Expression of glutathione and gamma-glutamylcysteine synthetase mRNA is Jun dependent.
The gene GLCLC encodes the catalytic subunit of gamma-glutamylcysteine synthetase (glutamate-cysteine ligase E.C. 188.8.131.52), the rate limiting enzyme for glutathione synthesis. When HepG2 cells were exposed to the serine/threonine phosphatase inhibitor okadaic acid (OA), increased expression of GLCLC was observed, as was the development of resistance to xenobiotic induced GSH depletion. Okadaic acid is known to activate both NF-kappaB and AP-1 activity. Inhibition of NF-kappaB activity by overexpression of an IkappaB alpha transdominant inhibitor or exposure to the protease inhibitor TLCK did not inhibit the OA mediated increase in GLCLC transcripts. Fibroblasts derived from a mouse containing a c-Jun null mutation exhibited diminished AP-1 binding activity, reduced levels of GLCLC message, and a correspondingly low GSH concentration compared to wild type cells. When the null cells, which express Jun B and Jun D, were exposed to OA, AP-1 binding activity increased, as did expression of GLCLC message. These results indicate that AP-1 transcription factors participate in the regulation of glutathione metabolism.


Quality of life, use of topical medications and socio-economic data in hand eczema: a Swedish nationwide survey.
Hand eczema is common and has an adverse impact on the lives of patients. There is a need for population-based surveys on the pharmacoepidemiological aspects, quality of life and impact of socioeconomic factors in hand eczema. The aim of this cross-sectional study was to investigate these factors. A questionnaire-based nationwide survey of health was performed, including questions on hand eczema, use of pharmaceuticals and socioeconomic factors. Quality of life was estimated with the generic instrument Short Form 36 (SF-36). The questionnaire was sent to 7,985 persons (age range 18-84 years), response rate 61.1% (n = 4,875). The 1-year prevalence of hand eczema in the study population was 7.5%. In this group, quality of life was lower. All dimensions of SF-36 were affected, most markedly general health and those dimensions reporting on mental health. In the group with self-reported hand eczema, 51% reported using topical pharmaceuticals. Hand eczema was more common among women (9.1%, n = 2,630) than among men (5.6%, n = 2,245) and in the age group below 65 years (8.5%, n = 3,274) compared with those aged 65 years and over (4.3%, n = 1,151). This survey clearly demonstrates the impact of hand eczema on several dimensions of life and also highlights age, gender and socioeconomic differences.


Identifying Heart Failure Symptoms and Poor Self-Management in Home Healthcare: A Natural Language Processing Study.
The goal of this natural language processing (NLP) study was to identify patients in home healthcare with heart failure symptoms and poor self-management (SM). The preliminary lists of symptoms and poor SM status were identified, NLP algorithms were used to refine the lists, and NLP performance was evaluated using 2.3 million home healthcare clinical notes. The overall precision to identify patients with heart failure symptoms and poor SM status was 0.86. The feasibility of methods was demonstrated to identify patients with heart failure symptoms and poor SM documented in home healthcare notes. This study facilitates utilizing key symptom information and patients' SM status from unstructured data in electronic health records. The results of this study can be applied to better individualize symptom management to support heart failure patients' quality-of-life.


[Comparative data on the tick-borne encephalitis virus infectiousness of hungry and satiated taiga ticks (based on the results of an immunoenzyme analysis)].
The results of individual investigation of 25,500 Ixodes persulcatus ticks from east Siberia are presented. The ticks were collected from grass, men, and animals before their sticking and after feeding at different intervals. The quantity of positive specimens was 6-11 times higher among the fed ticks hungry ones and averaged 10.36 and 1.85%, respectively, at the first stage due to the higher aggression of the infected females at the first stage and due to viral replication when the tick had feed and its better recognition as the titers increased. The proportion of ticks having high levels of tick-borne encephalitis virus antigen among the fed ticks was also considerably higher. The content of virus antigen increased in proportion with the duration of feeding. There is a moderate correlation (r = 0.59) of the infection index of hungry and fed ticks in different years and in various areas. The findings suggest that the fed ticks should be used as an additional marker for the features of tick-borne encephalitis virus circulation.


Technoeconomic data adopted for the development of a long-term electricity supply model for the Hashmite Kingdome of Jordan.
Electrical generation in Jordan currently relies on imported fossil fuels. In the past, most imported fossil fuels were subsidised by neighbouring countries through grants and aid. This has led to a regulated market, with subsidised low-cost electrical energy consumers, and the government being the sole buyer and seller of electricity. With the ageing of the national electrical infrastructure, political instability in the region, and lack of funds for direct investment, other options needed to be pursued. Long term Power Purchase Agreements (PPA) were granted to Independent Power Producers (IPP) to encourage investment in capacity and infrastructure. In addition, long-term fuel contracts were signed to secure steady flow of primary fuel sources. Over the past few years, renewable energy penetration has increased rapidly, but without proper planning or taking into consideration long term PPA and fuel contracts. Data in regard to the current infrastructure, renewable energy technology, signed energy commitments and system operation assumptions are described in this article, which may be used for modelling and analysis. The Data were collected from annual reports from the different energy related entities in Jordan.


Mechanically Guided Hierarchical Assembly of 3D Mesostructures.
3D, hierarchical micro/nanostructures formed with advanced functional materials are of growing interest due to their broad potential utility in electronics, robotics, battery technology, and biomedical engineering. Among various strategies in 3D micro/nanofabrication, a set of methods based on compressive buckling offers wide-ranging material compatibility, fabrication scalability, and precise process control. Previously reports on this type of approach rely on a single, planar prestretched elastomeric platform to transform thin-film precursors with 2D layouts into 3D architectures. The simple planar configuration of bonding sites between these precursors and their assembly substrates prevents the realization of certain types of complex 3D geometries. In this paper, a set of hierarchical assembly concepts is reported that leverage multiple layers of prestretched elastomeric substrates to induce not only compressive buckling of 2D precursors bonded to them but also of themselves, thereby creating 3D mesostructures mounted at multiple levels of 3D frameworks with complex, elaborate configurations. Control over strains used in these processes provides reversible access to multiple different 3D layouts in a given structure. Examples to demonstrate these ideas through both experimental and computational results span vertically aligned helices to closed 3D cages, selected for their relevance to 3D conformal bio-interfaces and multifunctional microsystems.


How has the brain disease model of addiction contributed to tobacco control?
Our paper evaluates the extent to which the brain disease model of addiction (BDMA) has contributed to reducing the prevalence of tobacco smoking and tobacco-related harm over the past 20 years. We discuss the ways in which genetic and neuroscience research on nicotine addiction have contributed to our understanding of tobacco smoking. We then examine the extent to which the BDMA has produced more effective treatments to assist smoking cessation. We also assess the degree to which the BDMA has contributed to the tobacco control policies that have produced substantial reductions in tobacco-related morbidity and mortality in the two decades since the model was first proposed by Alan Leshner. We also assess whether the BDMA has reduced the stigmatisation of people who smoke tobacco.


Impact of the c-MybE308G mutation on mouse myelopoiesis and dendritic cell development.
Booreana mice carrying the c-Myb308G point mutation were analyzed to determine changes in early hematopoiesis in the bone marrow and among mature cells in the periphery. This point mutation led to increased numbers of early hematopoietic stem and progenitor cells (HSPCs), with a subsequent reduction in the development of B cells, erythroid cells, and neutrophils, and increased numbers of myeloid cells and granulocytes. Myelopoiesis was further investigated by way of particular subsets affected. A specific question addressed whether booreana mice contained increased numbers of dendritic-like cells (L-DC subset) recently identified in the spleen, since L-DCs arise in vitro by direct differentiation from HSPCs co-cultured over splenic stroma. The non-lethal c-Myb mutation in booreana mice was associated with significantly lower representation of splenic CD8- conventional dendritic cells (cDCs), inflammatory monocytes, and neutrophils compared to wild-type mice. This result confirmed the bone marrow origin of progenitors for these subsets since c-Myb is essential for their development. Production of L-DCs and resident monocytes was not affected by the c-MybE308G mutation. These subsets may derive from different progenitors than those in bone marrow, and are potentially established in the spleen during embryogenesis. An alternative explanation may be needed for why there was no change in CD8+ cDCs in booreana spleen since these cells are known to derive from common dendritic progenitors in bone marrow.


Association between first oral examination characteristics and dental treatment needs in privately insured children: A claims data analysis.
Early childhood caries (ECC) remains the most common, preventable infectious disease among children in the United States. Screening is recommended after the eruption of the first tooth, but it is unclear how the age at first dental examination is associated with eventual restorative treatment needs. The authors of this study sought to determine how provider type and age at first dental examination are associated longitudinally with caries experience among children in the United States. Deidentified claims data were included for 706,636 privately insured children aged 0 through 6 years as part of the nationwide IBM Watson Health Market Scan (2012-2017). The authors used Kaplan-Meier survival analysis to describe the association between the age of first visit and restorative treatment needs. A total of 21% of this population required restorative treatment, and the average age at first dental examination was 3.6 years. A multivariable Cox proportional hazards model showed increased hazard for restorative treatment with age at first dental visit at 3 years (hazard ratio, 2.05; 95% CI, 1.97 to 2.13) and 4 years (hazard ratio, 3.99; 95% CI, 3.84 to 4.16). The high proportion of children requiring restorative treatment and late age at first dental screening show needed investments in educating general dentists, medical students, and pediatricians about oral health guidelines for pediatric patients. Communicating the importance of children establishing a dental home by age 1 year to parents and health care professionals may help reduce disease burden in children younger than 6 years.


Engineering Azobenzene Derivatives to Control the Photoisomerization Process.
In this work, we show how the structural features of photoactive azobenzene derivatives can influence the photoexcited state behavior and the yield of the trans/cis photoisomerization process. By combining high-resolution transient absorption experiments in the vis-NIR region and quantum chemistry calculations (TDDFT and RASPT2), we address the origin of the transient signals of three poly-substituted push-pull azobenzenes with an increasing strength of the intramolecular interactions stabilizing the planar trans isomer (absence of intramolecular H-bonds, methyl, and traditional H-bond, respectively, for 4-diethyl-4'-nitroazobenzene, Disperse Blue 366, and Disperse Blue 165) and a commercial red dye showing keto-enol tautomerism involving the azo group (Sudan Red G). Our results indicate that the intramolecular H-bonds can act as a "molecular lock" stabilizing the trans isomer and increasing the energy barrier along the photoreactive CNNC torsion coordinate, thus preventing photoisomerization in the Disperse Blue dyes. In contrast, the involvement of the azo group in keto-enol tautomerism can be employed as a strategy to change the nature of the lower excited state and remove the nonproductive symmetric CNN/NNC bending pathway typical of the azo group, thus favoring the productive torsional motion. Taken together, our results can provide guidelines for the structural design of azobenzene-based photoswitches with a tunable excited state behavior.


Comparison of the liquisolid technique and co-milling for loading of a poorly soluble drug in inorganic porous excipients.
Drug loading into mesoporous carriers may help to improve the dissolution of poorly aqueous-soluble drugs. However, both preparation method and carrier properties influence loading efficiency and drug release. Accordingly, this study aimed to compare two preparation methods: formulation into liquisolid systems (LSS) and co-milling for their efficiency in loading the poorly soluble model drug cyclosporine A (CyA) into mesoporous magnesium aluminometasilicate Neusilin® US2 (NEU) or functionalized calcium carbonate (FCC). Scanning electron microscopy was used to visualize the morphology of the samples and evaluate the changes that occurred during the drug loading process. The solid-state characteristics and physical stability of the formulations, prepared at different drug concentrations, were determined using X-ray powder diffraction. In vitro release of the drug was evaluated in biorelevant media simulating intestinal fluid. The obtained results revealed improved drug release profiles of the formulations when compared to the milled (amorphous) CyA alone. The dissolution of CyA from LSS was faster in comparison to the co-milled formulations. Higher drug release was achieved from NEU than FCC formulations presumably due to the higher pore volume and larger surface area of NEU.


Structural relationship between alpha 1-microglobulin from man, guinea-pig, rat and rabbit.
Rabbit alpha 1-microglobulin was purified from the urine of sodium-chromate-treated animals by the use of gel chromatography on Sephadex G-100, affinity chromatography on concanavalin-A--Sepharose and ion-exchange chromatography on DEAE-Sephadex. Rabbit alpha 1-microglobulin had a molecular mass of 25.6 kDa on SDS/polyacrylamide gel electrophoresis. Alpha 1-microglobulin has previously been purified from the urine of humans, guinea-pigs and rats by similar methods, and the molecular masses of the four homologues were compared by SDS/polyacrylamide gel electrophoresis and gel chromatography in a denaturing medium. By these two methods the human homologue was 6 kDa and 3 kDa larger, respectively, than the other three proteins. Endoglycosidase F digestion of alpha 1-microglobulin, followed by SDS/polyacrylamide gel electrophoresis, revealed three protein bands in the human alpha 1-microglobulin sample, and only two bands in guinea-pig, rat and rabbit alpha 1-microglobulin, with a gap between each band of 2.6--2.9 kDa. The amino-terminal amino acid sequences of the four homologues were determined and between 72% and 81% homology was seen. The five amino-terminal amino acids present in the other species were missing in guinea-pig alpha 1-microglobulin. Our results indicate that human alpha 1-microglobulin is substituted with two N-linked oligosaccharides, while only one is attached to each of the other alpha 1-microglobulins, and that the extra glycosylamine-linked oligosaccharide in the human protein is attached to asparagine in position 17. Finally it is shown that all four homologues inhibit antigen stimulation of human lymphocytes, a finding which is consistent with our previous suggestion that the N-linked oligosaccharides carry the immunosuppressive activity of alpha 1-microglobulin.


Challenge of predicting resting energy expenditure in children undergoing surgery for congenital heart disease.
To determine pre- and postoperative predictors of energy expenditure in children with congenital heart disease requiring open heart surgery; and to compare measured resting energy expenditure with current predictive equations. Prospective resting energy expenditure data were collected, using indirect calorimetry, for ventilated children admitted consecutively to the pediatric intensive care unit after surgery for congenital heart disease. A 30-min steady-state measurement was performed in suitable patients. Resting energy expenditure was compared to pre- and postoperative clinical variables, and to predicted energy expenditure, using currently used predictive equations. Pediatric intensive care unit at the Royal Brompton Hospital, London. Children ventilated in the pediatric intensive care unit post surgery for congenital heart disease. Measurement of energy expenditure by indirect calorimetry. Twenty-one mechanically ventilated children (n = 17 boys, 4 girls) were enrolled in the study. Mean +/- sd measured resting energy expenditure was 67.8 +/- 15.4 kcal/kg/day. Most children had inadequate delivery of nutrients compared with actual requirements. Cardiopulmonary bypass had a significant influence on energy expenditure after surgery; in patients who underwent cardiopulmonary bypass during surgery, mean resting energy expenditure was 73.6 +/- 14.45 kcal/kg/day vs. 58.3 +/- 10.29 kcal/kg/day in patients undergoing nonbypass surgery. Children who were malnourished preoperatively had greater resting energy expenditure postoperatively. There was also a significant difference between measured energy expenditure and the Schofield (p = .006), World Health Organization (p = .002), and pediatric intensive care unit-specific formula (p < .0001). However, energy expenditure or a relative energy deficit in the early postoperative period was not associated with severity or duration of organ dysfunction. Poor nutritional status preoperatively and cardiopulmonary bypass were associated with a greater energy expenditure post cardiac surgery. None of the current predictive equations predicted energy requirements within acceptable clinical accuracy.


Genome-wide association study on resistance of cultivated soybean to Fusarium oxysporum root rot in Northeast China.
Fusarium oxysporum is a prevalent fungal pathogen that diminishes soybean yield through seedling disease and root rot. Preventing Fusarium oxysporum root rot (FORR) damage entails on the identification of resistance genes and developing resistant cultivars. Therefore, conducting fine mapping and marker development for FORR resistance genes is of great significance for fostering the cultivation of resistant varieties. In this study, 350 soybean germplasm accessions, mainly from Northeast China, underwent genotyping using the SoySNP50K Illumina BeadChip, which includes 52,041 single nucleotide polymorphisms (SNPs). Their resistance to FORR was assessed in a greenhouse. Genome-wide association studies utilizing the general linear model, mixed linear model, compressed mixed linear model, and settlement of MLM under progressively exclusive relationship models were conducted to identify marker-trait associations while effectively controlling for population structure. The results demonstrated that these models effectively managed population structure. Eight SNP loci significantly associated with FORR resistance in soybean were detected, primarily located on Chromosome 6. Notably, there was a strong linkage disequilibrium between the large-effect SNPs ss715595462 and ss715595463, contributing substantially to phenotypic variation. Within the genetic interval encompassing these loci, 28 genes were present, with one gene Glyma.06G088400 encoding a protein kinase family protein containing a leucine-rich repeat domain identified as a potential candidate gene in the reference genome of Williams82. Additionally, quantitative real-time reverse transcription polymerase chain reaction analysis evaluated the gene expression levels between highly resistant and susceptible accessions, focusing on primary root tissues collected at different time points after F. oxysporum inoculation. Among the examined genes, only this gene emerged as the strongest candidate associated with FORR resistance. The identification of this candidate gene Glyma.06G088400 improves our understanding of soybean resistance to FORR and the markers strongly linked to resistance can be beneficial for molecular marker-assisted selection in breeding resistant soybean accessions against F. oxysporum.


Detection of chromosomal anomalies and c-myc gene amplification in the cribriform pattern of prostatic intraepithelial neoplasia and carcinoma by fluorescence in situ hybridization.
Grading of the cribriform pattern of prostate cancer is controversial, and the genetic changes are largely unknown. Furthermore, the pathogenetic relationship between the cribriform pattern of high-grade prostatic intraepithelial neoplasia (PIN) and cribriform carcinoma is poorly understood. We used fluorescence in situ hybridization with centromere-specific probes for chromosomes 7, 8, 10, 12, and Y and a region-specific probe for c-myc to evaluate genetic changes in matched foci of high-grade PIN (48 foci) and prostatic carcinoma (71 foci) in 25 whole-mount radical prostatectomy specimens from patients with metastatic cancer. These cases included 10 foci of cribriform PIN and 10 foci of cribriform cancer. Numeric chromosomal anomalies were found in 67 and 68% of the high-grade PIN and carcinoma foci, respectively. Extra copies of the c-myc gene were identified in 52 and 44% of the high-grade PIN and carcinoma foci, respectively. The cribriform pattern of cancer had a higher percentage of foci with gain of chromosomes 7, 12, and Y, loss of chromosome 8, and extra copies of c-myc gene than other Gleason Primary Patterns 3 and 4; there was no difference, however, for all paired comparisons of genetic changes between the cribriform pattern of cancer and Gleason Primary Pattern 5 cancer. Cribriform PIN and cribriform cancer generally exhibited similar anomalies, although the percentage of foci with gain of chromosomes 10 and 12 was higher in cribriform cancer. Our results indicate that the cribriform pattern of prostate cancer shares genetic changes with Gleason Primary Pattern 5 and that both contain more genetic changes than the cribriform pattern of PIN. These findings suggest that the cribriform pattern of prostate cancer has biologic similarity with Gleason Pattern 5 carcinoma and that the cribriform pattern of PIN is closely associated with the cribriform pattern of prostatic carcinoma.


Direct selective growth of ZnO nanowire arrays from inkjet-printed zinc acetate precursor on a heated substrate.
Inkjet printing of functional materials has drawn tremendous interest as an alternative to the conventional photolithography-based microelectronics fabrication process development. We introduce direct selective nanowire array growth by inkjet printing of Zn acetate precursor ink patterning and subsequent hydrothermal ZnO local growth without nozzle clogging problem which frequently happens in nanoparticle inkjet printing. The proposed process can directly grow ZnO nanowires in any arbitrary patterned shape, and it is basically very fast, low cost, environmentally benign, and low temperature. Therefore, Zn acetate precursor inkjet printing-based direct nanowire local growth is expected to give extremely high flexibility in nanomaterial patterning for high-performance electronics fabrication especially at the development stage. As a proof of concept of the proposed method, ZnO nanowire network-based field effect transistors and ultraviolet photo-detectors were demonstrated by direct patterned grown ZnO nanowires as active layer.


Characterization of the ceramide moieties of sphingoglycolipids from mouse brain by ESI-MS/MS: identification of ceramides containing sphingadienine.
Sphingoglycolipids (SGLs) are cell membrane constituents. As the ceramide structure influences the biological properties of the SGL, we characterized by electrospray ionization tandem mass spectrometry the molecular species of ceramides present in SGL of mouse brain. We report here for the first time the presence in mammalian brain of sphingadienine (d18:2). Sphingenine (d18:1) is present in all SGL species, in contrast to eicosasphingenine (d20:1), which is a constituent of only gangliosides. Sphingadienine is present in galactosylceramide and sulfatides. Free ceramides contain the three types of bases. Thus, there could be two separate pools of free ceramides (d18:1, d18:2 and d20:1, d18:1) as precursors of complex SGL.


Percutaneous transluminal coronary angioplasty. Six years experience.
In a six-year period (1982-1987), 248 patients were treated with 297 procedures (percutaneous transluminal coronary angioplasty, PTCA) on 282 vessels. Two hundred and fifty-nine (87.2%) of the procedures in 210 (76.7%) of the patients appeared successful angiographically. Thirty-eight procedures were unsuccessful due to failure to pass the stenosis in 18 patients, dissection or occlusion of the treated vessel in 11, and significant residual stenosis in 9 patients. Emergency operations were performed after 11 (3.7%) of the procedures. Two patients died postoperatively. Myocardial infarction was seen in 13 patients, of whom 4 developed pathologic Q-waves in their ECG. Restenosis occurred in 60 (28.6%) of the patients. In the last year of the study, the patients selected for PTCA were in a poorer state angiographically, but the results of PTCA were better, without any increase of the complication rate. Our results, which are in accordance with others, support the concept that PTCA is a relatively safe procedure with a primary success rate of almost 90 percent. However, approximately one third of the patients developed restenosis, which in most cases occurred within 3 months.


Cell wall, lignin and fatty acid-related transcriptome in soybean: Achieving gene expression patterns for bioenergy legume.
Increasing efforts to preserve environmental resources have included the development of more efficient technologies to produce energy from renewable sources such as plant biomass, notably through biofuels and cellulosic residues. The relevance of the soybean industry is due mostly to oil and protein production which, although interdependent, results from coordinated gene expression in primary metabolism. Concerning biomass and biodiesel, a comprehensive analysis of gene regulation associated with cell wall components (as polysaccharides and lignin) and fatty acid metabolism may be very useful for finding new strategies in soybean breeding for the expanding bioenergy industry. Searching the Genosoja transcriptional database for enzymes and proteins directly involved in cell wall, lignin and fatty acid metabolism provides gene expression datasets with frequency distribution and specific regulation that is shared among several cultivars and organs, and also in response to different biotic/abiotic stress treatments. These results may be useful as a starting point to depict the Genosoja database regarding gene expression directly associated with potential applications of soybean biomass and/or residues for bioenergy-producing technologies.


Diagnosis and management of cerebral malperfusion phenomena during aortic dissection repair by transesophageal Doppler echocardiographic monitoring.
Early diagnosis and surgical decision making are the key for survival in acute type A aortic dissection (AAD-A). As such, transesophageal echocardiography (TEE) is widely accepted tool in the diagnosis of AAD-A. We used TEE in 49 cases as the sole diagnostic examination of AAD-A since November 1989. It was particularly useful intraoperatively to detect cerebral malperfusion during AAD-A repair. We were able to accurately monitor the blood flow of the aortic arch by using TEE for all patients throughout the operation. Only two patients developed severe cerebral malperfusion after the distal anastomosis was finished under deep hypothermic circulatory arrest. TEE showed that the malperfusion after the bypass was re-established. In both cases the expanded false lumen blocked the true lumen. We immediately switched the perfusion cannula from the femoral artery to the ascending aortic graft to create antegrade flow. The subsequent TEE showed only the flow in the true lumen. One patient recovered without any complication while the other suffered mild, temporary neurological defects. Cerebral malperfusion is a potential catastrophic complication of AAD-A, which may exist before surgery or be caused by the operation itself. We recommend continuous intraoperative TEE to monitor aortic arch flow during these operations. This allows immediate detection of cerebral malperfusion and prompt action can be taken to prevent irreversible brain damage.


Chromosome-specific panels of tri- and tetranucleotide microsatellite markers for multiplex fluorescent detection and automated genotyping: evaluation of their utility in pathology and forensics.
A set of 391 microsatellite markers (Weber set 6), 85% of which consist of tri- and tetranucleotide repeat markers, were used to design chromosome-specific panels that allowed for a high degree of multiplexing with respect to the fragment size range and fluorophore (FAM, HEX, TET). This marker set has an average coverage of 10.5 cM, with the largest gap being 28.1 cM. The markers were divided into 49 panels, with a maximum degree of multiplexing of 15 markers per panel. The utility of the markers for analysis of DNA from blood, hair, and formalin-fixed archival tissue biopsies was evaluated with respect to amplification efficiency, product yield, and degree of preferential amplification of the shorter allele in heterozygotes. The amplification efficiency was inversely related to repeat length and amplicon length. Based on the analysis of DNA from formalin-fixed biopsies, 51 markers suitable for loss of heterozygosity (LOH) studies were identified. The utility of the marker set for genome scanning, LOH, and forensic analyses is discussed.


Stilbene Glucoside, a Putative Sleep Promoting Constituent from Polygonum multiflorum Affects Sleep Homeostasis by Affecting the Activities of Lactate Dehydrogenase and Salivary Alpha Amylase.
Chinese herbal medicine (CHM) has been used for treating insomnia for centuries. The most used CHM for insomnia was Polygonum multiflorum. However, the molecular mechanism for CHM preventing insomnia is unknown. Stilbene glucoside (THSG), an important active component of P. multiflorum, may play an important role for treating insomnia. To test the hypothesis, Kunming mice were treated with different dosages of THSG. To examine the sleep duration, a computer-controlled sleep-wake detection system was implemented. Electroencephalogram (EEG) and electromyogram (EMG) electrodes were implanted to determine sleep-wake state. RT-PCR and Western blot was used to measure the levels of lactate dehydrogenase (LDH) and saliva alpha amylase. Spearman's rank correlation coefficient was used to identify the strength of correlation between the variables. The results showed that THSG significantly prolonged the sleep time of the mice (p<0.01). THSG changed sleep profile by reducing wake and rapid eye movement (REM) period, and increasing non-REM period. RT-PCR and Western blot analysis showed that THSG could down-regulate the levels of LDH and saliva alpha amylase (p<0.05). The level of lactate and glucose was positively related with the activity of LDH and saliva alpha amylase (p<0.05), respectively. On the other hand, the activities of LDH and amylase were negatively associated with sleep duration (p<0.05). The levels of lactate and glucose affect sleep homeostasis. Thus, THSG may prevent insomnia by regulating sleep duration via LDH and salivary alpha amylase.


Structural and radiological parameters for the characterization of jawbone.
The aim of this study was to determine the Hounsfield values of selected bone sites on a computed tomography (CT) scan of the jaw and to investigate the relationship between this radiological parameter and structural parameters. A selection of 24 bone samples out of eight cadaver human jaws was made. The following parameters were measured: Hounsfield value in the jaw (HU1) determined by a first CT scan, Hounsfield value of the excised bone specimen (HU2) by a second CT scan, bone mineral density (BMD) by a dual-energy X-ray absorptiometry scan, bone volume (BV/TV) by the microfocus CT scan, first peak transmission time (T(US-1)) and first zero crossing transmission time (T(US-2)) by an ultrasound measurement and Young's modulus (E(MECH)) by a compression test. Thirteen specimens were composed of a mix of trabecular and a small amount of cortical bone, while another 11 specimens were composed of trabecular bone only. A good correlation was found between the HU value of the specimen (HU2) and BMD (rho = 0.99), BV/TV (rho = 0.97), T(US-1) (rho = -0.83), T(US-2) (rho = -0.87) and E(MECH) (rho = 0.83). For the pure trabecular bone specimens, the HU value of the excised bone specimen (HU2) was highly correlated (rho = 0.95) with the HU value of the total jaw scan (HU1). For mixed trabecular-cortical bone specimens, this relationship was weak (rho = 0.57). With the current CT scan technology, predictions of the mechanical properties of trabecular jaw bone based on Hounsfield values were only valid for jaws with a thin layer of cortical bone. For jaws with a thicker cortical layer, the prediction of the mechanical properties decreased significantly.


Membrane curvature allosterically regulates the phosphatidylinositol cycle, controlling its rate and acyl-chain composition of its lipid intermediates.
Signaling events at membranes are often mediated by membrane lipid composition or membrane physical properties. These membrane properties could act either by favoring the membrane binding of downstream effectors or by modulating their activity. Several proteins can sense/generate membrane physical curvature (i.e. shape). However, the modulation of the activity of enzymes by a membrane's shape has not yet been reported. Here, using a cell-free assay with purified diacylglycerol kinase ϵ (DGKϵ) and liposomes, we studied the activity and acyl-chain specificity of an enzyme of the phosphatidylinositol (PI) cycle, DGKϵ. By systematically varying the model membrane lipid composition and physical properties, we found that DGKϵ has low activity and lacks acyl-chain specificity in locally flat membranes, regardless of the lipid composition. On the other hand, these enzyme properties were greatly enhanced in membrane structures with a negative Gaussian curvature. We also found that this is not a consequence of preferential binding of the enzyme to those structures, but rather is due to a curvature-mediated allosteric regulation of DGKϵ activity and acyl-chain specificity. Moreover, in a fine-tuned interplay between the enzyme and the membrane, DGKϵ favored the formation of structures with greater Gaussian curvature. DGKϵ does not bear a regulatory domain, and these findings reveal the importance of membrane curvature in regulating DGKϵ activity and acyl-chain specificity. Hence, this study highlights that a hierarchic coupling of membrane physical property and lipid composition synergistically regulates membrane signaling events. We propose that this regulatory mechanism of membrane-associated enzyme activity is likely more common than is currently appreciated.


A portable device using a closed chamber system for measuring transepidermal water loss: comparison with the conventional method.
The measurement of transepidermal water loss (TEWL) is frequently used to evaluate the barrier function of the stratum corneum (SC) of diseased skin in vivo, as well as normal skin damaged by external injurious agents. For this purpose, most people use biophysical instruments, based on an open chamber system, measuring the water vapor pressure gradient produced above the skin surface by vapor passing through the SC barrier. Recently, we tested a new portable device (Model H4300 NIKKISO-YSI CO., LTD., Tokyo, Japan) based on a closed chamber system. Its measuring principle is based on a computed TEWL estimation, from the progressive increase in relative humidity inside a closed chamber, due to continuous water evaporation from the skin surface on which the chamber is placed. We conducted an in vivo comparative study between the new portable device and a conventional method, using the DermaLab (Cortex Technology, Hadsund, Denmark), on both healthy and diseased skin surfaces, under well-controlled, standardized, experimental conditions (21 degrees C and 50% relative humidity). We found that the sensitivity of the new device was similar to that of the DermaLab. For example, the poor SC barrier function of normal facial skin was clearly demonstrated by significantly higher TEWL values than those of the extremities (P<0.0001). The compromised barrier function of the lesional skin of atopic dermatitis and psoriasis was also demonstrated in a severity-dependent fashion. However, the TEWL values measured with the new device were much lower than those obtained with the DermaLab, probably reflecting differences in the methodological principles. Nevertheless, they correlated well with those of the latter method (P<0.001). For the instruments compared in the present study, we found the following equation useful for conversion: [corresponding value obtained with the DermaLab]= 1.42 x [the recorded value with Nikkiso-YSI Model H4300] + 1.892. We consider this closed chamber system practical because it can be carried easily and its portability enables TEWL measurements to be taken in any location, without ensuring a special environment against air turbulence. Moreover, it is far less expensive than other devices currently available.


The relationship between childhood BMI and adult serum cholesterol, LDL, and ankle brachial index.
Effects of childhood overweight may persist into adulthood. We assessed the effect of childhood overweight on cardiovascular disease high risk factor levels in the same participants as adults, after controlling for adult body mass index (BMI) status. A subset of participants in an observational study (Heartwatch) were contacted approximately 26-27 years after initial enrollment to participate in a follow-up study on the long-term effects of childhood overweight. During follow-up, BMI, waist:hip circumference (WHC), blood pressure (BP), serum lipids, and ankle brachial index (ABI) were measured; additional BMI measures throughout childhood were obtained as available from the electronic medical record. Primary outcomes were ABI and serum low density lipoprotein (LDL). The 1982 Heartwatch study was conducted with children participants living in Marshfield, Wisconsin; follow-up included original participants who were re-contacted and agreed to be enrolled. Participants were a stratified random sample of eligible participants in the original 1982 Heartwatch study. Of the original 3106 participants, 647 adult participants completed follow-up exams. Among males with 1982 BMI ≥ 85(th) percentile, adult BMI, WHC, (both P ≤ 0.001), ABI (P = 0.001), total cholesterol (P = 0.01), LDL (P = 0.003) and BP (P < 0.02) were higher in 2008-2009 as compared to males with 1982 BMI < 85(th) percentile. Among females, BMI, BP and WHC (all P < 0.001) were higher in 2008-2009. BMI in 1982 and 2008-2009 were correlated [r = 0.56 (males); 0.58 (females), P < 0.001]. 2008-2009 BMI was more strongly correlated with 2008-2009 measures of ABI (r = 0.16, P = 0.006, males) and high LDL [r = 0.18, P = 0.002 (males); r = 0.11, P = 0.046 (females)]. 1982 BMI was not independently associated with ABI or LDL after adjusting for adult BMI. In a cohort studying childhood and adult overweight, childhood BMI was associated with health outcomes relating to cardiovascular disease in adulthood. However, childhood BMI was not independently related to LDL-C or ABI levels in adulthood after accounting for adult BMI. Longitudinal measurements of BMI and other health risk factors were not found to improve accuracy of models for high cardiovascular disease risk factor levels.


Vitamin and mineral supplementation for preventing dementia or delaying cognitive decline in people with mild cognitive impairment.
Vitamins and minerals have many functions in the nervous system which are important for brain health. It has been suggested that various different vitamin and mineral supplements might be useful in maintaining cognitive function and delaying the onset of dementia. In this review, we sought to examine the evidence for this in people who already had mild cognitive impairment (MCI). To evaluate the effects of vitamin and mineral supplementation on cognitive function and the incidence of dementia in people with mild cognitive impairment. We searched ALOIS, the Cochrane Dementia and Cognitive Improvement Group's (CDCIG) specialised register, as well as MEDLINE, Embase, PsycINFO, CENTRAL, CINAHL, LILACs, Web of Science Core Collection, ClinicalTrials.gov, and the WHO Portal/ICTRP, from inception to 25 January 2018. We included randomised or quasi-randomised, placebo-controlled trials which evaluated orally administered vitamin or mineral supplements in participants with a diagnosis of mild cognitive impairment and which assessed the incidence of dementia or cognitive outcomes, or both. We were interested in studies applicable to the general population of older people and therefore excluded studies in which participants had severe vitamin or mineral deficiencies. We sought data on our primary outcomes of dementia incidence and overall cognitive function and on secondary outcomes of episodic memory, executive function, speed of processing, quality of life, functional performance, clinical global impression, adverse events, and mortality. We conducted data collection and analysis according to standard Cochrane systematic review methods. We assessed the risk of bias of included studies using the Cochrane 'Risk of bias' assessment tool. We grouped vitamins and minerals according to their putative mechanism of action and, where we considered it to be clinically appropriate, we pooled data using random-effects methods. We used GRADE methods to assess the overall quality of evidence for each comparison and outcome. We included five trials with 879 participants which investigated B vitamin supplements. In four trials, the intervention was a combination of vitamins B6, B12, and folic acid; in one, it was folic acid only. Doses varied. We considered there to be some risks of performance and attrition bias and of selective outcome reporting among these trials. Our primary efficacy outcomes were the incidence of dementia and scores on measures of overall cognitive function. None of the trials reported the incidence of dementia and the evidence on overall cognitive function was of very low-quality. There was probably little or no effect of B vitamins taken for six to 24 months on episodic memory, executive function, speed of processing, or quality of life. The evidence on our other secondary clinical outcomes, including harms, was very sparse or very low-quality. There was evidence from one study that there may be a slower rate of brain atrophy over two years in participants taking B vitamins. The same study reported subgroup analyses based on the level of serum homocysteine (tHcy) at baseline and found evidence that B vitamins may improve episodic memory in those with tHcy above the median at baseline.We included one trial (n = 516) of vitamin E supplementation. Vitamin E was given as 1000 IU of alpha-tocopherol twice daily. We considered this trial to be at risk of attrition and selective reporting bias. There was probably no effect of vitamin E on the probability of progression from MCI to Alzheimer's dementia over three years (HR 1.02; 95% CI 0.74 to 1.41; n = 516; 1 study, moderate-quality evidence). There was also no evidence of an effect at intermediate time points. The available data did not allow us to conduct analyses, but the authors reported no significant effect of three years of supplementation with vitamin E on overall cognitive function, episodic memory, speed of processing, clinical global impression, functional performance, adverse events, or mortality (five deaths in each group). We considered this to be low-quality evidence.We included one trial (n = 256) of combined vitamin E and vitamin C supplementation and one trial (n = 26) of supplementation with chromium picolinate. In both cases, there was a single eligible cognitive outcome, but we considered the evidence to be very low-quality and so could not be sure of any effects. The evidence on vitamin and mineral supplements as treatments for MCI is very limited. Three years of treatment with high-dose vitamin E probably does not reduce the risk of progression to dementia, but we have no data on this outcome for other supplements. Only B vitamins have been assessed in more than one RCT. There is no evidence for beneficial effects on cognition of supplementation with B vitamins for six to 24 months. Evidence from a single study of a reduced rate of brain atrophy in participants taking vitamin B and a beneficial effect of vitamin B on episodic memory in those with higher tHcy at baseline warrants attempted replication.


Mouthwashes for the control of supragingival biofilm and gingivitis in orthodontic patients: evidence-based recommendations for clinicians.
Properly performed daily mechanical biofilm control is the most important prevention strategy for periodontal diseases. However, proper mechanical biofilm control is not performed effectively by the majority of the population, mainly due to lack of motivation and of manual dexterity. Local biofilm retention factors may aggravate home oral hygiene quality. For this reason, patients wearing fixed orthodontic appliances comprise a group that may benefit from the daily use of mouthwashes. The purpose of this review was to perform a systematic search in the literature on antiseptics used to control supragingival biofilm and gingivitis in orthodontic patients. Six studies investigating the effect of chlorhexidine and 5 studies evaluating the effect of the daily use of antiseptics were found. Chlorhexidine showed better results in reducing plaque and gingivitis. However, because of its adverse effects after continuous use, it should not be indicated for long-term periods. Among the agents considered for daily use, the fixed combination of essential oils was the only one evaluated in a clinical trial, in which a comparative group presented a statistically significant clinical impact. There is no direct evidence supporting the indication of antiseptic agents for orthodontic patients other than chlorhexidine and essential oils. It can be concluded that, for patients undergoing orthodontic treatment, chlorhexidine should be considered for treating acute gingival inflammation, whereas essential oils should be indicated for long-term daily use in controlling supragingival biofilm.


Loci associated with genomic damage levels in chronic kidney disease patients and controls.
Chronic kidney disease (CKD) is a multifactorial disorder with an important genetic component, and several studies have demonstrated potential associations with allelic variants. In addition, CKD patients are also characterized by high levels of genomic damage. Nevertheless, no studies have established relationships between DNA damage, or genomic instability present in CKD patients, and gene polymorphisms. To fill in this gap, the potential role of polymorphisms in genes involved in base excision repair (OGG1, rs1052133; MUTYH, rs3219489; XRCC1, rs25487), nucleotide excision repair (ERCC2/XPD, rs1799793, rs171140, rs13181; ERCC4, rs3136166); phase II metabolism (GSTP1, rs749174; GSTO1, rs2164624; GSTO2, rs156697), and antioxidant enzymes (SOD1, rs17880135, rs1041740, rs202446; SOD2, rs4880; CAT, rs1001179; GPX1, rs17080528; GPX3, rs870406: GPX4, rs713041) were inquired. In addition, some genes involved in CKD (AGT, rs5050; GLO1, rs386572987; SHROOM3, rs17319721) were also evaluated. The genomic damage, the genomic instability, and oxidative damage were evaluated by using the micronucleus and the comet assay in 589 donors (415 CKD patients and 174 controls). Our results showed significant associations between genomic damage and genes directly involved in DNA repair pathways (XRCC1, and ERCC2), and with genes encoding for antioxidant enzymes (SOD1 and GPX1). GSTO2, as a gene involved in phase II metabolism, and MUTYH showed also an association with genomic instability. Interestingly, the three genes associated with CKD (AGT, GLO1, and SHROOM3) showed associations with both the high levels of oxidatively damaged DNA and genomic instability. These results support our view that genomic instability can be considered a biomarker of the CKD status.


Trusting information on cancer varies by source of information and political viewpoint.
This study investigated how trusting information on cancer varies by the source of information and political viewpoint. This study used cross-sectional survey data from the 2020 Health Information National Trends Survey (HINTS). The study comprised a sample of 2949 adults 18 years and older. The outcome variable was measured by assessing respondents' trust in cancer-related information from various sources, including religious organizations and leaders, government health agencies, charitable organizations, family or friends, and doctors. Political viewpoint was measured as liberal, moderate, and conservative. Multivariate linear probability models were estimated and adjusted for individual-level characteristics. Multivariate analysis found that conservatives (73%, 95% CI = 68-78%) were significantly less likely to trust information on cancer from government health agencies compared to liberals (84%, 95% CI = 80-88%). There was no statistically significant difference in trusting government health agencies between liberals and moderates (80%, 95% CI = 76-84%). Both moderates (27%, 95% CI = 21-34%) and conservatives (34%, 95% CI = 29-39%) were more likely to trust information on cancer from religious organizations and leaders compared to liberals (19%, 95% CI = 13-24%). The relationship between political viewpoint and trust of doctors, family or friends, and charitable organizations were not statistically significant. Compared to liberals, conservatives are more likely to trust information on cancer from religious organizations and leaders and less likely to trust government health agencies when adjusting for other covariates. This finding emphasizes the role of political viewpoint in shaping individuals' perceptions of information sources and cancer-related information.


Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses.
Thirteen clones of hybrid cells which synthesize antibodies directed against the Rous sarcoma virus (RSV) transforming protein, pp60src, were isolated. Mouse myeloma cells were fused with spleen cells from mice that had been immunized with purified pp60src from bacterial recombinants which direct the synthesis of the RSV src gene. The hybridomas which survived the selection medium were screened by immunoprecipitation of pp60src from 32P-labeled lysates of RSV-transformed cells. Monoclonal antibodies produced by subclones derived from 13 hybridomas recognized pp60src encoded by the Schmidt-Ruppin and Prague strains of RSV and the cellular homolog of pp60src. Antibody from clone 261 had a high affinity for the viral yes gene product, and antibodies from clones 443 and 463 recognized the transforming proteins encoded by viruses containing the related transforming genes fps and ros. Several other clones had a low affinity for the viral yes, fps, and ros gene products which could be detected by in vitro phosphorylation of the transforming proteins after immunoprecipitation with the monoclonal antibody. All of the monoclonal antibodies allowed phosphorylation of pp60src and casein in an immune complex-bound reaction.


German Cancer Consortium (DKTK) - A national consortium for translational cancer research.
The German Cancer Consortium ('Deutsches Konsortium für Translationale Krebsforschung', DKTK) is a long-term cancer consortium, bringing together the German Cancer Research Center (DKFZ), Germany's largest life science research center, and the leading University Medical Center-based Comprehensive Cancer Centers (CCCs) at seven sites across Germany. DKTK was founded in 2012 following international peer review and has positioned itself since then as the leading network for translational cancer research in Germany. DKTK is long term funded by the German Ministry of Research and Education and the federal states of each DKTK partner site. DKTK acts at the interface between basic and clinical cancer research, one major focus being to generate suitable multisite cooperation structures and provide the basis for including higher numbers of patients and facilitate effective collaborative forward and reverse translational cancer research. The consortium addresses areas of high scientific and medical relevance and develops critical infrastructures, for example, for omics technologies, clinical and research big data exchange and analysis, imaging, and clinical grade drug manufacturing. Moreover, DKTK provides a very attractive environment for interdisciplinary and interinstitutional training and career development for clinician and medical scientists.


The multi-configuration self-consistent field method within a polarizable embedded framework.
We present a detailed derivation of Multi-Configuration Self-Consistent Field (MCSCF) optimization and linear response equations within the polarizable embedding scheme: PE-MCSCF. The MCSCF model enables a proper description of multiconfigurational effects in reaction paths, spin systems, excited states, and other properties which cannot be described adequately with current implementations of polarizable embedding in density functional or coupled cluster theories. In the PE-MCSCF scheme the environment surrounding the central quantum mechanical system is represented by distributed multipole moments and anisotropic dipole-dipole polarizabilities. The PE-MCSCF model has been implemented in DALTON. As a preliminary application, the low lying valence states of acetone and uracil in water has been calculated using Complete Active Space Self-Consistent Field (CASSCF) wave functions. The dynamics of the water environment have been simulated using a series of snapshots generated from classical Molecular Dynamics. The calculated shifts from gas-phase to water display between good and excellent correlation with experiment and previous calculations. As an illustration of another area of potential applications we present calculations of electronic transitions in the transition metal complex, [Fe(NO)(CN)5](2-) in a micro-solvated environment. This system is highly multiconfigurational and the influence of solvation is significant.


Exposure to an incompatible virtual surgery impacts the null space components of the muscle patterns after re-adaptation but not the task performance.
Under the synergy hypothesis, novel muscle synergies may be required for motor skill learning. We have developed a "virtual surgery" experimental paradigm that alters the mapping of muscle activations onto virtual cursor motion during an isometric reaching task using myoelectric control. By creating virtual surgeries that are "incompatible" with the original synergies, we can investigate learning new muscle synergies in controlled experimental conditions. We have previously shown that participants are able to improve their task performance after an incompatible virtual surgery, using novel muscle patterns to overcome the perturbation. In this work, we investigated whether the activation of novel muscle patterns, that are required after an incompatible virtual surgery, affects task performance or the muscle patterns after re-adaptation to the unperturbed baseline mapping. We found that experiencing an incompatible virtual surgery did not affect the task performance during the baseline mapping. However, the adaptation to the incompatible virtual surgery resulted in changes in the null space components of the muscle patterns used in the unperturbed task.


TRPV4 channel participates in receptor-operated calcium entry and ciliary beat frequency regulation in mouse airway epithelial cells.
The rate of mucociliary clearance in the airways is a function of ciliary beat frequency (CBF), and this, in turn, is increased by increases in intracellular calcium. The TRPV4 cation channel mediates Ca(2+) influx in response to mechanical and osmotic stimuli in ciliated epithelia. With the use of a TRPV4-deficient mouse, we now show that TRPV4 is involved in the airways' response to physiologically relevant physical and chemical stimuli. Ciliary TRPV4 expression in tracheal epithelial cells was confirmed with immunofluorescence in TRPV4(+/+) mice. Ciliated tracheal cells from TRPV4(-/-) mice showed no increases in intracellular Ca(2+) and CBF in response to the synthetic activator 4alpha-phorbol 12,13-didecanoate (4alphaPDD) and reduced responses to mild temperature, another TRPV4-activating stimulus. Autoregulation of CBF in response to high viscosity solutions is preserved in TRPV4(-/-) despite a reduced Ca(2+) signal. More interestingly, TRPV4 contributed to an ATP-induced increase in CBF, providing a pathway for receptor-operated Ca(2+) entry but not store-operated Ca(2+) entry as the former mechanism is lost in TRPV4(-/-) cells. Collectively, these results suggest that TRPV4 is predominantly located in the cilia of tracheal epithelial cells and plays a key role in the transduction of physical and chemical stimuli into a Ca(2+) signal that regulates CBF and mucociliary transport. Moreover, these studies implicate the participation of TRPV4 in receptor-operated Ca(2+) entry.


Intratumoral aluminum hydroxide-anchored IL-12 drives potent antitumor activity by remodeling the tumor microenvironment.
IL-12 is a potent cytokine that can promote innate and adaptive anticancer immunity, but its clinical development has been limited by toxicity when delivered systemically. Intratumoral (i.t.) administration can expand the therapeutic window of IL-12 and other cytokines but is in turn limited by rapid drug clearance from the tumor, which reduces efficacy, necessitates frequent administration, and increases systemic accumulation. To address these limitations, we developed an anchored IL-12 designated ANK-101, composed of an engineered IL-12 variant that forms a stable complex with the FDA-approved vaccine adjuvant aluminum hydroxide (Alhydrogel). Following i.t. administration of murine ANK-101 (mANK-101) in early intervention syngeneic mouse tumors, the complex formed a depot that was locally retained for weeks as measured by IVIS or SPECT/CT imaging, while unanchored protein injected i.t. was cleared within hours. One or 2 i.t. injections of mANK-101 induced single-agent antitumor activity across a diverse range of syngeneic tumors, including models resistant to checkpoint blockade at doses where unanchored IL-12 had no efficacy. Local treatment with mANK-101 further induced regressions of noninjected lesions, especially when combined with systemic checkpoint blockade. Antitumor activity was associated with remodeling of the tumor microenvironment, including prolonged IFN-γ and chemokine expression, recruitment and activation of T and NK cells, M1 myeloid cell skewing, and increased antigen processing and presentation. Subcutaneous administration of ANK-101 in cynomolgus macaques was well tolerated. Together, these data demonstrate that ANK-101 has an enhanced efficacy and safety profile and warrants future clinical development.


Iodination of Escherichia coli lac repressor. Effect of tyrosine modification on repressor activity.
Treatment of Escherichia coli lac repressor with iodine (Kl3) at 4 degrees and pH 7.5 resulted in the rapid loss of repressor DNA binding activity. At a 30-fold molar excess of iodine to repressor, inactivation was complete within 15 sec. Inducer binding under the same conditions was only slightly affected. Iodinated repressor remained tetrameric, indicating that no gross structural alteration of the protein has taken place. Control experiments demonstrated that side products of the reaction did not contribute to the observed activity loss. Moreover, no restoration of binding activity was observed when iodinated repressor was assayed under a variety of assay conditions. Incubation of repressor with lac operator containing gamma plac DNA during the iodination reaction resulted in approximately 50% protection of binding activity. This protective effect was only partially operator specific, as gammaDNA lacking the operator binding site afforded roughly 25% protection under the same conditions. Incubation with either inducer and anti-inducer molecules during the iodination reaction did not protect repressor DNA binding activity. Iodination with K-131I-3 demonstrated that at complete inactivation virtually all of the bound iodine was recovered as iodotyrosine. A minor amount of cysteine oxidation to cysteinesulfonic acid was also detected. This oxidation encompassed no more than 30% of a single cysteine residue )tentatively identified as cysteine-107). Unstable intermediate oxidation products of cysteine did not appear to be involved in the loss of DNA binding activity. Modification of amino acids other than tyrosine and cysteine was not observed. Tryptic digestion of -131I-labeled repressor suggested that approximately 90% of the incorporated radioactivity was located in the repressor N-terminal tryptic peptide. Automated sequence analysis of iodinated repressor confirmed that at roughly 0.5 bound iodine atoms per repressor subunit (corresponding to approximately 5-10% activity loss) tyrosine residues 7, 12, and 17 were labeled in the ratios 1.0:0.5:0.8. Doubling the amount of bound iodine to 1.0 atom per subunit (corresponding to approximately 50-60% activity loss) did not significantly change the pattern of incorporation.


Farnesol induces hydrogen peroxide resistance in Candida albicans yeast by inhibiting the Ras-cyclic AMP signaling pathway.
Farnesol, a Candida albicans cell-cell signaling molecule that participates in the control of morphology, has an additional role in protection of the fungus against oxidative stress. In this report, we show that although farnesol induces the accumulation of intracellular reactive oxygen species (ROS), ROS generation is not necessary for the induction of catalase (Cat1)-mediated oxidative-stress resistance. Two antioxidants, alpha-tocopherol and, to a lesser extent, ascorbic acid effectively reduced intracellular ROS generation by farnesol but did not alter farnesol-induced oxidative-stress resistance. Farnesol inhibits the Ras1-adenylate cyclase (Cyr1) signaling pathway to achieve its effects on morphology under hypha-inducing conditions, and we demonstrate that farnesol induces oxidative-stress resistance by a similar mechanism. Strains lacking either Ras1 or Cyr1 no longer exhibited increased protection against hydrogen peroxide upon preincubation with farnesol. While we also observed the previously reported increase in the phosphorylation level of Hog1, a known regulator of oxidative-stress resistance, in the presence of farnesol, the hog1/hog1 mutant did not differ from wild-type strains in terms of farnesol-induced oxidative-stress resistance. Analysis of Hog1 levels and its phosphorylation states in different mutant backgrounds indicated that mutation of the components of the Ras1-adenylate cyclase pathway was sufficient to cause an increase of Hog1 phosphorylation even in the absence of farnesol or other exogenous sources of oxidative stress. This finding indicates the presence of unknown links between these signaling pathways. Our results suggest that farnesol effects on the Ras-adenylate cyclase cascade are responsible for many of the observed activities of this fungal signaling molecule.


Carboplatin with intravenous and subsequent oral administration of vinorelbine in resected non-small-cell-lung cancer in real-world set-up.
Adjuvant cisplatin-based chemotherapy is recommended for routine use in patients with Stage IIA, IIB or IIIA non-small cell lung cancer (NSCLC) after complete resection. Results obtained for Stage IB were not conclusive. While vinorelbine plus cisplatin is the preferred choice after resection, combining vinorelbine with carboplatin promises improved compliance and delivery of drugs due to lower toxicity. We evaluated the impact of this option on treatment compliance and survival under real-world conditions. A prospective, single-arm, multicenter, non-interventional study evaluated the tolerability, dose intensity and survival resulting from adjuvant use of intravenous carboplatin (AUC 5 on day 1) with vinorelbine administered both intravenously (25 mg/m2 on day 1) and orally (60 mg/m2 on day 8) within four cycles of 21 days each. A total of 74 patients with a median age of 64 years were observed. The mean number of accomplished cycles was 3.78, and 62 patients (83.7%) completed all four planned cycles. Relative dose intensity for carboplatin was 88.9%, for intravenous vinorelbine 93.1%, and for oral vinorelbine 83.2%. Median follow-up was 4.73 years. Median disease-specific survival (DSS) was 7.63 years, median overall survival (OS) was 5.90 years, median disease-free survival (DFS0) was 4.43 years, and five-year survival was 56.2%. TNM stage of disease significantly affected DSS and OS. Favorable survival was observed in females, nonsmokers, patients aged over 65 years, patient with prior lobectomy, patients with tumor of squamous histology, and those who finished the planned therapy, but the differences were non-significant. Adjuvant carboplatin with vinorelbine switched from intravenous to oral administration was shown to be a favorable regimen with regard to tolerability and safety. Compliance to therapy was high, and survival parameters were promising, showing that applied regimen can be another potential option for adjuvant chemotherapy in patients with NSCLC.


A young patient with history of Kawasaki disease presenting with triple vessel disease.
Kawasaki disease is primarily a condition that affects young children and it is associated with cardiac morbidity and mortality. This disease has been known to cause coronary artery aneurysms which occurs as a sequelae of vasculitis. The progression of triple vessel disease in adult which results from cardiac complications from Kawasaki disease is rare. We report a case of a young man with history of Kawasaki disease at infancy presenting with triple vessel disease requiring cardiac bypass surgery at the age of 20 years old.


Electron microscopic analysis of infection patterns for Venezuelan equine encephalomyelitis virus in the vector mosquito, Culex (Melanoconion) taeniopus.
The infectious process for Venezuelan equine encephalomyelitis virus in an enzootic vector mosquito, Culex (Melanoconion) taeniopus, was examined by transmission electron microscopy. Following large dose oral infection, virus was seen in the abdominal fat body and epidermis within 1 hr of engorgement. Replicated virus appeared to leave the mesenteron 3-4 hr post-infection. Dissemination to other organs occurred between days 1 and 2 of extrinsic incubation, when the hindgut and abdominal nerve ganglia were found to be infected. Virus reached the thoracic nerve ganglia, brain and salivary glands by 4 days post-infection, and flight muscles contained virus by day 6. Virus was never detected in the malphigian tubules or ovaries of infected mosquitoes within 21 days of extrinsic incubation. These results suggest that virus particles penetrate the mesenteron and accumulate in the abdominal fat body prior to replication within the vector. This pattern differs from that reported for other arbovirus-vector pairs.


Advancing paternal age and the risk of schizophrenia.
A major source of new mutations in humans is the male germ line, with mutation rates monotonically increasing as father's age at conception advances, possibly because of accumulating replication errors in spermatogonial cell lines. We investigated whether the risk of schizophrenia was associated with advancing paternal age in a population-based birth cohort of 87 907 individuals born in Jerusalem from 1964 to 1976 by linking their records to the Israel Psychiatric Registry. Of 1337 offspring admitted to psychiatric units before 1998, 658 were diagnosed as having schizophrenia and related nonaffective psychoses. After controlling for maternal age and other confounding factors (sex, ethnicity, education [to reflect socioeconomic status], and duration of marriage) in proportional hazards regression, we found that paternal age was a strong and significant predictor of the schizophrenia diagnoses, but not of other psychiatric disorders. Compared with offspring of fathers younger than 25 years, the relative risk of schizophrenia increased monotonically in each 5-year age group, reaching 2.02 (95% confidence interval, 1.17-3.51) and 2.96 (95% confidence interval, 1.60-5.47) in offspring of men aged 45 to 49 and 50 years or more, respectively. Categories of mother's age showed no significant effects, after adjusting for paternal age. These findings support the hypothesis that schizophrenia may be associated, in part, with de novo mutations arising in paternal germ cells. If confirmed, they would entail a need for novel approaches to the identification of genes involved in schizophrenia.


[Observations on the temporal sequence of reported pregnancy risks].
A prospective study was conducted with 383 pregnant women concerning the occurrence in respect of time, the duration, and the mutual interactions of pregnancy risks. 35 per cent of the women with hemorrhages during early pregnancy later had premature labour pain (p less than 0.01). The relative risk of premature labour is greater by the factor 2.2 in pregnant women with hemorrhages during early pregnancy than in women without haemorrhages. There was no difference between the investigated groups in respect of onset and duration of these two risk factors. 54 per cent of the pregnant women in whom circular sature of the cervix according to Shirodkar had been performed, also had premature labour pain (p less than 0.001). After the performance of the Shirodkar suture, premature labour followed in 40% of the pregnant women during the further course of pregnancy, corresponding to a relative risk of 2.3 compared with the group without Shirodkar. Premature labour with subsequent Shirodkar cerclage occurred earlier (21st week) and lasted for a longer time (13 weeks) than with the reverse sequence or if premature labour occurred alone. If cervical insufficiency occurred first, followed by premature labour, the time of birth was definitely much earlier (36 weeks) than with the controls. There was no evident connection between the risks "infections of urinary tract" and "premature labour". On the other hand, 32 per cent of the pregnant women with urinary tract infections suffered an EPH gestosis, corresponding to a relative risk of 2 (p less than 0.001). The results show that clear and specific pointers towards the possible occurrence of other risks can be derived from the presence of certain pregnancy risks based on specific findings.


APOE4 confers transcriptomic and functional alterations to primary mouse microglia.
Common genetic variants in more than forty loci modulate risk for Alzheimer's disease (AD). AD risk alleles are enriched within enhancers active in myeloid cells, suggesting that microglia, the brain-resident macrophages, may play a key role in the etiology of AD. A major genetic risk factor for AD is Apolipoprotein E (APOE) genotype, with the ε4/ε4 (E4) genotype increasing risk for AD by approximately 15 fold compared to the most common ε3/ε3 (E3) genotype. However, the impact of APOE genotype on microglial function has not been thoroughly investigated. To address this, we cultured primary microglia from mice in which both alleles of the mouse Apoe gene have been humanized to encode either human APOE ε3 or APOE ε4. Relative to E3 microglia, E4 microglia exhibit altered morphology, increased endolysosomal mass, increased cytokine/chemokine production, and increased lipid and lipid droplet accumulation at baseline. These changes were accompanied by decreased translation and increased phosphorylation of eIF2ɑ and eIF2ɑ-kinases that participate in the integrated stress response, suggesting that E4 genotype leads to elevated levels of cellular stress in microglia relative to E3 genotype. Using live-cell imaging and flow cytometry, we also show that E4 microglia exhibited increased phagocytic uptake of myelin and other substrates compared to E3 microglia. While transcriptomic profiling of myelin-challenged microglia revealed a largely overlapping response profile across genotypes, differential enrichment of genes in interferon signaling, extracellular matrix and translation-related pathways was identified in E4 versus E3 microglia both at baseline and following myelin challenge. Together, our results suggest E4 genotype confers several important functional alterations to microglia even prior to myelin challenge, providing insight into the molecular and cellular mechanisms by which APOE4 may increase risk for AD.


Glutathione S-transferase M1 genotypes and the risk of vulvar cancer: a population-based case-control study.
Glutathione S-transferases (GSTs) facilitate the excretion of a variety of potential carcinogens. Some 50-60% of Caucasians are homozygous for the null allele of GSTM1, a gene responsible for the presence of one of these enzymes. The authors examined whether women with the GSTM1 null genotype are at altered risk of vulvar cancer. They obtained peripheral blood specimens from 18- to 79-year-old residents of King, Pierce, and Snohomish counties of western Washington who were diagnosed with vulvar cancer between April 1991 and June 1994. Blood specimens were also obtained from controls identified via random digit telephone dialing of western Washington households. The authors determined the GSTM1 genotype of 137 cases (120 in situ and 17 invasive cases) and 248 controls. The frequency of the GSTM1 null genotype was 46.7% among cases and 57.3% among controls. The age-adjusted odds ratio associated with the GSTM1 null genotype was 0.7 (95% confidence interval: 0.4, 1.0). Among current smokers of cigarettes, the age-adjusted odds ratio associated with the GSTM1 null genotype was 0.5 (95% confidence interval: 0.2, 0.9), differing little between heavy and light smokers. Our data suggest that women with the GSTM1 null genotype are not at increased risk of vulvar cancer.


Joint Report of the Fourth International Bovine Lymphocyte Antigen (BoLA) Workshop, East Lansing, Michigan, USA, 25 August 1990.
Blood samples from 54 animals were exchanged between 15 laboratories in nine countries to improve and expand BoLA class I and class II typing. A total of 27 out of 33 (82%) of previously accepted BoLA-w specificities were represented within the cell panel. Seventeen new serum-defined BoLA specificities were accepted by the workshop participants, thus expanding the number of internationally recognized BoLA specificities to 50. The large number of new specificities detected resulted from the number of serological reagents used (n = 1139) and the genetic diversity of the cell panel. Confidence derived from the high percentage of agreement between the laboratories on antigen detection (97.3%; r = 0.84) permitted the removal of the workshop (w) notation from 23 BoLA-w specificities and their acceptance as full status BoLA-A antigens. Two new non-BoLA antigens were also detected, one completely included within the red blood cell factor S' (BoLy-S'), whereas a second (BoLy-w1) did not show any association with tested red blood cell factors. A comparison between serological, isoelectric focusing (IEF) and DNA typing for BoLA class II polymorphism was conducted with a subset of workshop cells. Correlation between the three methods was significant for three combinations of alleles. Three other serologically defined class II specificities were correlated with DR and/or DQ restriction fragment length polymorphism (RFLP) types, whereas six additional IEF types were correlated with DR and/or DQ RFLP types (r greater than or equal to 0.50). Several new IEF, DRB, DQA and DQB RFLP patterns were identified. In 46 animals that were typed for BoLA-DR and DQ genes by RFLP analysis, 46 different BoLA haplotypes were tentatively defined. These 46 haplotypes were distinguished by 31 serologically-defined BoLA-A alleles (and 2 'blanks'), 15 DRB RFLP types (plus up to 10 new DRB RFLP patterns) and 23 DQA-DQB haplotypes.


Quantification of urinary conjugates of bisphenol A, 2,5-dichlorophenol, and 2-hydroxy-4-methoxybenzophenone in humans by online solid phase extraction-high performance liquid chromatography-tandem mass spectrometry.
Urinary concentrations of phenols or their metabolites have been used as biomarkers to assess the prevalence of exposure to these compounds in the general population. Total urinary concentrations, which include both free and conjugated (glucuronide and sulfated) forms of the compounds, are usually reported. From a toxicologic standpoint, the relative concentrations of the free species compared with their conjugated analogs can be important because conjugation may reduce the potential biologic activity of the phenols. In this study, we determined the percentage of glucuronide and sulfate conjugates of three phenolic compounds, bisphenol A (BPA), 2,5-dichlorophenol (2,5-DCP), and 2-hydroxy-4-methoxybenzophenone (benzophenone-3, BP-3) in 30 urine samples collected between 2000 and 2004 from a demographically diverse group of anonymous adult volunteers. We used a sensitive on-line solid phase extraction-isotope dilution-high performance liquid chromatography-tandem mass spectrometry method. These three phenols were detected frequently in the urine samples tested. Only small percentages of the compounds (9.5% for BPA, and 3% for 2,5-DCP and BP-3) were excreted in their free form. The percentage of the sulfate conjugate was about twice that of the free compound. The glucuronide conjugate was the major metabolite, representing 69.5% (BPA), 89% (2,5-DCP), and 84.6% (BP-3) of the total amount excreted in urine. These results are in agreement with those reported before which suggested that BPA-glucuronide was an important BPA urinary metabolite in humans. To our knowledge, this is the first study describing the distribution of urinary conjugates of BP-3 and 2,5-DCP in humans.


Analysis of prices paid by low-income countries - how price sensitive is government demand for medicines?
Access to medicines is an important health policy issue. This paper considers demand structures in a selection of low-income countries from the perspective of public authorities as the evidence base is limited. Analysis of the demand for medicines in low-income countries is critical for effective pharmaceutical policy where regulation is less developed, health systems are cash constrained and medicines are not typically subsidised by a public health insurance system This study analyses the demand for medicines in low-income countries from the perspective of the prices paid by public authorities. The analysis draws on a unique dataset from World Health Organization (WHO) and Health Action International (HAI) using 2003 data on procurement prices of medicines across 16 low-income countries covering 48 branded drugs and 18 therapeutic categories. Variation in prices, the mark-ups over marginal costs and estimation of price elasticities allows assessment of whether these elasticities are correlated with a country's national income. Using the Ramsey pricing rule, the study's findings suggest that substantial cross-country variation in prices and mark-ups exist, with price elasticities ranging from -1 to -2, which are weakly correlated with national income. Government demand for medicines thus appears to be price elastic, raising important policy implications aimed at improving access to medicines for patients in low-income countries.


Nonmulberry silk proteins: multipurpose ingredient in bio-functional assembly.
The emerging field of tissue engineering and regenerative medicines utilising artificial polymers is facing many problems. Despite having mechanical stability, non-toxicity and biodegradability, most of them lack cytocompatibility and biocompatibility. Natural polymers (such as collagen, hyaluronic acid, fibrin, fibroin, and others), including blends, are introduced to the field to solve some of the relevant issues. Another natural biopolymer: silkworm silk gained special attention primarily due to its specific biophysical, biochemical, and material properties, worldwide availability, and cost-effectiveness. Silk proteins, namely fibroin and sericin extracted from domesticated mulberry silkwormBombyx mori, are studied extensively in the last few decades for tissue engineering. Wild nonmulberry silkworm species, originated from India and other parts of the world, also produce silk proteins with variations in their nature and properties. Among the nonmulberry silkworm species,Antheraea mylitta(Indian Tropical Tasar),A. assamensis/A. assama(Indian Muga), andSamia ricini/Philosamia ricini(Indian Eri), along withA. pernyi(Chinese temperate Oak Tasar/Tussah) andA. yamamai(Japanese Oak Tasar) exhibit inherent tripeptide motifs of arginyl glycyl aspartic acid in their fibroin amino acid sequences, which support their candidacy as the potential biomaterials. Similarly, sericin isolated from such wild species delivers unique properties and is used as anti-apoptotic and growth-inducing factors in regenerative medicines. Other characteristics such as biodegradability, biocompatibility, and non-inflammatory nature make it suitable for tissue engineering and regenerative medicine based applications. A diverse range of matrices, including but not limited to nano-micro scale structures, nanofibres, thin films, hydrogels, and porous scaffolds, are prepared from the silk proteins (fibroins and sericins) for biomedical and tissue engineering research. This review aims to represent the progress made in medical and non-medical applications in the last couple of years and depict the present status of the investigations on Indian nonmulberry silk-based matrices as a particular reference due to its remarkable potentiality of regeneration of different types of tissues. It also discusses the future perspective in tissue engineering and regenerative medicines in the context of developing cutting-edge techniques such as 3D printing/bioprinting, microfluidics, organ-on-a-chip, and other electronics, optical and thermal property-based applications.


Pharmacokinetics of a Novel, Transdermal Fentanyl Solution in Rhesus Macaques (Macaca mulatta).
Rhesus macaques (Macaca mulatta) are the most commonly used NHP biomedical model and experience both research and clinical procedures requiring analgesia. Opioids are a mainstay of analgesic therapy. A novel, transdermal fentanyl solution (TFS) has been developed as a long-acting, single-administration topical opioid and was reported to provide at least 4 d of effective plasma concentrations in beagles (Canis familiaris). To evaluate the pharmacokinetic profile of TFS in healthy adult rhesus macaques, we used a 2-period, 2-treatment crossover study of a single topical administration of 1.3 (25) and 2.6 mg/kg (50 μL/kg) TFS. TFS was applied to the clipped dorsal skin of adult rhesus macaques (n = 6; 3 male, 3 female) under ketamine sedation (10 mg/kg IM). We hypothesized that TFS in rhesus macaques would provide at least 4 d of effective plasma concentrations (assumed to be ≥ 0.2 ng/mL, based on human studies). Plasma fentanyl concentrations were determined by liquid chromatography-tandem mass spectrometry before drug administration and at 0, 0.5, 1, 2, 4, 8, 12, 24, 36, 48, 60, 72, 96, 120, 144, 168, 240, 336, 408, and 504 h afterward. Noncompartmental pharmacokinetic analysis was performed. For each dose (1.3 and 2.6 mg/kg), respectively, the maximal plasma concentration was 1.95 ± 0.40 and 4.19 ± 0.69 ng/mL, occurring at 21.3 ± 4.1 and 30.7 ± 8.7 h; the AUC was 227.3 ± 31.7 and 447.0 ± 49.1 h/ng/mL, and the terminal elimination half-life was 93.7 ± 7.1 and 98.8 ± 5.4 h. No adverse effects were noted after drug administration at either dose. Macaques maintained plasma fentanyl concentrations of 0.2 ng/mL or greater for at least 7 d after 1.3 mg/kg and at least 10 d after 2.6 mg/kg topical administration of TFS. A single TFS dose may provide efficacious analgesia to rhesus macaques and reduce stress, discomfort, and risk to animals and personnel.


Advances in neuro-oncology imaging.
Despite the fact that MRI has evolved to become the standard method for diagnosis and monitoring of patients with brain tumours, conventional MRI sequences have two key limitations: the inability to show the full extent of the tumour and the inability to differentiate neoplastic tissue from nonspecific, treatment-related changes after surgery, radiotherapy, chemotherapy or immunotherapy. In the past decade, PET involving the use of radiolabelled amino acids has developed into an important diagnostic tool to overcome some of the shortcomings of conventional MRI. The Response Assessment in Neuro-Oncology working group - an international effort to develop new standardized response criteria for clinical trials in brain tumours - has recommended the additional use of amino acid PET imaging for brain tumour management. Concurrently, a number of advanced MRI techniques such as magnetic resonance spectroscopic imaging and perfusion weighted imaging are under clinical evaluation to target the same diagnostic problems. This Review summarizes the clinical role of amino acid PET in relation to advanced MRI techniques for differential diagnosis of brain tumours; delineation of tumour extent for treatment planning and biopsy guidance; post-treatment differentiation between tumour progression or recurrence versus treatment-related changes; and monitoring response to therapy. An outlook for future developments in PET and MRI techniques is also presented.


Resveratrol activates duodenal Sirt1 to reverse insulin resistance in rats through a neuronal network.
Resveratrol improves insulin sensitivity and lowers hepatic glucose production (HGP) in rat models of obesity and diabetes, but the underlying mechanisms for these antidiabetic effects remain elusive. One process that is considered a key feature of resveratrol action is the activation of the nicotinamide adenine dinucleotide (NAD(+))-dependent deacetylase sirtuin 1 (SIRT1) in various tissues. However, the low bioavailability of resveratrol raises questions about whether the antidiabetic effects of oral resveratrol can act directly on these tissues. We show here that acute intraduodenal infusion of resveratrol reversed a 3 d high fat diet (HFD)-induced reduction in duodenal-mucosal Sirt1 protein levels while also enhancing insulin sensitivity and lowering HGP. Further, we found that duodenum-specific knockdown of Sirt1 expression for 14 d was sufficient to induce hepatic insulin resistance in rats fed normal chow. We also found that the glucoregulatory role of duodenally acting resveratrol required activation of Sirt1 and AMP-activated protein kinase (Ampk) in this tissue to initiate a gut-brain-liver neuronal axis that improved hypothalamic insulin sensitivity and in turn, reduced HGP. In addition to the effects of duodenally acting resveratrol in an acute 3 d HFD-fed model of insulin resistance, we also found that short-term infusion of resveratrol into the duodenum lowered HGP in two other rat models of insulin resistance--a 28 d HFD-induced model of obesity and a nicotinamide (NA)-streptozotocin (STZ)-HFD-induced model of mild type 2 diabetes. Together, these studies highlight the therapeutic relevance of targeting duodenal SIRT1 to reverse insulin resistance and improve glucose homeostasis in obesity and diabetes.


Psychoeducation in bipolar patients with comorbid personality disorders.
The co-occurrence of personality and bipolar disorders is quite common. Bipolar patients with personality disorders have been described as having poorer outcome than 'pure' bipolar patients. However, from a combined-approach point of view, a little has been done to improve the course of these patients. Psychoeducation has shown its efficacy in the prevention of relapses in the bipolar population but, to date, no data is available on its efficacy in the management of bipolar patients with personality disorders. The present study shows a subanalysis from a single-blind randomized prospective clinical trial on the efficacy of group psychoeducation in bipolar I patients. Bipolar patients fulfilling DSM-IV criteria for any personality disorder were randomized to either psychoeducational treatment or a non-structured intervention. There were 22 patients in the control group and 15 in the psychoeducation group. All patients received naturalistic pharmacological treatment as well. The follow-up phase comprised 2 years where all patients continued receiving naturalistic treatment without psychological intervention and were assessed monthly for several outcome measures. At the end of the follow-up phase (2 years), a 100% of control group patients fulfilled criteria for recurrence versus a 67% in the psychoeducation group (p < 0.005). Patients included in the psychoeducation group had a higher time-to-relapse and a significantly lower mean number of total, manic and depressive relapses. No significant differences regarding the number of patients who required hospitalization were found but the mean duration of days spent in the hospitalization room was significantly higher for the patients included in the control group. Psychoeducation may be a useful intervention for bipolar patients with comorbid personality disorders. Further studies should address the efficacy of specifically tailored interventions for this common type of patients.


Engagement of Gab1 and Gab2 in erythropoietin signaling.
Several signaling cascades are activated during engagement of the erythropoietin receptor to mediate the biological effects of erythropoietin. The members of the insulin receptor substrate (IRS) family of proteins play a central role in signaling for various growth factor receptors and cytokines by acting as docking proteins for the SH2 domains of signaling elements, linking cytokine receptors to diverse downstream pathways. In the present study we provide evidence that the recently cloned IRS-related proteins, Gab1 and Gab2, of the Gab family of proteins, are rapidly phosphorylated on tyrosine during erythropoietin treatment of erythropoietin-responsive cells and provide docking sites for the engagement of the SHP2 phosphatase and the p85 subunit of the phosphatidylinositol 3'-kinase. Furthermore, our data show that Gab1 is the primary IRS-related protein activated by erythropoietin in primary erythroid progenitor cells. In studies to identify the erythropoietin receptor domains required for activation of Gab proteins, we found that tyrosines 425 and 367 in the cytoplasmic domain of the erythropoietin receptor are required for the phosphorylation of Gab2. Taken together, our data demonstrate that Gab proteins are engaged in erythropoietin signaling to mediate downstream activation of the SHP2 and phosphatidylinositol 3'-kinase pathways and possibly participate in the generation of the erythropoietin-induced mitogenic responses.


Hsp90 chaperone complexes are required for the activity and stability of yeast protein kinases Mik1, Wee1 and Swe1.
The Wee1 protein kinase negatively regulates entry into mitosis by mediating the inhibitory tyrosine phosphorylation of Cdc2-cyclin B kinase. The stability and activity of Wee1 from the fission yeast Schizosaccharomyces pombe is critically dependent on functional Hsp90 chaperones. Here we identify two related tyrosine protein kinases, Mik1 from fission yeast and its Saccharomyces cerevisiae homolog Swe1, as Hsp90 substrates and show that the kinase domain is sufficient to mediate this interaction. Morphological and biochemical defects arising from overexpression of the kinases in fission yeast are suppressed in the conditional Hsp90 mutant swo1-26. A subset of all three kinases is associated with the Hsp90 cochaperones cyclophilin 40 and p23. Under conditions of impaired chaperone function or treatment with the Hsp90 inhibitory drug geldanamycin, intracellular levels of the kinases are reduced and the proteins become rapidly degraded by the proteasome machinery, indicating that Wee1, Mik1 and Swe1 require Hsp90 heterocomplexes for their stability and maintenance of function.


Penile cancer epidemiology and risk factors: a contemporary review.
Our objective is to present an overview of epidemiologic, clinical, and molecular risk factors with a focus on contemporary literature. Penile cancer is a rare and aggressive neoplasm that accounts for less than 1% of male malignancies in the United States. Geographical disparities in incidence of disease are evident with high rates concentrated in the developing world (2.8-6.8 per 100 000) where neonatal circumcision is low and socioeconomic conditions predispose patients to multiple risk factors. Western countries have a significantly lower incidence and can be as low as 0.3 per 100 000. Many risk factors have been identified including lack of circumcision, phimosis, balanitis, obesity, lichen sclerosus, smoking, and psoralen UV-A phototherapy. In addition, human papilloma virus (HPV) has been linked to nearly 40% of cases and molecular mediators continue to be investigated. Although Penile cancer can be a debilitating disease, several of the known risk factors are modifiable. Public health campaigns aimed to increase awareness, promote better hygiene, and deploy HPV vaccines have had varied success at decreasing disease burden. Focus should be placed on implementing such interventions in developing countries and at-risk populations.


Kinetic Uncertainty Relations for the Control of Stochastic Reaction Networks.
Nonequilibrium stochastic reaction networks are commonly found in both biological and nonbiological systems, but have remained hard to analyze because small differences in rate functions or topology can change the dynamics drastically. Here, we conjecture exact quantitative inequalities that relate the extent of fluctuations in connected components, for various network topologies. Specifically, we find that regardless of how two components affect each other's production rates, it is impossible to suppress fluctuations below the uncontrolled equivalents for both components: one must increase its fluctuations for the other to be suppressed. For systems in which components control each other in ringlike structures, it appears that fluctuations can only be suppressed in one component if all other components instead increase fluctuations, compared to the case without control. Even the general N-component system-with arbitrary connections and parameters-must have at least one component with increased fluctuations to reduce fluctuations in others. In connected reaction networks it thus appears impossible to reduce the statistical uncertainty in all components, regardless of the control mechanisms or energy dissipation.


Porcine γδ T lymphocytes can be categorized into two functionally and developmentally distinct subsets according to expression of CD2 and level of TCR.
Porcine γδ T cells have two levels of TCRγδ expression. Whereas TCRγδ(med) cells are mostly CD2(+)CD8(-) and CD2(+)CD8(+), TCRγδ(hi) cells are highly enriched for CD2(-)CD8(-). This distribution is independent of bacterial colonization and it is already established in the thymus prior to export of γδ cells to the periphery. Sorting and cultivation experiments revealed that CD2(-)CD8(-) γδ cells are unable to acquire CD2 and CD8, whereas CD2(+) subsets can gain or loose CD8. There is also differential susceptibility for proliferation between CD2(+) and CD2(-) γδ cells. Although CD2(-)CD8(-) almost do not proliferate, proliferation of CD2(+)CD8(-) and CD2(+)CD8(+) is substantial. Population of CD2(-) γδ cells is also absent in CD1(+) immature thymocytes. Additionally, subpopulations of CD2(+) and CD2(-) γδ cells in the thymus differ in expression of auxiliary surface molecules such as CD25, CD45RA/RC, and MHC class II. Moreover, TCRγδ(hi) cells can generate TCRγδ(med) cells but never the opposite. The only exception is the thymus, where a few TCRγδ(med) cells can be induced to TCRγδ(hi) but only under IL-2 influence. The repertoire of TCRδ is polyclonal in all subsets, indicating that there is the same extent of diversification and equal capability of immune responses. Results collectively indicate that CD2 expression determines two lineages of γδ cells that differ in many aspects. Because CD2(-) γδ cells are missing in the blood of humans and mice but are obvious in other members of γδ-high species such as ruminants and birds, our findings support the idea that circulating CD2(-) γδ T cells are a specific lineage.