is tunable in frequency, pulse duration, and intensity.


Future directions in lamellar corneal transplantation.
The concept of lamellar keratoplasty (LK) is that of targeted lamellar replacement of corneal tissue while retaining normal cornea. It involves replacing either anterior stroma [anterior lamellar keratoplasty (ALK)] or posterior replacement of deep stromal and endothelial layers [posterior lamellar keratoplasty (PLK) or endothelial keratoplasty]. Despite the distinct advantages of ALK surgery, penetrating keratoplasty (PK) remains the most common procedure-largely because lamellar surgery is more technically demanding and time consuming, and interface irregularity arising from manual lamellar dissection often results in suboptimal visual outcomes. Recent improvements of surgical technique and advances in instrumentation such as microkeratome-assisted lamellar transplantation have contributed to improved visual quality with LK surgery. Long-term graft survival rates and endothelial cell counts after PK continue to drop for many years after surgery, clearly showing the disadvantage of unnecessary replacement of healthy endothelium in anterior-stromal disorders. The aim of this review is to evaluate the shifting emphasis from penetrating to lamellar forms of corneal transplantation and to highlight some future directions in lamellar surgery including new techniques in both ALK and PLK and the use of microkeratomes and femtosecond lasers to enhance outcomes.


Evaluation of Treatment Models in the Treatment of Retinopathy of Prematurity.
This study aims to evaluate the treatment modalities applied for retinopathy of prematurity (ROP) and to determine the efficacy and results of treatment modalities. Premature babies, who needed treatment for ROP and followed-up in the Neonatal Intensive Care Unit (NICU) of our hospital or external centers, were retrospectively evaluated between January 2012 and January 2017. According to the criteria determined by the International ROP committee, the zones and stages of the cases were recorded. In this study, patients were evaluated in three groups. Group 1: plus disease with any stage in zone 1, group 2: plus disease in zone 2, together with stage 2 or 3, group 3: classified as aggressive posterior retinopathy (APROP). The birth weight, gestational age, treatment weeks and treatments that were administered were recorded. Regression in plus disease, macular dragging and retinal detachment did not develop were evaluated as successful treatment. 1746 preterm babies were examined. 65 (3.7%) preterm babies were included in this study, 31 female and 34 male. 126 eyes of preterm babies were intervened. The mean birth weight was 1159 (535-2200) grams, and the mean gestational age was 28.4±2.5 (24-34) weeks. Group 1 had 33 eyes (26.1%), group 2 had 71 eyes (56.3%), and group 3 had 22 eyes (17.4%). 94 eyes (74.6%) were treated once, 26 eyes (20.6%) were treated twice, 6 eyes (4.8%) received treatment three times. The first treatment was applied at 36±2.4 (32-41) weeks. The first treatment was performed with intravitreal bevacizumab (IVB) in 75.8% of group 1 and 95.5% of group 3, and with diode laser photocoagulation (LPC) in 78.9% of group 2. There was a significant correlation between birth week and birth weight and first treatment week. Re-treatment was applied to 32,8% in LPC group and 19.2% in the IVB group due to recurrence. 5 eyes which were applied LPC+IVB did not need any re-treatment. Stage 4a retinal detachment developed in both eyes of 1 patient from group 1. Macular traction was developed in 2 eyes of 1 patient in group 2. After the treatments, success in 122 eyes (96.8%) was obtained. ROP can be controlled by convenient and effective treatment. Although conventional LPC is still the first treatment option for ROP, IVB alone or combination with LPC is a highly effective treatment option for zone 1 disease and APROP. IVB reduces the number of ROP treatments.


A Novel Glycosyl Hydrolase Family 16 β-Agarase from the Agar-Utilizing Marine Bacterium Gilvimarinus agarilyticus JEA5: the First Molecular and Biochemical Characterization of Agarase in Genus Gilvimarinus.
The agarase gene gaa16a was identified from a draft genome sequence of Gilvimarinus agarilyticus JEA5, an agar-utilizing marine bacterium. Recently, three agarase-producing bacteria, G. chinensis, G. polysaccharolyticus, and G. agarilyticus, in the genus Gilvimarinus were reported. However, there have been no reports of the molecular characteristics and biochemical properties of these agarases. In this study, we analyzed the molecular characteristics and biochemical properties of agarases in Gilvimarinus. Gaa16A comprised a 1,323-bp open reading frame encoding 441 amino acids. The predicted molecular mass and isoelectric point were 49 kDa and 4.9, respectively. The amino acid sequence of Gaa16A showed features typical of glycosyl hydrolase family 16 (GH16) β-agarases, including a GH16 domain, carbohydrate-binding region (RICIN domain), and signal peptide. Recombinant Gaa16A (excluding the signal peptide and carbohydrate-binding region, rGaa16A) was expressed as a fused protein with maltose-binding protein at its N-terminus in Escherichia coli. rGaa16A had maximum activity at 55°C and pH 7.0 and 103 U/mg of specific activity in the presence of 2.5 mM CaCl₂. The enzyme hydrolyzed agarose to yield neoagarotetraose as the main product. This enzyme may be useful for industrial production of functional neoagaro-oligosaccharides.


PEGylation of a Peptide-Based Amphiphilic Delivery Agent and Influence on Protein Delivery to Cells.
The cell membrane is a restrictive biological barrier, especially for large, charged molecules, such as proteins. The use of cell-penetrating peptides (CPPs) can facilitate the delivery of proteins, protein complexes, and peptides across the membrane by a variety of mechanisms that are all limited by endosomal sequestration. To improve CPP-mediated delivery, we previously reported the rapid and effective cytosolic delivery of proteins in vitro and in vivo by their coadministration with the peptide S10, which combines a CPP and an endosomal leakage domain. Amphiphilic peptides with hydrophobic properties, such as S10, can interact with lipids to destabilize the cell membrane, thus promoting cargo internalization or escape from endosomal entrapment. However, acute membrane destabilization can result in a dose-limiting cytotoxicity. In this context, the partial or transient deactivation of S10 by modification with methoxy poly(ethylene glycol) (mPEG; i.e., PEGylation) may provide the means to alter membrane destabilization kinetics, thereby attenuating the impact of acute permeabilization on cell viability. This study investigates the influence of PEGylation parameters (molecular weight, architecture, and conjugation chemistry) on the delivery efficiency of a green fluorescent protein tagged with a nuclear localization signal (GFP-NLS) and cytotoxicity on cells in vitro. Results suggest that PEGylation mostly interferes with adsorption and secondary structure formation of S10 at the cell membrane, and this effect is exacerbated by the mPEG molecular weight. This effect can be compensated for by increasing the concentration of conjugates prepared with lower molecular weight mPEG (5 to ∼20 kDa) but not for conjugates prepared with higher molecular weight mPEG (40 kDa). For conjugates prepared with moderate-to-high molecular weight mPEG (10 to 20 kDa), partial compensation of inactivation could be achieved by the inclusion of a reducible disulfide bond, which provides a mechanism to liberate the S10 from the polymer. Grafting multiple copies of S10 to a high-molecular-weight multiarmed PEG (40 kDa) improved GFP-NLS delivery efficiency. However, these constructs were more cytotoxic than the native peptide. Considering that PEGylation could be harnessed for altering the pharmacokinetics and biodistribution profiles of peptide-based delivery agents in vivo, the trends observed herein provide new perspectives on how to manipulate the membrane permeabilization process, which is an important variable for achieving delivery.


Rapid diagnosis of M. tuberculosis using a piezoelectric immunosensor.
A piezoelectric immunosensor was developed for detecting Mycobacteria Tuberculosis (M. TB). Protein A was modified onto a silver-coated piezoelectric quartz-crystal sensor. Antibodies could be bond onto the crystal surface in an ordered orientation through protein A. An impendence analyzer was employed to record the admittance-frequency curve. The experimental results showed a successful and less-rigid bonding of protein A and antibodies. Compared with other traditional detection methods of TB, the method suggested in this paper was sensitive, selective and effective. The nonspecific response was limited by using control antibody-rabbit anti-honey bee venom (anti-HBV). A glycine-HCI buffer solution (pH = 2.4) was used to release antibodies from a crystal coated with protein A, and 0.5 mg/mL anti-TB was used to remove TB from a crystal that was bonded by an antibody. Good reusability was exhibited. Spike samples of sputum and saliva from normal people with and without adding M. tuberculosis were diagnosed using the proposed method. Good results were obtained.


The inhibition of neoplastic cell proliferation with human natural tumor necrosis factor.
Purified human natural tumor necrosis factor (n-TNF) was prepared by stimulating human leukemic B cell line (BALL-1) with Sendai virus. The colony formations of all of 18 human cancer-derived abnormal cell lines were suppressed by 10(1)-10(6) U/ml of n-TNF, while n-TNF was nontoxic to all human normal fibroblast cells. This in vitro inhibition of cell growth was reversible. In breast adenocarcinoma MCF7 cells treated with n-TNF a specific decrease of DNA synthesis was observed, and DNA histograms showed a block at G1 in the cell cycle. In vivo studies revealed that n-TNF suppressed the tumor growth of murine Meth A sarcoma, human renal adenocarcinoma (ACHN), malignant melanoma (SK-MEL-28) and glioblastoma (U-373 MG). Isobologram analysis showed that n-TNF synergistically inhibited cell growth in combination with human natural interferon (IFN)-a. In vivo synergism of n-TNF and IFN-a was also found in the U-373 MG tumor model implanted into nude mice.


Cortical Transcriptional Profiles in APOE4 Carriers with Alzheimer's Disease: Patterns of Protection and Degeneration.
Transcriptional profiling of postmortem Alzheimer's disease (AD) brain tissue has yielded important insights into disease. We recently described a novel approach to understand transcriptional changes in AD designed to identify both neuro-susceptibility and intrinsic neuroprotective factors in young non-AD E4 carriers. Here we extend our work to APOE4 related AD itself. In temporal cortex (BA 21), a region known to be vulnerable to AD pathology, we identified over 1400 transcripts that differed between APOE4 controls and APOE4 carriers diagnosed with AD. Results from somatosensory cortex (BA 1/2/3), a region relatively preserved in AD differed strikingly from temporal cortex in that differences were far fewer (37 vs. 1492). We also conducted another set of contrasts involving APOE3 AD cases and APOE4 AD cases to better understand what transcriptional differences were dependent on genotype, but independent of disease status and found 6 transcripts to differ. We also conducted detailed pathway analyses in BA 1/2/3 and found significant transcriptional upregulations in pro-survival gene networks (e.g., TNF and NFkB). In summary, our results indicate that many of the molecular changes identified in the brains of patients with AD reflect the non-specific consequences of neurodegeneration, rather than causative processes. Additionally, the molecular signatures specific to somatosensory cortex may make it uniquely resistant to AD pathology and thereby could provide important leads for treatment.


CsrA Participates in a PNPase Autoregulatory Mechanism by Selectively Repressing Translation of pnp Transcripts That Have Been Previously Processed by RNase III and PNPase.
Csr is a conserved global regulatory system that represses or activates gene expression posttranscriptionally. CsrA of Escherichia coli is a homodimeric RNA binding protein that regulates transcription elongation, translation initiation, and mRNA stability by binding to the 5' untranslated leader or initial coding sequence of target transcripts. pnp mRNA, encoding the 3' to 5' exoribonuclease polynucleotide phosphorylase (PNPase), was previously identified as a CsrA target by transcriptome sequencing (RNA-seq). Previous studies also showed that RNase III and PNPase participate in a pnp autoregulatory mechanism in which RNase III cleavage of the untranslated leader, followed by PNPase degradation of the resulting 5' fragment, leads to pnp repression by an undefined translational repression mechanism. Here we demonstrate that CsrA binds to two sites in pnp leader RNA but only after the transcript is fully processed by RNase III and PNPase. In the absence of processing, both of the binding sites are sequestered in an RNA secondary structure, which prevents CsrA binding. The CsrA dimer bridges the upstream high-affinity site to the downstream site that overlaps the pnp Shine-Dalgarno sequence such that bound CsrA causes strong repression of pnp translation. CsrA-mediated translational repression also leads to a small increase in the pnp mRNA decay rate. Although CsrA has been shown to regulate translation and mRNA stability of numerous genes in a variety of organisms, this is the first example in which prior mRNA processing is required for CsrA-mediated regulation. CsrA protein represses translation of numerous mRNA targets, typically by binding to multiple sites in the untranslated leader region preceding the coding sequence. We found that CsrA represses translation of pnp by binding to two sites in the pnp leader transcript but only after it is processed by RNase III and PNPase. Processing by these two ribonucleases alters the mRNA secondary structure such that it becomes accessible to the ribosome for translation as well as to CsrA. As one of the CsrA binding sites overlaps the pnp ribosome binding site, bound CsrA prevents ribosome binding. This is the first example in which regulation by CsrA requires prior mRNA processing and should link pnp expression to conditions affecting CsrA activity.


Scalable and massively parallel Monte Carlo photon transport simulations for heterogeneous computing platforms.
We present a highly scalable Monte Carlo (MC) three-dimensional photon transport simulation platform designed for heterogeneous computing systems. Through the development of a massively parallel MC algorithm using the Open Computing Language framework, this research extends our existing graphics processing unit (GPU)-accelerated MC technique to a highly scalable vendor-independent heterogeneous computing environment, achieving significantly improved performance and software portability. A number of parallel computing techniques are investigated to achieve portable performance over a wide range of computing hardware. Furthermore, multiple thread-level and device-level load-balancing strategies are developed to obtain efficient simulations using multiple central processing units and GPUs.


Hemorrhagic fever with renal syndrome: comparison of clinical course in Sweden and in the Western Soviet Union.
The severity of the clinical course in 355 Swedish and 603 Soviet patients with serologically confirmed hemorrhagic fever with renal syndrome (HFRS) was compared retrospectively. The overall frequency of hemorrhagic manifestations including also benign bleeding such as ecchymoses was higher among Soviet than among Swedish patients (37% vs. 10%). However, severe hemorrhagic complications (gastrointestinal and renal tract bleeding) were equally common in both countries. There were 2 fatal Soviet cases, whereas none of the Swedish patients died. We conclude that the clinical presentation of HFRS in Sweden and in the Western Soviet Union shows many similarities but there seems to be a higher frequency of life threatening complications among the patients in the Western USSR.


Evaluation of nonverbal emotion in face and voice: some preliminary findings on a new battery of tests.
This report describes some preliminary attributes of stimuli developed for future evaluation of nonverbal emotion in neurological populations with acquired communication impairments. Facial and vocal exemplars of six target emotions were elicited from four male and four female encoders and then prejudged by 10 young decoders to establish the category membership of each item at an acceptable consensus level. Representative stimuli were then presented to 16 additional decoders to gather indices of how category membership and encoder gender influenced recognition accuracy of emotional meanings in each nonverbal channel. Initial findings pointed to greater facility in recognizing target emotions from facial than vocal stimuli overall and revealed significant accuracy differences among the six emotions in both the vocal and facial channels. The gender of the encoder portraying emotional expressions was also a significant factor in how well decoders recognized specific emotions (disgust, neutral), but only in the facial condition.


[Outcome of childhood T-cell acute lymphoblastic leukemia: a report of 99 cases].
To explore the incidence, clinical characteristics and prognosis of childhood T-cell acute lymphoblastic leukemia (T-ALL) enrolled in protocol ALL-2005. Retrospective reviews of patients diagnosed as having T-ALL were studied in the period between May 2005 and August 2013 in Shanghai Children's Medical Center. Predictive values of early treatment responses, including prednisone response, bone marrow morphology on day 35 during induction chemotherapy, and minimal residual disease (MRD) monitored by flow cytometry on day 35 and 55, were analyzed. The Kaplan-Meier method was used to assess the survival rates. Ninety-nine evaluable patients aged 1-18 years with newly diagnosed T-ALL were enrolled in protocol ALL-2005. The median follow-up period for patients remaining alive in continuous remission was 60 months (24-109 months). The 5-year event free survival (EFS) and overall survival (OS) rates for these patients were 65% and 69%, respectively. Forty-three (43%) patients were older than 10 years at diagnosis. A total of 61 patients presented with initial white blood cell count (WBC)≥50×10(9)/L. Thirty-seven patients with mediastinal mass were found in this study. Seventy-one patients were defined as prednisone good responders (PGR) on day 8 of prednisone therapy. Ninty-four patients achieved complete remission (CR) on day 35 of induction therapy. MRD negative status (<0.01%) on day 35 were seen in 55% of the 42 assessed patients. MRD negative status on day 55 was seen in 78% of the 52 assessed patients. Patients with high MRD (>1%) on day 55 5-year EFS was 40% were associated with poor prognosis (P=0.03). Early T-cell precursor (ETP) subtype was not associated with treatment outcome in this study. Six patients abandoned therapy, 10 lost to follow-up, 22 had relapsed disease, 1 was diagnosed as having a second tumor and 20 patients died. Overall outcome for T-ALL on protocol ALL-2005 is favorable. MRD results on day 55 of induction therapy have important prognostic and therapeutic implications.


The effect of protein on the dissolution of phenytoin.
Ultrafiltration of a continuous flow system was used to follow the dissolution of phenytoin powder in buffered solutions of pH 1, 4 and 7.4. The presence of either human serum albumin (HSA) or casein caused changes in both the absolute solubility and the dissolution rate. Normal protein binding cannot be responsible for the effects noted. It would seem likely that within the diffusion layer a higher degree of protein binding occurs than is possible in the bulk aqueous solution. The drug solubilized in this way is then released in solution in a form that is at least metastable with respect to phenytoin in normal solution. The effect of protein on phenytoin dissolution may well affect the bioavailability of the drug.


Efficient upconversion fluorescence in a blue-emitting spirobifluorene-anthracene copolymer doped with low concentrations of Pt(II)octaethylporphyrin.
Upconversion-induced fluorescence in platinum-octaethylporphyrin (PtOEP)-doped thin films of a spirobifluorene-anthracene copolymer has been investigated. Upon exciting in the range of the absorption band (2.31 eV, 537 nm) of the guest molecules, blue fluorescence (2.75 eV, 450 nm) from the spirobifluorene host was observed. The intensity of the upconverted emission was found to be one order of magnitude higher than from a PtOEP doped but anthracene-free spirobifluorene copolymer and than previously reported for metallated porphyrin-doped polyfluorene samples. It is argued that the efficient upconversion originates from the triplet energy transfer from the phosphorescent dopant to the sensitive unit of the host polymer, followed by triplet-triplet annihilation and finally blue emission from the spirobifluorene host polymer backbone.