Pertussis toxin induces bronchopulmonary hyperresponsiveness in guinea-pigs while antagonizing the effects of formyl-L-methionyl-L-leucyl-L-phenylalanine.
Pertussis toxin injected i.v. at 0.8-20 micrograms/kg markedly enhanced bronchoconstriction induced by the i.v. administration of histamine or serotonin (5-HT) (0.5-16 micrograms/kg) to propranolol-treated guinea-pigs, under conditions where propranolol or pertussis toxin alone were poorly effective. In contrast, bronchoconstriction and the accompanying leukopenia induced by the i.v. administration of the secretagogue formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) were suppressed by pertussis toxin. Bronchoconstriction induced by histamine or 5-HT was not enhanced when perfused lungs from pertussis toxin-treated guinea-pigs were studied in vitro, under conditions where bronchoconstriction and thromboxane A2 release evoked by fMLP were suppressed. Pertussis toxin negatively modifies signal transduction in cells involved in the lung responses to fMLP both in vivo and in vitro, but positively and only in vivo it modifies signal transduction in cells involved in the lung responses to the direct constricting agents histamine and 5-HT. As hyperresponsiveness to histamine and 5-HT were exclusively found in vivo, the target for pertussis toxin is probably not the adrenergic nor the cholinergic systems, since neither hexamethonium, isoprenaline, atropine nor vagotomy were effective. In addition, since dexamethasone and nedocromil sodium were inactive and enrichment of bronchoalveolar lavage with inflammatory cells was not noted, despite lung invasion by neutrophils and lymphocytes, acute inflammation does not account for pertussis toxin-induced hyperresponsiveness.


Grisel syndrome, sigmoid sinus thrombosis and rheumatic carditis: Case report of a rare association.
Grisel's syndrome (GS) is a non traumatic atlanto-axial rotatory subluxation of C1-C2 joint. A six year old girl, 20 days after an episode of fever, developed a torticollis and a 3/6 heart murmur. The echocardiography showed a Rheumatic Carditis. The Brain and cervical spine Magnetic resonance imaging (MRI) and the Computerized Tomography (CT) showed rotary dislocation of C1-C2 vertebrae, compatible with GS, and cerebral venous thrombosis (CVT). An antibiotic therapy, Prednisone and a low molecular weight heparin for 7 days was prescribedfollowed by an oral anticoagulant for 6 months. After a month the MRI showed a reduction of the dislocation and cerebral venous recanalization.


Metabolism of 15N-labeled amino acids in rat lens.
Rat lenses were cultured 4-24 hr at 37 degrees C in balanced salt medium containing 5 mM [15N]-glutamate, [15N]-alanine, or amido-[15N]-glutamine. Free amino acids were extracted with 6% trichloracetic acid containing alpha-aminoisobutyrate as an internal standard, and trifluoroacetyl-n-butyl (TAB) derivatives were prepared. Amino acids were quantified by gas chromatography, and 15N enrichment in amino groups of several free amino acids was determined by mass spectrometry. Culture of lenses with either [15N]-glutamate or [15N]-alanine resulted in [15N]-labeling of the glutamate, aspartate, alanine, proline, serine and glycine pools. No detectable amino-15N (less than 5%) was observed in amino acids when lenses were cultured with amido-labeled [15N]-glutamine or with [15N]-ammonium chloride. It is concluded that the alpha-amino groups of several amino acids are actively involved in lens metabolism. In contrast, although glutamine may serve as the major source of glutamate for lens, the amido nitrogen of glutamine plays a minor role, if any, as a source of alpha-amino groups for amino acid metabolism.


Measuring Mobility in Chromatin by Intensity-Sorted FCS.
The architectural organization of chromatin can play an important role in genome regulation by affecting the mobility of molecules within its surroundings via binding interactions and molecular crowding. The diffusion of molecules at specific locations in the nucleus can be studied by fluorescence correlation spectroscopy (FCS), a well-established technique based on the analysis of fluorescence intensity fluctuations detected in a confocal observation volume. However, detecting subtle variations of mobility between different chromatin regions remains challenging with currently available FCS methods. Here, we introduce a method that samples multiple positions by slowly scanning the FCS observation volume across the nucleus. Analyzing the data in short time segments, we preserve the high temporal resolution of single-point FCS while probing different nuclear regions in the same cell. Using the intensity level of the probe (or a DNA marker) as a reference, we efficiently sort the FCS segments into different populations and obtain average correlation functions that are associated to different chromatin regions. This sorting and averaging strategy renders the method statistically robust while preserving the observation of intranuclear variations of mobility. Using this approach, we quantified diffusion of monomeric GFP in high versus low chromatin density regions. We found that GFP mobility was reduced in heterochromatin, especially within perinucleolar heterochromatin. Moreover, we found that modulation of chromatin compaction by ATP depletion, or treatment with solutions of different osmolarity, differentially affected the ratio of diffusion in both regions. Then, we used the approach to probe the mobility of estrogen receptor-α in the vicinity of an integrated multicopy prolactin gene array. Finally, we discussed the coupling of this method with stimulated emission depletion FCS for performing FCS at subdiffraction spatial scales.


Microwave-Assisted Palladium Acetate-Catalyzed C-P Cross-Coupling of Arylboronic Acids and >P(O)H Reagents in the Absence of the Usual Mono- and Bidentate P-Ligands: Mechanistic Insights.
A less-studied halogen-free variation of the Hirao reaction involving the coupling of arylboronic acids with >P(O)H reagents, such as diarylphosphine oxides, diethyl phosphite, and ethyl phenyl-H-phosphinate, was investigated in detail using Pd(OAc)2 as the catalyst precursor and applying some excess of the P-reagent to supply the ligand via its trivalent tautomeric (>P-OH) form. The optimum conditions (1.2 equiv of the P-reagent, 135-150 °C, and air) were explored for the synthesis of diaryl-phenylphosphine oxides, aryl-diphenylphosphine oxides, diethyl arylphosphonates, ethyl diphenylphosphinate, and two bisphosphinoyl derivatives. In the reaction of 4-chlorophenyl- or 3-chlorophenylboronic acid with Ph2P(O)H, triphenylphosphine oxide was also formed as a byproduct. Theoretical calculations suggested that the catalytic cycle of the P-C coupling of PhB(OH)2 with Ph2P(O)H is different from that of the usual cross-coupling reactions. It comprises the addition of a phenyl anion and then the tautomeric form >P-OH of the >P(O)H reagent to the Pd2+ catalyst complex. This is then followed by reductive elimination affording Ph3PO that is accompanied with the conversion of Pd2+ to Pd0. There is a need for a subsequent stoichiometric oxidation of Pd(0) by molecular oxygen. The spontaneous formation of the self-assembling ligands around the Pd2+ center from the >P(O)H reactant plays a crucial role in the mechanism and promotes the efficiency of the catalyst.


Painless Mini-Wellens Sign 5 Minutes After Exercise in a Man With Previously Undiagnosed Total Occlusion of the Left Anterior Descending and Right Coronary Arteries.
A 60-year-old man with no coronary artery history and a normal resting electrocardiogram came to the hospital with his second stroke and underwent a treadmill exercise test before carotid endarterectomy. He had no chest pain and stopped because of leg pain. Five minutes after exercise he developed terminal T-wave inversion in leads V3 and V4 that lasted 7 minutes. The T-wave pattern resembled the Wellens pattern that has usually been seen after intense preinfarction rest pain and has usually lasted hours, days, or even weeks. Coronary arteriography showed complete occlusion of the left anterior descending and right coronary arteries.


Nitrogen losses and cephalothin absorption in peritonitis treated by hourly peritoneal dialysis.
The effects of peritonitis on dialysate nitrogen losses were investigated. Eight patients who developed peritonitis while undergoing peritoneal dialysis were compared to seven noninfected dialysis patients. Dialysate protein losses increased during peritonitis, but nitrogen losses into the dialysate did not increase. These findings were caused by reduced urea nitrogen losses in the peritonitis group, which proportionately, more than made up for the increased nitrogen losses from protein. We speculate that the smaller loss of urea nitrogen was due to diminished intake of food. Cephalothin uptake from the peritoneal cavity was unaltered by peritonitis and a dialysate flow rate of 2 L/hour averaged 26 +/- 2 mL/minute and 27 +/- 2 mL/minute in the noninfected and infected groups, respectively.


Management strategies of sexual dysfunctions in Parkinson's disease.
Sexual dysfunctions (SD) are frequent and highly disabling nonmotor manifestations of Parkinson's disease (PD) but are also potentially treatable. Neurologists should actively discuss, recognize and treat sexual health issues as an integral part of the management of the disease. In this chapter, we provide recommendations for managing and treating both primary and secondary SD in PD. Many sexual problems can be, at least partially, improved by adjusting the treatment of motor, nonmotor symptoms and comorbidities. Although some treatments of primary SD are evidence-based, many therapeutic options have not been yet systematically studied in patients with PD. The development of new treatments and repurposing of existing remedies in patients with PD remain an unmet need.


[Cytochrome concentration at different times and low-temperature conditions of preservation of rat kidney cortical homogenates].
Content of cytochromes (a, c+c1) was decreased in rat kidney cortex homogenates, depending on periods and conditions of storage. Optimal conditions were chosen for prolonged storage of the biomaterial. Under these conditions concentrations of the cytochromes were altered only slightly.


A Therapeutic Trial Comparing Modified Autoinoculation, a Novel Approach with Topical Potassium Hydroxide Application in the Treatment of Molluscum Contagiosum.
Molluscum contagiosum (MC) is a common viral cutaneous infection. Despite multiple treatment options, there is no definitive treatment. In some cases, the lesions are severe, recurrent, and cosmetically odd. Modified autoinoculation (MAI) is a novel technique that induces cell-mediated immunity resulting in clearance of local as well as distant lesions. Potassium hydroxide (KOH) acts by dissolving the keratin and penetrating deeply destroys the hyperproliferative tissue. We would here like to compare MAI with topical KOH in the treatment of MC. The aim of this study was to assess the effectiveness of MAI in treatment of MC and to compare its response with topical KOH application. This was an open-labeled longitudinal therapeutic outcome study carried out at a tertiary care center over a period of 1 year. Hundred consenting MC patients attending the department of dermatology were randomized into Group A and Group B. Group A patients were subjected to MAI and Group B to topical application of 10% KOH. The continuous variables are presented as mean ± standard deviation (SD). The difference between the mean score was analyzed using Student's t test for independent variable and paired t test for paired results. At the end of 16 weeks, 91.48% showed complete clearance by MAI compared to 81.64% with topical 10% KOH solution. There was a significant reduction of mean score of lesions in patients treated by MAI compared to patients treated with KOH. MAI therapy provides a promising, easy, cost-effective, daycare option for MC infections.


Dendritic Cell-Activating Magnetic Nanoparticles Enable Early Prediction of Antitumor Response with Magnetic Resonance Imaging.
Cancer vaccines initiate antitumor responses in a subset of patients, but the lack of clinically meaningful biomarkers to predict treatment response limits their development. Here, we design multifunctional RNA-loaded magnetic liposomes to initiate potent antitumor immunity and function as an early biomarker of treatment response. These particles activate dendritic cells (DCs) more effectively than electroporation, leading to superior inhibition of tumor growth in treatment models. Inclusion of iron oxide enhances DC transfection and enables tracking of DC migration with magnetic resonance imaging (MRI). We show that T2*-weighted MRI intensity in lymph nodes is a strong correlation of DC trafficking and is an early predictor of antitumor response. In preclinical tumor models, MRI-predicted "responders" identified 2 days after vaccination had significantly smaller tumors 2-5 weeks after treatment and lived 73% longer than MRI-predicted "nonresponders". These studies therefore provide a simple, scalable nanoparticle formulation to generate robust antitumor immune responses and predict individual treatment outcome with MRI.


Evaluation of the new Micronaut-Candida system compared to the API ID32C method for yeast identification.
A new system, Micronaut-Candida, was compared to API ID32C to identify 264 yeast (Candida albicans, C. parapsilosis, C. tropicalis, C. krusei, C. inconspicua, C. norvegensis, C. lusitaniae, C. guilliermondii, C. dubliniensis, C. pulcherrima, C. famata, C. rugosa, C. glabrata, C. kefyr, C. lipolytica, C. catenulata, C. neoformans, Geotrichum and Trichosporon species, Rhodotorula glutinis, and Saccharomyces cerevisiae) clinical isolates. Results were in concordance in 244 cases. Eighteen out of the 20 of discordant results were correctly identified by Micronaut-Candida but not by API ID32C, as confirmed by PCR ribotyping.


Clinical perspectives of childhood tuberculosis in Taiwan.
Tuberculosis (TB) is an important public health issue in Taiwan and worldwide. Taiwan has made major progress in combating TB in the past 40 years. However, childhood TB still constitutes a significant challenge in disease control. From January to mid December 2011, 369 new cases of pediatric TB were confirmed. The relatively low case number and variable clinical presentations made it difficult for early detection. Latent TB infections in children also pose further complexity in clinical management. Knowledge of the clinical features of active and latent TB infection is crucial for efficient TB control.


Nonspecific in situ hybridization reaction in neuroendocrine cells and tumors of the gastrointestinal tract using oligonucleotide probes.
Oligonucleotides used in in situ hybridization (ISH), regardless of their sequence specificity, bind to neuroendocrine (NE) cells in normal gastrointestinal mucosa and tumors. This nonspecific binding, presumably related to the presence in NE cells of hidden NH2 groups of obscure origin, can be prevented by acetic anhydride treatment of the sections. This is a routine step in several ISH protocols but not in all. This study emphasizes the need to establish safe protocols and controls to check the specificity of ISH procedures.


Investigations of radicular dentin permeability and ultrastructural changes after irradiation with Er,Cr:YSGG laser and dual wavelength (2780 and 940 nm) laser.
The aim of this study was to assess the effectiveness of dual wavelength (2780 nm Er,Cr:YSGG, 940 nm diode) laser in elimination of smear layer comparing it with Er,Cr:YSGG laser in terms of radicular dentin permeability and ultrastructural changes of root canal walls. Fifty-one sound single-rooted extracted teeth were instrumented up to size F4 and divided into three groups: group Co, non-irradiated samples; group A, irradiated with Er,Cr:YSGG laser; group B, irradiated with the dual wavelength laser. Afterward, the roots were made externally impermeable, filled with 2% methylene blue dye, divided horizontally into three segments reflecting the cervical, middle, and apical thirds then examined under microscope. Using analytical software, the root section area and dye penetration area were measured, and then, the percentage of net dye penetration area was calculated. Additionally, scanning electron microscope investigations were accomplished. Analysis of variance (ANOVA) showed significant differences between all groups over the three root thirds. Dye permeation in dual wavelength laser group was significantly higher over the whole root length: cervical, middle, and apical compared to Er,Cr:YSGG laser group and non-irradiated samples (p < 0.001). Scanning electron micrographs of dual wavelength irradiated samples showed a distinctive removal of smear layer with preservation of the annular structure of dentinal tubules. Er,Cr:YSGG laser root canal irradiation produced uneven removal of smear layer, in efficient cleanliness especially in the apical third. There was no sign of melting and carbonization. Within the studied parameters, root canal irradiation with dual wavelength laser increased dentin permeability.


The efficacy of intralesional vitamin D3 injection in the treatment of cutaneous warts: A clinical therapeutic trial study.
The human papillomavirus that causes warts is usually harmless, and it can infect any part of the skin or mucous membranes. Despite the availability of several treatments, warts often return, and scarring, pigmentation changes, and recurrence are all possible side effects. Intralesional vitamin D3 was employed as an immunotherapy for cutaneous warts in the current investigation. In Al-Sadr Medical City in the city of Al-Najaf Al-Ashraf, a skin clinic conducted a therapeutic clinical experiment. A total of 204 cutaneous warts were examined in 40 patients (14 men and 26 females). Vitamin D3 solution of the dose (600 000 IU) was injected into the lesions' bases, with 0.2 mL per patient. Each session could only inject up to five warts. The injections were given every 2 weeks for a total of four times. Instructing patients to forego the use of any topical or oral medication was also a part of the protocol. Each patient was evaluated for recurrence risk before each therapy and then again 6 months later. There was a wide range of patient ages in this study, from 20 months to 52 years. About 65% of the cases included females. When looking at the many kinds of warts, the most prevalent was the common wart (71.6%). After four treatments, a positive response was considered to have occurred when all lesions had disappeared, a partial response when more than half of the lesions had disappeared, and no reaction when less than half of the lesions had disappeared. The final tally was 81.9% for those who responded in whole, 11.3% for those who responded partially, and 6.9% for those who did not respond at all. Next-session complete response rates were 12.7%, 29.9%, 54.9%, and 81.9%, respectively. Thirteen people experienced adverse symptoms, most noticeably minor swelling and itching. Within 6 months of follow-up, warts had completely disappeared for all patients with a partial or modest response except one who had no reaction. Vitamin D3 administered intralesional is an effective and low-cost treatment for cutaneous warts.


The allometry of leaf form in early plant ontogeny.
A general allometric model between metabolic rate and body size has been derived for early plant ontogeny. The scaling exponent is (2+N/6)/3, where N is the cell's degree of freedom of motion. For early plant ontogeny N=2, our prediction agrees well with Sack et al.'s observation [Sack, L., Maranon, T., Grubb, P.J., 2002. Science 295, 1923].


Effects of prazosin in patients with benign prostatic obstruction.
Prazosin, a selective alpha-1-adrenoceptor blocker, was used in a double-blind crossover study in 20 men with benign prostatic obstruction. Maximum and average flow rates increased, and residual volume and obstructive symptoms were reduced. Voiding pressure parameters, bladder capacity and irritative symptoms did not change significantly. No side effects were noted. We conclude that prazosin seems to be an effective therapeutic alternative in patients with benign prostatic obstruction.


Quantification of superparamagnetic iron oxide using inversion recovery balanced steady-state free precession.
Cellular and molecular MRI trafficking studies using superparamagnetic iron oxide (SPIO) have greatly improved non-invasive investigations of disease progression and drug efficacy, but thus far, these studies have largely been restricted to qualitative assessment of hypo- or hyperintense areas near SPIO. In this work, SPIO quantification using inversion recovery balanced steady-state free precession (IR-bSSFP) was demonstrated at 3T by extracting R2 values from a monoexponential model (P. Schmitt et al., 2004). A low flip angle was shown to reduce the apparent recovery rate of the IR-bSSFP time course, thus extending the dynamic range of quantification. However, low flip angle acquisitions preclude the use of traditional methods for combining RF phase-cycled images to reduce banding artifacts arising from off-resonance due to B0 inhomogeneity. To achieve R2 quantification of SPIO, we present a new algorithm applicable to low flip angle IR-bSSFP acquisitions that is specifically designed to identify on-resonance acquisitions. We demonstrate in this work, using both theoretical and empirical methods, that the smallest estimated R2 from multiple RF phase-cycled acquisitions correspond well to the on-resonance time course. Using this novel minimum R2 algorithm, homogeneous R2 maps and linear R2 calibration curves were created up to 100μg(Fe)/mL with 20° flip angles, despite substantial B0 inhomogeneity. In addition, we have shown this technique to be feasible for pre-clinical research: the minimum R2 algorithm was resistant to off-resonance in a single slice mouse R2 map, whereas maximum intensity projection resulted in banding artifacts and overestimated R2 values. With the application of recent advances in accelerated acquisitions, IR-bSSFP has the potential to quantify SPIO in vivo, thus providing important information for oncology, immunology, and regenerative medicine MRI studies.


Hepatitis B prophylaxis using lamivudine and individualized low-dose hepatitis B immunoglobulin in living donor liver transplantation.
Currently, most available experience concerning prophylaxis against hepatitis B virus (HBV) recurrence after living donor liver transplantation (LDLT) is limited to studies of small size and short follow-up. The objective of this study was to evaluate the efficacy of a prophylactic regimen using lamivudine and individualized low-dose intramuscular hepatitis B immunoglobulin (HBIG) in LDLT. We used a database of adult-to-adult right-lobe LDLT procedures performed from June 2002 to April 2012 at our center for HBV-related end-stage liver diseases. Patients were divided into 3 groups: group A, HBV-related decompensated liver cirrhosis; group B, fulminant hepatitis B; and group C, hepatocellular carcinoma (HCC). During a mean follow-up of 38.3 ± 28.9 months, 8 of 165 (4.8%) recipients developed HBV recurrences. The mean time for HBV reinfection was 15.8 + 11.0 months after transplantation. The overall 1-, 3-, and 5-year HBV recurrence rates were 3%, 7%, and 8.2%, respectively. Both patients with fulminant hepatitis B or HCC seemed to have higher rates of HBV recurrence than those with decompensated liver cirrhosis, albeit not significantly. The independent predictor of HBV recurrence was high HBV DNA level (≥10(5) copies/mL) at LDLT. Lamivudine and individualized low-dose intramuscular HBIG provides effective prophylaxis against HBV recurrence after LDLT. Pre-LDLT HBV DNA of ≥ 10(5) copies/mL was associated with HBV recurrence.


Effect of Single Dose Fluoxetine on Theophylline Serum Concentrations.
Fluoxetine has been previously reported to elevate plasma concentrations of several hepatically metabolized medications. Eight healthy male volunteers received intravenous aminophylline (approximately 6 mg kg(minus sign1)) on two occasions separated by 2 weeks. A 40-mg dose of oral fluoxetine was administered 8 h prior to the second administration of fluoxetine. Blood samples were collected and assayed for theophylline. Fluoxetine administration was not associated with any significant differences with respect to peak theophylline serum concentration, total body clearance, or half-life. Possible explanations include the fact that only a single dose of fluoxetine was administered and low fluoxetine serum concentrations were present at the time of aminophylline administration. The most likely explanation, however, is the fact that fluoxetine inhibits cytochrome P450 2D6 activity, whereas theophylline is predominately metabolized by cytochrome P450 1A2. In summary, a single 40-mg dose of fluoxetine failed to alter the plasma concentrations of theophylline when administered intravenously as aminophylline.


Abderrazak Hajjioui: shining a light on rehabilitation.
Abderrazak Hajjioui talks to Tatum Anderson about the need to prioritize multidisciplinary rehabilitation education, practice and research.


Typhoid fever in children of low and high socioeconomic strata: comparison of hygiene habits.
The relationship between the hygienic habits of children who had typhoid fever (TF) who had recently begun attending school and their family group, is assessed. It is supposed that children, independently of their SES, acquired TF because of inadequate habits which facilitated the oral-fecal cycle. The sample was formed of 40 child-mother dyads: 20 of low SES (group A) and 20 of high SES (group B), the child of each of which had had TF. Results showed that the hygienic habits of children with respect to the oral-fecal cycle, their perception of school toilet cleanliness as well as the mothers' explanation of their children's hygienic habits is very similar in the two groups. The importance of these results is that the SES is seem to be irrelevant in the case of TF but that the hygienic habits of the children are of importance. Public health policy should be modified to include the teaching of proper hygienic habits (oral-fecal cycle).


A dual-tracer approach using [11C]CH and [18F]FDG in HCC clinical decision making.
Early detection of recurrent or progressive HCC remains the strongest prognostic factor for survival. Dual tracer PET/CT imaging with [11C]CH and [18F]FDG can further increase detection rates as both tracers entail different metabolic pathways involved in HCC development. We investigated dual-tracer PET/CT in clinical decision making in patients suspected of recurrent or progressive HCC. All HCC patients who underwent both [11C]CH and [18F]FDG PET/CT in our institute from February 2018 to December 2021 were included. Both tracer PET/CT were within 4 weeks of each other with at least 6-month follow-up. Patients underwent dual tracer PET/CT because of unexplained and suspicious CT/MRI or sudden rise of serum tumour markers. A detected lesion was considered critical when the finding had prognostic consequences leading to treatment changes. Nineteen patients who underwent [11C]CH and [18F]FDG PET/CT were included of which all but six patients were previously treated for HCC. Dual-tracer critical finding detection rate was 95%, with [18F]FDG 68%, and [11C]CH 84%. Intrahepatic HCC recurrence finding rate was 65% for both tracers. [18F]FDG found more ablation site recurrences (4/5) compared to [11C]CH (2/5). Only [11C]CH found two needle tract metastases. Both tracers found 75% of the positive lymph nodes. Two new primary tumours were found, one by [18F]FDG and both by [11C]CH. Our study favours a dual-tracer approach in HCC staging in high-risk patients or when conventional imaging is non-conclusive.


Position statement on nurse-patient ratios in critical care.
The level of nurse-to-patient staffing ratios in critical care environments is a controversial issue. Tracy Pilcher and Mandy Odell review the literature and examine some of the main factors that influence staffing levels. The British Association of Critical Care Nurses' (BACCN) position statement is outlined.


Selective effects of Lipiodolized antitumor agents.
Lipiodol Ultra-Fluid (Lipiodol) remains selectively in the tumor for an extended time when applied through arteries feeding the tumor. Although lipophilic antitumor drugs are selective when combined with Lipiodol, wide application of common hydrophilic agents is limited, as these compounds are insoluble in oil. We propose "Lipiodolization" of water-soluble agents using as an intermediate Urografin, a water-soluble contrast medium. Thirteen patients with primary hepatocellular carcinoma were treated with this Lipiodol-Urografin system containing antitumor agents. Marked decrease in serum alpha-fetoprotein (AFP) levels, decrease in tumor size in the hepatic imaging, and histologic studies of the resected specimen revealed this mode of therapy to be effective in 10 of 13 patients (77%) with hepatocellular carcinoma. Lipiodolization of antitumor agents is a new approach to selective cancer chemotherapy.


Effects of elastic band resistance training and nutritional supplementation on muscle quality and circulating muscle growth and degradation factors of institutionalized elderly women: the Vienna Active Ageing Study (VAAS).
Regular resistance exercise training and a balanced diet may counteract the age-related muscular decline on a molecular level. The aim of this study was to investigate the influence of elastic band resistance training and nutritional supplementation on circulating muscle growth and degradation factors, physical performance and muscle quality (MQ) of institutionalized elderly. Within the Vienna Active Ageing Study, 91 women aged 83.6 (65.0-92.2) years were randomly assigned to one of the three intervention groups (RT, resistance training; RTS, resistance training plus nutritional supplementation; CT, cognitive training). Circulating levels of myostatin, activin A, follistatin, IGF-1 and GDF-15, as well as MQ and functional parameters were tested at baseline as well as after 3 and 6 months of intervention. MQ of lower extremities significantly increased in the RT group (+14 %) and RTS group (+12 %) after 6 months. Performance improved in the RT and RTS groups for chair stand test (RT: +18 %; RTS: +15 %). Follistatin increased only in the RT group (+18 %) in the latter phase of the intervention, accompanied by a decrease in the activin A-to-follistatin ratio (-7 %). IGF-1, myostatin and GDF-15 levels were not affected by the intervention. Our data confirm that strength training improves physical performance and MQ even in very old institutionalized women. This amelioration appears to be mediated by blocking muscle degradation pathways via follistatin rather than inducing muscle growth through the IGF-1 pathway. As plasma levels of biomarkers reflect an overall status of various organ systems, future studies of tissue levels are suggested.


[A study of the combination of vinorelbine and epirubicin as neoadjuvant chemotherapy regimen in the treatment of locally advanced breast cancer].
To evaluate the clinical efficacy and toxicity of vinorelbine (N) and epirubicin (E) as the neoadjuvant chemotherapy regimen in the treatment of locally advanced breast cancer (LABC). From September 2001 to December 2004, 158 patients with LABC were treated with NE chemotherapy before operation. Neoadjuvant chemotherapy containing vinorelbine (N), 25 mg/m(2) (days 1 and 8) and epirubicin (E), 60 mg/m(2) (days 1) was administered every 3 weeks for three cycles before local treatment. Response in the breast: the clinical objective response was 81.6% [23.4% (37/158) cCR and 58.2% (92/158) PR], 16.5% (26/158) SD and 1.9% (3/158) PD. Pathological complete response was found in 29 cases (18.3%). Eighteen cases (26.5%) who have positive FNA result in the axillary lymphnode before chemotherapy showed negative result in the surgery specimen. The most common toxicities were neutropenia, alopecia and nausea/vomiting. Neutropenia grade 3 - 4 was reported in 111 patients (70.3%) and there was no toxic deaths. The combination of vinorelbine and epirubicin is a very active and well-tolerated regimen as neoadjuvant chemotherapy for the LABC.


Myeloperoxidase in human peripheral blood lymphocytes: Production and subcellular localization.
Myeloperoxidase (MPO) is an important enzyme in the front-line protection against microorganisms. In peripheral blood, it is accepted that MPO is only produced by myeloid-lineage cells. Thus, MPO presence is unexpected in lymphocytes. We showed recently that B1-lymphocytes from mice have MPO. Here, we showed that subsets of human peripheral B, CD4(+) and CD8(+) T lymphocytes express MPO. The content of MPO in lymphocytes was very low compared to neutrophils/monocytes with a preferential distribution in the nucleus and perinuclear region. Also, we performed a MPO mRNA expression analysis from human blood cells derived from microarray raw data publicly available, showing that MPO is modulated in infectious disease. MPO was increased in CD4(+) T lymphocytes from HIV chronic infection and in CD8(+) T lymphocytes from HCV-positive patients. Our study points out MPO as a multifunctional protein due to its subcellular localization and expression modulation in lymphocytes indicating alternative unknown functions for MPO in lymphocytes.


Characterization of humoral responses in mice immunized with plasmid DNAs encoding SARS-CoV spike gene fragments.
The immunological characteristics of SARS-CoV spike protein were investigated by administering mice with plasmids encoding various S gene fragments. We showed that the secreting forms of S1, S2 subunits and the N-terminus of S1 subunit (residues 18-495) were capable of eliciting SARS-CoV specific antibodies and the region immediate to N-terminus of matured S1 protein contained an important immunogenic determinant for elicitation of SARS-CoV specific antibodies. In addition, mice immunized with plasmids encoding S1 fragment developed a Th1-mediated antibody isotype switching. Another interesting finding was that mouse antibodies elicited separately by plasmids encoding S1 and S2 subunits cooperatively neutralized SARS-CoV but neither the S1 nor S2 specific antibodies did, suggesting the possible role of both S1 and S2 subunits in host cell docking and entry. These results provide insights into understanding the immunological characteristics of spike protein and the development of subunit vaccines against SARS-CoV.


Persistence of translocations after accidental exposure to ionizing radiation.
To assess the validity of translocations for retrospective biodosimetry using FISH chromosome painting of peripheral lymphocytes in repeated samples of people accidentally exposed to radiation. Blood samples from five people exposed to mainly whole-body irradiation of gamma-rays from a radiation accident in Estonia in 1994 were taken at 2-4 month intervals. A total of eight samplings were carried out, including one directly after the accident. Fluorescence in situ hybridization with probes for chromosomes 1, 2 and 4 was performed on metaphase preparations from 48 h whole blood cultures; 1500 cells were scored from each individual per time point. Translocations remained relatively stable during 2 years after exposure in all subjects. A noticeable decrease in complete translocation yields and a significant decrease in incomplete translocations were observed in one person. In addition to whole-body exposure, he had also been exposed to partial-body irradiation. Due to the overall persistence of translocations, dose estimates were very similar throughout the 2 year period. A rapid decline in dicentric frequencies was noted during the first year after exposure. The results suggest that during 2 years the yield of translocations in peripheral lymphocytes remained at a constant level after whole-body exposure. This finding supports the use of translocations for retrospective dosimetry, at least within this relatively short period of follow-up. In the case of partial-body irradiation, however, the elimination of co-existing unstable aberrations reduced the translocation yield over time. Follow-up will be continued in order to determine the stability of translocations over longer times.


Correlation of lysozyme activity with proteoglycan biosynthesis in epiphyseal cartilage.
Pig epiphyseal cartilage (proximal ulna epiphysis) previously incubated into vitro in the presence of sodium [35S]sulfate or [3H]thymidine was either analyzed by autoradiography or separated into 9 morphologically defined consecutive layers and investigated for 35S-incorporation into the guanidinium chloride-extractable proteoglycans and for lysozyme activity. The lowest 35S incorporation and lysozyme activity were determined in the zone of resting cells, but there is a consecutive increase in the rate of proteoglycan synthesis and lysozyme activity toward the diaphyseal cartilage-bone junction, with the maximum at the lower columnar cell zone and a sharp reduction of both parameters at the hypertrophic zone. The maxima of 35S incorporation and [3H]thymidine incorporation do not coincide. The guanidinium chloride-soluble proteoglycans exhibit macromolecular polydispersity. Fractions excluded from as well as retarded by Sepharose 2B gel could be separated and were detected in all zones. The results indicate a correlation of proteoglycan biosynthesis and lysozyme activity in epiphyseal cartilage.


Efficacy of addition of fentanyl to epidural bupivacaine on postoperative analgesia after thoracotomy for lung resection in infants.
The authors evaluated the efficacy of adding fentanyl to epidural bupivacaine in infants up to 6 months of age after a thoracotomy in a prospective, randomized, double-blind study. The primary outcome was the total amount of rescue doses of intravenous nalbuphine in the first 24 h after surgery. Secondary outcomes included (1) time to first rescue dose of nalbuphine, (2) pain scores, and (3) behavior scores. Thirty-two infants were randomly assigned to receive an epidural infusion containing 0.1% bupivacaine (group B; n = 16) or 0.1% bupivacaine and 2 microg/ml fentanyl (group BF; n = 16). Patients were evaluated up to 24 h after surgery for pain; amount of analgesic rescues and time to first rescue; pain scores; behavior scores (five-item behavior score); and complications, including respiratory depression, oxygen requirement, vomiting, and urinary retention. The two groups had similar demographics. Nalbuphine consumption (P = 0.001) and pain scores (P < 0.001) in the first 24 h were significantly decreased in group BF compared with group B. The time to first analgesic rescue was significantly longer in group BF (P = 0.005). The five-item behavior score was significantly better in group BF than in group B (P = 0.01). The incidence of side effects, the time to first successful feeding, and the time to discharge were similar in both groups. Addition of 2 microg/ml epidural fentanyl to 0.1% bupivacaine results in improved postthoracotomy analgesia without any increase in side effects, compared with 0.1% bupivacaine, in infants up to 6 months of age.


The incidence and significance of early pericardial effusion after open heart surgery.
The presence and severity of postoperative pericardial effusions were studied echocardiographically in 114 consecutive patients (70 males, 44 females; mean age 56 +/- 10 years). An effusion was present in 35 patients at 3-5 days. An effusion was less common when a drainage tube was inserted for 24-36 h in the posterior as well as the anterior mediastinum than when only an anterior drain was used. Patients with effusions differed from those without in having more supraventricular arrhythmias, more wound infections, smaller total blood drainage and longer postoperative hospital stay. Three patients with posterior pericardial effusions developed cardiac tamponade 5-18 days postoperatively. The data show that pericardial effusions are associated with postoperative complications and suggest that effusion formation can be reduced by using posterior as well as anterior chest drains.


Characterizing the amplification of STR markers in multiplex polymerase chain displacement reaction using massively parallel sequencing.
Polymerase chain displacement reaction (PCDR) showed advantages in forensic low-template DNA analysis with improved amplification efficiency, higher allele detection capacity, and lower stutter artifact than PCR. However, characteristics of STR markers after PCDR amplification remain unclarified for the limited resolving power of capillary electrophoresis (CE). This issue can be addressed by massively parallel sequencing (MPS) technology with higher throughput and discriminability. Here, we developed a multiplex PCDR system including 24 STRs and amelogenin. In addition, a PCR reference was established for comparison. After amplification, products were subjected to PCR-free library construction and sequenced on the Illumina NovaSeq system. We implemented a sequence-matching pipeline to separate different amplicon types of PCDR products from the combination of primers. In the sensitivity test, the PCDR multiplex obtained full STR profiles with as low as 125 pg 2800M control DNA. Based on that, single-source DNA samples were tested. First, highly concordant genotypes were observed among the PCDR multiplex, the PCR reference, and CE-based STR kits. Next, read counts of different PCDR amplicon types were investigated, showing a relative abundance of 78:12:12:1 for the shortest amplicon S, the two medium amplicons M1 and M2, and the longest amplicon L. We also analyzed the stutter artifacts for distinct amplicon types, and the results revealed the reduction of N - 1 and N - 2 contraction stutters, and the increase of N + 1 and N + 2 elongation stutters in PCDR samples. Moreover, we confirmed the feasibility of PCDR for amplifying degraded DNA samples and unbalanced DNA mixtures. Compared to the previous proof of principle study, our work took a further step to characterize the complete profile of STR markers in the PCDR context. Our results suggested that the PCDR-MPS workflow is an effective approach for forensic STR analysis. Corresponding findings in this study may help the development of PCDR-based assays and probabilistic methods in future studies.


Application of soft computing to predict water quality in wetland.
Prediction of water quality is a critical issue because of its significant impact on human and ecosystem health. This research aims to predict water quality index (WQI) for the free surface wetland using three soft computing techniques namely, adaptive neuro-fuzzy system (ANFIS), artificial neural networks (ANNs), and group method of data handling (GMDH). Seventeen wetland points for a period of 14 months were considered for monitoring water quality parameters including conductivity, suspended solid (SS), biochemical oxygen demand (BOD), ammoniacal nitrogen (AN), chemical oxygen demand (COD), dissolved oxygen (DO), temperature, pH, phosphate nitrite, and nitrate. The sensitivity analysis performed by ANFIS indicates that the significant parameters to predict WQI are pH, COD, AN, and SS. The results indicated that ANFIS with Nash-Sutcliffe Efficiency (NSE = 0.9634) and mean absolute error (MAE = 0.0219) has better performance to predict the WQI comparing with ANNs (NSE = 0.9617 and MAE = 0.0222) and GMDH (NSE = 0.9594 and MAE = 0.0245) models. However, ANNs provided a comparable prediction and the GMDH can be considered as a technique with an acceptable prediction for practical purposes. The findings of this study could be used as an effective reference for policy makers in the field of water resource management. Decreasing variables, reduction of running time, and high speed of these approaches are the most important reasons to employ them in any aquatic environment worldwide.


Knee Arthrodesis Outcomes After Infected Total Knee Arthroplasty and Failure of Two-stage Revision With an Antibiotic Cement Spacer.
Reinfected total knee arthroplasty can be managed with a second two-stage exchange or a knee arthrodesis procedure. Twenty-three patients with knee arthrodesis after failed exchange arthroplasty for infection were reviewed. Patients were managed with a staged protocol of implant extraction, débridement, and implantation of an antibiotic spacer, with subsequent arthrodesis. Follow-up averaged 40.4 months, with a minimum of 1 year. Bony union with eradication of infection was achieved in 20/23 knees. Sixteen of the 20 patients were able to ambulate with minimal pain. The average time to union was 11.3 months, and the average leg length discrepancy was 4.85 cm. The average Knee Society Score after arthrodesis was 44, and the average visual analog scale pain score was 1.73. Three patients underwent above-knee amputation. Knee arthrodesis performed for persistent periprosthetic infection allowed for eradication of infection and union in 87% of the patients, creating a stable knee fusion.


The AU-rich sequences present in the introns of plant nuclear pre-mRNAs are required for splicing.
Plant cells do not in general process the introns of transcripts expressed from introduced vertebrate genes. By studying the processing of model introns in transfected plant protoplasts, we have investigated the special requirements for intron recognition by plant cells. Our results indicate that the requirements for intron recognition in plants are different from those of both metazoa and yeast. A synthetic intron of arbitrary sequence but incorporating splice site consensus sequences and a high proportion of U and A nucleotides, a characteristic feature of plant introns, was efficiently spliced in protoplasts. We have studied the effects of various sequence alterations and conclude that AU-rich sequences are necessary for intron recognition. In addition, we find that the criteria for branch site selection are relaxed, as they are in vertebrates, but a polypyrimidine tract is not necessary.


Chloroquine-mediated lysosomal dysfunction enhances the anticancer effect of nutrient deprivation.
To investigate the ability of chloroquine, a lysosomotropic autophagy inhibitor, to enhance the anticancer effect of nutrient deprivation. Serum-deprived U251 glioma, B16 melanoma and L929 fibrosarcoma cells were treated with chloroquine in vitro. Cell viability was measured by crystal violet and MTT assay. Oxidative stress, apoptosis/necrosis and intracellular acidification were analyzed by flow cytometry. Cell morphology was examined by light and electron microscopy. Activation of AMP-activated protein kinase (AMPK) and autophagy were monitored by immunoblotting. RNA interference was used for AMPK and LC3b knockdown. The anticancer efficiency of intraperitoneal chloroquine in calorie-restricted mice was assessed using a B16 mouse melanoma model. Chloroquine rapidly killed serum-starved cancer cells in vitro. This effect was not mimicked by autophagy inhibitors or LC3b shRNA, indicating autophagy-independent mechanism. Chloroquine-induced lysosomal accumulation and oxidative stress, leading to mitochondrial depolarization, caspase activation and mixed apoptotic/necrotic cell death, were prevented by lysosomal acidification inhibitor bafilomycin. AMPK downregulation participated in chloroquine action, as AMPK activation reduced, and AMPK shRNA mimicked chloroquine toxicity. Chloroquine inhibited melanoma growth in calorie-restricted mice, causing lysosomal accumulation, mitochondrial disintegration and selective necrosis of tumor cells. Combined treatment with chloroquine and calorie restriction might be useful in cancer therapy.


Chronic heat stress increases insulin-like growth factor-1(IGF-1) but does not affect IGF-binding proteins in growing pigs.
The insulin-like activity of insulin-like growth factor-1 (IGF-1) is heavily blunted by IGF binding proteins (IGFBPs, including IGFBP-1 and IGFBP-3). The effects of heat stress (HS) on the IGF-1 and IGFBPs in growing pigs are still not clear. This study aimed to investigate the alterations of IGF-1 and IGFBPs under chronic HS in growing pigs. Twenty-seven growing large white barrows with similar body weight were collected from nine litters and were assigned into three treatments. The litter effect is balanced in all treatments. Treatments were: 1) thermal-neutral (TN) group (23 °C), 2) chronic HS group (33 °C), and 3) pair-fed in TN condition (PFTN). The experiment lasted for 21 days. Compared with TN controls, decreased FI, lower average body weight gain, higher rectal temperature and increased respiration rates were observed in HS pigs. On D7, increased plasma insulin concentration and insulin:glucose ratio were observed in HS pigs compared to TN controls. A overall elevation of plasma IGF-1:IGFBP-3 ratio was detected in HS pigs compared with PFTN and TN counterparts. Besides, hepatic IGF-1 gene expression of HS pigs was 50% higher than TN counterparts. PFTN pigs, however, had no differences in plasma IGF-1:IGFBP-3 ratio and hepatic IGF-1 gene expression, compared with TN pigs. PFTN pigs increased plasma IGFBPs concentration and hepatic IGFBPs gene expression, compared with TN controls. However, no differences in plasma IGFBPs concentration and hepatic IGFBPs gene expression were observed between TN and HS group. Liver IRS-1 gene and protein expressions of HS pigs tended to be increased compared with TN controls, while PFTN pigs still kept liver IRS-1 gene and protein expressions in normal level. High temperature (33 °C), independent of feed intake reduction, increased IGF-1 but did not affect IGFBPs levels in growing pigs.


Discrimination of meat from fur-producing and food-providing animals using mass spectrometry-based proteomics.
Non-edible meat from fur-producing animals entering into meat consumption chain could pose a serious threat to public health. For the purpose of risk prevention and control of meat safety, in this study, marker peptides for discriminating non-edible meat of fur-producing animals (including fox, silver fox, blue fox, raccoon dog, ussuri raccoon dog, mink and American mink) from meat of food-providing animals (including pig, cattle, sheep and donkey) were explored by shot-gun proteomics and verified by target approach. Two mass spectrometry platforms combined with bioinformatic and chemometric tools were integratedly emloyed for method development. Meat samples were first subjected to in-solution protein digestion and the subsequently tryptic peptides were profiled and quantitated by ultra-high pressure liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF MS) with sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) mode. Candidate marker peptides screened by chemometric tools were further filtered for their biological specificity and detectability through bioinformatics analysis as well as multiple reaction monitoring (MRM) verification with UHPLC-triple quadrupole mass spectrometry (UHPLC-QQQ MS). As a result, 9 peptides, out of 104 candidates, were selected as markers for discriminating analysis, of which DQTLQEELAR was validated as primary indicator of non-edible meat from the concerned fur-producing animals. An MRM method based on the developed marker peptides for routine use was finally proposed for risk alarming of non-edible meat from fur-producing animals in food safety control.


Prenatal psychosocial stress exposure is associated with insulin resistance in young adults.
The objective of the study was to examine the association in humans between maternal psychosocial stress exposure during pregnancy and measures of glucose-insulin metabolism in the adult offspring. Healthy young adults whose mothers experienced major stressful life events during their pregnancy (n = 36, prenatal stress, PS group, mean age 25 +/- 5.14 [SD] years) and a comparison group (n = 22, CG, mean age 24 +/- 3.7 [SD] years) underwent an oral glucose tolerance test. Glucose levels were not significantly different across the groups; however, prenatally stressed subjects showed significantly elevated 2-hour insulin (P = .01) and C-peptide levels (P = .03). These differences were independent of other major risk factors for insulin resistance, including birth phenotype (birthweight, length of gestation), a family history of diabetes, gestational diabetes, body mass index, proinflammatory state, and smoking. Higher insulin responses reflect relative insulin resistance in these prenatally stressed young adults. This study is the first to provide evidence for a link in humans between prenatal psychosocial stress exposure and alterations in glucose-insulin metabolic function.


Effect of midbrain raphe lesion or 5,7-dihydroxytryptamine treatment on the prolactin-releasing action of quipazine and D-fenfluramine in rats.
The role of brain serotonin in regulating prolactin (PRL) secretion has been investigated by studying the effect of quipazine and D-fenfluramine, two serotonin-like drugs, on plasma PRL levels under various experimental conditions. Quipazine (5, 10 and 20 mg/kg i.p.) and D-fenfluramine (5, 7.5 and 10 mg/kg i.p.) induced dose-related increases in plasma PRL levels in male rats. Intraventricular injection of 5,7-dihydroxytryptamine (5,7-DHT) or electrolytic lesion of the nucleus raphe medianus (MR), which caused a marked and selective depletion of hypothalamic serotonin levels, significantly reduced the PRL-releasing effect of both quipazine and D-fenfluramine. These results suggest that the effect of these drugs on PRL release is mediated through a serotonergic mechanism in the brain.


The role of Sep (O-phosphoserine) tRNA: Sec (selenocysteine) synthase (SEPSECS) in proliferation, apoptosis and hormone secretion of trophoblast cells.
To investigate whether Sep (O-phosphoserine) tRNA: Sec (selenocysteine) synthase (SEPSECS), which plays an essential role in the synthesis of selenoprotein, affects proliferation, apoptosis and hormone secretion of human trophoblast cells. Human trophoblast JEG-3 cells were divided into four groups: control group, SEPSECS silenced-expression group, empty vector group and SEPSECS over-expression group. Over-expression and silenced-expression were achieved by transfection with plasmid DNA or RNA oligonucleotide, respectively. 3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT) and colony formation assays were performed to investigate cell proliferation, while apoptosis was tested by annexin V-FITC, PI double staining and caspases-3 activation assays, enzyme-linked immunosorbent assay (ELISA) was used to determine the level of progesterone (PG) and human chorionic gonadotropin (hCG). SEPSECS silenced-expression clearly inhibited proliferation of JEG-3 cells (p < 0.05), significantly induced cell apoptosis (p < 0.01) and reduced the production of PG and hCG (p < 0.05). On the contrary, SEPSECS over-expression significantly promoted both cell proliferation (p < 0.01) and secretion of PG and hCG (p < 0.05). SEPSECS significantly affects proliferation, apoptosis and hormone secretion of human trophoblast cells, suggesting that a potential relationship exists among SEPSECS, cell proliferation, apoptosis and hormone production of human placental trophoblast cells. Furthermore, this may provide a clue to uncover the relationship between selenium and human placental in association with an emphasis on the importance of selenium adequacy during pregnancy.


Complete Response of Chemo-Refractory Metastatic Metaplastic Breast Cancer to Paclitaxel-Immunotherapy Combination.
BACKGROUND Metaplastic breast cancer (MPBC) is a rare subtype of breast cancer that is difficult to manage and is resistant to therapy. Immunotherapy is becoming a promising option for care of several types of cancers including triple negative breast cancer. CASE REPORT We present a case of a patient with chemo-refractory metastatic metaplastic-type of breast cancer with failures to 3 active lines of chemotherapy. The patient achieved a complete and sustained response to chemo-immunotherapy combination with acceptable tolerability and minimal adverse events. The treatment was a combination of anti-PD-L1 antibody (durvalumab) immunotherapy in combination with the chemotherapeutic agent, paclitaxel. CONCLUSIONS This is the first case reporting the successful use of durvalumab and paclitaxel combination for treatment of triple negative breast cancer in general, and in metastatic MPBC in specific.


Soluble adhesion molecules in acute disseminated encephalomyelitis.
Soluble adhesion molecules are overexpressed in neuroinflammatory disorders. Their synthesis parallels that of their membrane-bound counterparts, which modulate lymphocyte transmigration through the blood-brain barrier. Blood-brain barrier cellular migration may be essential in the evolution of postinfectious inflammatory central nervous system disease. The serum levels of soluble intercellular adhesion molecule-1, soluble vascular adhesion molecule-1, and soluble E-selectin were measured in 12 children with acute disseminated encephalomyelitis and in two control groups (35 healthy and 35 affected by noninflammatory neurologic diseases) of similar age. In patients with acute disseminated encephalomyelitis, soluble E-selectin serum levels were significantly higher (median 113 ng/mL, range 54-144) than in both control groups (median 44, range 31-63, and median 58, range 43-89, respectively; one-way analysis of variance, P < 0.0001); a statistical trend for higher levels of soluble intercellular adhesion molecule-1 was observed in acute disseminated encephalomyelitis subjects, whereas soluble vascular adhesion molecule-1 titers did not differ between the three groups. The specific role played by each of these molecules in lymphocyte extravasation and the differential cytokine modulation of their expression might explain the result. Further larger studies are required including serial measurements and cerebrospinal fluid analysis.


Arsenite Oxidation by Dunaliella salina is Affected by External Phosphate Concentration.
Arsenic (As) contamination in terrestrial and aquatic environments is a well-known global environmental problem. The biooxidation of arsenite [As(III)] and subsequent arsenate [As(V)] removal have increasingly been used for remediation of As-polluted groundwater. However, little is known about As(III) oxidation by microalgae, especially those from saltwater environments. In this study, we investigated As(III) toxicity and oxidation in the marine microalga Dunaliella salina in the presence of different phosphate concentrations. The results of the As(III) toxicity experiments showed that D. salina was tolerant to As(III) (5.4 ± 0.31 mg As L-1 at 72 h of culture). The As(V) percentage in the P-enriched (11.2 mg L-1) medium was 7.2-fold greater than in the P-deficient one after 24-h exposure, indicating As(III) oxidation by D. salina was more pronounced with increased phosphate levels. Treatment of As(III) with and without 2,4-dinitrophenol (DNP) on the algal cells showed that As(III) oxidation occurred mainly on the cell surface and in the cytoplasm of D. salina. The results of this study suggest that transformation of As(III) into As(V) may be an important pathway of detoxification in D. salina and that phosphate plays a key role in this oxidation process.


Misoprostol to treat missed abortion in the first trimester.
Missed abortion in the first trimester is characterized by the arrest of embryonic or fetal development. The cervix is closed and there is no or only slight bleeding. Ultrasound examination shows an empty gestational sac or an embryo/fetus without cardiac activity. Based on a review of the published literature a single dose of 800 microg vaginal misoprostol may be offered as an effective, safe, and acceptable alternative to the traditional surgical treatment for this indication. Alternatively, 600 microg misoprostol can be administered sublingually. After administration of misoprostol, hospitalization is not necessary and the time to expulsion varies considerably. Bleeding may last for more than 14 days with additional days of light bleeding or spotting. The woman should be advised to contact a provider in case of heavy bleeding or signs of infection. A follow-up is recommended after 1 to 2 weeks.


Silver nanoparticles-mediated G2/M cycle arrest of renal epithelial cells is associated with NRF2-GSH signaling.
Silver nanoparticles (nAg) are known to evoke reactive oxygen species (ROS) generation and consequent cell damage. The transcription factor NF-E2-related factor 2 (NRF2) controls both the basal and inducible expression of multiple antioxidant genes. This study was aimed to investigate the role of NRF2 in nAg-induced renal epithelial cell damage. nAg treatment intensified DNA damage and G2/M cell cycle arrest by nAg in NRF2 knockdown HK-2 (NRF2i) compared with the control cells. As a signaling mechanism associated with nAg-mediated growth arrest, the levels of phospho-CDC25C and phospho-CDC2 were significantly increased in NRF2i. Target gene analysis revealed that nAg-mediated increase in γ-glutamate cysteine ligase expression is NRF2-dependent: nAg-treated NRF2i showed a reduction in glutathione (GSH) content and elevation in ROS level in comparison with the control cells. Additionally, pretreatment of N-acetylcystein in nAg-treated NRF2i alleviated ROS-mediated DNA damage and G2/M cell cycle arrest, while GSH depletion exacerbated DNA damage and cell cycle arrest in the control cells. Taken together, these results suggest that NRF2-mediated GSH increase plays a protective role in nAg-induced DNA damage and subsequent G2/M cell cycle arrest in human renal epithelial cells.


Chemoprevention of pancreatic cancer: a review of the molecular pathways involved, and evidence for the potential for chemoprevention.
Pancreatic cancer has a poor prognosis. The use of drugs or natural agents which inhibit or slow down tumour growth therefore has important potential in the development of future therapies. A literature search of the PubMed and ISI Web of Science databases was undertaken to review the current data available on the alterations in signalling pathways found in pancreatic carcinogenesis, in order to identify sites that could be targeted by chemopreventive agents. Several agents of particular relevance to pancreatic cancer were identified, and their possible mechanisms of action reviewed. Chemopreventive agents such as non-steroidal anti-inflammatory drugs, green tea constituents, and antioxidants have been shown to target various steps in intracellular signalling pathways, particularly those controlling cell proliferation and survival. Work on cell lines and animal models has shown that some of these agents may be able to modulate the growth of pancreatic tumours. Initial clinical trials of some chemopreventives in pancreatic cancer have been undertaken, and have yielded mixed results, prompting the need for further studies. As the molecular pathology of pancreatic cancer becomes better understood, sites of action of chemopreventive substances have been uncovered. Several agents have shown promising results by their ability to inhibit pancreatic carcinogenesis in laboratory studies. If these effects can be successfully translated into human studies then these agents may prove to be valuable adjuvant therapies in the future.


Uterine trauma associated with midtrimester abortion induced by intra-amniotic prostaglandin F2alpha with and without concomitant use of oxytocin.
Five of 80 (6.2%) nulliparous women sustained uterine trauma in association with midtrimester abortion induced by intra-amniotic prostaglandin F2alpha and intravenous oxytocin. All five women suffered cervical lacerations, one extending to the lower uterine segment of the corpus and another associated with myometrial necrosis caused by cornual sacculation and ischemia. No uterine trauma was observed among 95 parous women aborted in the same fashion during this study. The different mechanisms of cervical dilation in the parous woman and the nullipara are offered as an explanation for this difference. Thirty-nine other cases of uterine injury associated with the use of intraamniotic prostaglandin F2alpha from the literature were reviewed, and found to indicate that midtrimester abortion induced by intra-amniotic prostaglandin F2alpha is associated with a significant risk of uterine trauma in the nullipara. The risk seems to increase with the use of oxytocin and with increasing gestational age.


Why do only certain insects escape from a spider's web?
Only heavy insects and those that fly quickly can pass through a spider's web. When an insect is entangled in a web, permanent activity is the best way of getting free. Small wings are conducive to a successful escape, as is a special surface structure of the wings (scales, hairs, lipoid surface). The autotomy of legs, chewing mandibulae or an enzymatic lysis of single threads have nearly no effect. Experiments with approximately 40 taxa of the possible prey of web spiders explain their different behaviour in a spider's web.


Lovastatin: a new cholesterol-lowering agent.
Lovastatin is a potent new drug for lowering serum cholesterol through inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, the rate-limiting enzyme for cholesterol biosynthesis. Metabolic studies with lovastatin in healthy volunteers and patients with hypercholesterolemia suggest reduced synthesis of low-density lipoprotein cholesterol (LDL-C) as well as enhanced catabolism LDL-C mediated through LDL receptors as the principal mechanisms for lipid-lowering effects. Total cholesterol and LDL-C are reduced by 30% or more on average when added to baseline therapy, with the effects being more pronounced in nonfamilial than in familial hypercholesterolemia. Optimal dosing appears to be 20 mg given twice a day. The most common adverse effects are gastrointestinal, while the most serious are elevated transaminase levels and the potential for lens opacities. Lovastatin is the first of a new class of lipid-lowering agents, and is effective when added to diet therapy or in combination with other drugs.


Response of ionization chamber based pocket dosimeter to beta radiation.
Quantitative estimate of the response of ionization chamber based pocket dosimeters (DRDs) to various beta sources was performed. It has been established that the ionization chamber based pocket dosimeters do not respond to beta particles having energy (Emax)<1 MeV and same was verified using (147)Pm, (85)Kr and (204)Tl beta sources. However, for beta particles having energy >1 MeV, the DRDs exhibit measureable response and the values are ~8%, ~14% and ~27% per mSv for natural uranium, (90)Sr/(90)Y and (106)Ru/(106)Rh beta sources respectively. As the energy of the beta particles increases, the response also increases. The response of DRDs to beta particles having energy>1 MeV arises due to the fact that the thickness of the chamber walls is less than the maximum range of beta particles. This may also be one of the reasons for disparity between doses measured with passive/legal dosimeters (TLDs) and DRDs in those situations in which radiation workers are exposed to mixed field of gamma photons and beta particles especially at uranium processing plants, nuclear (power and research) reactors, waste management facilities and fuel reprocessing plants etc. The paper provides the reason (technical) for disparity between the doses recorded by TLDs and DRDs in mixed field of photons and beta particles.


Use of intramedullary polypropylene peg for joint stabilization in leprosy.
Joint arthrodesis is indicated when motion in a joint cannot be restored, and that joint requires stabilization in an optimal functional position. The Harrision Intramedullary peg has been used to advantage for this purpose in hand reconstruction in leprosy in a renal developing area. It is a method to be considered for selected cases requiring joint stabilization to improve total hand function.


Identification of a CD4 T-cell epitope in tumor rejection antigen RLakt on BALB/c radiation-leukemia RL male 1.
We have previously shown that the RLakt antigen was predominantly recognized by CD8 cytotoxic T lymphocytes (CTL) in RL male 1-bearing or -rejected syngeneic BALB/c mice. CD8 CTL were directed to the octamer pRL1a peptide IPGLPLSL of which recognition was H-2L(d)-restricted. In this study, we identified a CD4 T-cell epitope peptide in the tumor rejection antigen RLakt on BALB/c radiation-leukemia RL male 1. Analyses of the recognition of a bulk CD4 T-cell line using several recombinant RLakt proteins suggested the presence of multiple CD4 T-cell epitopes in the molecule. However, cloning from a bulk CD4 T-cell line resulted in only two clones from 200 wells seeded at three cells per well, and those two CD4 T-cell clones recognized the same epitope peptide in RLakt. The epitope peptide was 14-mer p12-25, AYREETLSIIPGLP, and its recognition was H-2IA(d)-restricted. This sequence overlapped with the CD8 T-cell epitope pRL1a in its N-terminal 5 amino acid residues. The relationship of the epitope to the pRL1a peptide predominantly recognized by CD8 CTL suggests that the 14-mer epitope is predominantly recognized by CD4 T-cells.


Role of urinary alpha1-antitrypsin in Padutin (kallikrein) inactivation.
An inverse relationship between proteolytic activity in the presence of kallikrein 0.4 IU and urinary alpha1-antitrypsin concentration has been demonstrated. This protease inhibitor can directly inactivate kallikrein activity. The inhibition was abolished by removal of urinary alpha1-antitrypsin by trypsin-sepharose treatment. Inhibition could be reversed by addition of purified alpha1-antitrypsin. These effects could not be demonstrated with inter-alpha-trypsin inhibitor or alpha2-macroglobulin. The inhibitory effect of alpha1-antitrypsin on kallikrein activity should be taken into account in studies in which kallikrein activity is estimated.


Executive summary of the KDIGO Controversies Conference on Supportive Care in Chronic Kidney Disease: developing a roadmap to improving quality care.
Patients with advanced chronic kidney disease (CKD) have a high burden of physical and psychosocial symptoms, poor outcomes, and high costs of care. Current paradigms of care for this highly vulnerable population are variable, prognostic and assessment tools are limited, and quality of care, particularly regarding conservative and palliative care, is suboptimal. The KDIGO Controversies Conference on Supportive Care in CKD reviewed the current state of knowledge in order to define a roadmap to guide clinical and research activities focused on improving the outcomes of people living with advanced CKD, including those on dialysis. An international group of multidisciplinary experts in CKD, palliative care, methodology, economics, and education identified the key issues related to palliative care in this population. The conference led to a working plan to address outstanding issues in this arena, and this executive summary serves as an output to guide future work, including the development of globally applicable guidelines.


Strategies of emergence in the chestnut weevil Curculio elephas (Coleoptera: Curculionidae).
In the chestnut weevil Curculio elephas, adult emergences spread over 3 or 4 years due to prolonged larval diapause in some individuals. Weevils with an extended diapause emerge, on the average, 1-10 days before those with simple diapause, but whatever the age of insects, emergences occur always from mid-August to early October. When the summer is dry, some adults cannot emerge because of the hardness of the soil. Emergence sucess of adults is smaller in females than in males. The result is that the sex ratio is female-biased before emergence and male-biased after. Summer drought cannot be predicted by the chestnut weevil, and when the soil is dry 27-78% of females cannot emerge and do not reproduce. The year after a summer drought, many reproducing females may emerge from larvae with prolonged diapause. These results suggest an evolutionary influence on the variability in diapause duration. Computer simulations and observations do not support the hypothesis that the main cause of variation in diapause length is the existence of several distinct genotypes within populations. On the contrary, our data strengthen the hypothesis for coin-flipping plasticity discussed in a previous paper.


The association of melioidosis with climatic factors in Darwin, Australia: A 23-year time-series analysis.
Melioidosis is an often fatal disease in humans and animals and endemic in Southeast Asia and northern Australia. It is caused by the environmental bacterium Burkholderia pseudomallei. We analysed weather and climate factors preceding new melioidosis cases in Darwin and compared the time between weather event and admission to hospital for severe and average wet season rainfall. In a time-series analysis from 1990 to 2013 we applied a boosted regression tree and a negative binomial model to investigate the association between melioidosis cases and weather events. Fitted Fourier terms controlled for long-term seasonal trends. We found a rise in the dew point, cloud cover, rainfall, maximum temperature and groundwater to be associated with an increased risk to acquire melioidosis. A shorter 'putative' incubation period was evident after severe rainfall events. Rainfall occurring early in the wet season was linked to more cases as was an increase in the local sea surface temperature reflecting local weather dynamics and precipitation. Our findings demonstrate a statistical association between frequency of recorded melioidosis cases and the nature and timing of rainfall related events and suggest a future rise in the sea surface and ambient temperature may lead to increased melioidosis.


Glutathione production by non-Saccharomyces yeasts and its impact on winemaking: A review.
Glutathione (GSH) is a non-protein thiol naturally present in grape berries and produced by yeasts during fermentation. It has a strong antioxidant activity; thus, the addition of pure GSH during winemaking is recommended to limit the oxidative phenomena of wine, preserving sensory characteristics and stability, ultimately promoting a healthier product by reducing the need for SO2 addition. A promising alternative approach considers the use of yeast starter cultures high producers of this compound in situ, during the fermentation process, in substitution of external GSH addition. Recent research showed that multistarter fermentations with non-Saccharomyces yeasts produce even higher concentrations of GSH compared to single Saccharomyces cerevisiae. Accumulation of GSH in yeast cells is also considered valuable during the growth and dehydration of biomass for starter production, aiding strains to overcome the stressful conditions of industrial process. Moreover, a current trend in oenology is the use during fermentation of inactivated dry yeasts preparations as a source of nutrients, and many of them contain GSH-enriched cells. The aim of this review was to assess the significance of GSH production for the exploitation of wine-related non-Saccharomyces yeasts, both in starter biomass production and during fermentations, which were until now studied in detail for S. cerevisiae. This compendium highlights an interesting new feature of non-conventional yeasts and upgrade the strategy of multistarter fermentation as a valuable tool to positively modulate wine composition.


Immunoprofiling in Neuroendocrine Neoplasms Unveil Immunosuppressive Microenvironment.
Checkpoint inhibitors have shown promising results in a variety of tumors; however, in neuroendocrine tumors (NET) and neuroendocrine carcinomas (NEC), low response rates were reported. We aimed herein to investigate the tumor immune microenvironment in NET/NEC to determine whether checkpoint pathways like programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) might play a role in immune escape and whether other escape mechanisms might need to be targeted to enable a functional antitumor response. Forty-eight NET and thirty NEC samples were analyzed by immunohistochemistry (IHC) and mRNA immunoprofiling including digital spatial profiling. Through IHC, both NET/NEC showed stromal, but less intratumoral CD3+ T cell infiltration, although this was significantly higher in NEC compared to NET. Expression of PD1, PD-L1, and T cell immunoglobulin and mucin domain-containing protein 3 (TIM3) on immune cells was low or nearly absent. mRNA immunoprofiling revealed low expression of IFNγ inducible genes in NET and NEC without any spatial heterogeneity. However, we observed an increased mRNA expression of chemokines, which attract myeloid cells in NET and NEC, and a high abundance of genes related to immunosuppressive myeloid cells and genes with immunosuppressive functions like CD47 and CD74. In conclusion, NET and NEC lack signs of an activation of the adaptive immune system, but rather show abundance of several immunosuppressive genes that represent potential targets for immunomodulation.


Evaluation of systemic chemotherapy with magnetic liposomal doxorubicin and a dipole external electromagnet.
The development of an active drug delivery system is an attractive approach to increase the targetability of anticancer agents. In the present study, we examined the efficiency of systemic chemotherapy with small magnetic liposomes containing doxorubicin (magnetic DOX liposomes) and an externally applied electromagnetic force in osteosarcoma-bearing hamsters. Syrian male hamsters inoculated with osteosarcoma, OS515, in the limb were studied 7 days after inoculation. The efficiency of this system was evaluated by measuring the tissue distribution and tumor-suppressing effects of DOX on primary tumor growth and lung metastases. A DC dipole electromagnet was used, and the hamster's tumor-bearing limb was placed between 2 poles after the i.v. administration of liposomes. The dose of DOX and the magnetic field strength were fixed at 5 mg/kg and 0.4 T, respectively. Administration of magnetic DOX liposomes followed by 60 min application of magnetic field produced a 3- to 4-fold higher maximum DOX concentration in the tumor. This newly designed systemic chemotherapy significantly suppressed primary tumor growth for at least 2 weeks, though other DOX treatments also suppressed compared to control. Histologic examination confirmed a greater antitumor effect of this systemic chemotherapy compared to standard methods. In addition, this approach significantly suppressed lung metastases measured at 3 weeks posttreatment. These results suggest that this systemic chemotherapy can effectively reduce primary tumor growth and suppress lung metastasis due to increased targeting of DOX. Such targeted drug delivery for anticancer agents would provide clinical advantages compared to current methods.


Lessons from case studies of metals: investigating exposure, bioavailability, and risk.
Since metals are widely distributed in environmental matrices, humans are exposed to them by either anthropogenic activities or inadvertently by necessity. Selected metals: arsenic, mercury, iron, tin, lead and chromium were chosen on the basis of their economic and public health importance to illustrate the diversity of exposure pathways and differences in factors governing bioavailability. Bioavailability is central to the toxicity of metals and this is discussed from the health risk paradigm standpoint of risk assessment and risk management and the impacts of including or excluding bioavailability in such assessment. The lessons from the case studies of arsenic, mercury, tin and chromium provide a unified concept of methods that can be used in investigating and controlling outbreaks due to metal poisoning in other similar situations.


Application of enhanced membrane bioreactor (eMBR) for the reuse of carwash wastewater.
Minimising the usage of potable water in industrial and cleaning processes is essential to conserve fresh water. Recycling treated wastewater will help to do so. However, high quality treated wastewater is required for reuse and recycling. This study evaluated the performance of an enhanced membrane bioreactor (eMBR) in treating car wash wastewater for the purpose of reuse. The eMBR consisted of an anaerobic tank, an anoxic tank, an aerobic membrane bioreactor (AMBR) and a UV disinfection unit. The effects of hydraulic retention time of the eMBR on the treated water quality parameters and operating parameters were evaluated. The eMBR produced high quality recyclable water (0.5-10.2 mg/L of COD, 0.18-0.83 NTU of turbidity, 0 org. of E. Coli/100 mL) meeting Class A recycle water standards. Decrease in the mixed liquor suspended solids concentration in the AMBR (from 294 to 117 mg/L) reduced the fouling of the membrane which increased the permeate flux (from 5.9 to 6.7 L/m2h). This is unique to the eMBR system used in this study. However, when the flux exceeded the critical flux, the trans-membrane pressure increased significantly.


Modulating molecular chaperone Hsp90 functions through reversible acetylation.
The molecular chaperone protein Hsp90 is a key regulator of approximately 100 'client' proteins crucial for numerous cell signaling processes. Consequently, understanding the molecular underpinnings that regulate Hsp90 activity is an important biological endeavor. Exciting new results now suggest that, at least for nuclear receptor activity, Hsp90 function is directly regulated by histone deacetylase 6 (HDAC6). These observations have consequences for various biological processes and potentially important implications for the development of cancer therapeutics.


End-to-end test for lung SBRT: An Italian multicentric pilot experience.
Set up a lung SBRT end-to-end (e2e) test and perform a multicentre validation. A group of medical physicists from four hospitals and the Italian Institute of Ionizing Radiation Metrology designed the present e2e test. One sub-group set up the test, while another tested its feasibility and ease of use. A satisfaction questionnaire was used to collect user feedback. Each participating centre (PC) received the ADAM breathing phantom, a microDiamond detector and radiochromic films. Following the e2e protocol, each PC performed its standard internal procedure for simulating, planning, and irradiating the phantom. Each PC uploaded its planning and treatment delivery data in a shared Google Drive. A single centre analyzed all the data. The e2e test was successfully performed by all PCs. Participants' comments indicated that ADAM was well suited to the purpose and the protocol well described. All PCs performed the test in static and dynamic modes. The ratio between measured and planned point dose obtained by PC1, PC2, PC3, PC4 was: 0.99, 0.96, 1.01 and 1.01 (static track) and 0.99, 1.02, 1.01 and 0.94 (dynamic track). The gamma passing rates (3 % global, 3 mm) between planned and measured dose maps were 98.5 %, 94.0 %, 99.1 % and 94.0 % (static track) and 99.5 %, 96.5 %, 86.0 % and 94.5 % (dynamic track) for PC1, PC2, PC3 and PC4, respectively. An e2e test for lung SBRT has been proposed and tested in a multicentre framework. The results and user feedback prove the validity of the proposed e2e test.


Insomnia: Personalized Diagnosis and Treatment Options.
Chronic insomnia is a clinical diagnosis fulfilled by criteria: (a) difficulty initiating or maintaining sleep, (b) inability to sleep despite having adequate opportunities, (c) having negative daytime effects due to lack of sleep, and (d) sleep difficulty not explained by other disorder-with symptoms at least three times per week during a period of 3 months. Cognitive behavioral therapy is considered a first-line treatment but can be supported with pharmacologic or digital therapeutics. When developing a patient's care plan, we should consider a "personomics" approach in which we personalize care plans as a form of sleep precision medicine.


A Novel Phosphoinositide-3-kinase Adapter Protein 1 Gene Missense Mutation in Familial Cirrhosis.
Familial cirrhosis is a condition that is associated with the presence of liver disease with genetic linkage among multiple family members in a generation or in multiple generations. With cirrhosis, most of these disease pathogeneses are related to a defect of an enzyme/transport protein leading to a deranged metabolic pathway with variable prevalence. Many studies and high-quality metanalyses have shed light on genetic linkage associated with nonalcoholic fatty liver disease and steatohepatitis such as the PNPLA3, MBOAT7, and TM6SF2 variants. In this report, we shed light on a novel missense mutation associated with cirrhosis in a family of brothers associated with phosphoinositide-3-kinase adapter protein 1 gene through high-output whole exosome gene sequencing methodology.


Design, synthesis fusidic acid derivatives alleviate acute lung injury via inhibiting MAPK/NF-κB/NLRP3 pathway.
Acute lung injury (ALI) refers to a series of lung lesions resulting from multiple lung injuries, even leading to morbidity and death, abundant previous reports have showed that anti-inflammatory as a key to treatment of ALI. Fusidic acid (FA) as an antibiotic has significant anti-bacterial activity and anti-inflammatory effects. In this study, we designed and synthesized 34 FA derivatives to identify new anti-inflammatory drugs. The anti-inflammatory activities of the derivatives were screened using lipopolysaccharide (LPS)-induced RAW264.7 cells to evaluate the anti-inflammatory activity of the compounds, we measured nitric oxide (NO) and interleukin-6 (IL-6). Most of compounds showed inhibitory effects on inflammatory NO and IL-6 in LPS-induced RAW264.7 cells. Based on the screening results, compound a1 showed the strongest anti-inflammatory activity. Compared with FA, the inhibition rate NO and IL-6 of compound a1 increased 3.08 and 2.09 times at 10 μM, respectively. We further measured a1 inhibited inflammatory factor NO (IC50 = 3.26 ± 0.42 μM), IL-6 (IC50 = 1.85 ± 0.21 μM) and TNF-α (IC50 = 3.88 ± 0.55 μM). We also demonstrated that a1 markedly inhibits the expression of certain immune-related cytotoxic factors, including cyclooxygenase-2 (COX-2) and inducible nitric-oxide synthase (iNOS). In vivo results indicate that a1 can reduce lung inflammation and NO, IL-6, TNF-α, COX-2 and iNOS in LPS-induced ALI mice. On the one hand, we demonstrated a1 inhibits the mitogen-activated protein kinase (MAPK) signaling pathway by down-regulating the phosphorylation of p38 MAPK, c-Jun N-terminal kinase (c-JNK) and extracellular signal-regulated kinase (ERK). Moreover, a1 also suppressing the phosphorylation of inhibitory NF-κB inhibitor α (IκBα) inhibits the activation of the nuclear factor-κB (NF-κB) signaling pathway. On the other hand, we demonstrated a1 also role in anti-inflammatory by inhibits nucleotide-binding domain (NOD)-like receptor protein 3 (NLRP3) inflammasome and further inhibits Caspase-1 and inflammatory factor interleukin-1β (IL-1β). In conclusion, our study demonstrates that a1 has an anti-inflammatory effect and alleviates ALI by regulating inflammatory mediators and suppressing the MAPK, NF-κB and NLRP3 inflammasome signaling pathways.


[Properties of bilayer lipid membranes formed from cholesterol and alkyltrimethylammonium bromides].
Flat bilayer lipid membranes (BLM) were obtained from cholesterol solution in n-octane in the presence of detergents of alkyltrimethyl ammonium bromides series in water medium. It has been shown that highly stable BLM are obtained only for the detergents of this series with the alkyl chain higher than C12. Changes in stability and electromechanical strength of the latter are conditioned by the peculiarities of stechiometric interaction between the molecules of cholesterol, detergent and solvent in the lipid bilayer.


The challenge of simultaneous major aortic surgery and pectus excavatum repair in a patient with Ehlers-Danlos type IV syndrome.
A unique case of a patient with Ehlers-Danlos type IV syndrome was referred with aortic root aneurysm and previous cosmetic primary repair of pectus excavatum (PE) through a solid silicone onlay prosthesis. The patient underwent successful removal of the silicone prosthesis and one-stage ascending aortic root replacement with concomitant Nuss procedure for repair of the PE.


Unmet needs: Research helps regulators do their jobs.
A plethora of innovative new medical products along with the need to apply modern technologies to medical-product evaluation has spurred seminal opportunities in regulatory sciences. Here, we provide eight examples of regulatory science research for diverse products. Opportunities abound, particularly in data science and precision health.


Inspiratory muscle training in patients with heart failure: A systematic review and meta-analysis.
To explore the effect of inspiratory muscle training (IMT) on patients with heart failure and further explore the impact of IMT on patients with heart failure with preserved ejection fraction. PubMed, EMBASE, Cochrane Library, CNKI, Wanfang and VIP databases were systematically searched. Randomized controlled trials of inspiratory muscle training in patients with heart failure were included. Revman 5.3 software was used to calculate the weighted mean difference (MD) of the combined effect size. The effects of IMT on the maximum oxygen uptake (peakVO2), maximum inspiratory pressure (PI max), ventilation efficiency (V E /VCO 2), six-minute walking distance (6MWD), forced expiratory volume (FEV1), forced vital capacity (FVC) and quality of life in patients with heart failure were compared and analyzed. After systematic retrieval and screening, 17 studies were included in this study, and the quality of the included studies was good. The results showed that IMT could increase peakVO2 (MD 2.53; 95% CI 1. 54, 3. 51; P < 0.0001) and PI max (MD 17.25; 95% CI 13. 75, 20. 75; P < 0.00001); improve the VE/VCO2 (MD -4.22; 95% CI -6.78, -1.66; P = 0.001) and significantly improve the quality of life in patients with heart failure (MD -13.34; 95% CI -20.42, -6.26; P = 0.0002). However, the effect of IMT on 6MWD in patients with heart failure was not statistically significant (MD 74.45; 95% CI -12.88,161.79; P = 0.09), and the effect on lung function (FEV1 and FVC) was also not statistically significant (P = 0.08; P = 0.86). IMT had a more significant positive effect on peakVO2 (MD 2.98; 95% CI 1.63, 4.34; P < 0.0001) and quality of life (MD -14.52; 95% CI -18.53, -10.52; P < 0.00001) in patients with heart failure with preserved ejection fraction. Descriptive analysis suggested that IMT may positively affect dyspnoea in patients with heart failure. In addition, the choice of evaluation scale may affect the evaluation results of quality of life and dyspnoea. IMT has a significant positive effect on respiratory status in patients with heart failure, but different dyspnoea and quality of life evaluation scales can affect the final evaluation results.


The outer membrane of plant mitochondria contains a calcium-dependent protein kinase and multiple phosphoproteins.
Highly purified mitochondria from potato (Solanum tuberosum L. cv. Bintje) tubers were subfractionated into a matrix fraction, an inner membrane fraction and an outer membrane fraction with minimal cross-contamination. When the matrix and inner membrane fractions were incubated with [gamma-32P]ATP only one and three prominent phosphoproteins were detected after SDS-PAGE and autoradiography, respectively. In contrast, more than 20 phosphoproteins could be labelled in the outer membrane fraction, the main ones at 12, 18, 26, 43, 58, 60, 65, 74 and 110 kDa. Only one band, at 18 kDa, was detectable when the labelling was done in the presence of EGTA. We conclude that the outer membrane of plant mitochondria contains at least one Ca(2+)-dependent protein kinase and more than 20 endogenous substrates.


Estimating daily PM2.5 and PM10 across the complex geo-climate region of Israel using MAIAC satellite-based AOD data.
Estimates of exposure to PM2.5 are often derived from geographic characteristics based on land-use regression or from a limited number of fixed ground monitors. Remote sensing advances have integrated these approaches with satellite-based measures of aerosol optical depth (AOD), which is spatially and temporally resolved, allowing greater coverage for PM2.5 estimations. Israel is situated in a complex geo-climatic region with contrasting geographic and weather patterns, including both dark and bright surfaces within a relatively small area. Our goal was to examine the use of MODIS-based MAIAC data in Israel, and to explore the reliability of predicted PM2.5 and PM10 at a high spatiotemporal resolution. We applied a three stage process, including a daily calibration method based on a mixed effects model, to predict ground PM2.5 and PM10 over Israel. We later constructed daily predictions across Israel for 2003-2013 using spatial and temporal smoothing, to estimate AOD when satellite data were missing. Good model performance was achieved, with out-of-sample cross validation R2 values of 0.79 and 0.72 for PM10 and PM2.5, respectively. Model predictions had little bias, with cross-validated slopes (predicted vs. observed) of 0.99 for both the PM2.5 and PM10 models. To our knowledge, this is the first study that utilizes high resolution 1km MAIAC AOD retrievals for PM prediction while accounting for geo-climate complexities, such as experienced in Israel. This novel model allowed the reconstruction of long- and short-term spatially resolved exposure to PM2.5 and PM10 in Israel, which could be used in the future for epidemiological studies.


Self-Organizing Human Induced Pluripotent Stem Cell Hepatocyte 3D Organoids Inform the Biology of the Pleiotropic TRIB1 Gene.
Establishment of a physiologically relevant human hepatocyte-like cell system for in vitro translational research has been hampered by the limited availability of cell models that accurately reflect human biology and the pathophysiology of human disease. Here we report a robust, reproducible, and scalable protocol for the generation of hepatic organoids from human induced pluripotent stem cells (hiPSCs) using short exposure to nonengineered matrices. These hepatic organoids follow defined stages of hepatic development and express higher levels of early (hepatocyte nuclear factor 4A [HNF4A], prospero-related homeobox 1 [PROX1]) and mature hepatic and metabolic markers (albumin, asialoglycoprotein receptor 1 [ASGR1], CCAAT/enhancer binding protein α [C/EBPα]) than two-dimensional (2D) hepatocyte-like cells (HLCs) at day 20 of differentiation. We used this model to explore the biology of the pleiotropic TRIB1 (Tribbles-1) gene associated with a number of metabolic traits, including nonalcoholic fatty liver disease and plasma lipids. We used genome editing to delete the TRIB1 gene in hiPSCs and compared TRIB1-deleted iPSC-HLCs to isogenic iPSC-HLCs under both 2D culture and three-dimensional (3D) organoid conditions. Under conventional 2D culture conditions, TRIB1-deficient HLCs showed maturation defects, with decreased expression of late-stage hepatic and lipogenesis markers. In contrast, when cultured as 3D hepatic organoids, the differentiation defects were rescued, and a clear lipid-related phenotype was noted in the TRIB1-deficient induced pluripotent stem cell HLCs. Conclusion: This work supports the potential of genome-edited hiPSC-derived hepatic 3D organoids in exploring human hepatocyte biology, including the functional interrogation of genes identified through human genetic investigation.


NO, hydrogen sulfide does not come first during tomato response to high salinity.
High salinity greatly impacts agriculture, particularly in tomato (Solanum lycopersicum), a crop that is a model to study this abiotic stress. This work investigated whether hydrogen sulfide (H2S) acts upstream or downstream of nitric oxide (NO) in the signaling cascade during tomato response to salt stress. An NO-donor incremented H2S levels by 12-18.9% while an H2S-donor yielded 10% more NO in roots. The NO accumulated in roots one-hour after NaCl treatment while H2S accumulation started two-hour later. The NO stimulated H2S accumulation in roots/leaves, but not the opposite (i.e H2S was unable to stimulate NO accumulation) two-hour post NaCl treatment. Also, NO accumulation was accompanied by an increment of transcript levels of genes that encode for H2S-synthesizing enzymes. Our results indicate that H2S acts downstream of NO in the mitigation of oxidative stress, which helps tomato plants to tolerate high salinity.


Evaluation of different magnetic resonance imaging techniques for the assessment of active left atrial emptying.
There is currently no agreement on the best method of assessing active left atrial (LA) emptying. This study evaluated the relative merits of cine- and velocity encoded (VENC) magnetic resonance imaging (MRI) for the assessment of active LA emptying. Total LA emptying volume (TLAEV) and active LA stroke volume (ALASV) were assessed in 107 consecutive patients using cine-MRI and transmitral flow measurements by VENC-MRI. The fraction of active LA emptying (ALAEF) was calculated as the ratio of ALASV to TLAEV. LA and left ventricular (LV) output were calculated by multiplying TLAEV and LV stroke volume by heart rate, respectively. Intra- and inter-observer variances were significantly larger for cine-MRI than for VENC-MRI measurements of ALASV (24.7 mL(2) vs. 3.7 mL(2) and 57.7 mL(2) vs. 4.2 mL(2); P < 0.0001). Biplane cine-MRI underestimated TLAEV (mean difference -57 ± 32 %; P < 0.0001) and ALASV (mean difference -24 ± 51 %; P < 0.0001) but overestimated ALAEF (mean difference 31 ± 54 %, P < 0.0001) compared with VENC-MRI. There was significantly better agreement between LV output and LA output measured by VENC-MRI compared with LA output measured by cine-MRI (mean difference 0.30 ± 1.12 L/min vs. -2.05 ± 1.44 L/min; P < 0.0001). VENC-MRI is the more appropriate method of assessing active LA emptying and its use should be favoured.


Correction to: Treatment of Open-Angle Glaucoma and Ocular Hypertension with Preservative-Free Tafluprost/Timolol Fixed-Dose Combination Therapy: The VISIONARY Study.
In the original article, the The VISIONARY Study Group Principal Investigator Jose Javier Garcia-Medina.