Structural analysis of the evolution of steroid specificity in the mineralocorticoid and glucocorticoid receptors.
The glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) evolved from a common ancestor. Still not completely understood is how specificity for glucocorticoids (e.g. cortisol) and mineralocorticoids (e.g. aldosterone) evolved in these receptors. Our analysis of several vertebrate GRs and MRs in the context of 3D structures of human GR and MR indicates that with the exception of skate GR, a cartilaginous fish, there is a deletion in all GRs, at the position corresponding to Ser-949 in human MR. This deletion occurs in a loop before helix 12, which contains the activation function 2 (AF2) domain, which binds coactivator proteins and influences transcriptional activity of steroids. Unexpectedly, we find that His-950 in human MR, which is conserved in the MR in chimpanzee, orangutan and macaque, is glutamine in all teleost and land vertebrate MRs, including New World monkeys and prosimians. Evolution of differences in the responses of the GR and MR to corticosteroids involved deletion in the GR of a residue corresponding to Ser-949 in human MR. A mutation corresponding to His-950 in human MR may have been important in physiological changes associated with emergence of Old World monkeys from prosimians.


Role of Coccidia in the occurrence of necrotic enteritis of chickens.
Clostridium perfringens type A, Eimeria acervulina, and Eimeria necatrix were used to produce necrotic enteritis in chickens. The disease was produced in all groups of birds that received feed contaminated with C. perfringens. Mortality due to necrotic enteritis was highest (53%) in birds infected with E. acervulina before infection with clostridia. There was a significant difference in mortality rates between birds infected with E. acervulina and birds infected with E. necatrix before infection with C. perfringens. Mortality rates also differed significantly between the group infected with E. necatrix and the group that received only feed contaminated with C. perfringens. It was concluded that under field conditions, coccidia can play a significant role in the occurrence of necrotic enteritis when a sufficient number of toxigenic strain of C. perfringens type A is present. The pathological changes induced by clostridia and coccidia are described.


Antisense targeting protein kinase C alpha and beta1 inhibits gastric carcinogenesis.
Protein kinase C (PKC) family, which functions through serine/threonine kinase activity, is involved in signal transduction pathways necessary for cell proliferation, differentiation, and apoptosis. Its critical role in neoplastic transformation and tumor invasion renders PKC a potential target for anticancer therapy. In this study, we investigated the effect of targeting individual PKCs on gastric carcinogenesis. We established gastric cancer cell lines stably expressing antisense PKCalpha, PKCbeta1, and PKCbeta2 cDNA. These stable transfectants were characterized by cell morphology, cell growth, apoptosis, and tumorigenicity in vitro and in vivo. PKCalpha-AS and PKCbeta1-AS transfectants showed a different morphology with flattened, long processes and decreased nuclear:cytoplasmic ratio compared with the control cells. Cell growth was markedly inhibited in PKCalpha-AS and PKCbeta1-AS transfectants. PKCalpha-AS and PKCbeta1-AS cells were more responsive to mitomycin C- or 5-fluorouracil-induced apoptosis. However, antisense targeting of PKCbeta2 did not have any significant effect on cell morphology, cell growth, or apoptosis. Furthermore, antisense inhibition of PKCalpha and PKCbeta1 markedly suppressed colony-forming efficiency in soft agar and in nude mice xenografts. Inhibition of PKCalpha or PKCbeta1 significantly suppressed transcriptional and DNA binding activity of activator protein in gastric cancer cells, suggesting that PKCalpha or PKCbeta1 exerts their effects on cell growth through regulation of activator protein activity. These data provide evidence that targeting PKCalpha and PKCbeta1 by antisense method is a promising therapy for gastric cancer.


BH4 as a Therapeutic Target for ADHD: Relevance to Neurotransmitters and Stress-Driven Symptoms.
Tetrahydrobiopterin (BH4) is a critical cofactor in a variety of metabolic pathways that have been linked to ADHD. There have been no previous studies utilizing BH4 as a supplement for ADHD. BH4 has been approved as a treatment for phenylketonuria (PKU). Individuals with PKU and ADHD appear to have low DA levels in common, suggesting that the hypodopaminergic state seen in both illnesses could be a relationship between the two. Clinical research involving supplementation of BH4 has shown low occurrence of adverse. In experiments, BH4 has also been found to have good blood-brain barrier permeability. BH4 also has the ability in scavenging ROS activity, which is an implication of stress and is seen in ADHD. BH4's significance in ADHD is reviewed in this paper because of its involvement in numerous neurodevelopmental metabolic pathways, and we anticipate that exogenous BH4 can be used to treat ADHD.


[Effect of N-acylethanolamines on adrenal cortex function].
The possibility of NAE to take part in the regulation of the function of adrenal glands was studied. It was shown that two times NAE (18:0) injection in a dose 5 mg/kg of weight increased the content of 11-hydroxysteroids in blood of intact male rats. NAE caused the raise of the blood hormone content by 4 times under the immobilization stress. It is apparent that augment of stress response under the influence of NAE in vivo is explained by the activation of hypophysis-adrenal cortex system. In vitro NAE lowered steroidogenesis by near 40%. One can suggest that this decrease is caused by membranotropic properties of NAE.


Evaluation of the Performance of the Turkish Regulatory Agency: Recommendations for Improved Patients' Access to Medicines.
This study was to evaluate the Turkish regulatory review process and timelines between 2016 and 2018 with a view to assess the changes that had taken place since the previous study, which evaluated the Turkish review processes and timelines 2013 to 2015. Data related to the Turkish Medicines and Medical Devices Agency (TİTCK) organizational structure and general information were collected from publicly available sources. A standard questionnaire was then used to collect data with the aim of identifying the TİTCK's review practices and key milestones for the marketing authorization process. Subsequently, a comparison with the previous study was conducted to identify the key changes and developments that had taken place from 2015- 2018. The TİTCK has made considerable efforts to improve its regulatory capacity since 2016, which has contributed to the overall decrease in the agency review times. The overall median approval time for new active substances; however, increased from 529 calendar days (2016) to 663 calendar days (2018), with the review time in the agency decreasing from 408 calendar days to 326 calendar days, while the company time increased from 137 to 268 calendar days, respectively, over this period. For the TİTCK to become an international reference agency, they will need to fully implement good review practices and a structured framework for benefit-risk assessment and decision making; consider implementing verification and abridged reviews based on a reliance model and expedite the PIC/S mutual recognition process as well as become a full member of the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH).


Structural requirements for active intestinal transport. Spatial and bonding requirements at C-3 of the sugar.
Analogues of d-glucose modified at C-3, and in some cases at a second position, were prepared and tested for active accumulation by everted segments of hamster intestine. Their relative affinity for the sugar carrier was measured by tissue/medium ratio, Michaelis-Menten kinetics and competitive inhibition of d-galactose or methyl alpha-d-glucoside transport. d-Glucose and its 3-deoxy-3-fluoro, 3-chloro-3-deoxy and to a smaller extent its 3-bromo-3-deoxy derivatives, bound and were transported more strongly than 3-deoxy-d-glucose and other sugars not containing an electronegative atom in the gluco configuration at C-3. 3-Deoxy-d-galactose, 3,6-dideoxy-d-glucose and d-gulose, which have two alterations from the d-glucose structure, were not, or only very weakly, transported. The results are interpreted as indicating the presence of a hydrogen bond from the carrier to the hydroxyl group at C-3 of d-glucose. Spatial requirements are also discussed. New syntheses are reported for 3-chloro-3-deoxy- and 3-bromo-3-deoxy-d-glucose and 3,6-dideoxy-d-glucose.


Identification of a 40 x 10(3) Mr centromere-associated protein in cultured mammalian cells.
Monoclonal antibodies were raised against a complex of proteins that was purified following the crosslinking of tubulin to the centromeres of CHO chromosomes using Lomant's reagent. One of the clones, hybridoma 32-9, produced antibodies that reacted with a 40 x 10(3) Mr protein present in the crosslinked complex. Furthermore, immunoblot analysis demonstrated that the 40 x 10(3) Mr antigen was present in various mammalian cell types from several different species. Indirect immunofluorescence using the antibody produced by clone 32-9 demonstrated that the 40 x 10(3) Mr antigen was associated with both spindle and cytoplasmic microtubules. In addition, centromere/kinetochore staining was detected in metaphase-arrested cells, while staining of prekinetochores in interphase nuclei was not observed. Unlike microtubule-associated proteins and microtubule-dependent ATPases, the 40 x 10(3) Mr protein did not copurify with microtubules when tubules were assembled from cellular homogenates using taxol and either GTP or GTP and AMP-PNP. Instead, the 40 x 10(3) Mr protein remained associated with the insoluble cellular material. The 40 x 10(3) Mr antigen could be released from the insoluble pelleted material by extraction with 1 M NaCl. Once solubilized, the 40 x 10(3) Mr protein was able to copurify with microtubules in assembly assays in vitro. This monoclonal antibody should serve as a valuable probe for studies of centromere/kinetochore structure and function.


Glucose transporter Glut-1 is detectable in peri-necrotic regions in many human tumor types but not normal tissues: Study using tissue microarrays.
The hypoxic tumor microenvironment is associated with malignant progression and poor treatment response. The glucose transporter Glut-1 is a prognostic factor and putative hypoxia marker. So far, studies of Glut-1 in cancer have utilized conventional immunohistochemical analysis in a series of individual biopsy or surgical specimens. Tissue microarrays, however, provide a rapid, inexpensive means of profiling biomarker expression. To evaluate hypoxia markers, tissue cores must show the architectural features of hypoxia; i.e. viable tissue surrounding necrotic regions. Glut-1 may be a useful biomarker to validate tissue microarrays for use in studies of hypoxia-regulated genes in cancer. In this study, we carried out immunohistochemical detection of Glut-1 protein in many tumor and normal tissue types in a range of tissue microarrays. Glut-1 was frequently found in peri-necrotic regions, occurring in 9/34 lymphomas, 6/12 melanomas, and 5/16 glioblastomas; and in 43/54 lung, 22/84 colon, and 23/60 ovarian tumors. Expression was rare in breast (6/40) and prostate (1/57) tumors, and in normal tissue, was restricted to spleen, tongue, and CNS endothelium. In conclusion, tissue microarrays enable the observation of Glut-1 expression in peri-necrotic regions, which may be linked to hypoxia, and reflect previous studies showing differential Glut-1 expression across tumor types and non-malignant tissue.


On transferring the grid technology to the biomedical community.
Natural scientists such as physicists pioneered the sharing of computing resources, which resulted in the Grid. The inter domain transfer process of this technology has been an intuitive process. Some difficulties facing the life science community can be understood using the Bozeman's "Effectiveness Model of Technology Transfer". Bozeman's and classical technology transfer approaches deal with technologies that have achieved certain stability. Grid and Cloud solutions are technologies that are still in flux. We illustrate how Grid computing creates new difficulties for the technology transfer process that are not considered in Bozeman's model. We show why the success of health Grids should be measured by the qualified scientific human capital and opportunities created, and not primarily by the market impact. With two examples we show how the Grid technology transfer theory corresponds to the reality. We conclude with recommendations that can help improve the adoption of Grid solutions into the biomedical community. These results give a more concise explanation of the difficulties most life science IT projects are facing in the late funding periods, and show some leveraging steps which can help to overcome the "vale of tears".


Serum copeptin and neuron specific enolase are markers of neonatal distress and long-term neurodevelopmental outcome.
The objective of this study was to evaluate the early changes in serial serum levels of copeptin and neuron-specific enolase (NSE) in neonates diagnosed with birth asphyxia, and to determine whether these biomarkers measured in the first 168 hours after birth are predictive of long-term neurodevelopmental outcome. Copeptin and NSE levels were measured from serum samples collected 6, 12, 24, 48, 72, and 168 hours after birth from 75 term neonates diagnosed with hypoxic-ischemic encephalopathy (HIE) and treated with therapeutic hypothermia for 72 hours. In addition, serum copeptin levels after birth were measured from 10 HIE diagnosed neonates, who were randomized to the normothermic arm of the TOBY cohort. All neonates underwent neurodevelopmental assessment using the Bayley Scales of Infant and Toddler Development-II at two years of age. Copeptin levels were highest at 6 hours after birth and steadily decreased, whereas the highest NSE levels were measured at 24 hours after birth. The biomarker levels correlated with blood-gas parameters (base excess, pH and lactate) at 6 and 12 hours after birth. Copeptin and NSE levels in the early postnatal period were significantly higher in neonates with poor outcome compared to those with favorable outcome at two years of age. Furthermore, in the TOBY cohort, copeptin levels were significantly lower in hypothermic compared to normothermic neonates. To conclude, copeptin and NSE measured in the early postnatal period are potential prognostic biomarkers of long-term neurodevelopmental outcome in term neonates diagnosed with HIE and treated with therapeutic hypothermia.


Influence of waves and currents on the growth rate of the kelp Laminaria digitata (Phaeophyceae).
The kelp Laminaria digitata growing in the low intertidal region along energetic coastlines are exposed to a range of hydrodynamic environments. Macroalgae in the intertidal zone can experience both waves and currents independently, but it is unknown how they influence growth rate. Relative growth rate of the meristematic region and the entire blade of L. digitata were measured to assess the influence of wave and current motion from three hydrodynamic environments: low current and low wave (LCLW), high current and low wave (HCLW), and high wave and low current (HWLC). Alongside hydrodynamic data, seawater nutrient concentrations and temperature were also collected and analyzed. Results suggest that differences in L. digitata relative growth rates were not attributed to the seawater nutrient concentrations or temperature, but attributed to the hydrodynamic environments. At the high current condition, kelp growth rate of the meristematic region was enhanced by 45% compared to the high wave condition. When including the entire blade growth rate, an average increase of 25% was observed between the high current and high wave condition. Potentially, the division in growth rate observed between the wave and current motion is related to the frequency and magnitude at which the hydrodynamic forces act. These findings highlight the complexity of the hydrodynamic environment and that forces associated with waves and currents may have a significant role on the productivity of kelp.


Tai Chi Exercise for Cancer-Related Fatigue in Patients With Lung Cancer Undergoing Chemotherapy: A Randomized Controlled Trial.
Tai Chi is a traditional Chinese health-promoting exercise. It has been shown to enhance mental health and improve psychological condition. We aimed to assess the effectiveness of Tai Chi exercise for cancer-related fatigue in patients with lung cancer undergoing chemotherapy. We conducted a randomized trial of Tai Chi exercise as compared with low-impact exercise as a control intervention. Exercises were practiced every other day, a one-hour session for 12 weeks for each of the study groups. The primary end point was a change in total score of the Multidimensional Fatigue Symptom Inventory-Short Form (MFSI-SF). Secondary end points were changes in five subscale scores of the MFSI-SF. All assessments were repeated at three time points, T0: before first course of chemotherapy; T1: before third course of chemotherapy; and T2: at the end of the fourth course of chemotherapy. Between January 2012 and December 2014, 96 patients were enrolled in this trial. At six and 12 weeks, the Tai Chi group had a lower MFSI-SF total score compared with the control group (59.5 ± 11.3 vs. 66.8 ± 11.9, P < 0.05; 53.3 ± 11.8 vs. 59.3 ± 12.2, P < 0.05). At six weeks, the Tai Chi group had lower MFSI-SF general subscale scores (18.1 ± 4.6 vs. 20.4 ± 4.5, P < 0.05) and physical subscale scores (17.5 ± 4.4 vs. 19.1 ± 4.5, P < 0.05), and higher MFSI-SF vigor subscale scores (14.5 ± 3.3 vs. 11.6 ± 3.4, P < 0.05), compared with the control group. But no significant differences were found in emotional subscale (20.2 ± 3.6 vs. 20.0 ± 3.5, P > 0.05) and mental subscale (18.2 ± 4.0 vs. 18.9 ± 3.9, P > 0.05) scores between the Tai Chi group and the control group. At 12 weeks, the MFSI-SF subscale scores showed the same trends as at six weeks. Tai Chi is an effective intervention for managing cancer-related fatigue in patients with lung cancer undergoing chemotherapy, especially for decreasing general fatigue and physical fatigue, and increasing vigor.


Prostacyclin production by the deendothelialized rabbit aorta.
The acute effect of in vitro deendothelialization on the production of prostacyclin (PGI2) by the rabbit aorta has been investigated. The effectiveness of removing endothelium by rubbing it against filter paper or scraping it with a scalpel was demonstrated by scanning electron microscopy and en face examination after silver staining. Endothelium removal produced an immediate stimulation of PGI2 release, resulting in 408% of the control after rubbing and 367% of the control after scraping, during the first 30-min period of incubation. This increased production of PGI2 gradually declined over time to reach values similar to the control after 2h. At that time, the deendothelialized aorta was totally unresponsive to the stimuli that increase PGI2 release in the intact aorta (acetylcholine, ADP, ionophore A23187, and arachidonic acid). The enhanced production of PGI2 in the deendothelialized aorta was associated with an increased release of free arachidonic acid (353% of the control): in contrast with PGI2, this stimulation was maintained for at least 150 min. A transient exposure of the deendothelialized aorta to ibuprofen (250 microM) was followed by a rebound of PGI2 production, which was also prolonged by BW-755C (3-10 microM). In conclusion, removal of the endothelium triggered an immediate and sustained mobilization of free arachidonic acid in the rabbit aorta: the resulting increase of PGI2 production was short-lived, probably as a consequence of cyclooxygenase self-inactivation. Our results indicate that the subendothelium has a significant capacity to produce PGI2, but that this capacity is expressed only briefly.


Upregulation of Bcl-2 enhances secretion of growth factors by adipose-derived stem cells deprived of oxygen and glucose.
There is an increasing recognition that beneficial effects of adipose-derived stem cell (ADSC) therapy may depend largely on the secretion of multiple growth factors. This study modified ADSCs with the Bcl-2 gene in order to increase the secretion of growth factors during oxygen-glucose deprivation (OGD). The phenotypes of human ADSCs that were passaged 4 times were analyzed using flow cytometry. Then, ADSCs were genetically modified with Bcl-2 and Bcl-2 gene transduction was verified with Western blotting. Proliferative capacity and multipotent differentiation properties were evaluated in Bcl-2-modified ADSCs. Secretion of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and basic fibroblast growth factor (bFGF) was evaluated using an enzyme-linked immunosorbent assay (ELISA) during OGD. Human ADSCs that were passaged 4 times expressed stem cell-associated markers but not a fibroblast marker or a hematopoietic stem cell marker. The Bcl-2 gene was efficiently transfected into ADSCs; Bcl-2 modification did not affect the proliferative and multilineage differentiation capacity of ADSCs. In addition, Bcl-2 overexpression enhanced the secretion of VEGF, bFGF, and HGF by 14.47%, 16.9%, and 91%, respectively, compared to ADSCs alone that were deprived of oxygen and glucose. These data suggest that Bcl-2 overexpression enhances secretion of angiogenic growth factors by ADSCs deprived of oxygen and glucose.


Bilateral abnormal anterior bellies of digastric muscles.
During dissection of the submental region of a male cadaver, we encountered an abnormal digastric muscle on both sides. Two muscle bundles, both of which fused with mylohyoid muscle close to the midline, were observed on the right side. The anterior one originated from the digastric fossa and its length and width were 27 and 9 mm, respectively. The posterior accessory belly originated from the right intermediate tendon and it was 32 mm in length and 11 mm wide. On the left side, there was a single accessory bundle that originated from the left intermediate tendon and inserted into the mylohyoid raphe at the fusion point of the right accessory bundles. The length and width of this belly were 29 and 9 mm, respectively. The remaining suprahyoid muscles of both sides were normal. Anatomical variations of digastric muscle have to be considered in the imaging procedures of the soft tissue masses in the submental region.


High-resolution carotid artery MRA. Comparison with fast dynamic acquistion and duplex ultrasound scanning.
To determine whether the diagnostic accuracy of contrast-enhanced MR angiography (CE-MRA) of the carotid arteries is improved by using a slow-injection, high-resolution technique. In 22 patients suspected to have internal carotid artery (ICA) stenosis at duplex ultrasound scanning (DUS), CE-MRA was performed both with a fast, dynamic (8 s/phase) and with a slower, high-resolution technique (scan time 2:20 min). There was conformity between the CE-MRA techniques regarding the degree of stenosis in 34/40 extracranial ICAs. In 3/6 discrepant cases, short occlusions were seen with the fast dynamic technique, whereas both the high-resolution CE-MRA technique and DUS showed patent vessels. There was an overall tendency toward higher stenosis grading with the dynamic technique. Overlying veins could be removed on a workstation in all high-resolution examinations. The high-resolution carotid CE-MRA technique proposed herein seems to improve the diagnostic accuracy, at least for differentiation between high-grade stenoses and occlusions.


What Is New in Amniotic Fluid Embolism?: Best Articles From the Past Year.
This month, we focus on current research in amniotic fluid embolism. Dr. Benson discusses four recent publications, and each is concluded with a "bottom line" that is the take-home message. The complete reference for each can be found in on this page, along with direct links to the abstracts.


Survival of Self-Replicating Molecules under Transient Compartmentalization with Natural Selection.
The problem of the emergence and survival of self-replicating molecules in origin-of-life scenarios is plagued by the error catastrophe, which is usually escaped by considering effects of compartmentalization, as in the stochastic corrector model. By addressing the problem in a simple system composed of a self-replicating molecule (a replicase) and a parasite molecule that needs the replicase for copying itself, we show that transient (rather than permanent) compartmentalization is sufficient to the task. We also exhibit a regime in which the concentrations of the two kinds of molecules undergo sustained oscillations. Our model should be relevant not only for origin-of-life scenarios but also for describing directed evolution experiments, which increasingly rely on transient compartmentalization with pooling and natural selection.


Phrenic nerve paralysis as the initial presentation in pleural sarcomatoid mesothelioma.
A 74-year-old man was referred to our hospital because of persistent cough. A chest radiograph revealed an elevation of the right diaphragm. Computed tomography (CT) images revealed a small nodule localized on the right mediastinum. Five months later, the nodule had grown and was diagnosed as malignant pleural mesothelioma (MPM) by a CT-guided needle biopsy. The patient underwent combined chemotherapy, but the disease progressed rapidly and he passed away. On autopsy, microscopic findings and immunohistological examinations supported the diagnosis of sarcomatoid mesothelioma. Therefore, we diagnosed this rare case as localized sarcomatoid MPM showing phrenic nerve paralysis as an initial presentation.


Effect of photobiomodulation on mitochondrial dynamics in peripheral nervous system in streptozotocin-induced type 1 diabetes in rats.
There is no effective treatment to halt peripheral nervous system damage in diabetic peripheral neuropathy. Mitochondria have been at the center of discussions as important factors in the development of neuropathy in diabetes. Photobiomodulation has been gaining clinical acceptance as it shows beneficial effects on a variety of nervous system disorders. In this study, the effects of photobiomodulation (904 nm, 45 mW, 6.23 J/cm2, 0.13 cm2, 60 ns pulsed time) on mitochondrial dynamics were evaluated in an adult male rat experimental model of streptozotocin-induced type 1 diabetes. Results presented here indicate that photobiomodulation could have an important role in preventing or reversing mitochondrial dynamics dysfunction in the course of peripheral nervous system damage in diabetic peripheral neuropathy. Photobiomodulation showed its effects on modulating the protein expression of mitofusin 2 and dynamin-related protein 1 in the sciatic nerve and in the dorsal root ganglia neurons of streptozotocin-induced type 1 diabetes in rats.


The genetics and neuropathology of Parkinson's disease.
There has been tremendous progress toward understanding the genetic basis of Parkinson's disease and related movement disorders. We summarize the genetic, clinical and pathological findings of autosomal dominant disease linked to mutations in SNCA, LRRK2, ATXN2, ATXN3, MAPT, GCH1, DCTN1 and VPS35. We then discuss the identification of mutations in PARK2, PARK7, PINK1, ATP13A2, FBXO7, PANK2 and PLA2G6 genes. In particular we discuss the clinical and pathological characterization of these forms of disease, where neuropathology has been important in the likely coalescence of pathways highly relevant to typical PD. In addition to the identification of the causes of monogenic forms of PD, significant progress has been made in defining genetic risk loci for PD; we discuss these here, including both risk variants at LRRK2 and GBA, in addition to discussing the results of recent genome-wide association studies and their implications for PD. Finally, we discuss the likely path of genetic discovery in PD over the coming period and the implications of these findings from a clinical and etiological perspective.


Proximal contact areas of the maxillary anterior dentition.
The goal of this study was to quantify the apicoincisal extent of the proximal contact area (PCA) between the eight maxillary anterior teeth. A total of 140 PCA sites and 160 crown lengths were measured in 20 healthy patients. The percentage ratio of PCA to clinical crown length was computed and defined as the proximal contact area proportion (PCAP). Mean PCA dimensions between central incisors (CI/CI), central and lateral incisors (CI/LI), lateral incisors and canines (LI/CA), and canines and first premolars (CA/PM) were 4.2, 2.9, 2.0, and 1.5 mm, respectively. Mesial mean PCAPs were 41%, 32%, 20%, and 18%, respectively. The paired sample t test demonstrated significant differences between all PCAs (P < .0001), except for CA/PM sites (P = .24). Contact areas, not contact points, were observed between neighboring maxillary anterior teeth. Natural PCAPs emerged as well defined in the maxillary anterior dentition bilaterally. Therefore, PCAPs should be taken into consideration for clinical anterior restorations since they determine the papillary and incisal embrasures.


Workplace safety and health programs, practices, and conditions in auto collision repair businesses.
This article describes the results of a pre-intervention safety assessment conducted in 49 auto collision repair businesses and owners' commitments to specific improvements. A 92-item standardized audit tool employed interviews, record reviews, and observations to assess safety and health programs, training, and workplace conditions. Owners were asked to improve at least one-third of incorrect, deficient, or missing (not in compliance with regulations or not meeting best practice) items, of which a majority were critical or highly important for ensuring workplace safety. Two-thirds of all items were present, with the highest fraction related to electrical safety, machine safety, and lockout/tagout. One-half of shops did not have written safety programs and had not conducted recent training. Many had deficiencies in respiratory protection programs and practices. Thirteen businesses with a current or past relationship with a safety consultant had a significantly higher fraction of correct items, in particular related to safety programs, up-to-date training, paint booth and mixing room conditions, electrical safety, and respiratory protection. Owners selected an average of 58% of recommended improvements; they were most likely to select items related to employee Right-to-Know training, emergency exits, fire extinguishers, and respiratory protection. They were least likely to say they would improve written safety programs, stop routine spraying outside the booth, or provide adequate fire protection for spray areas outside the booth. These baseline results suggest that it may be possible to bring about workplace improvements using targeted assistance from occupational health and safety professionals.


Shedding of collagen XVII/BP180: structural motifs influence cleavage from cell surface.
Collagen XVII/BP180, an epithelial adhesion molecule, belongs to the group of collagenous transmembrane proteins, which are characterized by ectodomain shedding. We recently showed that ADAMs can cleave collagen XVII, but also that furin participates in this process (Franzke, C. W., Tasanen, K., Schäcke, H., Zhou, Z., Tryggvason, K., Mauch, C., Zigrino, P., Sunnarborg, S., Lee, D. C., Fahrenholz, F., and Bruckner-Tuderman, L. (2002) EMBO J. 21, 5026-5035). To define the cleavage region in the juxtamembranous NC16A linker domain and assess its structure and requirements for shedding, we constructed deletion mutants of the NC16A domain, expressed them in COS-7 cells, and analyzed their structural integrity and shedding behavior. A mutant lacking the furin consensus sequence was shed in a normal manner, demonstrating that furin does not cleave collagen XVII but rather activates ADAMs (a disintegrin and metalloproteinase). Large deletions of the NC16A domain prevented shedding, and analysis of defined smaller deletions pointed to the stretch of amino acid residues 528-547 as important for sheddase recognition and cleavage. Secondary protein structure predictions showed that deletion of this stretch resulted in an NC16A domain with a positive net charge and an amphipathic alpha-helix, which can cause conformational changes in the collagen XVII homotrimer. Assessment of triple-helix folding of the mutants revealed a lower thermal stability of all non-shed variants than of wild-type collagen XVII or the shed mutants. In contrast, deletion of the putative nucleation site for triple-helix folding of collagenous transmembrane proteins did not affect folding of collagen XVII. The data indicate that the conformation of the NC16A domain and steric availability of the cleavage site influence shedding and is important for folding of collagen XVII.


Pleistocene glacial cycles drove lineage diversification and fusion in the Yosemite toad (Anaxyrus canorus).
Species endemic to alpine environments can evolve via steep ecological selection gradients between lowland and upland environments. Additionally, many alpine environments have faced repeated glacial episodes over the past two million years, fracturing these endemics into isolated populations. In this "glacial pulse" model of alpine diversification, cycles of allopatry and ecologically divergent glacial refugia play a role in generating biodiversity, including novel admixed ("fused") lineages. We tested for patterns of glacial pulse lineage diversification in the Yosemite toad (Anaxyrus [Bufo] canorus), an alpine endemic tied to glacially influenced meadow environments. Using double-digest RADseq on populations densely sampled from a portion of the species range, we identified nine distinct lineages with divergence times ranging from 18 to 724 thousand years ago (ka), coinciding with multiple Sierra Nevada glacial events. Three lineages have admixed origins, and demographic models suggest these fused lineages have persisted throughout past glacial cycles. Directionality indices supported the hypothesis that some lineages recolonized Yosemite from east of the ice sheet, whereas other lineages remained in western refugia. Finally, refugial niche reconstructions suggest that low- and high-elevation lineages have convergently adapted to similar climatic niches. Our results suggest glacial cycles and refugia may be important crucibles of adaptive diversity across deep evolutionary time.


Accumulation of albumin-linked and free-form pentosidine in the circulation of uremic patients with end-stage renal failure: renal implications in the pathophysiology of pentosidine.
Pentosidine is an advanced glycation end product and its formation is shown to be closely related to oxidative processes. Recent studies have shown that pentosidine levels are increased not only in plasma and matrix proteins from diabetic patients, but also markedly in nondiabetic hemodialysis patients. Currently, the mechanism of accumulation and kinetics of pentosidine formation in hemodialysis patients remain unknown. Gel filtration of uremic plasma revealed that plasma pentosidine exists in the albumin fraction (approximately 90%) and, interestingly, in free form (approximately 5%) as well. Plasma free pentosidine was undetectable in subjects with normal renal function. There was a significant correlation between the plasma levels of albumin-linked and free pentosidine in hemodialysis patients. Kinetic studies indicated that dietary pentosidine was absorbed into the circulation and that, after either oral or intravenous administration of pentosidine to intact or nephrectomized rats, the plasma free pentosidine level was closely linked to the level of renal function. These findings demonstrate that: (1) Pentosidine accumulates as albumin-linked and in free form in the circulation of uremic patients; (2) dietary pentosidine can be absorbed into the circulation, thus being one possible origin of circulating free pentosidine; (3) free pentosidine may accumulate as a result of decreased glomerular filtration; and (4) the mechanism of accumulation of albumin-linked pentosidine is not related to high glucose levels. It suggests the simultaneous accumulation, during renal failure, of either unknown pentosidine precursor(s) or catalyst(s) of glycoxidation, independent of glucose.


Dual action of sonic hedgehog on chondrocyte hypertrophy: retrovirus mediated ectopic sonic hedgehog expression in limb bud micromass culture induces novel cartilage nodules that are positive for alkaline phosphatase and type X collagen.
Members of the vertebrate hedgehog gene family (HH) are involved in patterning and modulation of differentiation. Recently it has been shown that ectopic expression of HH gene family members in vivo blocks chondrocyte maturation through activation of a parathyroid hormone related peptide (PTHrP) dependent negative regulatory loop in the perichondrium. However, the direct effect of HH on chondrocyte maturation has not been tested. Here, we studied the effect of retroviral overexpression of the chicken sonic hedgehog gene (Shh) on the growth and maturation of limb bud cells in micromass cultures. Shh is neither expressed nor required for the initiation of cellular condensation in normal micromass cultures. With Shh over-expression, micromass cultures developed novel tightly whorled nodules in addition to the normal Alcian Blue positive cartilage nodules. We characterized the new nodules and showed that they are strongly positive for alkaline phosphatase, enriched in type X collagen and weakly positive for Alcian Blue staining. Shh overexpression also increased cell proliferation, but this cannot account for the formation of the new nodules. This current study shows that misexpression of Shh in in vitro chondrogenic cultures promotes characteristics of hypertrophic chondrocytes. Thus HH has two complementary functions; a direct positive effect on chondrocyte hypertrophy in the absence of PTHrP pathway, and an indirect negative feedback loop through PTHrP to prevent other less differentiated chondrocytes from becoming hypertrophic. These two complementary actions of HH coordinate the progression of cartilage maturation.


Psychosis associated with epilepsy: significance of the laterality of the epileptogenic lesion.
The psychiatric histories in a group of patients who had previously undergone temporal lobectomy for intractable epilepsy, were analysed for the occurrence of psychosis. The data suggest that epileptic patients with left temporal lobe lesions are especially disposed to develop a schizophrenic-like psychosis.


Toxicokinetics of Brominated Azo Dyes in the Early Life Stages of Zebrafish (Danio rerio) Is Prone to Aromatic Substituent Changes.
Brominated azo dyes (BADs) have been identified as predominant indoor brominated pollutants in daycare dust; thus, their potential health risk to children is of concern. However, the toxicities of BADs remain elusive. In this study, the toxicokinetics of two predominant BADs, Disperse Blue 373 (DB373) and Disperse Violet 93 (DV93), and their suspect metabolite 2-bromo-4,6-dinitroaniline (BDNA) was investigated in embryos of zebrafish (Danio rerio). The bioconcentration factor of DV93 at 120 hpf is 6.2-fold lower than that of DB373. The nontarget analysis revealed distinct metabolism routes between DB373 and DV93 by reducing nitro groups to nitroso (DB373) or amine (DV93), despite their similar structures. NAD(P)H quinone oxidoreductase 1 (NQO1) and pyruvate dehydrogenase were predicted as the enzymes responsible for the reduction of DB373 and DV93 by correlating time courses of the metabolites and enzyme development. Further in vitro recombinant enzyme and in vivo inhibition results validated NQO1 as the enzyme specifically reducing DB373, but not DV93. Global proteome profiling revealed that the expression levels of proteins from the "apoptosis-induced DNA fragmentation" pathway were significantly upregulated by all three BADs, supporting the bioactivation of BADs to mutagenic aromatic amines. This study discovered the bioactivation of BADs via distinct eukaryotic enzymes, implying their potential health risks.


Recombinant factor VIIa is effective for bleeding and surgery in patients with Glanzmann thrombasthenia.
Recombinant activated factor VII (rFVIIa) was found to be effective and safe in treating 24 bleeding episodes and to prevent bleeding during one bilateral herniorrhaphy in four children with Glanzmann thrombasthenia. One of the patients had alloantibodies to platelet membrane glycoprotein (GP) IIb/IIIa and was refractory to platelet transfusion. rFVIIa was administered at 89 to 116 microg/kg per injection every 2 hours, in association with antifibrinolytic drugs. Bleeding stopped in all cases, but platelet transfusion was required in one. Two bleeding episodes recurred 36 and 63 hours after discontinuation of rFVIIa, but were successfully treated with additional doses. No adverse effects of rFVIIa were observed. Although the number of patients is small, our study suggests that rFVIIa may be an alternative to platelet transfusions in patients with a severe congenital thrombocytopathy.


Electronic transitions of molecules: vibrating Lewis structures.
Since the conception of the electron pair bond, Lewis structures have been used to illustrate the electronic structure of a molecule in its ground state. But, for excited states, most descriptions rely on the concept of molecular orbitals. In this work we demonstrate a simple and intuitive description of electronic resonances in terms of localized electron vibrations. By partitioning the 3N-dimensional space of a many-electron wavefunction into hyper-regions related by permutation symmetry, chemical structures naturally result which correspond closely to Lewis structures, with identifiable single and double bonds, and lone pairs. Here we demonstrate how this picture of electronic structure develops upon the admixture of electronic wavefunctions, in the spirit of coherent electronic transitions. We show that π-π* transitions correspond to double-bonding electrons oscillating along the bond axis, and n-π* transitions reveal lone-pairs vibrating out of plane. In butadiene and hexatriene, the double-bond oscillations combine with in- and out-of-phase combinations, revealing the correspondence between electronic transitions and molecular normal mode vibrations. This analysis allows electronic excitations to be described by building upon ground state electronic structures, without the need for molecular orbitals.


DBPSK and DQPSK crosstalk in single-span WDM systems using DRA.
A closed-loop formula is derived for the nonlinear crosstalk degradation due to self-phase modulation and cross-phase modulation in a wavelength-division multiplexing system. The crosstalk is investigated in differential binary phase-shift keying (DBPSK) and differential quadrature phase-shift keying (DQPSK) schemes. The system under consideration is standard single-mode fiber with a single-span distributed Raman amplifier. The backward DBPSK shows a better performance with minimum crosstalk performance of 71%, as compared to backward DQPSK.


Herpes simplex virus variants restraint to high concentrations of acyclovir exist in clinical isolates.
Acyclovir (ACV) has been shown to inhibit the replication of herpes simplex virus (HSV) in vitro. We examined a wide variety of HSV clinical isolates for the presence of naturally occurring ACV-resistant (ACVr) variants. Although the ACV doses that inhibited 50% of these isolates were within the range of doses inhibiting 50% of the ACV-susceptible wild-type strains, we successfully isolated variants resistant to high ACV concentrations (25 to 75 microM) from each virion population even in the absence of prior drug exposure. Furthermore, we demonstrated, by fluctuation analysis of two encephalitis strains, that the ACVr variants were clonally distributed in the virus populations before exposure to ACV and did not result from rapid adaptation to ACV. All variants isolated after a single exposure to a high dose of ACV were true ACVr variants, as demonstrated by their plating efficiencies in the presence of ACV. We found that 36 and 50% of the ACVr variants of the two strains examined in detail displayed plating efficiencies in phosphonoacetic acid of greater than 0.1, possibly indicating that many of the ACVr variants contained alterations in the DNA polymerase gene locus. Because the distribution of ACVr variants in natural populations is relatively high (10(-4), these results suggest that selection of ACVr strains during ACV therapy is possible.


Nursing medication administration and workflow using computerized physician order entry.
The benefits of computerized physician order entry systems have been described widely; however, the impact of computerized physician order entry on nursing workflow and its potential for error are unclear. The purpose of this study was to determine the impact of a computerized physician order entry system on nursing workflow. Using an exploratory design, nurses employed on an adult ICU (n = 36) and a general pediatric unit (n = 50) involved in computerized physician order entry-based medication delivery were observed. Nurses were also asked questions regarding the impact of computerized physician order entry on nursing workflow. Observations revealed total time required for administering medications averaged 8.45 minutes in the ICU and 9.93 minutes in the pediatric unit. Several additional steps were required in the process for pediatric patients, including preparing the medications and communicating with patients and family, which resulted in greater time associated with the delivery of medications. Frequent barriers to workflow were noted by nurses across settings, including system issues (ie, inefficient medication reconciliation processes, long order sets requiring more time to determine medication dosage), less frequent interaction between the healthcare team, and greater use of informal communication modes. Areas for nursing workflow improvement include (1) medication reconciliation/order duplication, (2) strategies to improve communication, and (3) evaluation of the impact of computerized physician order entry on practice standards.


Validity and Reliability of Questionnaires That Assess Barriers and Facilitators of Sedentary Behavior in the Pediatric Population: A Systematic Review.
We systematically reviewed the literature about the validity and reliability of barriers and facilitators of sedentary behavior questionnaires for children and adolescents, considering accelerometers as the reference method. We included studies that assessed the agreement between the barriers and facilitators of sedentary behavior through a questionnaire and an objective measure (e.g., accelerometry). We searched four electronic databases (MEDLINE/PubMed, CINAHL, Web of Science, and SCOPUS): these databases were searched for records from inception to 5 March 2021, and updated to November 2022. The search strategy used the following descriptors: children and adolescents; barriers or facilitators; questionnaires; accelerometers; and validation or reliability coefficient. Studies identified in the search were selected independently by two reviewers. The inclusion criteria were: (i) population of children and adolescents, (ii) original studies, (iii) subjective and objective measurement methods, (iv) studies that report validity or reliability, and (v) population without specific diseases. Seven studies were eligible for our review. The main exclusion reasons were studies that did not report validity or reliability coefficients (56.6%) and non-original studies (14.5%). The participants' ages in the primary studies ranged from 2 to 18 years. Cronbach's alpha coefficient was the most reported reliability assessment among the eligible articles, while Pearson and Spearman's coefficients were prevalent for validity. The reliability of self-report questionnaires for assessing sedentary behavior ranged from r = 0.3 to 1.0. The validity of the accelerometers ranged from r = -0.1 to 0.9. Family environment was the main factor associated with sedentary behavior. Our findings suggest that questionnaires assessing the barriers and facilitators of sedentary behavior are weak to moderate. PROSPERO Registration (CRD42021233945).


Diffuse large B-cell lymphoma arising in a composite lymphoma with biclonality by flow cytometry and one monoclonal band by PCR.
Composite lymphoma (CL) refers to the presence of two or more distinct types of lymphomas in a single organ or tissue. CL is an infrequent finding and may be due to the existence of two genetically related neoplasms, i.e. transformation of a single lymphoma into another lymphoma, or be due to the presence of two clonally unrelated lymphomas. CL composed of more than two lymphomas is even rare. Herein we describe a case of diffuse large B-cell lymphoma (DLBCL) arising in a CL of follicular lymphoma (FL) and small lymphocytic lymphoma (SLL) in an inguinal lymph node of an 85 year old woman. The three lymphomas were morphologically and immunophenotypically distinct while flow cytometry detected two monoclonal B-cell populations. Karyotyping and Polymerase Chain Reaction (PCR) for B-cell clonality each detected a single monoclonal B-cell population. The morphology findings may suggest DLBCL being transformed from FL while Richter transformation from SLL appears to be less likely in our case. Due to the single clone by chromosome study and PCR study, the precise relationships of the three lymphomas are unknown.


Genomic dissection of the cytokine-controlled STAT5 signaling network in liver.
Growth hormone (GH) controls the physiology and pathophysiology of the liver, and its signals are conducted by two members of the family of signal transducers and activators of transcription, STAT5A and STAT5B. Mice in which the Stat5a/b locus has been inactivated specifically in hepatocytes display GH resistance, the sex-specific expression of genes associated with liver metabolism and the cytochrome P-450 system is lost, and they develop hepatosteatosis. Several groups have shown by global gene expression profiling that a cadre of STAT5A/B target genes identify genetic cascades induced by GH and other cytokines. Evidence is accumulating that in the absence of STAT5A/B GH aberrantly activates STAT1 and STAT3 and their downstream target genes and thereby offers a partial explanation of some of the physiological alterations observed in Stat5a/b-null mice and human patients. We hypothesize that phenotypic changes observed in the absence of STAT5A/B are due to two distinct molecular consequences: first, the failure of STAT5A/B target genes to be activated by GH and second, the rerouting of GH signaling to other members of the STAT family. Rerouting of GH signaling to STAT1 and STAT3 might partially compensate for the loss of STAT5A/B, but it certainly activates biological programs distinct from STAT5A/B. Here we discuss the extent to which studies on global gene expression profiling have fostered a better understanding of the biology behind cytokine-STAT5A/B networks in hepatocytes. We also explore whether this wealth of information on gene activity can be used to further understand the roles of cytokines in liver disease.


Multi-glycoside of Tripterygium wilfordii Hook. f. attenuates glomerulosclerosis in a rat model of diabetic nephropathy by exerting anti-microinflammatory effects without affecting hyperglycemia.
Multi-glycoside of Tripterygium wilfordii Hook. f. (GTW) has been proven to be clinically effective in relieving microinflammation in patients with early diabetic nephropathy (DN). However, the therapeutic mechanisms involved in vivo remain unclear. In the process of early DN, microinflammation and activation of p38 mitogen‑activated protein kinase (MAPK) and canonical nuclear factor (NF)-κB signaling pathways are the important mechanisms by which hyperglycemia contributes to glomerulosclerosis (GS). Therefore, this study aimed to examine the ameliorative effects of GTW on GS, and then to clarify its anti‑microinflammatory mechanisms by inhibiting p38 MAPK and NF-κB signaling activities in the kidney. All rats were divided into 4 groups: the sham group, the sham + GTW group, the vehicle group and the GTW group. The suitable dose of GTW and vehicle were daily administered for 8 weeks after the induction of DN by unilateral nephrectomy combined with intraperitoneal injections of streptozotocin (STZ). The general status of the rats, biochemical parameters, renal histological changes and macrophages in glomeruli, as well as expression of the key proteins in the p38 MAPK and canonical NF-κB signaling pathways and inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-1β and transforming growth factor (TGF)-β1 in the kidney were examined, respectively. The results revealed that, GTW improved the general cond-ition and biochemical parameters of the rats, but did not lower blood glucose; GTW attenuated GS and suppressed glomerular microinflammation including the infiltration of ED1+ cells in glomeruli and the protein overexpression of TNF-α, IL-1β and TGF-β1 in the kidney; GTW inhibited the protein overexpression of key signaling molecules of p38 MAPK and canonical NF-κB pathways in the kidney including phosphorylated p38 MAPK, phosphorylated inhibitor protein IκB and NF-κB (p65). On the whole, we expounded that GTW, as a natural regulator in vivo, alleviates GS without affecting hyperglycemia, by exerting anti-microinflammatory effects, including reducing macrophage infiltration in glomeruli, suppressing TNF-α, IL-1β and TGF-β1 overexpression in the kidney and inhibiting p38 MAPK and NF-κB signaling activities.


Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer.
The scavenger receptor, CD163, is a macrophage-specific marker. Recent studies have shown that CD163 expression in breast and rectal cancer cells is associated with poor prognosis. This study was conducted to evaluate the relationship between CD163 expression as a macrophage trait in cancer cells, and macrophage infiltration and its clinical significance in colorectal cancer. Immunostaining of CD163 and macrophage infiltration were evaluated in paraffin-embedded specimens, earlier analyzed for CD31, D2-40 and S-phase fraction, from primary tumors and normal colorectal mucosa of 75 patients with colorectal carcinoma. The outcomes were analyzed in relation to clinical-pathological data. CD163 expression was positive in cancer cells in 20 % of colorectal cancer patients and was related to advanced tumor stages (P = 0.008) and unfavorable prognosis (p = 0.001). High macrophage infiltration was related to shorter survival and positive CD163 expression in tumor cells. The prognostic impact of macrophage infiltration was independent of tumor stage and CD163 expression in cancer cells (p = 0.034). The expression of macrophage phenotype in colorectal cancer cells is associated with macrophage density in tumor stroma and lower survival rates. Macrophage infiltration has an independent prognostic impact on mortality in colorectal cancer. In accordance with previous experimental studies, these findings provide new insights into the role of macrophages in colorectal cancer.


Surface modification of dense membranes using radical graft polymerization enhanced by monomer filtration.
Surface graft polymerization is a promising way to modify membranes for improved performance. Redox-initiated graft polymerization of vinyl monomers is a facile and inexpensive method carried out at room temperature in aqueous media; however, its use is often limited by slow kinetics, low surface specificity, and excessive consumption of chemicals on undesired homopolymerization. It is shown that in the case of RO or NF membranes these drawbacks may be eliminated by utilizing the selectivity of the membranes toward monomers and carrying out the polymerization while applying pressure, i.e., under filtration conditions. Concentration polarization that ensues raises the concentration of reagents near the membrane surface and thereby drastically increases the rate of reaction and preferentially directs it towards surface grafting. Grafting experiments using 2-hydroxyethyl methacrylate and other monomers and characterization of modified membranes using permeability measurements, ATR-FTIR, AFM, XPS, and contact angle demonstrate that the required monomer concentrations can be drastically reduced, particularly when a small fraction of a cross-linker is added. As an additional benefit, this approach enables broadening the spectrum of utilizable monomers to sparingly soluble hydrophobic, charged, and macro-monomers, as was demonstrated using sparingly soluble ethyl methacrylate and 2-ethoxyethyl methacrylate and other monomers. Even though the kinetics of the process is substantially complicated by evolution and concentration polarization of oligomeric and polymeric species, especially in the presence of a cross-linker, it is well offset by the benefits of higher rate, specificity, and reduced monomer consumption.


Diffusion tensor imaging of the kidney with parallel imaging: initial clinical experience.
To evaluate the clinical feasibility of diffusion tensor imaging (DTI) of the kidney in volunteers and patients with renal diseases. Ten volunteers and 22 patients (mean age, 56 +/- 14.3) with renal masses and renal artery stenosis underwent breath-hold coronal fat-saturated echo-planar DTI (as provided by the manufacturer, 6 diffusion directions, diffusion weightings b = 0 and 300 s/mm2, repetition time 730 ms/echo time 72 ms; 5 slices; slice thickness, 6 mm; inplane resolution, 2.1 x 2.1 mm2; acquisition time, 26 seconds) of the kidneys at 1.5 T (MAGNETOM Avanto, Siemens Medical Solutions, Erlangen, Germany). The parallel imaging technique, generalized autocalibrating partially parallel acquisitions with an acceleration factor 2, was applied. Using the commercially available Syngo DTI task card software, regions of interests were placed in the cortex, medulla, and in renal masses if present. Fractional anisotropy (FA) and apparent diffusion coefficients (ADC) were determined, and tractography was used to visualize the renal diffusion properties. Statistical analysis was performed using the Wilcoxon signed-rank sum test and paired t tests. In all volunteers, FA was significantly (P < 0.01) higher in the medulla (0.36 +/- 0.03) than in the cortex (0.21 +/- 0.02), whereas the ADC was significantly (P < 0.01) higher in the cortex (2.43 +/- 0.19) than in the medulla (2.16 +/- 0.22). Tractography typically revealed a radial preferred direction of medullary diffusion basically reflecting medullary flow.FA/ADC of simple renal cysts (n = 8) was 0.14 +/- 0.05/2.86 +/- 0.15. Renal cell carcinoma (n = 10) showed a wide FA range from 0.11 to 0.56. Using tractography, the structural organization of renal cell carcinoma such as pseudocapsules could be visualized.In 1 patient with unilateral high-grade renal artery stenosis, the cortical ADC of the affected kidney was lower than on the contralateral side (1.77/2.27) and the FA was increased (0.33/0.18). The FA of the medulla was increased (0.70/0.41) and the ADC decreased (1.43/1.90). Using parallel imaging, DTI measurements of the kidneys are feasible within a single breath-hold with good discrimination between cortex and medulla. Parallel imaging allows more slices and a superior resolution. DTI measurements of the kidney allows visualization of medullary flow, in pathology ADC and FA were altered. Further investigations will be required to evaluate the role of DTI for studying and monitoring renal ultrastructure.


Sustained Survival Benefit in Recurrent Medulloblastoma by a Metronomic Antiangiogenic Regimen: A Nonrandomized Controlled Trial.
Medulloblastoma recurrence in patients who have previously received irradiation has a dismal prognosis and lacks a standard salvage regimen. To evaluate the response rate of pediatric patients with medulloblastoma recurrence using an antiangiogenic metronomic combinatorial approach (Medulloblastoma European Multitarget Metronomic Anti-Angiogenic Trial [MEMMAT]). This phase 2, investigator-initiated, multicenter nonrandomized controlled trial assessed 40 patients with relapsed or refractory medulloblastoma without a ventriculoperitoneal shunt who were younger than 20 years at original diagnosis. Patients were enrolled between April 1, 2014, and March 31, 2021. Treatment consisted of daily oral thalidomide, fenofibrate, celecoxib, and alternating 21-day cycles of low-dose (metronomic) oral etoposide and cyclophosphamide, supplemented by intravenous bevacizumab and intraventricular therapy consisting of alternating etoposide and cytarabine. The primary end point was response after 6 months of antiangiogenic metronomic therapy. Secondary end points included progression-free survival (PFS), overall survival (OS), and quality of life. Adverse events were monitored to assess safety. Of the 40 patients (median [range] age at treatment start, 10 [4-17] years; 25 [62.5%] male) prospectively enrolled, 23 (57.5%) achieved disease control after 6 months of treatment, with a response detected in 18 patients (45.0%). Median OS was 25.5 months (range, 10.9-40.0 months), and median PFS was 8.5 months (range, 1.7-15.4 months). Mean (SD) PFS at both 3 and 5 years was 24.6% (7.9%), while mean (SD) OS at 3 and 5 years was 43.6% (8.5%) and 22.6% (8.8%), respectively. No significant differences in PFS or OS were evident based on molecular subgroup analysis or the number of prior recurrences. In patients demonstrating a response, mean (SD) overall 5-year PFS was 49.7% (14.3%), and for patients who remained progression free for the first 12 months of treatment, mean (SD) 5-year PFS was 66.7% (16.1%). Treatment was generally well tolerated. Grade 3 to 4 treatment-related adverse events included myelosuppression, infections, seizures, and headaches. One heavily pretreated patient with a third recurrence died of secondary acute myeloid leukemia. This feasible and well-tolerated MEMMAT combination regimen demonstrated promising activity in patients with previously irradiated recurrent medulloblastoma. Given these results, this predominantly oral, well-tolerated, and outpatient treatment warrants further evaluation. ClinicalTrials.gov Identifier: NCT01356290.


Time Pressure and In-group Favoritism in a Minimal Group Paradigm.
Based on the group heuristic model and the model of intuitive cooperation, we hypothesized that in-group favoritism would be conspicuously shown through an intuitive process. To test this hypothesis, we utilized a minimal group paradigm, which is traditionally used in social psychological studies, and manipulated decision time in a one-shot prisoner's dilemma game to compare the cooperative contribution level toward in-group and out-group members under three conditions: intuitive, empathic deliberation, and rational deliberation. Our findings confirmed that in-group favoritism was clearly shown in the intuitive condition only, suggesting that the intuitive cooperation model may only be valid in the context of social exchange with in-group members. Additional analysis also showed that in-group favoritism disappeared for participants who had been forced into empathic or rational deliberation for decision making. The theoretical implications of the findings are discussed.


Long-term multicentre experience of adjuvant radiotherapy for pN3 squamous cell carcinoma of the penis.
To present the long-term adjuvant radiotherapy outcomes of patients with pN3 squamous cell carcinoma of the penis (SCCp) treated at two UK centres. We conducted a retrospective audit of all pN3 SCCp patients, deemed suitable for adjuvant therapy by a specialist multidisciplinary team at St George's and Leeds Hospitals, who received adjuvant radiotherapy. Primary outcomes were recurrence-free survival (RFS), cancer-specific survival (CSS) and overall survival (OS). Secondary outcomes were time to adjuvant treatment, frequency of in-field recurrence, site and side of recurrence, and dose and schedule of radiotherapy. A total of 146 patients were included: 121 completed radiotherapy, 4 did not complete radiotherapy and 21 did not start it. The median (interquartile range [IQR]) age was 59 (54-70)years. The 5-year RFS was 51%, CSS was 51% and OS was 44%. Adjuvant radiotherapy was started at a median (IQR) of 75 (48-106) days. A dose of 45 Gy in 20 fractions was most commonly used. Of the 125 patients who started adjuvant treatment, 55 relapsed. Of these relapses, 30 occurred in an inguinal or pelvic nodal station and 26 of the 30 were in a radiation field. Relapses in 18 of the 55 cases were in visceral sites only and seven were in both nodal (non-irradiated sites) and visceral sites. Doses of <50 Gy were used more commonly before 2013 and higher doses (>50 Gy) were more commonly used after 2013. Application of a standard radiotherapy protocol within a centralized supra-network setting has achieved survival outcomes that would appear better than those previously documented for either radiotherapy or chemotherapy in a cohort with solely pN3 disease. The addition of adjuvant chemotherapy may improve these outcomes further. These data suggest that adjuvant radiotherapy has a role to play in the management of men with pN3 SCCp.


Tools for primary care management of inflammatory bowel disease: do they exist?
Healthcare systems throughout the world continue to face emerging challenges associated with chronic disease management. Due to the likely increase in chronic conditions in the future it is now vital that cooperation and support between specialists, generalists and primary health care physicians is conducted. Inflammatory bowel disease (IBD) is one such chronic disease. Despite specialist care being essential, much IBD care could and probably should be delivered in primary care with continued collaboration between all stakeholders. Whilst most primary care physicians only have few patients currently affected by IBD in their caseload, the proportion of patients with IBD-related healthcare issues cared for in the primary care setting appears to be widespread. Data suggests however, that primary care physician's IBD knowledge and comfort in management is suboptimal. Current treatment guidelines for IBD are helpful but they are not designed for the primary care setting. Few non-expert IBD management tools or guidelines exist compared with those used for other chronic diseases such as asthma and scant data have been published regarding the usefulness of such tools including IBD action plans and associated supportive literature. The purpose of this review is to investigate what non-specialist tools, action plans or guidelines for IBD are published in readily searchable medical literature and compare these to those which exist for other chronic conditions.


A selective ER-phagy exerts procollagen quality control via a Calnexin-FAM134B complex.
Autophagy is a cytosolic quality control process that recognizes substrates through receptor-mediated mechanisms. Procollagens, the most abundant gene products in Metazoa, are synthesized in the endoplasmic reticulum (ER), and a fraction that fails to attain the native structure is cleared by autophagy. However, how autophagy selectively recognizes misfolded procollagens in the ER lumen is still unknown. We performed siRNA interference, CRISPR-Cas9 or knockout-mediated gene deletion of candidate autophagy and ER proteins in collagen producing cells. We found that the ER-resident lectin chaperone Calnexin (CANX) and the ER-phagy receptor FAM134B are required for autophagy-mediated quality control of endogenous procollagens. Mechanistically, CANX acts as co-receptor that recognizes ER luminal misfolded procollagens and interacts with the ER-phagy receptor FAM134B. In turn, FAM134B binds the autophagosome membrane-associated protein LC3 and delivers a portion of ER containing both CANX and procollagen to the lysosome for degradation. Thus, a crosstalk between the ER quality control machinery and the autophagy pathway selectively disposes of proteasome-resistant misfolded clients from the ER.


The student journey: Living and learning following traumatic brain injury.
Primary objective: In this research we set out to gain further understanding of the experiences of students participating in secondary and tertiary education following TBI: exploring academic and non-academic factors, as well as changes in experiences over time.Methods and procedures: A longitudinal, qualitative investigation was completed. 12 students (17-32 years) completed up to three in-depth interviews over a period of 4-15 months, capturing atotal of 30 time points. Data were analyzed using grounded theory methods.Main outcomes and results: Students' participation experiences were unique and varied with different timelines and outcomes, however they shared many similar critical points. We interpreted their experiences as a student journey traveling through four significant landscapes, "Choosing to study", "Studying", "Deciding what to do", "Making the next step." Journeys involved complex processes of living and learning. Moving along the pathway was not always smooth or straightforward.Conclusions: Students' experiences of returning to study following TBI can be interpreted as a complex journey of living and learning. Four important stages of the journey provide clinicians and educators with landscape features that can provide a structure for exploring supports to address both academic and non-academic factors to assist students in their study journey.


The performance of magnetic resonance imaging in early cervical carcinoma: a long-term experience.
We report our long-term experience of performance of magnetic resonance imaging (MRI) in localizing cervical tumor, assessing tumor size, staging, and lymph node infiltration in patients with early cervical cancer. MRI of 150 patients with early carcinoma between 1995-2005 was retrospectively reviewed. Tumor location, size, tumor distance from internal os, parametrial invasion, myometrial invasion, lymph node size, and location were documented. All patients underwent surgery, pelvic lymphadenectomy, and histological correlation of MRI findings. For staging, MRI and histopathology had kappa value of 0.89. For parametrial invasion, MRI had specificity, negative predictive value (NPV) of 97% and 100%, respectively. For tumor size, MRI and histology had mean difference of -0.9 mm with 95% limits of agreement between -12.6 to +13 mm. In tumors greater than 10 mm, mean difference was 0.3 mm and limits of agreement were -7.5 to +7.9 mm. For internal os involvement sensitivity, specificity, positive predictive value (PPV) and NPV were 90%, 98%, 86%, 98%. respectively. For myometrial invasion sensitivity, specificity, NPV, and PPV were 100%, 99%, 88%, 100%, respectively. Incidence of nodal metastases was 2.9%. On a per-patient basis, sensitivity, specificity for nodal involvement was 37% and 92% and on node-by-node basis, sensitivity and specificity of MRI was 27% and 99%, respectively. Our study confirms MRI is highly accurate in localizing cervical tumor, excluding parametrial invasion, confirming myometrial and internal os invasion. MRI is therefore useful in selecting patients for surgery and mandatory in patients for fertility-preserving surgery. Using accepted size criteria for nodal involvement, MRI is insensitive and currently will not avoid need for pelvic lymphadenectomy.


[Investigation of subalin effect on urogenital microflora of pregnant women].
Bacteriological examination of 30 women with incomplete pregnancy has been conducted. Representatives of 18 genera of aerobic and anaerobic bacteria as well as yeast-like fungi of Candida genus were found. It has been established that spore-forming bacteria of the Bacillus genus from subalin preparation have antagonistic activity in respect of the strains of pathogenic and opportunistic bacteria with characteristic zones of growth depression of 12-15 mm. This demonstrates that the agents of infectional urogenital process are effectively suppressed by subalin.


Electrophysiology of GABA-mediated synaptic transmission and possible roles in epilepsy.
Epileptogenic conditions come about from a disequilibrium between excitatory and inhibitory mechanisms, creating a state of neuronal hypersynchrony. From experimental studies in animal models of epilepsy it appears that several mechanisms, alone or in combination, could be responsible for this imbalance. An alteration of GABA-mediated inhibition has long been considered to be one of the most likely candidates. We review recent data on the synaptic physiology of GABA-mediated inhibition, with emphasis on GABAA and GABAB receptors and their conductances. We describe the integrative role of GABAergic local-circuit neurons in the normal control of recurrent excitation. We then discuss possible alterations in GABAA-mediated inhibition in two chronic animal models of epilepsy, the kindled rat and the kainate-treated rat. Finally, we review studies on GABA inhibition in human epileptic cortex resected for the treatment of intractable epilepsy.


Centrosomal aggregates and Golgi fragmentation disrupt vesicular trafficking of DAT.
Lewy bodies containing the centrosomal protein γ-tubulin and fragmentation of Golgi apparatus (GA) have been described in nigral neurons of Parkinson's disease (PD) patients. However, the relevance of these features in PD pathophysiology remains unknown. We analyzed the impact of proteasome inhibition in the formation of γ-tubulin-containing aggregates as well as on GA structure. SH-SY5Y cells were treated with the proteasome inhibitor Z-Leu-Leu-Leu-al (MG132) to induce centrosomal-protein aggregates. Then, microtubules (MTs) and Golgi dynamics, as well as the vesicular transport of dopamine transporter (DAT) were evaluated both in vitro and in living cells. MG132 treatment induced γ-tubulin aggregates which altered microtubule nucleation. MG132-treated cells containing γ-tubulin aggregates showed fragmentation of GA and perturbation of the trans-Golgi network. Under these conditions, the DAT accumulated at the centrosomal-Golgi region indicating that the vesicular transport of DAT was disrupted. Thus, centrosomal aggregates and fragmentation of GA are 2 closely related processes that could result in the disruption of the vesicular transport of DAT toward the plasma membrane in a model of dopaminergic neuronal degeneration.


Bolus magnesium infusion in humans is associated with predominantly unfavourable changes in platelet aggregation and certain haemostatic factors.
The purpose of this study was an attempt to extrapolate favorable observations on the effects of magnesium on platelets and haemostasis from animal models to humans. Intravenous magnesium in the treatment of acute myocardial infarction has been tested in several large clinical trials and remains controversial. The mechanism for the cardioprotective properties of magnesium is unknown. Based on experimental studies, several different hypotheses have been advanced to explain the benefits of magnesium including it's effects on platelets and haemostasis. However, few studies have analysed such a relationship in humans. This project was designed to assess the effect of bolus magnesium infusion on ex vivo platelet aggregation and certain haemostatic parameters in healthy volunteers. One gram of magnesium was diluted in 50 ml of D5W and infused over a period of 15 min. The changes of platelet aggregability, plasma antithrombin-III, Protein C, total Protein S, fibronectin, endothelin-1, as well as the metabolites of thromboxane and prostacyclin were determined prior to and immediately after magnesium infusion. Serum magnesium concentration increased from 2.10 +/- 0.08 at baseline to 3.12 +/- 0.13 mg dl-1 (P = 0.0002) post-infusion. Magnesium infusion was associated with a significant elevation in ADP (+19.3%); ristocetin (+13.6%); and collagen-induced platelet aggregation (+14.2%); an increase in plasma antithrombin-III (+10.3%) and thromboxane (+49.4%) when compared to pre-infusion levels. Against this, total Protein S (-20.7%), Protein C (-11.2%) and ET-1 (-26.3%) plasma concentrations were markedly decreased. There were no significant differences in plasma fibronectin and prostacyclin levels. Contrary to expectations, magnesium infusion in human volunteers is associated with platelet activation and mostly small unfavourable changes in certain haemostatic factors. However, the observed changes were small and for the most part remained within the normal physiologic range.


Combined chemo-immunotherapy as a prospective strategy to combat cancer: a nanoparticle based approach.
The prime objective of this study was to develop a combined chemo-immunotherapeutic formulation which could directly kill cancer cells as well as activate the immunosuppressed tumor microenvironment to mount a robust antitumor immune response. Paclitaxel (PTX) and SP-LPS (nontoxic derivative of lipopolysaccharide) were selected as anticancer drug and immunostimulant respectively. Poly(lactic-co-glycolic acid) (PLGA) based PTX and SP-LPS containing nanoparticles (TLNP) were prepared by the double-emulsion method (w/o/w) and characterized in terms of size, zeta potential and transmission electron microscopy (TEM). The release behavior of PTX and SP-LPS from the TLNP exhibited a biphasic pattern characterized by an initial burst followed by slow continuous release. In vitro anticancer activity of TLNP was found to be higher compared to PTX when studied in a tumor cell-splenocyte coculture system. TLNP activated murine monocytes induced the secretion of various proinflammatory cytokines. After iv administration of TLNP in tumor bearing C57BL/6 mice, the amount of PTX in the tumor mass was found to be higher in TLNP treated mice as compared to commercial Taxol group at all time points studied. In vitro studies suggest that nanoparticles containing PTX and SP-LPS have both direct cytotoxicity and immunostimulatory activity. Hence this might have potential as a chemo-immunotherapeutic formulation against cancer with advantage over present day chemotherapy with Taxol, in terms of tumor targeting, less toxicity and immunostimulation.


Oblimersen: Augmerosen, BCL-2 antisense oligonucleotide - Genta, G 3139, GC 3139, oblimersen sodium.
Oblimersen is an antisense oligonucleotide developed by Genta for systemic use as an injection. It comprises a phosphorothioate backbone linking 18 modified DNA bases. Oblimersen targets the first six codons of Bcl-2 mRNA to form a DNA/RNA complex. The duplex is subsequently recognised as a foreign message and is cleaved enzymatically, thereby destroying the Bcl-2 message. The Bcl-2 protein, which is a potent inhibitor of apoptosis, is overexpressed in many cancers, including follicular lymphomas, breast, colon and prostate cancers, and intermediate-/high-grade lymphomas. By reducing the amount of Bcl-2 protein in cancer cells, oblimersen may enhance the effectiveness of conventional anticancer treatments. Genta has reported results from randomised phase III trials of oblimersen in four different indications: malignant melanoma, chronic lymphocytic leukaemia (CLL), multiple myeloma and acute myleoid leukaemia (AML). A negative opinion has been issued for the company's MAA for the product in the treatment of malignant melanoma in the EU; the EMEA has indicated an additional confirmatory trial is needed in this indication for approval. An NDA for CLL was deemed non-approvable by the US FDA; the company is appealing this decision. The phase III trials in multiple myeloma and AML did not meet their primary endpoints. Phase I and II trials are also underway or have been completed for a range of other cancer types. Genta and sanofi-aventis (formerly Aventis) entered into a collaboration agreement in 2002; however, this agreement was terminated by sanofi-aventis in May 2005. Genta became solely responsible for all costs relating to oblimersen at this time. Genta expanded its Cooperative Research and Development Agreement (CRADA) with the National Cancer Institute in November 2001. The expanded collaboration was to investigate the use of oblimersen in combination with standard anticancer therapy in a broad range of cancers. This expansion occurred following the Gensynergy project, which showed that oblimersen was synergistic with other anticancer therapies. Genta signed a 5-year manufacturing agreement with Avecia Ltd in December 2002 to supply it with oblimersen. Genta's NDA was submitted to the FDA in December 2005 and accepted for review in March 2006. The application was based on data from a phase I/II trial (NCT00021749) of oblimersen alone in approximately 40 patients and a phase III study (NCT00024440) of 241 patients who received fludarabine and cyclo-phosphamide with or without oblimersen. Genta received a Special Protocol Assessment (SPA) from the FDA in October 2006 for a randomised, pivotal, clinical trial of oblimersen in CLL. The trial will be conducted in patients who have not received prior chemotherapy and who would be randomised to receive fludarabine and rituximab with or without oblimersen. This trial has not yet begun.Fast-track status was given to oblimersen for CLL in June 2003 by the FDA. Oblimersen previously obtained orphan drug status in the US and EU for CLL in September 2001. Genta previously submitted the MAA under the centralised licensing procedure and Spain and France were assigned as rapporteur and co-rapporteur countries, respectively. It was supported by an extended 24-month follow-up of patients from a phase III study (NCT00016263) of oblimersen plus dacarbazine. The EMEA validated the MAA for review in January 2006. Genta received a number of scientific questions from the EMEA in June 2006, which the company responded to. Genta intends to file a formal complaint and a request for correction of information with the FDA under the Federal Data Quality Act. The complaint is related to a key statistical analysis of the company's data for oblimersen in the treatment of melanoma used by the FDA at the Oncology Drug Advisory Committee (ODAC) in May 2004. Genta believes that analysis sought to discredit the finding that treatment with oblimersen significantly increased progression-free survival; ODAC previously agreed this endpoint would support full approval in the absence of a survival improvement in patients with advanced melanoma.A rolling NDA submission was submitted to the FDA in the third quarter of 2003; however, Genta and Aventis withdrew the NDA after the application failed to gain marketing approval from the FDA's Oncology Drug Advisory Committee (ODAC). In May 2004, ODAC voted that phase III trial results did not provide substantial evidence of effectiveness to outweigh toxicity of oblimersen treatment in patients with metastatic melanoma. Genta has the option to resubmit this application. The FDA gave oblimersen orphan drug status for malignant melanoma in August 2000. In October 1999, fast-track status was given to oblimersen by the FDA for malignant melanoma when used in combination with dacarbazine. In addition, oblimersen received orphan drug status for malignant melanoma in Australia in October 2006.A phase III study (NCT00016263) of oblimersen in combination with dacarbazine was conducted in patients with malignant melanoma. The combination treatment did not significantly increase overall survival time, but did significantly increase progression-free survival time, compared with dacarbazine treatment alone. The phase III trial enrolled 771 patients at 140 sites in 12 different countries. Patients were randomly assigned to receive dacarbazine alone or in combination with oblimersen. The primary endpoint of this trial was to compare the overall survival between the two treatment arms. Secondary endpoints included comparative analyses of progression-free survival and tumour response. Genta will conduct another phase III study of oblimersen in patients with advanced melanoma. The trial is designed to provide additional safety and efficacy evidence of the drug, in combination with dacarbazine, in patients who have not previously received chemotherapy. Approximately 300 patients are expected to be enrolled in the trial, which is planned to begin during mid-2007, at sites throughout Europe, Australia, and North and South America. Genta is conducting a phase I clinical trial (NCT00409383) to evaluate the combination of oblimersen, ABI 007, and temozolomide in chemotherapy-naive patients with advanced melanoma. This trial was initiated in November 2006 and is the first follow-on study to Genta's phase III trial of oblimersen plus dacarbazine. Oblimersen received orphan drug status in the US and EU for multiple myeloma in September 2001. In addition, fast-track designation was given to oblimersen by the FDA in the same month.A phase I/II clinical study (NCT00062244) of oblimersen was conducted by the NCI in patients with Waldenstrom's macroglobulinaemia, a disease that is similar to multiple myeloma. The study results indicated that oblimersen may be a useful treatment in this group of patients (all had high levels of Bcl-2 expression). In June 2003, Genta and Aventis announced the presentation of clinical data from a phase II trial of oblimersen in combination with docetaxel injection concentrate for patients with advanced HRPC. Researchers reported that these findings validated progression into phase III trials. Genta has licensed eight US patents relating to oblimersen and its backbone chemistry and these expire between 2008 and 2015. Genta has two pending US patent applications that relate to oblimersen. Corresponding patent applications have been filed in Canada, Europe and Japan. Genta owns three US patents relating to methods of using oblimersen that will expire in 2020, and also has approximately 45 corresponding foreign patent applications.


Mycosis fungoides associated with B-cell malignancies.
The coexistence of mycosis fungoides, a peripheral T-cell lymphoma, and B-cell malignancies or Hodgkin's lymphoma in the same patient is unusual. Most descriptions are isolated case reports and case series are strikingly sparse. To detect cases of mycosis fungoides associated with B-cell malignancies or Hodgkin's lymphoma and to analyse the characteristics of and the interplay between the lymphoproliferative neoplasms. Patients with mycosis fungoides who had B-cell malignancies or Hodgkin's lymphoma were selected from among 398 patients either treated or followed up in two tertiary medical centres during a 7-year period. Eleven patients with mycosis fungoides and B-cell malignancy were detected (seven of non-Hodgkin's lymphoma, three of chronic lymphocytic leukaemia, one of multiple myeloma). No case of Hodgkin's lymphoma was found. In seven patients the mycosis fungoides preceded the B-cell malignancy whereas in four it was the B-cell malignancy which occurred first. The time elapsed between onset of the two malignancies ranged from 4 to 22 years (average: 12 years). Patients who had mycosis fungoides as the first neoplasm presented with earlier stages of mycosis fungoides (four of seven: IA, three of seven: IB) than those who had mycosis fungoides as their second neoplasm (of four, one: IB, one: folliculotropic, two: IIB). Among the four patients in whom the appearance of mycosis fungoides followed the B-cell malignancy, three had been treated with multiagent chemotherapy. Two patients who presented with early-stage mycosis fungoides (IA) as the first lymphoma developed mycosis fungoides tumours after becoming immunosuppressed. In two patients infiltrates composed of both malignant T- and B-cell populations were found in a single biopsy. One showed two distinct populations of the malignant cells in the skin tumour, thus constituting a classical composite lymphoma of mycosis fungoides and chronic lymphocytic leukaemia, while in the other patient the two malignant populations of marginal B-cell lymphoma and mycosis fungoides (as evidenced by both phenotypic and genotypic findings) were intermingled. This case series indicates that while the coexistence of Hodgkin's lymphoma and mycosis fungoides is extremely rare, the association of mycosis fungoides and B-cell malignancies is not as rare as reflected in the literature, with non-Hodgkin's lymphoma constituting the most common associated B-cell malignancy. In this series as well as in the cases reported in the literature mycosis fungoides usually preceded the development of B-cell malignancies, which may be in accordance with previous reports of an increased risk of developing a second haematological neoplasm. The importance of a competent immune system for patients with mycosis fungoides is well demonstrated in these cases. It is suggested that for greater precision the criteria for diagnosis of composite lymphoma of the skin should include both phenotypic and genotypic features.


Pyruvate kinase type M2 is upregulated in colorectal cancer and promotes proliferation and migration of colon cancer cells.
Pyruvate kinase type M2 (PKM2) has been reported to be involved in aerobic glycolysis and cell growth in various tumors. However, the expression pattern of PKM2 in colorectal cancer (CRC) and the correlation between PKM2 expression and CRC remains unclear. The aim of this study is to investigate PKM2 expression and its possible role in CRC. We found that expression of PKM2 was increased in CRC and the increased PKM2 expression was associated with later stage and lymph metastasis of the tumors. Knockdown of PKM2 suppressed the aerobic glycolysis and decreased lactate production of colon cancer RKO cells. Knockdown of PKM2 repressed proliferation and migration of the cells. Inhibition of PKM2 suppressed xenograft tumor growth of RKO cells in vivo. These results suggest that the expression of PKM2 plays a critical role in development of CRC, and it may provide a growth advantage for colon cancer cells. Thus, PKM2 might be a potential therapeutic target for CRC.


Becoming organisms: the organisation of development and the development of organisation.
Despite the radical importance of embryology in the development of organicism, developmental biology remains philosophically underexplored as a theoretical and empirical resource to clarify the nature of organisms. This paper discusses how embryology can help develop the organisational definition of the organism as a differentiated, functionally integrated, and autonomous system. I distinguish two conceptions of development in the organisational tradition that yield two different conceptions of the organism: the life-history view claims that organisms can be considered as such during their whole ontogeny; the constitutive view distinguishes two periods in the life history, a period of generation and a period of self-maintenance of a constitutive organisation. Arguing in favour of the constitutive view, it will be claimed that the organisational criteria for the definition of organism (i.e., differentiation, functional integration, and autonomy) can only be applied to the developmental system when it has entered the period of self-maintenance of a constitutive organisation. Under the light of current research in developmental biology, it is possible to make explicit how organisms come to be as organisms. To this end, I explore key ontogenetic events that help us clarify the core aspects of animal organisation and allow us to identify the developmental stage that marks the ontological transition between an organism in potency and an organism in actuality. The structure of this ontogenetic unfolding parallels the conceptual structure of the very notion of organism; the generation of the being of a particular organism parallels its definition.


Comparisons of audio and audiovisual measures of stuttering frequency and severity in preschool-age children.
To determine whether measures of stuttering frequency and measures of overall stuttering severity in preschoolers differ when made from audio-only recordings compared with audiovisual recordings. Four blinded speech-language pathologists who had extensive experience with preschoolers who stutter measured stuttering frequency and rated overall severity from audio-only and audiovisual recordings of 36 preschool children who were stuttering. Stuttering frequency (percentage of syllables stuttered [%SS]) was based on counts of perceptually unambiguous stutterings, made in real time, and overall severity was measured using a 9-point rating scale. Stuttering frequency was statistically significantly lower by around 20% when made from audio-only recordings. This was found to be directly attributable to differences in the counts of stuttered syllables, rather than to differences in the total numbers of syllables spoken. No significant differences were found between recording modalities for the ratings of overall severity. Correlations between %SS scores in the 2 modalities and severity rating scores in the 2 modalities were high, indicating that observers agreed on data trends across speech samples. Measures of %SS made from audio-only recordings may underestimate stuttering frequency in preschoolers. Although audio-only %SS measures may underestimate stuttering frequency at the start of a clinical trial to a clinically significant extent, posttreatment scores at or below 1.0%SS are likely to underestimate by 0.2%SS or less, which is clinically insignificant.


Yoga improves occupational performance, depression, and daily activities for people with chronic pain.
Chronic pain is a complex accumulation of physical, psychological, and social conditions, thus interventions that address pain and promote occupational performance are needed. A holistic intervention, with mind and body components, is likely necessary to best treat the complexities of chronic pain. Thus, we developed and tested a yoga intervention for people with chronic pain. In a randomized control trial (RCT), participants with chronic pain were randomized to a yoga intervention or usual care group. Between and within group differences for pre-and post-outcome measure scores were assessed for: occupational performance, completion of activities, and depression. Pilot RCT with participant allocation to 8 weeks of yoga or usual care. Both groups received ongoing monthly self-management programming. Data were collected before and after the 8-week intervention. Participants were randomized to yoga or usual care after baseline assessments. Demographics were collected and measures included: Canadian Occupational Performance Measure (COPM) to assess occupational performance; the 15-item Frenchay Activities Index (FAI)(activities); and the 9-item Patient Health Questionnaire (PHQ-9) for depression. Independent t-tests were used to assess differences between groups. Paired t-tests were used to assess differences between pre- and post 8-week intervention for both the yoga and the usual care groups. Percent change scores and effect sizes were calculated. 83 people were recruited for the study and completed baseline assessments; 44 individuals were randomized to yoga and 39 to the control group. The average age of all participants was 51.4±10.5 years, 68% were female; and 60% had at least some college education. There were no significant differences in demographics or outcome measures between groups at baseline or 8 weeks; however, the study was not powered to see such differences. Individuals randomized to the control group did not significantly improve in any outcome measure over the 8 weeks. There were significant improvements in COPM performance and COPM satisfaction scores for individuals randomized to the yoga group; both scores significantly improved. COPM performance improved by 27% with a moderate to large effect size (3.66±1.85 vs 4.66±1.93, p < 0.001, d = 0.76). COPM satisfaction significantly improved by 78% (2.14±2.31 vs. 3.80±2.50, p < 0.001) and had a large effects size (d = 1.02). FAI scores improved, indicating increased activity or engagement in daily occupation during the 8-week intervention. Scores increased by 5% (38.13±8.48 vs. 39.90±8.57, p = 0.024) with a small effect size (d = 0.37). Depression significantly decreased from 13.21±5.60 to 11.41±5.82, p = 0.041, with a small effect size. Data from this pilot RCT indicate yoga may be an effective therapeutic intervention with people in chronic pain to improve occupational performance, increase engagement in activities, and decrease depression. Occupational therapy practitioners may consider adding yoga as a treatment intervention to address the needs of people with pain.


Genome-wide patterns of single-feature polymorphism in Arabidopsis thaliana.
We used hybridization to the ATH1 gene expression array to interrogate genomic DNA diversity in 23 wild strains (accessions) of Arabidopsis thaliana (arabidopsis), in comparison with the reference strain Columbia (Col). At <1% false discovery rate, we detected 77,420 single-feature polymorphisms (SFPs) with distinct patterns of variation across the genome. Total and pair-wise diversity was higher near the centromeres and the heterochromatic knob region, but overall diversity was positively correlated with recombination rate (R(2) = 3.1%). The difference between total and pair-wise SFP diversity is a relative measure contrasting diversifying or frequency-dependent selection, similar to Tajima's D, and can be calibrated by the empirical genome-wide distribution. Each unique locus, centered on a gene, has a diversity and selection score that suggest a relative role in past evolutionary processes. Homologs of disease resistance (R) genes include members with especially high levels of diversity often showing frequency-dependent selection and occasionally evidence of a past selective sweep. Receptor-like and S-locus proteins also contained members with elevated levels of diversity and signatures of selection, whereas other gene families, bHLH, F-box, and RING finger proteins, showed more typical levels of diversity. SFPs identified with the gene expression array also provide an empirical hybridization polymorphism background for studies of gene expression polymorphism and are available through the genome browser http://signal.salk.edu/cgi-bin/AtSFP.