Correlation between serial changes in left-sided heart chambers and atrial natriuretic peptide and N-terminal pro atrial natriuretic peptide after a first myocardial infarction. An echocardiographic study.
Dilatation of the left ventricle predicts morbidity and mortality after acute myocardial infarction. We compared serial echocardiographic examinations of the left atrium and ventricle with measurements of plasma atrial natriuretic peptide (ANP) and plasma N-terminal pro atrial natriuretic peptide (NT-proANP) in 22 patients during the first 6 months following a first myocardial infarction. ANP, but not NT-proANP, was found to be significantly correlated to the diastolic/systolic size of the left atrium (r = 0.58/0.60) and the systolic size of the left ventricle (r = 0.43) in the acute phase 2-4 days after infarction. At 10-12 days after the infarction, we found a significant correlation between all sizes of the left-sided chambers and ANP, whereas NT-proANP only correlated with the left atrial sizes. Three months after the infarction, all sizes of the left-sided chambers correlated with both ANP and NT-proANP, with the exception of a non-significant correlation between NT-proANP and the left atrial diastolic size. After 6 months only the area of the diastolic and systolic left atrium correlated with plasma ANP and only the systolic size of the left atrium correlated with NT-proANP. The percentage change in the size of the left atrium, but not the left ventricle, correlated significantly with the percentage change in both ANP (r = 0.57) and NT-proANP (r = 0.70). We conclude that the distension of the left atrium rather than the dilatation of the left ventricle is related to the concentration of ANP and NT-proANP after an acute myocardial infarction.


Ultrastructure and immunocytochemical distribution of GABA in layer III of the rat medial entorhinal cortex following aminooxyacetic acid-induced seizures.
Layer III of the entorhinal cortex (EC) is lesioned in patients with temporal lobe epilepsy (TLE). A similar neuropathology is also present in different animal models of TLE. For example, injection of the "indirect" excitotoxin aminooxyacetic acid (AOAA) into the EC of rats causes behavioral seizures and preferential loss of neurons in layer III of the medial EC. The animals also develop hyperexcitability of the EC and the hippocampal region CA1. To further explore the neuropathological changes within the EC, the ultrastructure and distribution of GABA-like immunoreactivity were assessed in layer III, 28 days after an intraentorhinal AOAA injection. At this time point, light microscopic preparations revealed that a large proportion of pyramidal (putative excitatory) neurons in layer III of the medial EC had degenerated, whereas GABA-immunoreactive neurons had survived. In immunogold-labeled ultrathin sections, the lesioned neuropil was found to contain morphologically intact GABA-containing neurons and nerve terminals. Pathologically swollen dendrites and electron-dense neuronal profiles were present in the lesioned sector as well. The majority of the electron-dense profiles was identified as degenerating dendritic spines that were closely apposed to strongly glutamate-immunopositive axon terminals. Thus, the entorhinal chemoarchitecture is dramatically altered following an episode of AOAA-induced epileptic seizures. One possible consequence of this pathology is a reduced "drive" of the surviving layer III GABA neurons, which in turn may cause hyperexcitability of the EC and the hippocampus. These findings may be of relevance for the genesis and spread of temporal lobe seizures.


TRPA1 involved in miR-141-5p-alleviated neuropathic pain induced by oxaliplatin.
Oxaliplatin (OXA) is widely used to treat advanced colorectal cancer, but it can induce severe peripheral neuropathy. Accumulating evidence has shown that microRNAs (miRNAs) are closely linked to neuropathic pain induced by sciatic nerve lesion and spinal cord injury. However, the study on the role of miRNAs in OXA-induced neuropathic pain is rare and needs to be further investigated. The study is aiming to investigate the effects of miR-141-5p on OXA-induced neuropathic pain and its underlying mechanisms. The neuropathic pain rat model was built through intraperitoneal injection of OXA. Mechanical withdrawal threshold and tail withdrawal latency were measured. The expressions of miR-141-5p and TRPA1 in dorsal root ganglion were detected by qRT-PCR, western blot, and immunohistochemistry. The results indicated that OXA down-regulated the expression of miR-141-5p. By contrast, OXA significantly up-regulated the expression of TRPA1 mRNA and protein. Besides, intrathecal injection of miR-141-5p mimic attenuated OXA-induced neuropathic pain and reduced the expression of TRPA1, a predicted target of miR-141-5p. Collectively, the results suggest that TRPA1 may mediate miR-141-5p-alleviated neuropathic pain induced by OXA. Our findings provide a potential therapeutic target for OXA-induced neuropathic pain.


[3H]phorbol 12,13-dibutyrate/PKC binding in thoracic spinal cord: no change in amyotrophic lateral sclerosis.
[3H]phorbol 12,13-dibutyrate ([3H]PDBu), a selective ligand for various protein kinase C isozymes (PKC), was used to investigate the distribution of [3H]PDBu/PKC binding sites in thoracic spinal cords of patients who died with amyotrophic lateral sclerosis (ALS) and subjects free of neurological disease. In controls, binding of [3H]PDBu was mostly concentrated in the substantia gelatinosa and to a lesser extent in other dorsal horn regions (laminae III and IV). [3H]PDBu binding sites were also present in the ventral horn and laminae X but in somewhat lower quantities. The distribution of [3H]PDBu binding in thoracic spinal cords of patients who died with ALS was unchanged compared to controls and no significant differences were observed in the amount of specific binding in ALS. The present results on the distribution of [3H]PDBu binding sites in human spinal cord are consistent with previous studies of [3H]PDBu binding in rodents indicating that high levels of PKC are present in the dorsal horn. The results also suggest that levels of PKC isozymes are not altered in ALS spinal cord.


Utility of hyaluronic acid in pleural fluid for differential diagnosis of pleural effusions: likelihood ratios for malignant mesothelioma.
The level of hyaluronic acid (HA) was determined in the pleural fluid of 99 patients, including 19 with malignant mesothelioma, 27 with lung cancer, 1 with breast cancer, 1 with mediastinal tumor and 51 with non-malignant diseases. With a cut-off level at 100 micrograms/ml, the pleural fluid concentration of HA was high in 36.8% of patients (7 of 19) with malignant mesothelioma and 1.3% of patients (1 of 80) with lung cancer and other malignant and non-malignant diseases. The mean concentration of pleural fluid HA was significantly higher in patients with mesothelioma than in those with lung cancer and other malignant and non-malignant diseases. The pre-test probability of MM was 5.9% in this series. The LRs for > or = 100, 50-99 and < or = 49 micrograms/ml are 28.3, 3.3 and 0.5, respectively; these put the post-test probabilities at 64, 17 and 3%, respectively. Indeed, in cases of uncommon disease such as MM, the post-test probability is low even if the cut-off level of HA is > or = 100 micrograms/ml. The discrimination between malignant mesothelioma and lung cancer needs special attention. In these two diseases, the LRs of MM for pleural fluid CEA > 30, 10-30 and < 10 ng/ml were 0.2, 1.9 and 2.4, respectively. The pre-test probability of MM for HA > or = or 100 micrograms/ml is 64%. Furthermore, because the LR for CEA is < 10 ng/ml, the post-test probability is 81%. When the combination of two markers is considered, the high level of HA and the low level of CEA may be useful for the differential diagnosis of MM from pleuritis carcinomatosa.


Long-term persistence of oligoclonal serum IgM repertoires in patients treated with allogeneic bone marrow transplantation (BMT).
Immunoglobulin gene rearrangements in patients treated with BMT have restricted repertoire diversity. Clonal variability remains low for 3 months and reconstitution of the humoral immune system appears to follow a wave-like pattern. In the present study we analysed serum IgM and IgG repertoires in 44 patients from 1 week to 3 years after transplantation. We applied a quantitative immunoblot technique in combination with a newly developed method for estimation of repertoire diversity in complex mixtures of antibodies. Our results demonstrate that 60% of BMT patients have severely reduced diversity in the IgM repertoire during and after the first year post-BMT, compared with healthy controls. In contrast, the majority of patients have a polyclonal IgG repertoire, similar to that of healthy controls. Serum IgM repertoires remain oligoclonal even though the serum concentration of total IgM is within normal range around 6 months post-BMT. During the first years after transplantation IgM as well as IgG repertoires are less diverse in patients receiving a BM graft from a sibling donor compared with those receiving a graft from an HLA-matched unrelated donor. Patients in the latter group show a higher incidence of infections and minor antigen mismatches which may promote the development of a diverse immunoglobulin repertoire post-BMT.


Antigen-presenting-cell function of interferon gamma-treated human gingival fibroblasts.
The present study was carried out to examine the antigen-presenting cell (APC) functions of human gingival fibroblasts (HGF) elicited with IFN gamma. Stimulation of HGF with IFN gamma clearly induced HLA-DR expression and enhanced expression of intercellular adhesion molecule-1 (ICAM-1) on HGF. Despite the phenotypical resemblance of IFN gamma-treated HGF to so-called APC, HLA-DR positive HGF were unable to induce proliferation of allo-reactive peripheral blood T cells (PBT) isolated from different donors. The failure of IFN gamma-treated HGF to stimulate unprimed allo-reactive PBT was not due to the lack of production of IL-1 or the immunosuppressive effect of PGE2 from HGF. On the other hand, the fact that detectable expression of CD80, ligand for CD28, was not found on IFN gamma-treated HGF may at least in part explain the ineffective function of HGF as APC. Interestingly, IFN gamma-treated HGF induced proliferation of primed allo-reactive CD4+ T cells in a HLA-DR dependent manner, suggesting that IFN gamma-treated HGF may have the ability to stimulate pre-activated T cells. We then confirmed that high levels of IFN gamma mRNA were detectable in inflamed gingival tissue. Although it cannot be concluded from this study that HGF are incapable of effectively presenting antigenic peptides to autologous T cells bearing appropriate T cell receptors, present results suggest that HGF may be affected by locally-secreted IFN gamma and that the IFN gamma-stimulated HGF may play a role in regulating immune responsiveness in inflammatory periodontal lesions.


Preparation of pure and intact Plasmodium falciparum plasma membrane vesicles and partial characterisation of the plasma membrane ATPase.
In host erythrocytes, the malaria parasite must contend with ion and drug transport across three membranes; its own plasma membrane, the parasitophorous membrane and the host plasma membrane. Isolation of pure and intact Plasmodium falciparum plasma membrane would provide a suitable model to elucidate the possible role played by the parasite plasma membrane in ion balance and drug transport. This study describes a procedure for isolating parasite plasma membrane from P. falciparum-infected erythrocytes. With this method, the trophozoites released by saponin treatment were cleansed of erythrocyte membranes using anti-erythrocyte antibodies fixed to polystyrene beads. These trophozoites were then biotinylated and the parasite plasma membrane was disrupted by nitrogen cavitation. This process allows the membranes to reform into vesicles. The magnetic streptavidin beads bind specifically to the biotinylated parasite plasma membrane vesicles facilitating their recovery with a magnet. These vesicles can then be easily released from the magnetic beads by treatment with dithiotreithol. The parasite plasma membrane showed optimal ATPase activity at 2 mM ATP and 2 mM Mg2+. It was also found that Ca2+ could not substitute for Mg2+ ATPase activity in parasite plasma membranes whereas activity was completely preserved when Mn2+ was used instead of Mg2+. Other nucleoside triphosphates tested were hydrolysed as efficiently as ATP, while the nucleoside monophosphate AMP was not. We have described the successful isolation of intact P. falciparum plasma membrane vesicles free of contaminating organelles and determined the experimental conditions for optimum ATPase activity.


Estimation of sound power of baffled planar sources using radiation matrices.
A technique is presented in this paper for the analysis of acoustic radiation of baffled planar sources. This method is based on the concept of the radiation matrix which is a representation of the sound field radiated by complex sources. Both sound power and far-field pressure can be estimated using this method. Numerical simulation and experimental investigation were carried out to verify the proposed method. In comparison with the conventional pressure-based method ISO 3745, the proposed technique produced more accurate estimation of sound power. With comparable number of measuring positions as the conventional methods, the proposed approach does not require special measuring environments. Computation complexity can be significantly reduced if only dominant radiation modes in the low-frequency range are retained. Estimation of far-field pressure can also be obtained using the proposed method.


An optimized method of extracting and quantifying active Neutrophil serine proteases from human whole blood cells.
Neutrophil serine proteases (NSPs) are implicated in numerous inflammatory diseases. Thus, a robust methodology to monitor and quantify NSPs is important to study disease progression and evaluate the effect of pharmacological interventions. A comparison of the various methods used to extract NSPs from neutrophil granulocytes has not been published, providing the impetus to conduct this method optimization and comparison study. Two NSP recovery methodologies were evaluated on samples from five human donors: zymosan stimulation and cell pellet extraction. For the zymosan stimulation method, 1 mL donor blood was added to zymosan and samples were incubated at 37°C for 30 min while shaking. Samples were then centrifuged, and the plasma was collected for quantitation of NSP activity. For the cell pellet extraction procedure, 2 mL whole blood samples were centrifuged into white blood cell pellets following red blood cell lysis. To each pellet, three sequential lysis steps were performed using either 0.05% Nonidet P-40 Substitute (NP40) or 0.02% Triton X-100 lysis buffers under agitation followed by centrifugation. NSP activities were quantified using an exogenous peptide substrate specific to each of the three NSPs being analyzed: neutrophil elastase, cathepsin G, and proteinase 3. The zymosan stimulation method resulted in lower recovery of active NSPs and was unable to stimulate significant release of active cathepsin G. In contrast, the NP40 pellet extraction method showed consistent inter-donor NSP release with greater recoveries of active NSPs than the Triton method or the zymosan stimulation method. Overall, the pellet extraction procedure provided 13.3-fold greater recovery of active neutrophil elastase, 283-fold greater recovery of active cathepsin G, and 2.9-fold greater recovery of active proteinase 3 than the zymosan method. The NP40 cell pellet extraction method resulted in greater extraction of active NSPs compared to the other methods investigated here, which may allow for a more accurate and complete biomarker profile when evaluating human clinical samples.


Anatomical organization of MCH connections with the pallidum and dorsal striatum in the rat.
Neurons producing the melanin-concentrating hormone (MCH) are distributed in the posterior hypothalamus, but project massively throughout the forebrain. Many aspects regarding the anatomical organization of these projections are still obscure. The present study has two goals: first to characterize the topographical organization of neurons projecting into the cholinergic basal forebrain (globus pallidus, medial septal complex), and second to verify if MCH neurons may indirectly influence the dorsal striatum (caudoputamen) by innervating afferent sources to this structure. In the first series of experiments, the retrograde tracer fluorogold was injected into multiple sites in the pallidal and medial septal regions and the distribution of retrogradely labeled neurons were analyzed in the posterior lateral hypothalamus. In the second series of experiments, fluorogold was injected into the caudoputamen, and the innervation by MCH axons of retrogradely labeled cells was analyzed. Our results revealed that the MCH system is able to interact with the basal nuclei in several different ways. First, MCH neurons provide topographic inputs to the globus pallidus, medial septal complex, and substantia innominata. Second, striatal projecting neurons in the cortex, thalamus, and substantia nigra presumably receive only sparse inputs from MCH neurons. Third, the subthalamic nucleus is heavily innervated by MCH projections, thus, presumably serves as one important intermediate station to mediate MCH influence on other parts of the basal nuclei.


Dollars and sense. Is there a better way to determine private surgical fees in New Zealand?
In New Zealand, private insurers reimburse surgeons on a fee-for-service basis. Ideally, the level of reimbursements should reflect competitive market prices. Due to concerns such a market does not exist, other countries have adopted Relative Value Scales (RVS) to estimate a fair reimbursement level for different procedures. No such scale exists in New Zealand for surgeons, but it does for anaesthetists. This study compares reimbursements to surgeons and anaesthetists from private insurers using data from 3186 procedures performed between 1996 and 2002. We calculate an implicit hourly rate of reimbursement and compare the level of reimbursement between procedures and the variance of reimbursements within procedures for surgeons and anaesthetists. The results suggest that there are significantly greater deviations in average reimbursements between procedures for surgeons than for anaesthetists. Furthermore, the variability of reimbursements is greater for reimbursements to surgeons within specific procedures. While the results do not necessarily imply that surgical reimbursements are inconsistent with underlying market rates, the results are consistent with the hypothesis that anaesthetist's fees show greater stability because of the existence of a RVS. We conclude by discussing what would be required to implement a RVS for surgical fees in New Zealand.


Defining and Regulating Acute Inflammatory Lesion Formation during the Pathogenesis of Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis.
The primary pathology of the human central nervous system disease multiple sclerosis (MS) and the animal counterpart experimental autoimmune encephalomyelitis (EAE) includes immunological and inflammatory events. Immune system involvement in MS has been widely debated but the role of inflammation has received less attention. Classic acute inflammation features vasculitis, resident tissue macrophage and mast cell participation plus the involvement of circulatory-derived neutrophils and platelets. Pre-lesion development in MS incorporates cerebral vasculitis, activated resident microglia in normal appearing white matter together with infiltrating cell types and factors indicative of an acute inflammatory reaction. Similarly, the formation of perivascular lesions during early EAE includes characteristic neurovasculitis, the participation of central nervous system microglial phenotypes plus haemopoietic cells and mediators, signifying an ongoing acute inflammatory response. EAE has been extensively used as a screen to select drugs for MS treatment but has been criticised as unrepresentative of the human condition due to fundamental differences in disease induction and pathogenesis. The review provides compelling evidence for a distinct acute inflammatory phase in MS lesion formation that is convincingly reproduced in early EAE pathology. Moreover, consideration of drug efficacy studies undertaken during initial EAE validates the occurrence of an acute inflammatory phase in disease pathogenesis. Critical appraisal, recognition and acceptance of the mutual acute inflammatory components inherent in the primary pathology of MS and EAE reveals new targets and encourages confident and reliable employment of the animal model in the assessment of novel compounds for the control of key primary pathological events in human demyelinating disease.


Constitutive tyrosine and serine phosphorylation of STAT4 in T-cells transformed with HTLV-I.
STAT4 is a critical mediator of IL-12-stimulated gene regulation in T-helper type 1 (Th1) cell. IL-12 activates the Janus family tyrosine kinases JAK2 and Tyk2, which in turn phosphorylate STAT4 on tyrosine 693. The p38 mitogen-activated protein kinase (MAPK) signaling pathway is also activated in response to IL-12, followed by phosphorylation of STAT4 on serine 721, which is required for STAT4 full transcriptional activity. In the present study, we demonstrated constitutive activation of STAT4 in HTLV-I-transformed T-cell lines MT-2, MT-4 and HUT102 by immunoprecipitation, Western blotting and electrophoretic mobility shift assay (EMSA). In HTLV-I-transformed T-cell lines, STAT4 was constitutively phosphorylated not only on tyrosine 693 but also on serine 721, and formed a heterodimer with STAT3. Constitutive phosphorylation of its upstream activators, JAK2, Tyk2 and p38 MAPK was also observed in the cells. EMSA and transient transfection studies further showed that the high-affinity sis-inducible element (hSIE) preferentially binds the STAT3/STAT4 heterodimer and is constitutively transactivated in MT-2 cells in the absence of exogenous cytokine stimulation. When STAT4 expression vector was cotransfected along with STAT3 expression vector into MT-2 cells, STAT4 significantly synergized with STAT3 to transactivate hSIE, showing the functional importance of heterodimer formation between STAT4 and STAT3.


Metastatic tumor antigen in hepatocellular carcinoma: golden roads toward personalized medicine.
Hepatocellular carcinoma (HCC), a prototype of hypervascular tumors, is one of the most common malignancies in the world, especially hyperendemic in the Far East where chronic hepatitis B virus (HBV) infection is highly prevalent. It is characterized by the clinical feature of a poor prognosis or a high mortality due to its already far advanced stages at diagnosis. It is so multifactorial that hepatocarcinogenesis cannot be explained by a single molecular mechanism. To date, a number of pathways have been known to contribute to the development, growth, angiogenesis, and even metastasis of HCC. Among the various factors, metastatic tumor antigens (MTAs) or metastasis-associated proteins have been vigorously investigated as an intriguing target in the field of hepatocarcinogenesis. According to recent studies including ours, MTAs are not only involved in the HCC development and growth (molecular carcinogenesis), but also closely associated with the post-operative recurrence and a poor prognosis or a worse response to post-operative anti-cancer therapy (clinical significance). Herein, we review MTAs in light of their essential structure, functions, and molecular mechanism in hepatocarcinogenesis. We will also focus in detail on the interaction between hepatitis B x protein (HBx) of HBV and MTA in order to clarify the HBV-associated HCC development. Finally, we will discuss the prognostic significance and clinical application of MTA in HCC. We believe that this review will help clinicians to understand the meaning and use of the detection of MTA in order to more effectively manage their HCC patients.


Short-term effects of resistance training frequency on body composition and strength in middle-aged women.
Although a dose-response relationship between resistance training frequency and strength has been identified, there is limited research regarding the association between frequency and body composition. This study evaluated the effects of 3 vs. 4 d·wk(-1) of resistance training on body composition and strength in middle-aged women. Twenty-one untrained women (age 47.6 ± 1.2 years) completed 8 weeks of resistance training either 3 nonconsecutive days of the week using a traditional total-body protocol (RT3) or 4 consecutive days of the week using an alternating split-training protocol (RT4). The RT3 completed 3 sets of 8 exercises, whereas RT4 completed 3 sets of 6 upper body exercises or 6 sets of 3 lower body exercises. Both groups completed 72 sets per week of 8-12 repetitions at 50-80% 1 repetition maximum. Weekly training volume load was calculated as the total number of repetitions × load (kg) completed per week. Body composition was measured using air displacement plethysmography. At baseline and after 8 weeks of resistance training, there were no significant between-group differences. Both protocols resulted in significant increases in absolute lean mass (1.1 ± 0.3 kg; p = 0.001), body weight (1.02 ± 0.3 kg; p = 0.005), body mass index (0.3 ± 0.1 kg·m(-2); p = 0.006), strength (p < 0.001), and weekly training volume load (p < 0.001). Correlation analysis revealed that weekly training volume load was strongly and positively related to gains in lean mass (r = 0.56, p = 0.05) and strength (r = 0.60, p = 0.006). In these untrained, middle-aged women, initial short-term gains in lean mass and strength were not influenced by training frequency when the number of training sets per week was equated.


Spontaneous renal tubular carcinoma in Fischer-344 rat littermates.
Two of 632 Fischer-344 rats in a food restriction study had spontaneous, bilateral, multicentric renal tubular cell carcinomas. Although there were 104 litters represented in this study, both rats that developed this rare neoplasm were from the same litter. The littermates, one male and one female, were in the food-restricted treatment groups (60% of ad libitum intake) and were 550 and 447 days old, respectively, at death. The probability that the two rare bilateral renal neoplasms occurred by a chance event in littermates is approximately 0.8%. The apparent familial predisposition for development of specific types of neoplasms emphasizes the importance of randomization of individuals into treatment groups and consideration of lineage for rare tumors.


LDL Apheresis: an effective and safe treatment for refractory hypercholesterolemia.
Through the efforts of Edward H. Ahrens, LDL apheresis became available for the treatment of patients, often with familial hypercholesterolemia, who have no alternative therapy for severely elevated LDL cholesterol levels. In the U.S., the FDA has approved this treatment for individuals on maximum diet and drugs with an LDL cholesterol greater than 300 mg/dL or greater than 200 mg/dL with coronary artery disease. Unlike plasmapheresis, apolipoprotein B-containing lipoproteins (LDL, Lp(a), and VLDL) are selectively removed by heparin precipitation or columns containing dextran sulfate cellulose or antibodies to apolipoprotein B. The acute lowering of LDL-cholesterol by a typical 2 - 3 h treatment is up to 80%, and the time-averaged lowering in the 1 to 2 week interval between treatments is up to 50%, with very few side effects. The lowering of LDL-cholesterol and other cardioprotective effects of LDL apheresis have reduced chest pain, prevented new disability and prolonged life. Whole blood compatible columns in development offer the possibility of simpler and less expensive treatments.


Chloroplast DNA Sequence Homologies among Vascular Plants.
The extent of sequence conservation in the chloroplast genome of higher plants has been investigated. Supercoiled chloroplast DNA, prepared from pea seedlings, was labeled in vitro and used as a probe in reassociation experiments with a high concentration of total DNAs extracted from several angiosperms, gymnosperms, and lower vascular plants. In each case the probe reassociation was accelerated, demonstrating that some chloroplast sequences have been highly conserved throughout the evolution of vascular plants. Only among the flowering plants were distinct levels of cross-reaction with the pea chloroplast probe evident; broad bean and barley exhibited the highest and lowest levels, respectively. With the hydroxylapatite assay these levels decreased with a decrease in probe fragment length (from 1,860 to 735 bases), indicating that many conserved sequences in the chloroplast genome are separated by divergent sequences on a rather fine scale. Despite differences observed in levels of homology with the hydroxylapatite assay, S1 nuclease analysis of heteroduplexes showed that outside of the pea family the extent of sequence relatedness between the probe and various heterologous DNAs is approximately the same: 30%. In our interpretation, the fundamental changes in the chloroplast genome during angiosperm evolution involved the rearrangement of this 30% with respect to the more rapidly changing sequences of the genome. These rearrangements may have been more extensive in dicotyledons than in monocotyledons. We have estimated the amount of conserved and divergent DNA interspersed between one another.From the reassociation experiments, determinations were made of the percentage of chloroplast DNA in total DNA extracts from different higher plants; this value remained relatively constant when compared with the large variation in the diploid genome size of the plants.


Chitosan-Platelet-Rich Plasma Implants Improve Rotator Cuff Repair in a Large Animal Model: Pivotal Study.
The purpose of this study was to assess the safety and efficacy of chitosan-platelet-rich plasma (PRP) hybrid implants used as an adjunct to surgical rotator cuff repair in a pivotal Good Laboratory Practice (GLP)-compliant study. The infraspinatus tendon was transected in 48 skeletally mature ewes and repaired with a transosseous-equivalent (TOE) technique. In the two treatment groups, a chitosan-PRP solution was injected at the footprint between the tendon and the bone and on top of the repaired site (2 mL or 3 mL doses, n = 12 per group). To further assess chitosan safety, a chitosan-water solution was injected at the same sites (3 mL, n = 12). Outcome measures included Magnetic Resonance Imaging (MRI) assessment and clinical pathology at 3 months and 6 months and histopathology at 6 months. The tendon gap was decreased at 3 months on MRI images and certain histopathological features were improved at 6 months by chitosan-PRP treatment compared to controls. The group treated with chitosan-water was not different from controls. Chitosan-PRP treatment induced no negative effects in the sheep, which suggests high safety. This study provides further evidence on the safety and efficacy of chitosan-PRP for rotator cuff repair augmentation, which could eventually be used in a clinical setting.


Blue light-modulation of chlorophyll a fluorescence transients in guard cell chloroplasts.
Chlorophyll a fluorescence transients from mesophyll and single guard cell pairs of Vicia faba were measured by microspectrofluorometry. In both chloroplast types, fluorescence induction (O to P) was similar under actinic blue and green light. In slow transients from mesophyll cell chloroplasts, blue and green light induced identical, typical rapid quenching from P to S, and the M peak. In contrast, the P to S transient from guard cell (GC) chloroplasts irradiated with blue light showed a much slower quenching rate, and the P to T transition showed no M peak. Actinic green light induced mesophyll-like transients in GC chloroplasts, including rapid quenching from P to S and the M peak. Detection of these transients in single pairs of GC and isolated protoplasts ruled out mesophyll contamination as a signal source. Green light induced a rapid quenching and the M peak in GC chloroplasts from several species. The effect of CO(2) concentration on the fluorescence transients was investigated in the presence of HCO(3) (-) at pH 6.8 and 10.0. In transients induced by green light in both chloroplast types, a pH increase concomitant with a reduction in CO(2) concentration caused an increase in the initial rate of quenching and the elimination of the M peak. Actinic blue light induced mesophyll-like transients from GC chloroplasts in the presence of 10 micromolar KCN, a concentration at which the blue light-induced stomatal opening is inhibited. Addition of 100 to 200 micromolar phosphate also caused large increases in fluorescence quenching rates and a M peak. These results indicate that blue light modulates photosynthetic activity in GC chloroplasts. This blue light effect is not observed in the absence of transduction events connected with the blue light response and in the presence of high phosphate concentrations.


Characterization and Risk Factor Analysis of Osteoporosis in a Large Cohort of Patients with Chronic Graft-versus-Host Disease.
The National Institutes of Health Chronic Graft-versus-Host Disease (cGVHD) Consensus Project Ancillary and Supportive Care Guidelines recommend annual assessment of bone mineral density (BMD) to monitor bone health. The study of osteoporosis in patients with cGVHD has been limited to small numbers of patients, and the guidelines are based on experience with other chronic diseases and expert opinion. We hypothesized that the prevalence of osteoporosis is high in a cohort of 258 patients with moderate to severe cGVHD because of prolonged exposure to risk factors for osteoporosis after allogeneic hematopoietic stem cell transplantation. We defined osteoporosis using BMD criteria (T-score ≤-2.5) at 3 anatomic sites-the femoral neck (FN), lumbar spine (LS), and total hip (TH)-and characterized risk factors through univariate and multivariate analyses. We found that low body weight (FN, P < .0001; LS, P = .0002; TH, P < .0001), malnutrition (FN, P = .0002; LS, P = .03; TH, P = .0076), higher platelet count (FN, P = .0065; TH, P = .0025), higher average National Institutes of Health organ score (FN, P = .038), higher prednisone dose (LS, P = .032), lower complement component 3 (LS, P = .0073), and physical inactivity (FN, P = .01) were associated with osteoporosis in at least 1 site. T-scores were significantly lower in the FN compared with the LS or TH (P < .0001 for both). The prevalence of osteoporosis and osteopenia was high (17% and 60%, respectively), supporting current recommendations for frequent monitoring of BMD. The association of higher platelet count in patients with cGVHD and osteoporosis has not been reported previously and represents a new area of interest in the study of osteoporosis after allogeneic hematopoietic stem cell transplantation.


Controlled transport of timolol maleate through artificial membranes under passive and iontophoretic conditions.
The passive and iontophoretic permeability of timolol maleate (TM) through porous and dense artificial membranes was investigated in order to select the most optimal membrane for a transdermal drug delivery system. For the meso-porous membranes (pore diameter 2-50 nm), the TM permeability for passive diffusion and iontophoresis was practically the same. For the micro-porous membranes (pore diameter<2 nm), a significant transport contribution of iontophoresis was observed, which was more pronounced when higher current densities were applied. The electrical resistance of all the porous membranes was lower than the electrical resistance of human skin. For dense membranes, passive and iontophoretic TM permeability was significantly lower than for porous membranes and in most cases their electrical resistance was comparable or even higher than the resistance of human skin. For most of the membranes studied the average adsorption of TM at 37 degrees C was low (0.02-0.33 mg/cm(2)) and independent of the TM concentration. For the meso-porous mixed cellulose acetate-cellulose nitrate membrane the TM adsorption was significantly higher and increased with the TM concentration. Based on our results, the optimum membrane for an iontophoretic transdermal TM delivery system is the LFC 1 micro-porous membrane because it mainly controls the TM delivery (TM iontophoretic permeability: 0.86 x 10(-6) cm/s), has very low electrical resistance (0.9-1.5 komega cm(2)) and the TM adsorption to it is low (0.15 mg/cm(2)). The therapeutic plasma TM concentration is achievable by application of this membrane in realistic sizes (5-64 cm(2)) and by application of current densities between 0.13 and 0.5 mA/cm(2).


Are district nurses well placed to provide equitable end-of-life care to individuals who are homeless?
This literature review aimed to answer the focus question: are district nurses well placed to provide equitable end of life care (EOL) for homeless individuals? It focused on 10 primary research studies, from which two themes emerged and subsequently formed the basis of the discussion: (1) the difficulty in predicting disease trajectory in people who are homeless and (2) the gaps in existing systems. The main findings from these themes were a lack of education on the recognition of the dying and a general lack of knowledge of the complex challenges faced by and health needs of homeless people, which cause stigma from both the general public and health professionals towards these marginalised individuals. Further, there is certainly a lack of suitable places to deliver palliative and EOL care for people who are homeless. Available services are inflexible and have no tolerance for substance misuse, which creates an access barrier for homeless people in need of EOL care.


Data evaluation of laminar flow diffusion chamber nucleation experiments with different computational methods.
In order to evaluate the experimental data from laminar flow diffusion chamber (LFDC) experiments on homogeneous nucleation, an extensive postmeasurement computational analysis is required. The present work investigates the influence of the used computational methodology on the derived nucleation curves. To this end a reanalysis is made of previous LFDC experiments of 1-butanol nucleation in helium [D. Brus et al., J. Chem. Phys. 122, 214506 (2005)] using two different methods. The first method is based on single fluid heat and vapor transport in the carrier gas ignoring the aerosol processes, as commonly made in LFDC data evaluations. The second method is more comprehensive as is based on multidimensional computational fluid-particle dynamics. The calculations are made under the usual simplification of one-way coupling between fluid flow and particles, which is a valid approximation in most practical aerosols, while full aerosol dynamical effects are accommodated. Similar results were produced by the two methods. This finding corroborates the usual practice of omitting aerosol calculations in LFDC experimental data evaluation.


Computation of a tetrahedral mesh for striated muscle deformation simulation.
Competing concepts exist regarding surgery for instance of the cleft lip and palate to date. Morphology-based simulations at histological scale may one day be used to help the surgeon predict the possible outcome of a variety of approaches. It however can be a challenge to generate volume meshes that are applicable to the mathematical modelling of three-dimensional spatial modifications. Computation of surface meshes may be considered less delicate. The aim of this study is to design and evaluate a novel algorithm that supports finite element methods. Images of histological serial sections of a striated muscle were segmented. Results of the three-dimensional reconstruction of multiple layers of the polygonal segmentation data characterized the hull of the muscle. The corresponding surface mesh was then converted into a tetrahedral mesh to generate volume. This was achieved by mapping multiple template types onto neighbouring intersection polygons. Muscle contraction was subsequently simulated by mesh deformation. The technique successfully generated volumes and was able to provide data on contraction directions. The mesh supported a novel approach to simulate representations of contraction. However, several drawbacks were evident. Mathematical modelling of scenarios with more than one striated muscle will require considerable modification of the currently presented approach. Future studies need to then evaluate the applicability of volume meshes to represent arrays of three-dimensional biological objects. Surface mesh based mathematical modelling of cleft lip and palate surgery and its results are therefore not yet in reach.


Dose escalation studies of cytarabine, daunorubicin, and etoposide with and without multidrug resistance modulation with PSC-833 in untreated adults with acute myeloid leukemia younger than 60 years: final induction results of Cancer and Leukemia Group B Study 9621.
P-glycoprotein (Pgp) is strongly inhibited by PSC-833. A chemotherapy dose-escalation study was performed with PSC-833 in patients younger than 60 years with untreated acute myeloid leukemia. Clinical rather than pharmacokinetic end points were used to develop two induction therapies containing drugs susceptible to Pgp-mediated efflux and associated with comparable toxicities at the maximum-tolerated doses. A total of 410 patients were enrolled. Fifteen induction regimens containing variable doses of daunorubicin (DNR) and etoposide (ETOP) and fixed doses of cytarabine were evaluated with (ADEP) or without (ADE) a fixed dose of PSC-833. Doses selected for phase III testing were DNR 90 mg/m(2) and ETOP 100 mg/m(2) in ADE, and DNR and ETOP each 40 mg/m(2) in ADEP. Intolerable mucosal toxicity occurred at higher doses of ADEP. Although the design of this study precludes direct comparisons, there was an apparent advantage for receiving ADEP with respect to disease-free and overall survival in patients < or = 45 years old, despite the significantly lower doses of DNR and ETOP given in ADEP compared with ADE. A large clinical data set was used to develop induction regimens containing two drugs susceptible to Pgp-mediated efflux, with and without an inhibitor of Pgp function. The chosen doses have comparable antileukemia activity and toxicity, making them suitable for use in a phase III comparative study of induction chemotherapy for patients with acute myeloid leukemia younger than 60 years. That trial will also clarify whether patients < or = 45 years old are especially likely to benefit from Pgp inhibition during induction therapy.


Manganese-enhanced magnetic resonance imaging (MEMRI) of rat brain after systemic administration of MnCl₂: hippocampal signal enhancement without disruption of hippocampus-dependent behavior.
Manganese (Mn(2+))-enhanced magnetic resonance (MR) imaging (MEMRI) in rodents offers unique opportunities for the longitudinal study of hippocampal structure and function in parallel with cognitive testing. However, Mn(2+) is a potent toxin and there is evidence that it can interfere with neuronal function. Thus, apart from causing adverse peripheral side effects, Mn(2+) may disrupt the function of brain areas where it accumulates to produce signal enhancement and, thereby, Mn(2+) administration may confound cognitive testing. Here, we examined in male adult Lister hooded rats if a moderate systemic dose of MnCl₂ (200 μmol/kg; two intraperitoneal injections of 100 μmol/kg separated by 1 h) that produces hippocampal MR signal enhancement would disrupt hippocampal function. To this end, we used a delayed-matching-to-place (DMP) watermaze task, which requires rapid allocentric place learning and is highly sensitive to interference with hippocampal function. Tested on the DMP task 1 h and 24 h after MnCl₂ injection, rats did not show any impairment in indices of memory performance (latencies, search preference) or any sensorimotor effects. However, MnCl₂ injection caused acute peripheral effects (severe ataxia and erythema, i.e. redness of paws, ears, and nose) which subsided over 30 min. Additionally, rats injected with MnCl₂ showed reduced weight 1 day after injection and failed to reach the normal weight-growth curve of control rats within the 16 days monitored. Our results indicate that 200 μmol/kg MnCl₂ produces hippocampal MR signal enhancement without disrupting hippocampus-dependent behavior on a rapid place learning task, even though attention must be paid to peripheral side effects.


Laforin, a dual specificity phosphatase that dephosphorylates complex carbohydrates.
Laforin is the only phosphatase in the animal kingdom that contains a carbohydrate-binding module. Mutations in the gene encoding laforin result in Lafora disease, a fatal autosomal recessive neurodegenerative disorder, which is diagnosed by the presence of intracellular deposits of insoluble complex carbohydrates known as Lafora bodies. We demonstrate that laforin interacts with proteins known to be involved in glycogen metabolism and rule out several of these proteins as potential substrates. Surprisingly, we find that laforin displays robust phosphatase activity against a phosphorylated complex carbohydrate. Furthermore, this activity is unique to laforin, since several other phosphatases are unable to dephosphorylate polysaccharides. Finally, fusing the carbohydrate-binding module of laforin to the dual specific phosphatase VHR does not result in the ability of this phosphatase to dephosphorylate polysaccharides. Therefore, we hypothesize that laforin is unique in its ability to utilize a phosphorylated complex carbohydrate as a substrate and that this function may be necessary for the maintenance of normal cellular glycogen.


Patients with borderline personality disorder who are chronically suicidal: therapeutic alliance and therapeutic limits.
Therapeutic work with patients who are chronically suicidal and have borderline personality disorder (BPD) is challenging, and clinicians often resort to setting firm limits or excessively cautious interventions in efforts to prevent manipulation, regression, or over-dependence. Litigation and malpractice fears reinforce these stances, and reduced compensation for additional time and energy devoted to patients adds further disincentives to sole providers. However, elements of the working alliance and therapeutic limits are within the therapist's control. A case vignette illustrates an individual therapist's modification of usual therapeutic limits while working with a chronically suicidal patient with BPD within a dialectical behavior therapy (DBT) framework over a 16-week period. Discussions regarding the case, interventions used, DBT, and legality concerns follow.


Implementation of the YMCA Diabetes Prevention Program throughout an Integrated Health System: A Translational Study.
HealthSpan Physicians (HSP), an integrated medical system in Northeast Ohio, partnered with the Young Men's Christian Association (YMCA) of Greater Cleveland to implement a referral system for the evidence-based Diabetes Prevention Program (DPP) throughout HSP. The YMCA of USA employs a cost-effective, customized version of the original DPP in which coaches take the place of in-house clinical staff. Efficacy of the YMCA DPP was shown earlier in the DEPLOY Study. To improve outcomes of metrics used in the DEPLOY Study. Observational study focusing on engagement, persistence, recruitment, and adherence to the DPP. In August 2014, HSP mailed an invitation to 2200 patients identified as both Medicare eligible and at risk of prediabetes to attend no-obligation information sessions about the DPP. After these sessions, YMCA staff called interested participants and asked them to enroll in and to commit to the program. Motivation and reinforcement were provided to patients through YMCA-provided signs, brochures, and posters; the HSP Web site; and in-person conversations with primary care physicians. Average weight loss at the end of 16 weeks in the program and average retention through Session 9. Of the 2200 patients contacted, 351 (16.0%) responded by attending the information session, and 228 enrolled in the YMCA DPP (11.3%) and persisted through at least Week 9. This result is an improvement over the 1.7% of eligible enrollees who responded to the DEPLOY Study's mailing. A marketing approach to implementing the YMCA DPP in an integrated medical system results in excellent outcomes.


Immunohistochemical organization patterns of the follicular dendritic cells, myofibroblasts and macrophages in the human spleen--new considerations on the pathological diagnosis of splenectomy pieces.
There is reliable information about how changes in spleen histology are influenced by the relationship among B and T lymphocytes, macrophages, dendritic cells and myofibroblasts. Moreover, if it can be applied in the day-by-day pathology laboratory. This work intends to elucidate morpho-functional aspects of relationships of these cells in the different spleen compartments, how they are influenced by pathological conditions and how basic immunohistochemical techniques could optimize the histopathological diagnosis. We analyzed the usefulness of the monoclonal antibodies CD45RO, CD20, CD21, CD35, CD68, caldesmon, the smooth muscle alpha-actin type 1 (SMA-1) in 91 specimens. CD21(+) CD35(+) follicular dendritic cells were organized into three patterns in agreement with the immune condition of the lymphoid follicle. Smooth muscle alpha-actin type 1(+)and caldesmon(+)myofibroblasts draw two double rings: marginal-perifollicular and germinal-marginal. The latter is closely related to T-cells. CD68(+)red pulp macrophages had clear and linear configuration. The interruption of this CD68(+) linear pattern in splenic marginal zone lymphoma cases could be a criterion to differentiate it from reactive hyperplasia. CD45RO, CD20, CD21, CD68 and SMA-1 provide a basic and quality immunohistochemical battery for a better comprehension of the human spleen and could improve its histopathological diagnosis.


The role of ethylene in the senescence of oat leaves.
The evolution of ethylene, both from the endogenous source and from added 1-aminocyclopropane-1-carboxylic acid (ACC), has been followed in close relationship with the senescent loss of chlorophyll from seedling oat leaves. In white light, where chlorophyll loss is slow, the ethylene evolution increases slowly at first, but when the loss of chlorophyll becomes more rapid, ethylene evolution accelerates. CoCl(2) inhibits this increase and correspondingly maintains the chlorophyll content, with an optimum concentration of 10 micromolar. The rapid rate of chlorophyll loss in the dark is slightly decreased by 3-aminoethoxyvinyl glycine (AVG), by cobalt, and slightly stimulated by ACC. The slower chlorophyll loss in white light, however, is almost completely inhibited by silver ions, greatly decreased by cobalt and by AVG, and strongly increased by ACC. Since the chlorophyll loss is accompanied by proteolysis, it represents true senescence. Chlorophyll loss in light is also strongly antagonized by CO(2), 1% CO(2) giving almost 50% chlorophyll maintenance in controls, while in the presence of added ACC or ethylene gas, the chlorophyll loss is 50% reversed by about 3% CO(2). The ethylene system in leaves is thus more sensitive to CO(2) than that in fruits. Indoleacetic acid also clearly decreases the effect of ACC. It is shown that kinetin, CO(2), Ag(+), and indoleacetic acid, all of which oppose the effect of ethylene, nevertheless increase the evolution of ethylene by the leaves, and it is suggested that ethylene evolution may, in many instances, mean that its hormonal metabolism is being prevented.Abscisic acid somewhat increases ethylene evolution also, but its action in promoting senescence in light is antagonized only partially by Ag(+), Co(2+), or AVG. For this and a number of other reasons it is concluded that ethylene and abscisic acid both independently control leaf senescence in the light.


Bone Regeneration of a 3D-Printed Alloplastic and Particulate Xenogenic Graft with rhBMP-2.
This study aimed to evaluate the bone regeneration capacity of a customized alloplastic material and xenograft with recombinant human bone morphogenetic protein-2 (rhBMP-2). We prepared hydroxyapatite (HA)/tricalcium phosphate (TCP) pure ceramic bone blocks made using a 3D printing system and added rhBMP-2 to both materials. In eight beagle dogs, a total of 32 defects were created on the lower jaws. The defective sites of the negative control group were left untreated (N group; 8 defects), and those in the positive control group were filled with particle-type Bio-Oss (P group; 12 defects). The defect sites in the experimental group were filled with 3D-printed synthetic bone blocks (3D group; 12 defects). Radiographic and histological evaluations were performed after healing periods of 6 and 12 weeks and showed no significant difference in new bone formation and total bone between the P and 3D groups. The 3D-printed custom HA/TCP graft with rhBMP-2 showed bone regeneration effects similar to that of particulate Bio-Oss with rhBMP-2. Through further study and development, the application of 3D-printed customized alloplastic grafts will be extended to various fields of bone regeneration.


Season-induced variation in lipid composition is associated with semen quality in Holstein bulls.
Season-induced variation in fatty acid and cholesterol composition in bovine semen has been associated with semen quality. Given the specific roles of the various semen compartments (seminal fluids, sperm head, and sperm tail) in fertilization, we hypothesized that environmental-stress-induced alterations in the lipid composition of a specific compartment might impair semen quality and sperm function. Semen samples were collected from five mature Holstein-Friesian bulls during the summer (August to September) and winter (December to January). Semen was evaluated by computerized sperm-quality analyzer, calibrated for bulls' semen, and centrifuged to separate the spermatozoa from the seminal fluids. The spermatozoal fraction was sonicated to separate the sperm head and tail compartments. Cold lipid extraction was performed with chloroform:methanol (2:1, vol/vol). Lipids were identified and quantified by gas chromatography. Seasonal variation was found in both physiological and structural parameters. The proportion of spermatozoa defined as morphologically normal was higher in the winter, with higher motility, progressive motility, and velocity relative to summer samples. Lipid composition within fractions varied between seasons with prominent impairment in the tail compartment, characterized by high saturated fatty acid, low polyunsaturated fatty acid, and low cholesterol concentrations during the summer. Given the association between alterations in lipid composition and reduced sperm motility and velocity during the summer, it is suggested that lipid composition might serve to predict sperm quality.


Characterization of ATPase activity of recombinant human Pif1.
Saccharomyces cerevisiae Pif1p helicase is the founding member of the Pif1 subfamily that is conserved from yeast to human. The potential human homolog of the yeast PIF1 gene has been cloned from the cDNA library of the Hek293 cell line. Here, we described a purification procedure of glutathione S-transferase (GST)-fused N terminal truncated human Pif1 protein (hPif1deltaN) from yeast and characterized the enzymatic kinetics of its ATP hydrolysis activity. The ATPase activity of human Pif1 is dependent on divalent cation, such as Mg2+, Ca2+ and single-stranded DNA. Km for ATP for the ATPase activity is approximately 200 microM. As the ATPase activity is essential for hPif1's helicase activity, these results will facilitate the further investigation on hPif1.


[Rupture of the renal artery aneurysm in a patient with symptoms of a systemic connective tissue disorder--A case review].
The renal arteries aneurysms are relatively rare, however, their rupture has a high mortality rate. The authors present a case-review of a 38-year old female, hospitalized with a pain in the left lumbar region, lasting for 24 hours, which spread into the mesogastrium and the left groin. Upon angiographic examination, the left renal artery aneurysm was detected. Embolization was conducted, aiming to save the kidney, however, it proved unsuccessful and was complicated by a rupture of the aneurysm. Consequently, urgent nephrectomy was completed. Prior to the procedure, massive haemorrhaging from the right groin following angiography, was managed. Relapsing haemorrhaging from the right groin and development of a major retroperitoneal haematoma, complicated the postoperative course and required repetitive postoperative revisions. The accidental finding of a minor aneurysm of the lienal artery, the patient's constitution, her anamnestic data and the complicated course of hospitalization, suggested the systemic connective tissue disorder. Furthermore, the authors discuss contemporary diagnostic and therapeutic approach to the renal arteries aneurysms.


Prognostic Value of MicroRNAs in Preoperative Treated Rectal Cancer.
Patients with locally advanced rectal cancer are treated with preoperative chemoradiotherapy followed by surgical resection. Despite similar clinical parameters (uT2-3, uN+) and standard therapy, patients' prognoses differ widely. A possible prediction of prognosis through microRNAs as biomarkers out of treatment-naïve biopsies would allow individualized therapy options. Microarray analysis of 45 microdissected preoperative biopsies from patients with rectal cancer was performed to identify potential microRNAs to predict overall survival, disease-free survival, cancer-specific survival, distant-metastasis-free survival, tumor regression grade, or nodal stage. Quantitative real-time polymerase chain reaction (qPCR) was performed on an independent set of 147 rectal cancer patients to validate relevant miRNAs. In the microarray screen, 14 microRNAs were significantly correlated to overall survival. Five microRNAs were included from previous work. Finally, 19 miRNAs were evaluated by qPCR. miR-515-5p, miR-573, miR-579 and miR-802 demonstrated significant correlation with overall survival and cancer-specific survival (p < 0.05). miR-573 was also significantly correlated with the tumor regression grade after preoperative chemoradiotherapy. miR-133b showed a significant correlation with distant-metastasis-free survival. miR-146b expression levels showed a significant correlation with nodal stage. Specific microRNAs can be used as biomarkers to predict prognosis of patients with rectal cancer and possibly stratify patients' therapy if validated in a prospective study.


Multiple chemotactic factors: fine control or redundancy?
Recruitment of leukocytes either during development or to the site of injury is crucial. The molecular regulation of cell trafficking is complex but well orchestrated by the temporal and spatial expression of a multitude of chemokines (chemoattractant cytokines) and chemokine receptors. The chemokines, classified primarily into four classes (CC, C, CXC and CX3C) bind either to specific or shared receptors and often exhibit intra-class redundancy. At the cellular level, the biological activity of chemokines is finely controlled by varied receptor display, binding affinity and/or varied signal-transducing mechanisms. Perturbation in these finely regulated processes leads to acute or chronic inflammatory conditions, developmental abnormalities and tumorigenesis/angiogenesis.


Transcranial Doppler quantification of residual shunt after percutaneous patent foramen ovale closure. Comparison of two devices.
Recurrent paradoxical embolism after catheter-based closure of right-to-left shunt (RLS) can be related to residual RLS. To improve closure success, we need a better understanding of the anatomic and device-related factors associated with closure efficacy. Patients with cryptogenic neurologic events and severe RLS (Valsalva Spencer transcranial Doppler [TCD] grade 5/5+) who underwent patent foramen ovale (PFO) closure by either central pin (Amplatzer(®) PFO [A-PFO]) or central occluding (Amplatzer(®) SO [A-SO]) devices were evaluated for residual shunt by quantitative TCD evaluation at 3 months. The findings were correlated with atrial septal aneurysm (ASA), device type, and device size. We closed 628 consecutive patients with either the A-PFO (n = 327) or A-SO (n = 301) device. The frequency of large defects, small defects, and ASA was 12%, 88%, and 44% of cases, respectively. Severe residual shunt was detected in 13% of A-PFO and 7% of A-SO recipients (P = 0.005). This difference was attributable to a much higher frequency of severe residual shunt among patients with large defects closed with the A-PFO compared to the A-SO device (12 out of 29 [41%] vs. 3 out of 42 [7%], respectively; P < 0.001). There was no significant difference in device failure frequency for small defects. The presence of ASA increased the frequency of severe residual shunt compared to those without this feature (36 out of 275 [13%] vs. 28 out of 353 [8%], respectively; P = 0.046) but did not influence device-related differences. (1) Both noncentering and central occluding closure devices effectively reduce RLS after PFO closure. (2) Large PFO defects with or without ASA have lower residual shunt grades at 3 months when closed by central occluding devices.


Gentle exercise of 40 minutes with dietary counseling is effective in treating metabolic syndrome.
Obese people may succeed in reducing their weight temporarily, but most of them regain the lost weight within a few years. We, therefore, recommend moderate long-term changes in habit rather than a strict temporary program. The purpose of this study was to examine whether physical activity of 40 minutes, mainly walking, was effective in improving or preventing metabolic syndrome when combined with dieting. Participants, all of whom had a waist circumference >or= 85 cm (men) or >or= 90 cm (women), were assigned into one of two groups: the control group (10 men and 11 women, age: 64.2 +/- 3.4 [SD] years) and the intervention group (10 men and 12 women, age: 62.4 +/- 4.2 years). Participants (n = 22) enrolled in a 52-week program; their diets were assessed using a 3-day dietary record and daily steps were measured with a pedometer. No significant change was seen in their dietary records, but the mean number of daily steps increased from 5,806 to 8,000-9,000. In the intervention group, BMI, waist circumference, and systolic and diastolic blood pressures decreased significantly by week 12 (all p < 0.001) and remained low until week 52. In fact, no significant change was seen between weeks 12 and 52. The control group showed no significant changes in all variables. These results indicate that gentle exercise of about 40 minutes in combination with dieting is effective in preventing metabolic syndrome among a sedentary population, although its effect may be limited.


Paraferritin: a protein complex with ferrireductase activity is associated with iron absorption in rats.
Recent studies reported that iron salts were absorbed in the duodenum utilizing a pathway involving membrane-associated integrin and a cytosolic protein named mobilferrin. In addition, a large molecular weight cytoplasmic complex was labeled with radioiron during mucosal uptake of iron in the duodenum. The molecular mass of this protein was 520 000 daltons, slightly larger than ferritin. On denaturing SDS-PAGE, the purified protein complex appeared to consist of at least four polypeptides, closely associated with each other. This complex was called paraferritin because its hydrodynamic volume resembled ferritin. In the present work, antibody studies demonstrate the presence of integrin, mobilferrin, and flavin monooxygenase in the water-soluble complex. Biochemical studies demonstrate the presence of a NADPH-dependent flavin monooxygenase ferrireductase activity that reduces Fe(III) to Fe(II). Antibodies against either integrin or mobilferrin inhibit monooxygenase activity. Inhibition of monooxygenase activity decreases radioiron uptake by tissue culture intestinal cells. Thus, we postulated that paraferritin plays a role in the mucosal uptake and transport of inorganic iron in small intestinal absorptive cells and is a mechanism for both the internalization of integrin from membranes to cellular cytosol and the delivery of iron to cellular constituents in an appropriate redox state.


Elevated APE1/Ref-1 Levels of Synovial Fluids in Patients with Rheumatoid Arthritis: Reflection of Disease Activity.
There is growing evidence that apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) regulates inflammatory responses. Rheumatoid arthritis (RA) is an autoimmune disease, which is characterized with synovitis and joint destruction. Therefore, this study was planned to investigate the relationship between APE1/Ref-1 and RA. Serum and synovial fluid (SF) were collected from 46 patients with RA, 45 patients with osteoarthritis (OA), and 30 healthy control (HC) patients. The concentration of APE1/Ref-1 in serum or SF was measured using the sandwich enzyme-linked immunosorbent assay (ELISA). The disease activity in RA patients was measured using the 28-joint disease activity score (DAS28). The serum APE1/Ref-1 levels in RA patients were significantly increased compared to HC and OA patients (0.44 ± 0.39 ng/mL for RA group vs. 0.19 ± 0.14 ng/mL for HC group, p < 0.05 and vs. 0.19 ± 0.11 ng/mL for OA group, p < 0.05). Likewise, the APE1/Ref-1 levels of SF in RA patients were also significantly increased compared to OA patients (0.68 ± 0.30 ng/mL for RA group vs. 0.31 ± 0.12 ng/mL for OA group, p < 0.001). The APE1/Ref-1 concentration in SF of RA patients was positively correlated with DAS28. Thus, APE1/Ref-1 may reflect the joint inflammation and be associated with disease activity in RA.


Teaching in the clinical setting: factors influencing residents' perceptions, confidence and behaviour.
Because residents are expected to assume a significant share of the teaching responsibilities of peers and students, the authors surveyed first-, second- and third-year paediatric residents at the Children's Hospital National Medical Center in July 1979 to examine those factors influencing their perceptions, confidence and behaviour with regard to teaching. The experimental group received 8 hours of instruction on teaching and learning, which included workshops and feedback sessions. The results indicate that a limited amount of formal instruction in teaching can effect a change in the teaching attitudes and behaviour of residents in addition to their level of confidence and the way in which teachers, students and peers evaluate them.


Kava hepatotoxicity--a clinical review.
This review critically analyzes the clinical data of patients with suspected kava hepatotoxicity and suggests recommendations for minimizing risk. Kava is a plant (Piper methysticum) of the pepper family Piperaceae, and its rhizome is used for traditional aqueous extracts in the South Pacific Islands and for commercial ethanolic and acetonic medicinal products as anxiolytic herbs in Western countries. A regulatory ban for ethanolic and acetonic kava extracts was issued in 2002 for Germany on the basis of reports connecting liver disease with the use of kava, but the regulatory causality assessment was a matter of international discussions. Based on one positive reexposure test with the kava drug, it was indeed confirmed that kava is potentially hepatotoxic. In subsequent studies using a structured, quantitative and hepatotoxicity specific causality assessment method in 14 patients with liver disease described worldwide, causality for kava +/- comedicated drugs and dietary supplements including herbal ones was highly probable (n = 1), probable (n = 4) or possible (n = 9) regarding aqueous extracts (n = 3), ethanolic extracts (n = 5), acetonic extracts (n = 4), and mixtures containing kava (n = 2). Risk factors included overdose, prolonged treatment, and comedication with synthetic drugs and dietary supplements comprizing herbal ones in most of the 14 patients. Hepatotoxicity occurred independently of the used solvent, suggesting poor kava raw material quality as additional causative factor. In conclusion, in a few individuals kava may be hepatotoxic due to overdose, prolonged treatment, comedication, and probably triggered by an unacceptable quality of the kava raw material; standardization is now required, minimizing thereby hepatotoxic risks.


Use of disk phosphoroscopes.
With the help of operational calculus, an effort is made to give the best conditions for the use of disk phosphoroscopes for measuring phosphorescence decays as well as for spectral measurements. This method, which up to now was only used for monomolecular processes, can be applied to any kind of process. We have been able to define limits both for the use and validity of several photometric and photographic experiments. Our results show what kind of apparatus is the most appropriate and how each measurement can be used in relation to the quantity to be found, whether it be the population of traps or the photometric efficiency of phosphorescence processes.


Comparision of nirS, cnorB, MCR genes against water quality parameters to monitor uncontrolled landfills.
This study was conducted to investigate the possible relationship between a molecular biological investigation and water quality parameters in monitoring groundwater pollution at the immediate boundary of uncontrolled landfills and their downgradient aquifers, which may consequently facilitate unbiased monitoring for the sites. Two closed landfills, Jicksan and Taejang in Korea, were chosen for this study, where the diversity of the microbial community was characterized and three specific genes, i.e. nirS (nitrite reductase coding gene), cnorB (nitric oxide reductase coding gene) and MCR (methyl coenzyme M reductase coding gene), were quantified. The quantified genes were then compared with conventional water quality parameters. From the analyzed DNA sequences, Proteobacteria phylum was most dominantly observed. A quantitative analysis revealed that the copy numbers (gene abundance) of denitrification enzyme coding genes, i.e. nirS gene and cnorB gene in Jicksan (J) site, are seven and four times, respectively, higher than Taejang (T) site. This simply implied that denitrification was possibly higher in J site than T site. In addition, a methane production enzyme coding gene, i.e. MCR, in a J1 bore immediately bordering the sources in the J site showed the greatest concentration, but it was precipitously decreased in the downgradient direction toward the outer boundary of landfill. A comparative investigation between the copy numbers of three genes, i.e. nirS, cnorB, and MCR, and conventional monitoring parameters, i.e. Cl-, alkalinity, TOC, NH3-N, and NO2-N, showed that they had overall correlation as given by more than 0.99 of the squared correlation coefficient (R2) for almost all of the concerned bores. It was concluded that the comparison between the molecular biological investigation and the conventional groundwater monitoring parameters showed good relationship between them, so that both tools could be more efficiently used for assessing the levels of contamination and prediction of the fate of pollutants, rather than being applied separately.


Alternative splicing of its 5'-UTR limits CD20 mRNA translation and enables resistance to CD20-directed immunotherapies.
Aberrant skipping of coding exons in CD19 and CD22 compromises responses to immunotherapy for B-cell malignancies. Here, we show that the MS4A1 gene encoding human CD20 also produces several mRNA isoforms with distinct 5' untranslated regions (5'-UTR). Four variants (V1-4) were detectable by RNA-seq in distinct stages of normal B-cell differentiation and B-lymphoid malignancies, with V1 and V3 being the most abundant by far. During B-cell activation and Epstein-Barr virus infection, redirection of splicing from V1 to V3 coincided with increased CD20 positivity. Similarly, in diffuse large B-cell lymphoma only V3, but not V1, correlated with CD20 protein levels, suggesting that V1 might be translation-deficient. Indeed, the longer V1 isoform was found to contain upstream open reading frames (uORFs) and a stem-loop structure, which cooperatively inhibited polysome recruitment. By modulating CD20 isoforms with splice-switching Morpholino oligomers, we enhanced CD20 expression and anti-CD20 antibody rituximab-mediated cytotoxicity in a panel of B-cell lines. Furthermore, reconstitution of CD20-knockout cells with V3 mRNA led to the recovery of CD20 positivity, while V1-reconstituted cells had undetectable levels of CD20 protein. Surprisingly, in vitro CD20-directed CAR T cells were able to kill both V3- and V1-expressing cells, but the bispecific T cell engager mosunetuzumab was only effective against V3-expressing cells. To determine whether CD20 splicing is involved in immunotherapy resistance, we performed RNA-seq on four post-mosunetuzumab follicular lymphoma relapses and discovered that in two of them downregulation of CD20 was accompanied by the V3-to-V1 shift. Thus, splicing-mediated mechanisms of epitope loss extend to CD20-directed immunotherapies. In normal & malignant human B cells, CD20 mRNA is alternatively spliced into four 5'-UTR isoforms, some of which are translation-deficient.The balance between translation-deficient and -competent isoforms modulates CD20 protein levels & responses to CD20-directed immunotherapies. We discovered that in normal and malignant B-cells, CD20 mRNA is alternatively spliced to generate four distinct 5'-UTRs, including the longer translation-deficient V1 variant. Cells predominantly expressing V1 were still sensitive to CD20-targeting chimeric antigen receptor T-cells. However, they were resistant to the bispecific anti-CD3/CD20 antibody mosunetuzumab, and the shift to V1 were observed in CD20-negative post-mosunetuzumab relapses of follicular lymphoma.


Decline and preservation of reversal learning abilities and acquisition in the course of senescence.
Different types of learning and memory functions decrease at different rates in senescence. The present study examines which types of mental functions show a relatively early decline and which learning abilities are relatively preserved in late senescence by investigating different types of learning abilities in water maze tests. Two groups of senescent male Wistar rats aged 24 months (group W24) and 30 months (group W30), respectively, were compared to adult rats (12 months, group W12). Group W24 represents 'senescent' and group W30 'late-senescent' rats. Whereas acquisition showed a relatively late decline (in group W30), reversal learning was impaired relatively early (group W24).


Myofibroma of the mandible. Clinicopathologic study and review of the literature.
A case of mandibular myofibroma in a 2-month-old boy is presented. Including this case, 24 pediatric and 11 adult patients with maxillofacial myofibroma have been reported since 1981. Of the 24 pediatric patients, 15 (62.5%) had lesions affecting the mandible. The adult cases had no mandibular involvement. Histologic evaluation of the tissue specimen revealed an interlacing pattern of spindle-shaped cells with long oval nuclei. Tissue immunohistochemical staining found it to be reactive for antibodies directed against vimentin and alpha-smooth muscle actin, but not desmin, S-100 protein, neuron-specific enolase, or myoglobin. Electron microscopy examination revealed the following cells: myofibroblast-like cells, fibroblast-like cells, and intermediate cells that were similar to the fibroblast-like cells except for the presence of a few microfilaments. Myoblast-like cells were not seen.


Sentence context influences the interpretation of word meaning by Alzheimer patients.
This study examined whether, in Alzheimer patients, verbal context selectively activates elements of a word's meaning that are relevant to the context. Subjects were shown a sentence ending in a target noun, followed by a question about an attribute of that target. The sentence primed either the attribute in the question, a different attribute, or no attribute. Both normals and AD patients answered the question faster when the target had appeared in a context priming that particular attribute. These results suggest that Alzheimer patients retain knowledge of semantic attributes and that these attributes are susceptible to contextual activation.


Effect of different intravenous nutrients on upper gastrointestinal secretion in rats.
Total parenteral nutrition has been advocated for nutritional support of patients with proximal small bowel fistula. Data on the direct effects of different nutrients on upper gastrointestinal secretion are controversial. Therefore, we examined the effects of glucose, amino acid, fat and mixed parenteral solutions on gastric juice secretion, hydrochloric acid and pepsin content, duodenal juice secretion, and pancreatic protein, amylase, bilirubin and bicarbonate content in thirty rats with chronic gastric and duodenal fistulas. Gastric secretion was increased by glucose solution (D10W) and by amino acid solution (5% AA) (13% and 12% increase respectively) but not to significant degree by fat emulsion (2% F). Both D10W and 5% AA caused comparable increase in hydrochloric acid secretion (D10W: +34%; 5% AA: +28%) and the pepsin secretion (D10W: +37%). The single infusion of hypertonic glucose, amino acids and fat were associated with decreases in duodenal fistula volume (p less than 0.05) or content output. Mixed nutrients decreased both gastric secretion (volume: -28%; HCl: -19%; pepsin: -65%) and duodenal juice volume (p less than 0.05), protein and amylase content (p less than 0.05). The elevated bilirubin in reduced volume of duodenal juice caused by mixed nutrients may be responsible for bile sludge formation. This study indicates that the use of parenteral nutrition can provide full nutritional support while decreases the fistula output.


On variables that affect estimates of the true sizes and densities of radioactively labeled cell nuclei.
Tritiated thymidine has been widely used as a nuclear marker of cell birth. The true diameters and packing densities (nuclei/microns 3) of such radioactively labeled nuclei cannot be measured directly from tissue sections. Here we show that existing stereological corrections cannot be applied to data from radioactively labeled nuclei. We empirically measured the number of silver grains exposed by nuclei containing tritiated thymidine. The nuclei were separated from the photographic emulsion by known thicknesses of fixed, embedded avian telencephalon. The results of this experiment were used to develop an equation that estimates the number of silver grains exposed by a cell nucleus of any given diameter, containing a given amount of radioactive label, and located at any given distance from the photographic emulsion. The equation also allows one to calculate the probability that a label-containing nucleus will be correctly classified as labeled. Simulations of the equation revealed that not all label-containing nuclei are correctly classified by using commonly employed identification procedures and that larger nuclei are less likely to be correctly classified than smaller nuclei, given the same amount of label. The equation can be used to modify one class of existing stereological equations so as to be applicable to measurements of radioactively labeled nuclei. Finally, we discuss the assumptions and limitations of this modification.


Evaluating the clarity of research reports written for research subjects.
It is not simple to report research results to the people who participated in the research as subjects of study. Few evaluations have examined subjects' understanding of written research findings. In two recent studies of solvent exposure, subjects received brief summaries of the research. Before they were sent, the summaries were evaluated by research, health, and communication specialists as well as by representatives of the target audience. For one of the studies, interviews were conducted with subjects before and after they received the reports. Although the subjects said they understood the summary, their answers to questions about its content indicated otherwise. The results suggest that researchers not rely on subjects' self-assessments of their understanding but instead design methods to test subjects' knowledge directly. Those tests would be most valuable if applied in formative evaluations, when the opportunity exists to improve the research summaries.


Potent antimalarial activity of clotrimazole in in vitro cultures of Plasmodium falciparum.
The increasing resistance of the malaria parasite Plasmodium falciparum to currently available drugs demands a continuous effort to develop new antimalarial agents. In this quest, the identification of antimalarial effects of drugs already in use for other therapies represents an attractive approach with potentially rapid clinical application. We have found that the extensively used antimycotic drug clotrimazole (CLT) effectively and rapidly inhibited parasite growth in five different strains of P. falciparum, in vitro, irrespective of their chloroquine sensitivity. The concentrations for 50% inhibition (IC(50)), assessed by parasite incorporation of [(3)H]hypoxanthine, were between 0.2 and 1.1 microM. CLT concentrations of 2 microM and above caused a sharp decline in parasitemia, complete inhibition of parasite replication, and destruction of parasites and host cells within a single intraerythrocytic asexual cycle (approximately 48 hr). These concentrations are within the plasma levels known to be attained in humans after oral administration of the drug. The effects were associated with distinct morphological changes. Transient exposure of ring-stage parasites to 2.5 microM CLT for a period of 12 hr caused a delay in development in a fraction of parasites that reverted to normal after drug removal; 24-hr exposure to the same concentration caused total destruction of parasites and parasitized cells. Chloroquine antagonized the effects of CLT whereas mefloquine was synergistic. The present study suggests that CLT holds much promise as an antimalarial agent and that it is suitable for a clinical study in P. falciparum malaria.


Hypolipidemic effects of Hypericum Scabrum extract on the serum lipid profile and obesity in high-fat diet fed rats.
Diets included high-fat (HFD) and high calories intake is correlated with greater risk of obesity and oxidative stress, which lead to increase the risk of related diseases such as cardiovascular and metabolic disease. In the present study, we have examined the hypolipidemic activity of Hypericum Scabrum extract on HFD fed rats. Fifty-four male Wistar rats divided into six groups: 1) control, 2) H. Scabrum extract (100 mg/kg gavage per day), 3) H. Scabrum extract (300 mg/kg), 4) HFD, 5) HFD and H. Scabrum extract (100 mg/kg), 6) HFD and H. Scabrum extract (300 mg/kg). The groups were fed their diet and treatment for 3 months. Biochemical analysis showed elevated lipid serum profile in HFD rats compared to control group. H. Scabrum extract supplementation significantly ameliorated triglyceride, total cholesterol and LDL-cholesterol. H. Scabrum extract supplementation leading to increase HDL-cholesterol in HFD treated groups. This experiment showed that H. Scabrum extract decreased HFD complications and might be beneficial herbal drug for treatment of hyperlipidemia and obesity.


[Minimizing the prehospital delay in patients with acute myocardial infarct].
Despite the known importance of early intervention, delays between symptoms onset and manifestation of acute myocardial infarction (AMI) in the hospital are common. Most of the prehospital delay consists of the interval between the onset of symptoms and the decision to seek medical help. The aim of this paper is to review the factors associated with the long duration of patient delay (patient time delay). The media campaigns designed to reduce the delay between the onset of AMI have so far not been proved to be worthwhile and it is not certain that further campaigns will be more successful. Physicians and other health care professionals play an important role in reducing the delay to treatment in patients with evolving AMI. Patients with some cardiovascular disease are at high risk for AMI. High-risk patients have to be educated about the symptoms they may develop during a coronary occlusion, what steps to take if symptoms occur, and the importance of contacting emergency medical services immediately. These instruction need to be reviewed frequently and reinforced with appropriate printed material. The situational and psychological variables are important determinants of the length of decision delays in response to symptoms of AMI. No single intervention, no matter how carefully designed and implemented, will alter the patient's propensity to delay. However, consistent, regularly delivered information may be helpful. Family members should participate in all instructions because they play an important role in increasing or decreasing the delay to treatment in patients with AMI.


Ecotoxicity and genotoxicity relating to fish in wastewaters discharged from the Vilnius treatment plant.
The toxicity and genotoxicity of untreated raw (RWW) and treated wastewaters (TWW) samples from Vilnius wastewater treatment plant was assessed using fish (rainbow trout) at different stages of development. The survival of larvae and fish exposed to RWW in short-term and longterm tests reduced, whereas gill ventilation frequency, heart rate and relative body mass increase of larvae decreased significantly. The long-term exposure of fish to TWW induced significant decreases in white blood cell count and significant increases in micronuclei in blood of treated Oncorhynchus mykiss. The physical, chemical analysis of oil products (C₁₄-C₂₈), benzo(a)pyrene, suspended solids, and heavy metals (Pb, Cd, Zn, Cu, Hg) in RWW demonstrated that the concentrations of xenobiotics and some heavy metals did not exceed their Maximum Permissible Concentrations in the sewerage system and concentrations of substances in TWW corresponded to their criteria for effluents discharged into receiving waters.


Consequences of p53 gene expression by adenovirus vector on cell cycle arrest and apoptosis in human aortic vascular smooth muscle cells.
p53 shows its tumor suppresser activity by inducing cell cycle arrest and/or apoptosis of tumor cells and these activities are in part mediated by p21 cyclin-dependent kinase inhibitor (also called as WAF1, Cip1 and SDI1). Using human aortic vascular smooth muscle cells, here we demonstrate that adenovirus vector expressing p53-induced p21, cell cycle arrest at G1 and G2/M boundary, and accumulation of cells in G1 subgroup. However, adenovirus vector expressing p21 induced only G1 cell cycle arrest. The adenovirus vector expressing p53 was 200 times more cytotoxic to human aortic vascular smooth muscle cells than adenovirus vector expressing p21. These results suggest that adenovirus expressing p53 induces cytotoxicity in human vascular smooth muscle cells by apoptosis and this cytotoxicity can not be fully accounted by p21 induction.


The Rules of Cerebellar Learning: Around the Ito Hypothesis.
As a tribute to Masao Ito, we propose a model of cerebellar learning that incorporates and extends his original model. We suggest four principles that align well with conclusions from multiple cerebellar learning systems. (1) Climbing fiber inputs to the cerebellum drive early, fast, poorly-retained learning in the parallel fiber to Purkinje cell synapse. (2) Learned Purkinje cell outputs drive late, slow, well-retained learning in non-Purkinje cell inputs to neurons in the cerebellar nucleus, transferring learning from the cortex to the nucleus. (3) Recurrent feedback from Purkinje cells to the inferior olive, through interneurons in the cerebellar nucleus, limits the magnitude of fast, early learning in the cerebellar cortex. (4) Functionally different inputs are subjected to plasticity in the cerebellar cortex versus the cerebellar nucleus. A computational neural circuit model that is based on these principles mimics a large amount of neural and behavioral data obtained from the smooth pursuit eye movements of monkeys.


The influenza A virus protein PB1-F2: killing two birds with one stone?
PB1-F2 is a 90 amino acid protein that is expressed from the +1 open reading frame in the PB1 gene of some influenza A viruses. The PB1-F2 protein has been shown to contribute to viral pathogenicity, but the molecular mechanisms for mediating virulence have been unclear. Previous reports demonstrate that PB1-F2 promotes cell death, causes immunopathology and increases pro-inflammatory responses. Our group has identified a single point mutation from asparagine (N) to serine (S) at position 66 in the PB1-F2 protein that dramatically increases the virulence of highly pathogenic avian H5N1 influenza viruses and of the 1918 pandemic strain. In search for the mechanism by which PB1-F2 N66S increases pathogenicity, we have identified and characterized a novel function of PB1-F2, i.e. interferon antagonism, both in vitro and in the mouse model. Here, we discuss a hypothesis for a possible molecular link between the pro-apoptotic and anti-interferon functions of PB1-F2.


Nanofilled Resin Composite Properties and Clinical Performance: A Review.
The aim of this review was to compile recent evidence related to nanofilled resin composite materials regarding the properties and clinical performance. Special attention was given to mechanical properties, such as strength, hardness, abrasive wear, water sorption, and solubility. The clinical performance of nanocomposite materials compared with hybrid resin composites was also addressed in terms of retention and success rates, marginal adaptation, color match, and surface roughness. A search of English peer-reviewed dental literature (2003-2017) from PubMed and MEDLINE databases was conducted using the terms "nanocomposites" or "nanofilled resin composite" and "clinical evaluation." The list was screened, and 82 papers that were relevant to the objectives of this work were included in the review. Mechanical properties of nanocomposites are generally comparable to those of hybrid composites but higher than microfilled composites. Nanocomposites presented lower abrasive wear than hybrids but higher sorption values. Their clinical performance was comparable to that of hybrid composites.


Clinically Relevant Mechanisms of Lipid Synthesis, Transport, and Storage.
Lipids not only are fundamental nutrients but also serve as basic structural components of cells and as multifunctional signaling molecules. Lipid metabolism pathways underlie basic processes in health and disease and are the targets of novel therapeutics. In this review, we explore the molecular control of lipid synthesis, trafficking, and storage, with a focus on clinically relevant pathways. To illustrate the clinical relevance of molecular lipid regulation, we highlight how these biochemical processes contribute to the pathogenesis of nonalcoholic fatty liver disease, a component of the metabolic syndrome and a paradigmatic example of lipid dysregulation.


TIGR/MYOC gene sequence alterations in individuals with and without primary open-angle glaucoma.
To discover sequence alterations in the TIGR/MYOC gene associated with primary open-angle glaucoma (POAG) in Chinese subjects. Two hundred one unrelated Chinese patients with POAG and 291 unrelated individuals without glaucoma, aged 50 years or more, were screened for sequence alterations in the TIGR/MYOC gene by polymerase chain reaction, conformation sensitive gel electrophoresis, and DNA sequencing. Up to 111 more control subjects were screened for some of the alterations. Fourteen sequence variants that lead to amino acid changes were identified. Seven were novel: Pro16Leu, Ala17Ser, Leu95Pro, Leu215Pro, Glu300Lys, Glu414Lys, and Tyr471Cys. Of these, Glu300Lys and Tyr471Cys were found only in POAG. Arg46Stop was found in 4 patients with POAG (2.0%) and 9 of 402 control subjects (2.2%); one control subject was homozygous. IOP showed a trend (P = 0.11) toward a decrease of 1.5 mm Hg among the control subjects, with Arg46Stop compared with matched control subjects without Arg46Stop. Gly12Arg occurred four times as frequently in control subjects as in patients, but the difference was not statistically significant. Gly12Arg might be negatively associated with POAG, suggesting a protective effect. Three patients with POAG had a sequence change not found in control subjects, for a frequency of possible disease-causing TIGR/MYOC mutations of 1.5% (95% confidence interval [CI] = 0.3%-4.3%). Arg46Stop occurred with similar frequency in patients with POAG and control subjects, suggesting that the reduced amount of TIGR/MYOC predicted to result from this truncation does not dramatically increase or decrease risk of glaucoma.


[Hereditary chondrocalcinosis in Tunisia. Apropos of 3 families].
Hereditary chondrocalcinosis was reported in three Tunisian families. The clinical manifestations appeared early in life and extensive radiologic involvement was apparent. The mode of inheritance seemed to be autosomal with incomplete penetrance. In one family HLA typing revealed that all affected subjects had the haplotype A1 B12 DR3.


Comparison of clinical course of patients with severe acute respiratory syndrome among the multiple generations of nosocomial transmission.
Severe acute respiratory syndrome (SARS) is characterized by both an atypical pneumonia and efficient nosocomial transmission. However, it remains unknown whether the infectivity and the virulence of the pathogen will change throughout the successive transmission. This study was conducted to compare the clinical features and management regimens of patients with SARS among the multiple generations from nosocomial transmission initiated by a super-spreader. The clinical data of 84 epidemiologically-linked SARS patients from a hospital outbreak were retrospectively studied. All patients, in whom a clear-cut transmission generation could be noted, had a direct or indirect exposure to the index patient and the epidemic successively propagated through the multiple generations of cases within a short period of time. There were 66 women and 18 men with mean age of (29.2 +/- 10.3) years in this cluster; and 96.4% of whom were health care workers. Detailed contact tracing identified 35 (41.7%) first-generation cases, 34 (40.5%) second-generation cases, and 15 (17.8%) third-generation cases. No statistical differences among the multiple generations of transmission were found in terms of age, gender, incubation period and length of hospital stay. With the advanced transmission generations, the initial temperature lowered, the number of cases with dry cough decreased. There were no statistical differences in the peak temperature and duration of fever, other accompanying symptoms, leucopenia; however, the time from initial pulmonary infiltrates to radiographic recovery shortened (P < 0.05). No differences were found in maximum number of lung fields involved, duration from the onset of fever to the occurrence of pulmonary infiltrates and time from the initial pulmonary infiltrate to its peak among the multiple transmission generations (P > 0.05). No statistical differences were found in modes of oxygen therapy and sorts of antibiotics prescribed among the various transmission generations (P > 0.05); however, as with the advanced transmission generations, the number of cases prescribed with methylprednisolone, human gamma-globulin, interferon-alpha, antiviral drugs (oral ribavirin or oseltamivir) increased (P < 0.05) and time from admission to starting these medication shortened (P < 0.05). There is no evidence that SARS infection will evolve or transmit within a fashion that permits it to become less powerful throughout the successive transmission within a short time.


[Optimization of Preparation, and Stability of Indomethacin Mouth Wash for Oral Mucositis Caused by Cancer Therapy].
Severe oral mucositis caused by chemo- and radio-therapy is a common adverse event in patients with cancer. In this study, we investigated the development of an indomethacin mouth wash(IM-MW)as a novel approach to treat pain due to oral mucositis. We examined the appropriate preparation methods for IM-MW with suitable stability. IM- MW was made from bulk IM, controlled release IM capsules, and IM capsules. Dissolution in water was tested at water temperatures of 70℃, 90℃, and 98℃(n=3), and with a shaking time of 30 or 60s(n=3). We determined the IM concentration in IM-MW by HPLC-UV analysis(n=5)at time points between just after preparation and day 7, to estimate the shelf- life at 4℃ and 25℃. At 70℃, bulk IM did not dissolve, but at 90℃ and 98℃, bulk IM, controlled release IM capsules, and IM capsules all dissolved effectively. Shaking times of 30 and 60s were sufficient to dissolve bulk IM, controlled release IM capsules, and IM capsules. The stability of IM in IM-MW was 98.6±2.8%(bulk), 99.2±6.0%(controlled release capsule), and 98.5±6.0%(capsule)over 7 days at 4℃. However, at 25℃, IM stability in IM-MW decreased to 95.3±1.8% (bulk), 86.1±4.8%(controlled release capsule), and 83.6±1.6%(capsule). In this study, we identified the most suitable method for the preparation of IM-MW(90℃, shaking time of over 30s). IM-MW was stable when stored at 4℃ for at least 7 days after preparation.


Purification, crystallization and preliminary X-ray diffraction analysis of yeast nucleosome-assembly factor Cia1p.
Yeast Cia1p is a homologue of human CIA (CCG1-interacting factor A), which possesses nucleosome-assembly activity and interacts with the human TFIID subunit CCG1 and the C-terminal domain of histone H3. The yeast Cia1p without the C-terminal polyanionic stretch has been expressed in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using PEG 8000 as precipitant. The protein was crystallized in orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 106.70, b = 46.92, c = 40.60 A and one molecule in the asymmetric unit. The crystal diffracted beyond 2.95 A resolution using synchrotron radiation.