Automated criteria-based selection and analysis of fluorescent synaptic puncta.
The use of fluorescent probes such as FM 1-43 or synapto-pHluorin to study the dynamic aspects of synaptic function has dramatically increased in recent years. The analysis of such experiments is both labor intensive and subject to potentially significant experimenter bias. For our analysis of fluorescently labeled synapses in cultured hippocampal neurons, we have developed an automated approach to punctum identification and classification. This automatic selection and processing of fluorescently labeled synaptic puncta not only reduces the chance of subjective bias and improves the quality and reproducibility of the analyses, but also greatly increases the number of release sites that can be rapidly analyzed from a given experiment, increasing the signal-to-noise ratio of the data. An important innovation to the automation of analysis is our method for objectively selecting puncta for analysis, particularly important for studying and comparing dynamic functional properties of a large population of individual synapses. The fluorescence change for each individual punctum is automatically scored according to several criteria, allowing objective assessment of the quality of each site. An entirely automated and thus unbiased analysis of fluorescence in the study of synaptic function is critical to providing a comprehensive understanding of the cellular and molecular underpinnings of neurotransmission and plasticity.


A logistic model for the prediction of endometriosis.
To develop a model that uses individual and lesion characteristics to help surgeons choose lesions that have a high probability of containing histologically confirmed endometriosis. Secondary analysis of prospectively collected information. Government research hospital in the United States. Healthy women 18-45 years of age, with chronic pelvic pain and possible endometriosis, who were enrolled in a clinical trial. All participants underwent laparoscopy, and information was collected on all visible lesions. Lesion data were randomly allocated to a training and test data set. Predictive logistic regression, with the outcome of interest being histologic diagnosis of endometriosis. After validation, the model was applied to the complete data set, with a sensitivity of 88.4% and specificity of 24.6%. The positive predictive value was 69.2%, and the negative predictive value was 53.3%, equating to correct classification of a lesion of 66.5%. Mixed color; larger width; and location in the ovarian fossa, colon, or appendix were most strongly associated with the presence of endometriosis. This model identified characteristics that indicate high and low probabilities of biopsy-proven endometriosis. It is useful as a guide in choosing appropriate lesions for biopsy, but the improvement using the model is not great enough to replace histologic confirmation of endometriosis.


Inducible nitric oxide synthase activity induced by sodium chloride solution prolongs luminal pH elevation in rat and mouse stomachs.
Sodium chloride (NaCl) is a strong promoter of gastric cancer. We hypothesized that inducible nitric oxide synthase (iNOS) induced by NaCl may be involved in its promoting effects. We investigated iNOS expression by hypertonic NaCl solutions and its pathophysiological roles in the gastric mucosa of rats and mice. iNOS mRNA and protein expressions were examined in the rat and mouse gastric mucosa after intragastric administration of NaCl solution by northern blot, reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The effect on luminal pH by iNOS activity was assessed using aminoguanidine, a potent iNOS inhibitor, and iNOS gene deficient (iNOS-/-) mice. iNOS expression was detected at concentrations higher than 1.7 M, mainly in the cells infiltrating the damaged mucosa of rats. Administration of a higher than 3.4 M NaCl solution elevated luminal pH of the rat stomach remarkably, enabling bacterial overgrowth and dramatically increasing iNOS expression (n = 5 per concentration). Pretreatment with ampicillin (p.o), an antibiotic, attenuated the iNOS expression in duplicate experiments. Co-treatment with aminoguanidine (s.q) accelerated recovery of elevated luminal pH at 8 h and 16 h or 24 h after administration of 3.4 M (n = 8) and 5.2 M NaCl solution (n = 5), respectively. iNOS expression and luminal pH elevation were also observed in mice stomachs after administration of 3.4 M NaCl solution. The elevated luminal pH of iNOS-/- mice stomachs after the administration of NaCl solution was significantly lower at 6 h (n = 7) and at 9 h (n = 11), compared to that of wild type mice (n = 9 and 10, respectively). Hypertonic NaCl solutions induced iNOS expression in the gastric mucosa. iNOS activity prolonged the elevation of the luminal pH, potentially leading to bacterial overgrowth, which in turn enhanced iNOS expression. This vicious cycle might be related to the promoting effect of NaCl.


Epitope mapping of Grapevine virus A capsid protein.
Previously characterized monoclonal antibodies (MAbs) to Grapevine virus A (GVA) showed a differential reactivity against intact or partially destabilized virus particles [2]. In the present study, this differential reactivity was confirmed and several peptides reacting with a panel of four different antibodies were identified by the PEPSCAN method of epitope mapping. Oligopeptide sequences comprised between coat protein residues 61 (V) and 72 (T) were recognized by all the antibodies tested. One of these peptides (VGPKASK) was also reactive when expressed on recombinant phage particles as a fusion protein with protein pVIII. The specificity of this sequence for antibody binding was also demonstrated by competitive-ELISA using one of the GVA MAbs. The results of this study suggest that GVA particles carry a highly structured epitope centered on a common peptide region of the coat protein sequence.


Differential accumulation of radiopaque contrast material in acute myocardial infarction.
The differential accumulation of radiographic contrast materials in ischemically damaged and normal myocardium was assessed with direct measurement (fluorescent excitation analysis) of the iodine content of tissue samples from dogs with 48 hour old myocardial infarctions. Tissue samples were obtained 10, 30, 60 and 180 minutes after the intravenous administration of 2 ml/kg body weight of diatrizoate meglumine and sodium (Renografin-76). At all time intervals, the iodine concentration of infarcted tissue was at least threefold greater than that of normal myocardium. At 180 minutes the ratio between iodine concentration in infarcted myocardium and that in normal myocardium was 8.5 and between that in infarcted myocardium and that in blood was 2.6. The iodine concentration in the liver was similar to or greater than that in the infarcted area at time intervals after 10 minutes. These results suggest that the intravenous administration of contrast material may facilitate the identification of acutely infarcted myocardium with computerized X-ray transmission tomography.


[Effects of different penetration enhancers on percutaneous absorption of lappaconitine gel in vitro].
To study the effects of different penetration enhancers on the transcutaneous permeability of lappaconitine gel in vitro and therefore to screen out the effective accelerator to enhance the permeation rate of lappaconitine. Using improved Franz-type diffusion cell and excised big mouse skin in vitro as transdermal barrier, the kinetics parameters such as cumulative permeation quantity, permeation rate and permeation lagged time were determined by HPLC. The enhancement ability of four different enhancers such as azone (Azone), propylene glycol (PG), oleic acid (OA) and lauryl alcohol (LA), was investigated when used either uniquely or combinatively each other at random. When used combinatively, Azone + PG, LA + PG, OA + PG can enhance the cumulative permeation quantity, OA + PG was the best one among them. The selection of the best penetration enhancers provided reference for lappaconitine transdermal delivery.


Influence of shear on microbial adhesion to PEO-brushes and glass by convective-diffusion and sedimentation in a parallel plate flow chamber.
Microbial adhesion to surfaces often occurs despite high wall shear rates acting on the adhering microorganisms. In this paper, we compare the wall shear rates needed to prevent microbial adhesion to bare glass and poly(ethylene oxide) (PEO)-brush coated glass in a parallel plate flow chamber. Initial microbial deposition rates were determined for different wall shear rates between 4 and 1600 s(-1) on the top and bottom plates of the flow chamber. Deposition efficiencies alpha(SL), based on the Smoluchowski-Levich approach, for Pseudomonas aeruginosa D1, Escherichia coli O2K2 and Candida tropicalis GB 9/9 decreased with increasing wall shear rates and were lower for PEO-brush coated glass than for bare glass. Characteristic shear rates preventing adhesion to the bottom plate were around 10 and 1.0 s(-1) for the bacteria on glass and the PEO-brush and 36 and 3.4s(-1) for the yeast strain on glass and the PEO-brush, respectively. This demonstrates that the adhesive forces between microorganisms and a PEO-brush are comparatively weak, although some strains may have the ability to adhere to a PEO-brush under low shear conditions. Microbial deposition efficiencies alpha(SL) were much larger, however, than unity for bottom plate deposition, but could be reduced to realistic values by averaging the deposition rates found for the top (negative contribution of sedimentation) and bottom (positive contribution of sedimentation) plates.


Intervention targeted at nurses to improve venous thromboprophylaxis.
To assess the effectiveness of an intervention targeting both physicians and nurses vs. physicians only in improving venous thromboprophylaxis for older patients. Cluster randomized trial. Fifty hospital-based post-acute care departments in France. Patients aged 65 years or older. A multifaceted intervention to implement a clinical practice guideline addressing venous thromboprophylaxis. The effectiveness outcomes were elastic stocking use, ambulation or mobilization under the supervision of a physical therapist and anticoagulant-based prophylaxis. Patient outcomes included deep vein thrombosis and anticoagulant-related adverse events. One department allocated to the intervention targeted at physicians only and seven departments allocated to the intervention targeted at both physicians and nurses dropped out of the study. Compared with the intervention targeted at physicians only (n = 497 patients), the intervention targeted at both physicians and nurses (n = 315 patients) was associated with a higher rate of mobilization (62 vs. 37%, P < 0.001) and comparable levels of elastic stocking (32 vs. 39%, P = 0.74) and anticoagulant (55 vs. 48%, P = 0.36) use. The rates of deep vein thrombosis (15 vs. 13%, P = 0.50), bleeding (1 vs. 1%, P = 0.99) and thrombocytopaenia (0 vs. 0.2%, P = 0.99) did not differ between the two groups. A multifaceted intervention targeting nurses in addition to physicians can increase the frequency of mobilization of older patients to prevent venous thromboembolism but does not alter the use of elastic stockings and anticoagulant. A differential drop-out of departments might have contributed to creating imbalances in baseline characteristics and outcomes in this study.


Novel gene signature reveals prognostic model in acute lymphoblastic leukemia.
Acute lymphoblastic leukemia (ALL) is a type of hematological malignancy and has a poor prognosis. In our study, we aimed to construct a prognostic model of ALL by identifying important genes closely related to ALL prognosis. We obtained transcriptome data (RNA-seq) of ALL samples from the GDC TARGET database and identified differentially expressed genes (DEGs) using the "DESeq" package of R software. We used univariate and multivariate cox regression analyses to screen out the prognostic genes of ALL. In our results, the risk score can be used as an independent prognostic factor to predict the prognosis of ALL patients [hazard ratio (HR) = 2.782, 95% CI = 1.903-4.068, p < 0.001]. Risk score in clinical parameters has high diagnostic sensitivity and specificity for predicting overall survival of ALL patients, and the area under curve (AUC) is 0.864 in the receiver operating characteristic (ROC) analysis results. Our study evaluated a potential prognostic signature with six genes and constructed a risk model significantly related to the prognosis of ALL patients. The results of this study can help clinicians to adjust the treatment plan and distinguish patients with good and poor prognosis for targeted treatment.


Primary structure of allophycocyanin from the unicellular rhodophyte, Cyanidium caldarium. The complete amino acid sequences of the alpha and beta subunits.
The complete amino acid sequences of the alpha and beta subunits of allophycocyanin from the unicellular rhodophyte, Cyanidium caldarium, were determined by automated Edman degradation of the proteins and peptides derived from them by chemical and enzymatic cleavages. The sequence of the alpha subunit was determined from the sequences of tryptic, endoproteinase lysine-C, and cyanogen bromide peptides and carboxypeptidase A and Y digestion of the protein. The sequence of the beta subunit was determined from the sequences of tryptic, endoproteinase lysine-C, Staphylococcus aureus V8 protease, and cyanogen bromide peptides and in addition, a peptide derived from acid cleavage of an aspartyl-prolyl bond. The carboxyl-terminal sequence of the protein was determined by digestion with carboxypeptidase A. The alpha subunit contains 160 amino acids, one phycocyanobilin chromophore attached at residue 80 by a cysteinyl-thioether linkage, and the Mr calculated from the sequence is 18,160. The beta subunit contains 161 amino acids, one phycocyanobilin chromophore attached at residue 81 by a cysteinyl-thioether linkage, and the Mr calculated from the sequence is 18,125. The amino acid sequences of the alpha and beta subunits of allophycocyanin from C. caldarium are the first complete amino acid sequences of an allophycocyanin from a eukaryotic red alga. A matrix comparison of the alpha and beta subunits of C. caldarium allophycocyanin and phycocyanin (Offner, G.D., Brown-Mason, A.S., Ehrhardt, M. M., and Troxler, R. F. (1981) J. Biol. Chem. 256, 12167-12175; Troxler, R. F., Ehrhardt, M. M., Brown-Mason, A. S., and Offner, G. D. (1981) J. Biol. Chem. 256, 12176-12184) shows homology ranging from 26 to 39%. Comparison of the sequences of alpha and beta subunits of C. caldarium allophycocyanin with the sequences of the corresponding subunits of allophycocyanin from two prokaryotic cyanobacteria (Sidler, W., Gysi, J., Isker, E., and Zuber, H. (1981) Hoppe-Seyler's Z. Physiol. Chem. 362, 611-628; DeLange, R. J., Williams, L. C. and Glazer, A. N. (1981) J. Biol. Chem. 256, 9558-9566) shows homology ranging from 81 to 85%. The significance of this with respect to phycobiliprotein structure and function is discussed.


Identity domains capture individual differences from across the behavioral repertoire.
Personality traits can offer considerable insight into the biological basis of individual differences. However, existing approaches toward understanding personality across species rely on subjective criteria and limited sets of behavioral readouts, which result in noisy and often inconsistent outcomes. Here we introduce a mathematical framework for describing individual differences along dimensions with maximum consistency and discriminative power. We validate this framework in mice, using data from a system for high-throughput longitudinal monitoring of group-housed male mice that yields a variety of readouts from across the behavioral repertoire of individual animals. We demonstrate a set of stable traits that capture variability in behavior and gene expression in the brain, allowing for better-informed mechanistic investigations into the biology of individual differences.


[Malignant hyperthermia. Report on a case successfully treated by procaine infusion (author's transl)].
A case of malignant hyperthermia with muscle stiffness is described. An early diagnosis, along with procaine infusion, and an energetic unspecific treatment helped to cure this patient.


Reconstruction of the left ventricle with autologous pericardium.
Autologous pericardium was used to reconstruct different parts of the left ventricle in 25 desperately ill patients. Fourteen patients had intractable sepsis resulting from infective endocarditis and myocardial abscess and 10 patients had noninfectious disorders. Of the patients with infections, 12 had valvular endocarditis with periannular abscess and three had interventricular septal abscess. The noninfected patients had acute rupture of the ventricular wall after mitral valve replacement (one patient) heavily calcified or surgically absent mitral anulus (three patients), or rupture of the interventricular septum after acute myocardial infarction (six patients). The interventricular septum, the posterior wall of the left ventricle, and the periannular areas were reconstructed by suturing appropriately tailored pericardial patches directly to the endocardium. In patients who also required valve replacement, the prosthetic valve was partially or completely secured to the pericardial patch. There were three operative deaths. All three patients were in either septic or cardiogenic shock before operation and in none of them was the death related to the pericardial patch. All 22 survivors have been observed from 3 to 34 months, an average of 14 months. There has been no case of patch dehiscence, patch aneurysm, prosthetic valve dehiscence, or recurrent endocarditis. Autologous pericardium appears to be safe for reconstruction of the left ventricle. It is easy to handle and problems with suture line bleeding are practically nonexistent.


Histone Deacetylase Activity and Phytotoxic Effects Following Exposure of Duckweed (Lemna pausicostata L.) to Apicidin and HC-Toxin.
ABSTRACT The effects of two cyclic tetrapeptide fungal toxins, apicidin (from Fusarium spp.) and HC-toxin (from Cochliobolus carbonum), on duckweed (Lemna pausicostata L.) were examined. Both toxins inhibited histone deacetylase (HD) activity from duckweed plantlets; the effective concentration (EC(50)) for inhibition of HD was 5.6 and 1.1 muM for apicidin and HC-toxin, respectively. Approximately 65 and 85% of in vitro HD activity was inhibited by 50 muM apicidin or HC-toxin, respectively. Exposing duckweed for 72 h to apicidin or HC-toxin (25 or 50 muM) enhanced cellular leakage, impaired chlorophyll synthesis, and inhibited growth (cell division). At equivalent concentrations, the effects of HC-toxin were more pronounced than those of apicidin. In fronds, 72 h of exposure to 50 muM apicidin resulted in chloroplast deterioration indicated by loss of orientation and excess starch accumulation. In roots, a 72-h treatment with 50 muM apicidin resulted in the loss of the root cap and increased vacuolization and starch accumulation in plastids.


Investigation of changes in the lipid content of guinea-pig lung after anaphylaxis.
A detailed analysis of the lipid content of guinea-pig lung following anaphylaxis in vivo induced by aerosolized antigen showed a significant reduction in all fractions. Anoxia induced by nitrogen produced reductions in the partial glycerides and ethanolamine phospholipid. Exposure to an aerosol of histamine caused a reduction in all but the choline phospholipid and sphingolipid fractions. It was concluded that the losses of choline phospholipid and sphingolipid result from the anaphylactic reaction and not from subsequent changes.


The Force is Strong with This Epigenome: Chromatin Structure and Mechanobiology.
All life forms sense and respond to mechanical stimuli. Throughout evolution, organisms develop diverse mechanosensing and mechanotransduction pathways, leading to fast and sustained mechanoresponses. Memory and plasticity characteristics of mechanoresponses are thought to be stored in the form of epigenetic modifications, including chromatin structure alterations. These mechanoresponses in the chromatin context share conserved principles across species, such as lateral inhibition during organogenesis and development. However, it remains unclear how mechanotransduction mechanisms alter chromatin structure for specific cellular functions, and if altered chromatin structure can mechanically affect the environment. In this review, we discuss how chromatin structure is altered by environmental forces via an outside-in pathway for cellular functions, and the emerging concept of how chromatin structure alterations can mechanically affect nuclear, cellular, and extracellular environments. This bidirectional mechanical feedback between chromatin of the cell and the environment can potentially have important physiological implications, such as in centromeric chromatin regulation of mechanobiology in mitosis, or in tumor-stroma interactions. Finally, we highlight the current challenges and open questions in the field and provide perspectives for future research.


Modelling the Effect of Vaccination Program and Inter-state Travel in the Spread of COVID-19 in Malaysia.
A modified version of the SEIR model with the effects of vaccination and inter-state movement is proposed to simulate the spread of COVID-19 in Malaysia. A mathematical analysis of the proposed model was performed to derive the basic reproduction number. To enhance the model's forecasting capabilities, the model parameters were estimated using the Nelder-Mead simplex method by fitting the model outputs to the observed data. Our results showed a good fit between the model outputs and available data, where the model was then able to perform short-term predictions. In line with the rapid vaccination program, our model predicted that the COVID-19 cases in the country would decrease by the end of August. Furthermore, our findings indicated that relaxing travel restrictions from a highly vaccinated region to a low vaccinated region would result in an epidemic outbreak.


Dose-dependent benzalkonium chloride toxicity imparts ocular surface epithelial changes with features of dry eye disease.
Inclusion of the preservative benzalkonium chloride (BAC) in ophthalmic solutions is prevalent, despite the noted potential for exacerbating dry eye disease (DED). Whilst studies incorporating BAC have assessed its' effects as a mouse model of DED, the impact on limbal epithelia is under-studied. Our investigation aimed to comprehensively assess the impact of different BAC dosing regimens and their suitability as a mouse model of DED. C57BL/6J mice (n = 72) were administered topical BAC (0.05-0.2%) over 7 days. Fluorescein staining, corneal smoothness index, and immuno-histological analyses were applied to determine architectural and cellular changes on the ocular surface following BAC treatment. The effect of BAC (0.0001-0.01%) on cultivated primary mouse corneo-limbal epithelial cells (CLECs) (n = 6) was examined using morphological and functional assays. Whilst 0.2% BAC induced severe corneal epithelial defects, 0.1% BAC dispensed once daily over 7 days, induced punctate fluorescein staining without detriment to corneal smoothness. Histochemical staining revealed disorganized basal corneal epithelial cells with enlarged cytoplasmic halos. Furthermore, PAS+ goblet cells were decreased. BAC treatment also modulated K14 expression and distribution within the limbus. In cultured CLEC, BAC triggered cell contraction and vacuolation, increased LDH release and elevated cell necrosis by 4.1-fold. Concentrations of BAC as low as 0.0001% decreased colony formation. This study describes how exposing C57BL/6 mice to BAC induce some clinicopathological features of DED seen in humans, and therefore provides the foundations to explore the consequences on the ocular surface, particularly on limbal epithelia and its' stem cells.


Effective training of nursing staff enhances hospital information system efficiency.
As more hospitals use computerized information systems, the need to address related nursing issues and to effectively train nursing personnel has become imperative. This article discusses the challenges met by a hospital's nursing administration to educate its staff members in computer use and evaluates the training program and its outcome after the system was started.


Partial deletion of the human host range gene in the attenuated vaccinia virus MVA.
The genome of a strongly attenuated vaccinia virus strain, MVA, was investigated by Southern blot and sequence analyses. Three major deletions, relative to the WR strain, were localized in MVA DNA. The deletions occurred near both ends of the viral genome and one of them affected a 55 K as well as the 32 K human host range gene. Although more than two thirds of the host range gene were eliminated from the MVA, the virus could still multiply in certain human cells.


[Degradation of the EDTA and EDTA complexes with metals by immobilized cells of Chelativorans oligotrophicus LPM-4 bacteria].
Enzymatic oxidative degradation of EDTA and EDTA complexes with metals has been investigated using immobilized cells of Chelativorans oligotrophicus LPM-4. A polarographic method, which makes it possible to register oxygen consumption by cells, has been used. For the first time, it has been indicated that the Cd-EDTA and Ni-EDTA complexes undergo degradation by the bacteria under study.


How Carvedilol activates β2-adrenoceptors.
Carvedilol is among the most effective β-blockers for improving survival after myocardial infarction. Yet the mechanisms by which carvedilol achieves this superior clinical profile are still unclear. Beyond blockade of β1-adrenoceptors, arrestin-biased signalling via β2-adrenoceptors is a molecular mechanism proposed to explain the survival benefits. Here, we offer an alternative mechanism to rationalize carvedilol's cellular signalling. Using primary and immortalized cells genome-edited by CRISPR/Cas9 to lack either G proteins or arrestins; and combining biological, biochemical, and signalling assays with molecular dynamics simulations, we demonstrate that G proteins drive all detectable carvedilol signalling through β2ARs. Because a clear understanding of how drugs act is imperative to data interpretation in basic and clinical research, to the stratification of clinical trials or to the monitoring of drug effects on the target pathway, the mechanistic insight gained here provides a foundation for the rational development of signalling prototypes that target the β-adrenoceptor system.


[Brown-Séquard syndrome as the initial clinical manifestation ofmultiple sclerosis].
Multiple sclerosis is a disorder with different forms of clinical presentation. The Brown-Séquard syndrome has occasionally been described in association with multiple sclerosis. We present the case of a patient whose initial clinical presentation of multiple sclerosis was the Brown-Séquard syndrome.


Alpha 1-antitrypsin deficiency and PI typing in patients with chronic urticaria.
A group of 281 patients with chronic urticaria was classified into various subtypes and compared with 357 healthy controls for PI types of alpha1-antitrypsin. Total alpha 1-antitrypsin was measured by the Mancini technique, and PI types were determined by isoelectric focusing. The MZ phenotype and the Z gene frequency were significantly more frequent in the urticaria group. The Z gene predominated in the groups with cold urticaria and acquired angio-oedema. The total group of chronic urticaria patients showed a significant decrease in total alpha 1-antitrypsin compared with the control group (P less than 0.03). Deficiencies in alpha 1-antitrypsin may predispose to the development of certain types of urticaria and angio-oedema.


HIV vector production mediated by Rev protein transduction.
HIV-1-based vectors are promising tools for gene therapy because of their ability to integrate into nondividing cells. Their safety in clinical applications remains a major concern. Recombination events occurring among plasmid constructs during vector production could potentially lead to the generation of replication-competent viruses. The safety of HIV-1-based vectors can be improved by removing all regions of the viral genome that are not absolutely required for vector production or function. In this study, we demonstrate that the HIV-1 rev gene is dispensable for the production of HIV-1-based vectors if the vector-producing cells are supplied with purified Rev protein. We compared the efficiency of vector production among Rev, TAT-Rev (Rev fused to the protein transduction domain of the HIV TAT protein), and Rev/Pep-1 (Rev complexed with the carrier peptide Pep-1). Our results showed that 293T cells efficiently internalized TAT-Rev and Rev/Pep-1 and high-titer vector preparations were obtained with this approach. Vectors generated by such an approach showed little difference in their efficiencies of transduction of established cell lines and primary cells compared with vectors generated by standard plasmid cotransfection. Eliminating the requirement for the HIV-1 rev gene during vector production should improve the safety of applying HIV vectors in human clinical trials.


Juvenile-hormone-dependent interaction of actin and spectrin is crucial for polymorphic differentiation of the larval honey bee ovary.
Programmed cell death in the worker ovary of Apis mellifera reduces the number of ovarioles during metamorphosis from 150-200 primordia to less than 10. In contrast, practically all ovarioles in the ovary of queens survive to the adult stage. The correct formation and persistence of polyfusomes has been suggested as a critical factor for ovariole survival. We have analyzed the developmental dynamics of F-actin and alpha-spectrin in fusomes of queen and worker larvae, and in juvenile-hormone-treated worker larvae. Small fusomes containing actin and spectrin can be detected in the ovaries of fourth instar larvae in both castes. After molting to the fifth instar, the actin-spectrin association persists in the enlarged fusomes of queen ovarioles. In workers, actin dissociates from the fusomal and cortical alpha-spectrin. Coinciding with the appearance of apoptosis markers, large agglomerates of actin are detectable in worker ovarioles. Treatment of fourth-instar worker larvae with juvenile hormone rescues ovarioles from apoptosis and maintains the actin-spectrin association. Juvenile-hormone-dependent actin-spectrin interaction is thus one of the earliest steps in the differentiation of a polymorphic ovary. Plasticity in ovariole numbers as a result of hormone-dependent fusome formation may be a more widespread phenomenon in insects, extending beyond caste polymorphism in highly eusocial Hymenoptera.


Single agent efficacy of the HDAC inhibitor DATS in preclinical models of glioblastoma.
Glioblastoma (GB) remains incurable despite aggressive chemotherapy, radiotherapy, and surgical interventions; immunotherapies remain experimental in clinical practice. Relevant preclinical models that can accurately predict tumor response to therapy are equally challenging. This study aimed to validate the effect of the naturally occurring agent diallyl trisulfide (DATS) in human GB in relevant pre-clinical models. Ex vivo slice culture, in vivo cell line derived orthotopic xenograft and patient-derived orthotopic xenograft (PDX) animal models of GB were utilized to assess efficacy of treatment with DATS. Our results showed 72-h treatments of 25 µM DATS induced cell death in ex vivo human GB slice culture. We treated U87MG orthotopic xenograft models (U87MGOX) and patient-derived orthotopic xenograft models (PDX) with daily intraperitoneal injections of DATS for 14 days. Magnetic resonance (MR) imaging of mice treated with DATS (10 mg/kg) demonstrated reduced tumor size at 5 weeks when compared with saline-treated U87MGOX and PDX controls. Hematoxylin (H&E) staining demonstrated dose-dependent reduction in gross tumor volume with decreased proliferation and decreased angiogenesis. Western blotting showed that DATS was associated with increases in histone acetylation (Ac-Histone H3/H4) and activated caspase-3 in this novel preclinical model. Histological assessment and enzyme assays showed that even the highest dose of DATS did not negatively impact hepatic function. DATS may be an effective and well-tolerated therapeutic agent in preventing tumor progression and inducing apoptosis in human GB.


Endovascular aneurysm repair (EVAR).
Endovascular repair of an abdominal aortic aneurysm (AAA) is an evolving technology which continues to be practiced in more and more centres worldwide in an effort to avoid major abdominal surgery and its associated morbidity and mortality. As with conventional open surgery, the objective of endovascular management is to provide a durable repair maintaining antegrade flow in the graft whilst simultaneously excluding flow in the aneurysm. Endovascular treatment offers a less invasive alternative to standard surgical repair with the potential of reduced hospitalisation, morbidity and mortality.


Design of study without drugs--a Surinamese school-based drug-prevention program for adolescents.
The aim of this study was to design the content and accompanying materials for a school-based program--Study without Drugs--for adolescents in junior secondary schools in Suriname based on the starting points and tasks of the fourth step of the Intervention Mapping protocol (which consists of six steps). A program based on this protocol should include a combination of theory, empirical evidence, and qualitative and quantitative research. Two surveys were conducted when designing the program. In Survey I, teachers and students were asked to complete a questionnaire to determine which school year they thought would be most appropriate for implementing a drug-prevention program for adolescents (we completed a similar survey as part of previous research). An attempt was made to identify suitable culturally sensitive elements to include in the program. In Survey II, the same teachers were asked to complete a questionnaire to determine the programs' scope, sequence, structure, and topics as well as the general didactic principles to serve as a basis for program design. After outlining the program plan, lessons, and materials, we conducted a formative pretest evaluation among teachers, students, and parents. That evaluation included measures related to the program's attractiveness, comprehensibility, and usefulness. The resulting lessons were presented to the teachers for assessment. The drug-prevention program we developed comprises 10 activities and lasts 2-2.5 months in an actual school setting. The activities take place during Dutch, biology, physical education, art, religion, and social studies lessons. We based the structure of the lessons in the program on McGuire's Persuasion Communication Model, which takes into account important didactic principles. Evaluations of the program materials and lesson plans by students, teachers, and parents were mostly positive. We believe that using the fourth step of the Intervention Mapping protocol to develop a drug-prevention intervention for adolescents has a produced promising, feasible program.


Three states for the formyl peptide receptor on intact cells.
Three distinct states of the formyl peptide receptor have been described. These are: 1) the ternary complex of ligand, receptor, and G protein (LRG); 2) the rapidly dissociating occupied receptor (ligand-receptor complex (LR]; and 3) a desensitized slowly dissociating guanine nucleotide-insensitive receptor (desensitized ligand-receptor complex ("LRX"]. During cell activation there is a rapid interconversion among receptor states from a rapidly dissociating form (t 1/2 approximately 10 s) to a slowly dissociating form (t 1/2 greater than or equal to 2 min). Neither the dynamics of the states nor their interconversion is influenced by ribosylation of G protein in the presence of pertussis toxin. In contrast to ribosylation, treatment of cells with either 2-deoxyglucose or fluoride ion, both of which lead to a loss of adenine and guanine nucleotides, causes a time-dependent change in ligand dissociability. After short periods of treatment (5-15 min) rapid dissociation is observed; after longer times (30-60 min), slow dissociation is once again detected. When intact cells are first ribosylated and then energy-depleted, only a rapidly dissociating receptor is detected. These results are discussed in terms of a model with the following elements: 1) intact cell dynamics during cell activation are dominated by an energy-dependent interconversion from LR to LRX; 2) under activation conditions, LRG appears and disappears too rapidly to be detected; 3) in cells depleted of energy and guanine nucleotide, LRG is stabilized; 4) in cells both ribosylated and depleted of energy, LR is stabilized.


[PMSG dosage precision in young and old sows in the course of biotechnical ovulation synchronization. 3. Fertility and litter effects].
One single PMSG batch, Pregmagon-Dessau, was compared with other PMSG doses on 2,126 adult sows and 1,700 gilts from five farms. Additional comparisons were made with other Pregmagon or Prolosan batches on 1,178 adult sows and 1,026 gilts. All experiments were made in the context of a programme for ovulation synchronisation and term-oriented insemination. Doses of 700, 800, 900, and 1,000 IU were tested on adult sows and 600, 800, and 1,00 on gilts, with the mean dose of 800IU being found to be sufficient or superior to other doses for fertility results. Doses for adult sows could be adjusted and related to present and previous litter numbers, with the dosage being between 700IU (treatment for third litter) and 700 to 800IU (treatment following average to high previous litter. The preferential dose for gilts was found to be 800IU. Higher PMSG doses (up to 1,000IU) should not be used in synchronisation of ovulation and term-oriented insemination unless extraordinary conditions were safely established.


The effects of O6-benzylguanine and hypoxia on the cytotoxicity of 1,3-bis(2-chloroethyl)-1-nitrosourea in nitrosourea-resistant SF-763 cells.
O6-Alkylguanine-DNA alkyltransferase (AGT) activity is associated with resistance of brain tumor cell lines to the cytotoxic effects of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). SF-763 cells exhibit high AGT activity and are resistant to BCNU. In this study, we compared the effects of the AGT inhibitor O6-benzylguanine (BG) on the cytotoxicity of BCNU in oxic and hypoxic SF-763 cells; we also measured AGT activity, ornithine decarboxylase (ODC) activity, and polyamine levels to determine if there was any correlation with cell survival as determined by colony-forming efficiency assay. Exponentially growing monolayer cells were pretreated with 10 microM BG for 2 h under oxic or hypoxic (95% nitrogen/5% CO2) conditions and then exposed to graded concentrations of BCNU for 1 h. BG significantly lowered AGT activity but had no cytotoxic effect in oxic or hypoxic cells; hypoxia alone was not cytotoxic. The cytotoxicity of BCNU was 4 times higher in BG-treated hypoxic cells than in oxic cells treated with BCNU alone; the BCNU doses required for a 1-log cell kill were 75 and 300 microM, respectively. ODC activity was lowered by hypoxia alone but was not significantly affected by BG in either hypoxic or oxic cells. Polyamine levels were not significantly affected by hypoxia or BG. These results indicate that pretreatment with BG dramatically lowers AGT activity and increases the cytotoxicity of BCNU in both oxic and hypoxic SF-763 cells. The mechanism of this enhanced cytotoxicity is apparently unrelated to ODC activity or polyamine levels.


ME3738 protects from concanavalin A-induced liver failure via an IL-6-dependent mechanism.
ME3738 is a new compound that attenuates liver disease in several models of acute and chronic liver inflammation. We used the concanavalin A (Con A) model to elucidate the molecular mechanisms of ME3738 to block liver cell damage. Pretreatment of BALB/c mice with ME3738 prior to Con A injection resulted in a significant reduction in liver injury. The protective effect of ME3738 prior to Con A injection was associated with a reduction in IL-6 serum levels and NF-kappaB DNA binding in liver nuclear extracts. However, STAT3 DNA binding was induced via ME3738 prior to Con A injection. Further analysis showed that ME3738 induces IL-6 serum levels and activates STAT3 DNA binding and target gene transcription. The relevance of this finding was assessed in IL-6(-/-) mice. In these animals, ME3738 induced no increase in IL-6 serum expression, and activation of IL-6-dependent pathways was not found. In addition, ME3738 did not protect IL-6(-/-) animals from Con A-induced liver failure, while IL-6 injection was still effective. Therefore, we demonstrate that ME3738 triggers IL-6 expression, which activates pathways that are relevant to protect from Con A-induced liver failure.


Discordance between lung function of chinese university students and 20-year-old established norms.
We examined the validity of the 20-year-old established Asian norms for pulmonary function in a contemporary cohort of Hong Kong Chinese university students. Pulmonary function testing was conducted in university students (n = 805). A university campus in Hong Kong. Parameters recorded included gender, age, height, weight, standard lung function variables (ie, FEV1, FVC, and peak expiratory flow rate [PEFR]), and exhaled carbon monoxide (CO) level. Subjects completed a questionnaire on pulmonary health, smoking history, and their dietary and exercise habits within 3 months of the study. Data were compared with the established norms for lung function for Chinese persons from Hong Kong. On average, subjects were taller than those reported in the original cohort, on whom the established norms are based; however, FEV1, FVC, and PEFR were lower. As predicted, the exhaled CO level was higher in smokers. Those who exercised regularly had a higher FEV1 and FVC, and reported fewer respiratory complaints. Our findings support the idea that lung function norms not only differ across ethnic groups, but that they may be susceptible to change over a single generation within an ethnic group living in the same geographic region. Assuming the equivalence of our testing methods and those on which established norms are based, our findings shed further insight into the dynamic nature of lung function, and have implications regarding the definition of normal pulmonary function and its variance over the short term.


Assessing Coagulation Parameters in Healthy Asian Elephants (Elephas maximus) from European and Thai Populations.
The Asian elephant population is continuously declining due to several extrinsic reasons in their range countries, but also due to diseases in captive populations worldwide. One of these diseases, the elephant endotheliotropic herpesvirus (EEHV) hemorrhagic disease, is very impactful because it particularly affects Asian elephant calves. It is commonly fatal and presents as an acute and generalized hemorrhagic syndrome. Therefore, having reference values of coagulation parameters, and obtaining such values for diseased animals in a very short time, is of great importance. We analyzed prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen concentrations using a portable and fast point-of-care analyzer (VetScan Pro) in 127 Asian elephants from Thai camps and European captive herds. We found significantly different PT and aPTT coagulation times between elephants from the two regions, as well as clear differences in fibrinogen concentration. Nevertheless, these alterations were not expected to have biological or clinical implications. We have also sequenced the coagulation factor VII gene of 141 animals to assess the presence of a previously reported hereditary coagulation disorder in Asian elephants and to investigate the presence of other mutations. We did not find the previously reported mutation in our study population. Instead, we discovered the presence of several new single nucleotide polymorphisms, two of them being considered as deleterious by effect prediction software.


Endoscopic treatment of vesicoureteral reflux in children.
We treated 35 children with different grades of vesicoureteral reflux with endoscopic injection of polytetrafluoroethylene (Teflon) behind the intravesical ureter. A total of 51 refluxing ureters was treated. In all cases absence of reflux was noted on a cystogram performed immediately after the procedure. At 3 to 12-month followup reflux was absent in 32 ureters and it recurred in 4, including 3 ureters with low grade reflux. A second injection of polytetrafluoroethylene in 3 ureters resulted in its absence, while in 1 case reimplantation was performed. The procedure is simple and reliable. An operation is avoided, hospital stay is reduced and no complications have occurred.


Risk assessment of adenoviruses detected in treated drinking water and recreational water.
Human adenoviruses (HAds) have previously been detected in sewage and polluted river and dam water, as well as treated drinking water. The 51 serotypes of HAds cause a wide range of infections with clinical manifestations associated with the gastrointestinal, respiratory and urinary tracts, and the eyes. Water may play a meaningful role in the transmission of many of these HAd serotypes, specifically the enteric HAds which are transmitted via the faecal-oral route. The presence of these viruses in water used for drinking and recreational purposes is considered to constitute a potential health risk. In this study, the risk of infection by the group of HAds previously detected over a period of 1 year in selected drinking water supplies, as well as river and dam water used for recreational purposes, was assessed. Adenoviruses were previously detected in nine of 204 (4.41%) samples of two drinking water supplies (A and B) treated and disinfected according to international specifications, in four of 51 (7.8%) samples of river water and nine of 51 (17.7%) samples of dam water. Application of these previously published results in an exponential risk assessment model indicated an annual risk of infection of 1.01 x 10(-1) and 1.7 x 10(-1) for drinking water supplies A and B, respectively, assuming a daily consumption of 2 l day(-1). The daily risk of infection constituted by HAds in the river water was calculated as 1.71 x 10(-4), and in the dam water as 3.12 x 10(-5), assuming a consumption of 30 ml of water per day. The risk of infection exceeded the tolerable risk of one infection per 10 000 consumers per year proposed for drinking water. However, the results for river and dam water used for recreational purposes were within the tolerable risk of one infection per 1000 bathers per day proposed for environmental waters used for recreational purposes. The study showed that the risk of HAd infection calculated for the drinking water supplies and the recreational water may overestimate the actual risk of infection, as conservative values were assumed for some of the variables. For a more accurate assessment of the potential risk of infection research should at least include a thorough investigation of the water consumption of individuals in South Africa, and the efficiency of recovery of the glass wool adsorption-elution method.


Hemoglobinuria in the Early Poststem-Cell-Transplant Period: Risk Factors and Association with Outcomes.
Information on risk factors of hemoglobinuria after hematopoietic stem-cell transplant (HSCT) and its association with AKI, mortality, and engraftment is limited. We conducted a retrospective cohort study on all consecutive adults that underwent HSCT from January 6, 1999, to November 6, 2017. The study included 6039 patients that underwent bone marrow transplantation (BMT), umbilical cord blood, and peripheral blood stem-cell transplantation (PBSCT). Early post-HSCT, AKI occurred in 393 (7%) patients, and 52 (0.9%) patients had post-HSCT hemoglobinuria. Post-HSCT hemoglobinuria was associated with graft type (BMT+Cord), underlying disease (lymphoma, acute leukemia), and fludarabine-based conditioning regimen. Post-HSCT hemoglobinuria was associated with early (48-72 hours) post-HSCT AKI. Graft type (BMT+Cord) was associated with AKI among patients with hemoglobinuria. AKI in patients with hemoglobinuria was associated with delayed platelet engraftment and delayed WBC engraftment but not 100-day mortality. Close monitoring is recommended in this patient group to facilitate a good engraftment outcome.


Antioxidant activities and structural characterization of flavonol O-glycosides from seeds of Japanese horse chestnut (Aesculus turbinata BLUME).
We attempted to evaluate the contents and distribution of antioxidants in the whole seeds, seed shells, and peeled seeds of the Japanese horse chestnut. The seed shells exhibited the highest antioxidant activities due to the presence of highly polymeric proanthocyanidins as we have reported recently. On the other hand, the peeled seeds predominantly contained flavonols such as quercetin and kaempferol at a high level of 66.7% of total polyphenols, also contributing to the predominant antioxidant activities. The instrumental analysis of the extract from the whole seeds revealed the identification of eight flavonol O-glycosides, including six compounds with quercetin and two species with kaempferol as aglycones. The isolated species exhibited different antioxidant activities depending on the types of aglycones, glycosides, and acylated moieties. The results indicate that the peeled seeds are a good source of flavonol O-glycosides serving as antioxidants to be used for food additives and dietary supplements.


Transient activation of midbrain dopamine neurons by reward risk.
Dopamine neurons of the ventral midbrain are activated transiently following stimuli that predict future reward. This response has been shown to signal the expected value of future reward, and there is strong evidence that it drives positive reinforcement of stimuli and actions associated with reward in accord with reinforcement learning models. Behavior is also influenced by reward uncertainty, or risk, but it is not known whether the transient response of dopamine neurons is sensitive to reward risk. To investigate this, monkeys were trained to associate distinct visual stimuli with certain or uncertain volumes of juice of nearly the same expected value. In a choice task, monkeys preferred the stimulus predicting an uncertain (risky) reward outcome. In a Pavlovian task, in which the neuronal responses to each stimulus could be measured in isolation, it was found that dopamine neurons were more strongly activated by the stimulus associated with reward risk. Given extensive evidence that dopamine drives reinforcement, these results strongly suggest that dopamine neurons can reinforce risk-seeking behavior (gambling), at least under certain conditions. Risk-seeking behavior has the virtue of promoting exploration and learning, and these results support the hypothesis that dopamine neurons represent the value of exploration.


Learning and memory deficits in Notch mutant mice.
Notch is a critical component of evolutionarily conserved signaling mechanisms that regulate development and may contribute to plasticity-related processes, including changes in neurite structure and maintenance of neural stem cells. Deficits in the Notch pathway are responsible for Alagille and Cadasil syndromes, which are associated with mental retardation and dementia. Additionally, in postmitotic neurons, Notch proteins interact with presenilins and with beta-amyloid precursor protein and could therefore have a role in the memory deficits associated with familial and sporadic Alzheimer's disease. To test if alterations in Notch signaling can lead to learning and memory deficits, we studied mice with mutations in this pathway. Here, we show that null heterozygous mutations in Notch1 result in deficits in spatial learning and memory without affecting other forms of learning, motor control, or exploratory activity. We also show that null heterozygous mutations in the downstream cofactor RBP-J result in similarly specific spatial learning and memory deficits. These data indicate that a constitutive decrease in Notch signaling can result in specific learning and memory deficits and suggest that abnormalities in Notch-dependent transcription may contribute to the cognitive deficits associated with Alzheimer's disease and Alagille and Cadasil syndromes.


Northern Ireland's home care services at breaking point.
THE HEALTH service in Northern Ireland is failing to meet the needs of older people, according to a senior official in the ruling Assembly.


Acute postprandial effect of yacon syrup ingestion on appetite: A double blind randomized crossover clinical trial.
Yacon syrup is a rich source of fructooligosaccharides (FOS); however, its diet supplementation effect on subjective sensation and appetite biomarkers in human is still unknown. This study aimed to evaluate the acute postprandial effect of yacon syrup ingestion on appetite. The double-blind crossover clinical trial was carried out with 40 adult women: 20 eutrophic and 20 obese grade I. On each day, the first blood collection was performed after a 12-h fast. Then, the volunteers ingested either intervention A (breakfast + 40 g of placebo) or intervention B (breakfast + 40 g of yacon syrup, containing 14 g of FOS). New aliquots of blood were collected at 45, 60, 90, 120, and 180 min. Appetite was assessed by estimating ghrelin and glucagon-like peptide-1 (GLP-1) levels and by assessing subjective appetite sensation. Analysis was performed using two-way ANOVA, followed by Bonferroni's multiple comparison test. No effect of yacon syrup was observed on postprandial ghrelin and GLP-1 levels at all times evaluated. Similar observations were made after stratifying the analysis by BMI (body mass index) (eutrophic and obese). The effect of yacon syrup on postprandial subjective sensations of hunger, satiety, fullness, and desire to eat was not evident in the total group of women evaluated and even after BMI stratification. We concluded that yacon syrup had no effect on postprandial ghrelin and GLP-1 levels and on the subjective appetite sensation in young adult women.


Elevated pulmonary artery pressure predicts poor outcome after cardiac resynchronization therapy.
Pulmonary artery hypertension is correlated with poor clinical prognosis in patients with chronic heart failure. However, there is a paucity of data concerning the impact of baseline pulmonary artery systolic pressure (PASP) on clinical outcome after cardiac resynchronization therapy (CRT). The aim of the study is to evaluate the association of baseline PASP with CRT response. One hundred eighty-seven consecutive patients undergoing CRT were screened for retrospective analysis. Patients were stratified into three groups based on preoperative PASP which was determined by echocardiogram (group I, PASP < 45 mmHg; group II, 45 mmHg ≤ PASP < 70 mmHg; and group III, PASP ≥ 70 mmHg). Clinical and echocardiographic improvements, as well as the response rate, were assessed 6 months after CRT. Long-term prognosis, measured as transplantation- and hospitalization-free survival, was also compared across the three groups. One hundred eighty-three patients were evaluated with a mean age of 60.7 ± 11.2 years. At 6-month follow-up, patients in group I demonstrated greater clinical improvements (p < 0.05) and higher response rate (p < 0.01) than groups II and III. Compared with group I, death or transplantation was significantly more often noted in group II (hazard ratio, 4.89; 95% CI, 1.53-15.60; p = 0.007) and group III (hazard ratio, 5.91; 95% CI, 1.25-27.94; p = 0.025) as was heart failure readmission for group II (hazard ratio, 2.48; 95% CI, 1.24-4.98; p = 0.011) and group III (hazard ratio, 6.39; 95% CI, 2.26-18.06; p < 0.001). Patients with elevated PASP derive less benefit from CRT. Baseline PASP could serve as an independent predictor for long-term prognosis.


Maleic acid treatment of biologically detoxified corn stover liquor.
Elimination of microbial and enzyme inhibitors from pretreated lignocellulose is critical for effective cellulose conversion and yeast fermentation of liquid hot water (LHW) pretreated corn stover. In this study, xylan oligomers were hydrolyzed using either maleic acid or hemicellulases, and other soluble inhibitors were eliminated by biological detoxification. Corn stover at 20% (w/v) solids was LHW pretreated LHW (severity factor: 4.3). The 20% solids (w/v) pretreated corn stover derived liquor was recovered and biologically detoxified using the fungus Coniochaeta ligniaria NRRL30616. After maleic acid treatment, and using 5 filter paper units of cellulase/g glucan (8.3mg protein/g glucan), 73% higher cellulose conversion from corn stover was obtained for biodetoxified samples compared to undetoxified samples. This corresponded to 87% cellulose to glucose conversion. Ethanol production by yeast of pretreated corn stover solids hydrolysate was 1.4 times higher than undetoxified samples, with a reduction of 3h in the fermentation lag phase.


The influence of WeChat use on driving behavior in China: A study based on the theory of planned behavior.
Driving has become common, and distracted driving, especially that caused by WeChat use, is a significant cause of traffic crashes. Based on the theory of planned behavior, this study analyzes self-reports from a sample of 286 drivers from China to explore the influence of different WeChat functions on driving behavior. The analyses reveal that the intention to use WeChat while driving can substantially predict the use of WeChat while driving. Moreover, drivers' attitudes can effectively predict whether they will send texts, listen to voice messages, and send and browse pictures on WeChat while driving. However, drivers' attitudes cannot effectively predict whether they will read texts or send voice messages on WeChat while driving. In recent years, WeChat has become a popular messaging software, and many drivers use it. Therefore, it is important and necessary to raise awareness among drivers about the dangers of using WeChat while driving.


No phobias about PhoB activation.
Structures of the inactive and activated forms of the receiver domain of PhoB reported in this issue of Structure (Bachhawat et al., 2005) suggest that the OmpR/PhoB subclass of transcription factors becomes active by dimerization about a symmetric axis utilizing the alpha4-beta5-alpha5 surface.


Impairment in a T-maze task following unilateral lateral hypothalamic lesions.
Rats with unilateral LH lesions showed great difficulty performing a contraversive turn to escape shock in a T-maze. Rats that were required to turn to the side ipsilateral to the side of their lesion did not show this performance deficit. The impairment in contraversive responding was shown to dissipate over time. It is proposed that animals with LH lesions do not suffer learning disabilities in escape-avoidance tasks, but can not activate motor behavior directed toward the side contralateral to their lesion.


Caveolin-1 promotes glioma progression and maintains its mitochondrial inhibition resistance.
Glioma is a lethal brain cancer and lacking effective therapies. Challenges include no effective therapeutic target, intra- and intertumoral heterogeneity, inadequate effective drugs, and an immunosuppressive microenvironment, etc. Deciphering the pathogenesis of gliomas and finding out the working mechanisms are urgent and necessary for glioma treatment. Identification of prognostic biomarkers and targeting the biomarker genes will be a promising therapy. From our RNA-sequencing data of the oxidative phosphorylation (OXPHOS)-inhibition sensitive and OXPHOS-resistant cell lines, we found that the scaffolding protein caveolin 1 (CAV1) is highly expressed in the resistant group but not in the sensitive group. By comprehensive analysis of our RNA sequencing data, Whole Genome Bisulfite Sequencing (WGBS) data and public databases, we found that CAV1 is highly expressed in gliomas and its expression is positively related with pathological processes, higher CAV1 predicts shorter overall survival. Further analysis indicated that (1) the differentiated genes in CAV1-high groups are enriched in immune infiltration and immune response; (2) CAV1 is positively correlated with tumor metastasis markers; (3) the methylation level of CAV1 promoters in glioma group is lower in higher stage than that in lower stage; (4) CAV1 is positively correlated with glioma stemness; (5) higher expression of CAV1 renders the glioma cells' resistant to oxidative phosphorylation inhibitors. Therefore, we identified a key gene CAV1 and deciphered its function in glioma progression and prognosis, proposing that CAV1 may be a therapeutic target for gliomas.


A randomized crossover trial comparing three plateletpheresis machines.
The aim of the study was to compare three different apheresis machines with the same donors regarding the processing time required to obtain a 3.5 x 10(11) platelet (PLT) dose and acceptance by donors. A randomized crossover trial was performed to evaluate the differences between the Amicus Crescendo (Baxter Biotech Corp.), the MCS Plus (Haemonetics Corp.), and the Trima Accel (Gambro BCT). Donations from 51 donors were compared for time adjusted to obtain a standard 3.5 x 10(11) PLT dose (TSD3.5), efficiency, adverse reactions, yield, leukodepletion, machine accuracy, and donor preferences. Processing times were measured by chronometer. The same vein access was used during all three processes in each donor. In the statistical analysis, to take into account the nonindependence of several measurements from the same donor, generalized estimating equations were used with an autoregressive correlation matrix. The Accel produced a TSD3.5 (mean +/- SEM) of 47.9 +/- 1.0 min; the Amicus, 60.3 +/- 1.0 min; and the MCS Plus, 66.7 +/- 1.0 (p < 0.0001). The Amicus presented the greatest efficiency (87.5%; p < 0.0028). The MCS Plus demonstrated the highest capacity for leukodepletion (p < 0.0002) despite one process presenting more than 1 x 10(6) white blood cells per unit. The MCS Plus also measured the processing time with the greatest accuracy. No severe adverse effects were observed. The donors preferred the Accel (61%) followed by the Amicus (35%) and the MCS Plus (4%; p < 0.0001) and the processing speed was the most highly valued measure (55%). The Accel is the fastest and, because of this advantage, the machine preferred by donors. The Amicus was the most efficient and the MCS Plus was the only one not to underestimate the processing time.


Inhibitory effect of naked neural BC1 RNA or BC200 RNA on eukaryotic in vitro translation systems is reversed by poly(A)-binding protein (PABP).
Regulated protein biosynthesis in dendrites of neurons might be a key mechanism underlying learning and memory. Neuronal dendritic BC1 RNA and BC200 RNA and similar small untranslated RNAs inhibit protein translation in vitro systems, such as rabbit reticulocyte lysate. Likewise, co-transfection of these RNAs with reporter mRNA suppressed translation levels in HeLa cells. The oligo(A)-rich region of all active small RNAs were identified as the RNA domains chiefly responsible for the inhibitory effects. Addition of recombinant human poly(A)-binding protein (PABP) significantly compensated the inhibitory effect of the small oligo(A)-rich RNA. In vivo, all BC1 RNA appears to be complexed with PABP. Nevertheless, in the micro-environment of dendritic spines of neuronal cells, BC1 RNPs or BC200 RNPs might mediate regulatory functions by differential interactions with locally limited PABP and/or directly or indirectly, with other translation initiation factors.


Oral steroid treatment for hearing improvement in Ménière's disease and endolymphatic hydrops.
To determine the effect of oral steroid treatment on hearing in unilateral Ménière's disease and endolymphatic hydrops patients. Retrospective chart review. Tertiary referral center. All patients presenting during the 2010 calendar year with confirmed unilateral Ménière's disease or endolymphatic hydrops. Those with a first visit and second visit audiogram (n = 58) were included in the analysis of oral steroid treatment effect. Steroid treatment for hearing loss. Change in hearing, as defined by change in affected ear threshold values or speech discrimination score from pretreatment visit to posttreatment visit. Hearing (threshold, speech discrimination score) in patients' affected ear did not significantly change from first visit to second visit after treatment with steroids relative to patients who did not receive steroid treatment. The results of this and other studies would indicate that a Ménière's disease or endolymphatic hydrops patient is unlikely to experience an improvement in hearing from a short course of oral steroid. Clinically observed temporary improvement did not sustain over several months. Further work to elucidate the mechanisms underlying hearing loss in hydrops, perhaps focusing on the dendrite damage noted in animal models of hydrops, is warranted.


Donor plasmapheresis: a comparative study using four different types of equipment.
From September 1985 to May 1986, 106 healthy volunteers underwent automated plasmapheresis in our Institution (CT-AVIS, PAVIA) using four different types of equipment. The aim of this study was to verify the feasibility of harvesting plasma from a single donor and to ascertain: the acceptability of the procedure to donors; the speed, economy and safety of the different types of equipment; the quality of the plasma. The equipment employed in this study was: PCS (Plasma Collection System-Haemonetics) FILTRA (Dideco) PLASMAPUR (Organon Teknika). AUTOPHERESIS CTM-(Hemascience). All four used a single-needle technique. Procedures were well tolerated by donors; no significant changes were observed in pre- and post-procedure laboratory values.


Glycine therapy in isovaleric acidemia.
The therapeutic efficacy of oral glycine was tested in a 3-year-old girl with isovaleric acidemia. An oral leucine load (25 mg/kg) caused a rise of the blood levels of isovaleric, lactic, and pyruvic acids as well as an increase of urinary excretion of the ketone bodies. These changes did not occur when oral glycine (250 mg/kg) was given with the leucine. Glycine supplementation favored the formation of isovalerylglycine, a nontoxic conjugate of isovaleric acid which is excreted rapidly. Excretion of isovalerylglycine rose threefold when leucine and glycine were administered simultaneously. Chronic glycine therapy was tolerated well and may have prevented one episode of ketoacidosis.


Hilar granule cells of the mouse dentate gyrus: effects of age, septotemporal location, strain, and selective deletion of the proapoptotic gene BAX.
The dentate gyrus (DG) principal cells are glutamatergic granule cells (GCs), and they are located in a compact cell layer. However, GCs are also present in the adjacent hilar region, but have been described in only a few studies. Therefore, we used the transcription factor prospero homeobox 1 (Prox1) to quantify GCs at postnatal day (PND) 16, 30, and 60 in a common mouse strain, C57BL/6J mice. At PND16, there was a large population of Prox1-immunoreactive (ir) hilar cells, with more in the septal than temporal hippocampus. At PND30 and 60, the size of the hilar Prox1-ir cell population was reduced. Similar numbers of hilar Prox1-expressing cells were observed in PND30 and 60 Swiss Webster mice. Prox1 is usually considered to be a marker of postmitotic GCs. However, many Prox1-ir hilar cells, especially at PND16, were not double-labeled with NeuN, a marker typically found in mature neurons. Most hilar Prox1-positive cells at PND16 co-expressed doublecortin (DCX) and calretinin, markers of immature GCs. Double-labeling with a marker of actively dividing cells, Ki67, was not detected. These results suggest that, surprisingly, a large population of cells in the hilus at PND16 are immature GCs (Type 2b and Type 3 cells). We also asked whether hilar Prox1-ir cell numbers are modifiable. To examine this issue, we conditionally deleted the proapoptotic gene BAX in Nestin-expressing cells at a time when there are numerous immature GCs in the hilus, PND2-8. When these mice were examined at PND60, the numbers of Prox1-ir hilar cells were significantly increased compared to control mice. However, deletion of BAX did not appear to change the proportion that co-expressed NeuN, suggesting that the size of the hilar Prox1-expressing population is modifiable. However, deleting BAX, a major developmental disruption, does not appear to change the proportion that ultimately becomes neurons.


Metastatic involvement of the humerus: a retrospective study of 51 cases.
The authors performed a retrospective review of 53 patients with 56 completed or impending pathological fractures of the humerus. Sixteen cases were treated with a proximal and one with a distal humeral prosthesis, 38 with an intramedullary device with cement and one with plate fixation. Forty-eight patients with 51 humeral metastases were available to follow-up. No or only occasional pain was observed in 92% of the operated extremities and adequate postoperative function in 90%. No major complications occurred, except one rod migration.


Cell-surface co-receptors: emerging roles in signaling and human disease.
Extracellular signals are transmitted to cells through two classes of cell-surface receptors: signaling receptors that directly transduce signals and signaling co-receptors that bind ligand but that, traditionally, have not been thought to signal directly. Signaling co-receptors modulate the ligand binding and signaling of their respective signaling receptors. In recent years, roles for co-receptors have expanded to include essential functions in morphogen gradient formation, localizing signaling, signaling independently, regulating cell adhesion and orchestrating the signaling of several pathways. The importance of signaling co-receptors is demonstrated by their ubiquitous expression, their conservation during evolution, their prominent role in signaling cascades, their indispensable role during development and their frequent mutation or altered expression in human disease.


In vivo optophysiology reveals that G-protein activation triggers osmotic swelling and increased light scattering of rod photoreceptors.
The light responses of rod and cone photoreceptors have been studied electrophysiologically for decades, largely with ex vivo approaches that disrupt the photoreceptors' subretinal microenvironment. Here we report the use of optical coherence tomography (OCT) to measure light-driven signals of rod photoreceptors in vivo. Visible light stimulation over a 200-fold intensity range caused correlated rod outer segment (OS) elongation and increased light scattering in wild-type mice, but not in mice lacking the rod G-protein alpha subunit, transducin (Gαt), revealing these responses to be triggered by phototransduction. For stimuli that photoactivated one rhodopsin per Gαt the rod OS swelling response reached a saturated elongation of 10.0 ± 2.1%, at a maximum rate of 0.11% s-1 Analyzing swelling as osmotically driven water influx, we find the H2O membrane permeability of the rod OS to be (2.6 ± 0.4) × 10-5 cm⋅s-1, comparable to that of other cells lacking aquaporin expression. Application of Van't Hoff's law reveals that complete activation of phototransduction generates a potentially harmful 20% increase in OS osmotic pressure. The increased backscattering from the base of the OS is explained by a model combining cytoplasmic swelling, translocation of dissociated G-protein subunits from the disc membranes into the cytoplasm, and a relatively higher H2O permeability of nascent discs in the basal rod OS. Translocation of phototransduction components out of the OS may protect rods from osmotic stress, which could be especially harmful in disease conditions that affect rod OS structural integrity.


Implementation of a patient-held urinary catheter passport to improve catheter management, by prompting for early removal and enhancing patient compliance.
Over the past few years a number of strategic initiatives to improve catheter management and reduce associated infections have been introduced. This paper details the introduction of a patient-held catheter passport and an improved documentation record using the PDSA (Plan, Do, Study, Act) cycle of change implementation in one large acute National Health Service (NHS) trust and local health economy (NHS Institute for Innovation and Improvement, 2008).


Ayurvedic Drug Triphala in Combination with Providone Iodine Mitigates Radiation-Induced Mucositis in Head and Neck Cancer Patients without Affecting the Tumor Response.
Radiation-induced mucositis is a dose-limiting concern in the treatment of head and neck cancers (HNC). This study was conducted to determine the effectiveness of the Ayurvedic drug Triphala in reducing radiation-induced mucositis and influencing tumour control when combined with providone iodine. Data from patient files of HNC patients who received Triphala in conjunction with iodine or iodine alone over the course of curative radiotherapy (> 60 Gy) from May 2013 to February 2015 were extracted for this retrospective chart based study. Data was subjected to statistical analysis, X2 and unpaired t test using the Statistical Package for Social Sciences (SPSS), version 17 (IBM, Chicago, USA). When compared to iodine alone, the group that utilised Triphala gargling was very efficient in delaying mucositis, the extent of weight loss (p = 0.038), the incidence (p = 0.03), and the number (p = 0.02) of treatment breaks. However, it had no influence on the radiation-induced tumour response. According to the observations, Triphala coupled with iodine was more successful in preventing radiation mucositis, and without affecting the killing of tumour cells than iodine gargle alone. According to the authors, this is the first observation to demonstrate the value of combining providone iodine with Triphala in preventing radiation-induced oral mucositis.


Gross hematuria and urinary retention among men from a nationally representative survey in Sierra Leone.
To estimate the prevalence of gross hematuria and urinary retention among men in Sierra Leone and report on barriers to care and associated disability. Gross hematuria and urinary retention are classic urologic complaints that require medical attention for significant underlying pathology, but their burden has not been quantified in a developing country. A cluster randomized, cross-sectional household survey was administered in Sierra Leone using the Surgeons OverSeas Assessment of Surgical need tool as a verbal head-to-toe examination. A total of 2 respondents in each of 25 households in 75 clusters were surveyed to assess surgical needs. Data on questions related to blood from the penis and the inability to urinate for men>12 years were included in the present analysis to determine the period and point prevalence of hematuria and urinary retention. From 3645 total respondents, 1054 (28.9%) were men>12 years included in the analysis. Period and point prevalence of gross hematuria were 21.8 per 1000 (95% confidence interval [CI] 13.0-30.7) and 12.3 per 1000 (95% CI 5.7-19.0), respectively, and for urinary retention, they were 19.9 per 1000 (95% CI 11.5-28.4) and 4.7 per 1000 (95% CI 0.5-8.9), respectively. Lack of financial resources was the major barrier to care. Disability assessment showed 19.1% were not able to work as a result of urinary retention, and 34.8% felt ashamed of their gross hematuria. The results provide a prevalence estimate of gross hematuria and urinary retention for men in Sierra Leone. Accessible medical and surgical care will be critical for early intervention and management.


Phosphorylation stoichiometry determination in plant photosynthetic membranes.
This chapter describes different strategies for the study of phosphorylation dynamics and stoichiometry in photosynthetic membranes. Detailed procedures for the detection, large-scale identification, and quantification of phosphorylated proteins optimized for plant thylakoid proteins are given.


Discovery of extended product structural space of the fungal dioxygenase AsqJ.
The fungal dioxygenase AsqJ catalyses the conversion of benzo[1,4]diazepine-2,5-diones into quinolone antibiotics. A second, alternative reaction pathway leads to a different biomedically important product class, the quinazolinones. Within this work, we explore the catalytic promiscuity of AsqJ by screening its activity across a broad range of functionalized substrates made accessible by solid-/liquid-phase peptide synthetic routes. These systematic investigations map the substrate tolerance of AsqJ within its two established pathways, revealing significant promiscuity, especially in the quinolone pathway. Most importantly, two further reactivities leading to new AsqJ product classes are discovered, thus significantly expanding the structural space accessible by this biosynthetic enzyme. Switching AsqJ product selectivity is achieved by subtle structural changes on the substrate, revealing a remarkable substrate-controlled product selectivity in enzyme catalysis. Our work paves the way for the biocatalytic synthesis of diverse biomedically important heterocyclic structural frameworks.


Double burden of nutritional disorders in young Iranian children: findings of a nationwide screening survey.
To assess the national prevalence of short stature, underweight, overweight and obesity in 6-year-old Iranian children before school entry. Cross-sectional nationwide survey. Mandatory national screening programme before entrance to elementary school in 2008 in Iran. All Iranian children entering public and private elementary schools. The study population comprised 862,433 children (48.4% girls, 77.2% urban resident). Overall, 6.5% of children had short stature, 19.1% were underweight, 12.8% were overweight and 3.4% were obese. There was no significant difference in terms of gender, but considerably larger differences were documented among various provinces. The double burden of nutritional disorders among young children warrants a multi-faceted national policy with evidence-based local programmes. Such planning needs a comprehensive surveillance system and centralized data registry for children's growth.


Intercalation-assisted longitudinal unzipping of carbon nanotubes for green and scalable synthesis of graphene nanoribbons.
We have demonstrated an effective intercalation of multi-walled carbon nanotubes (MWCNTs) for the green and scalable synthesis of graphene nanoribbons (GNRs) using an intercalation-assisted longitudinal unzipping of MWCNTs. The key step is to introduce an intercalation treatment of raw MWCNTs with KNO3 and H2SO4, making it promising to decrease the strong van der Waals attractions in the MWCNTs bundles and between the coaxial graphene walls of CNTs. Systematic micro Raman, X-ray photoelectron spectroscopy (XPS), and X-ray diffraction (XRD) characterizations suggest that potassium, nitrate, and sulfate ions play an important role in the CNT intertube and intratube intercalations during the pretreatment. Detailed scanning electron microscopy (SEM), transmission electron microscopy, XRD, and micro Raman characterizations indicate that the developed methodology possesses the ability to synthesis GNRs effectively with an improved CNT concentration in H2SO4 of 10 mg/ml at 70 °C, which is amenable to industrial-scale production because of the decreased amount of strong acid. Our work provides a scientific understanding how to enhance the GNR formation by accelerating the CNT longitudinal unzipping via suitable molecular intercalation.


[How a self-managed team can improve cohesion and communication during the patient's journey in a radiotherapy department: Experience of centre Jean-Bernard (Le Mans, France)].
Efficient communication between professionals is of upmost importance in improving treatment safety in a radiotherapy department, and is also necessary to enhance the quality of work life. Taking as example the organizations in industry, a self-managed team centred on patients with head and neck cancers treated with radiation has been implemented in 2018 in centre Jean-Bernard (Le Mans, France). After over a year's experience, a real benefice has been found and validates the plan to extend this model to other departments.


Acute myocardial infarction and ischemia in the left anterior descending artery territory in a patient with single coronary artery.
This report describes a 77-year-old woman with a single coronary artery who suffered an acute anterior wall myocardial infarction. The single coronary artery arose from the right coronary artery through the transverse trunk, and there were no other cardiovascular anomalies. Coronary angiography did not reveal significant coronary artery stenosis in the left anterior descending artery. The patient was treated medically in the acute phase. She developed typical angina and evidence of myocardial ischemia, and underwent successful coronary artery bypass grafting in the chronic phase with anterior chest pain.


Large and growing environmental reservoirs of Deca-BDE present an emerging health risk for fish and marine mammals.
Polybrominated diphenyl ethers (PBDEs) have been the subject of intense scientific and regulatory scrutiny during recent years. Of the three commercial forms (Penta, Octa and Deca) of PBDEs that have been widely used as flame retardants in textiles, furniture upholstery, plastics, and electronics, only Deca-BDE remains on the general market in North America, while a recent ruling of the European Court spells an impending end to its use in Europe. We review here highlights of aquatic research documenting the rapid emergence of PBDEs as a high priority environmental concern in Canada. PBDEs are being introduced in large quantities to the aquatic environment through sewage discharge and atmospheric deposition. In certain environmental compartments, the single congener BDE-209, the main ingredient in the Deca-BDE formulation, has surpassed the legacy PCBs and DDT as the top contaminant by concentration. Limited biomagnification of BDE-209 in aquatic food webs reflects its high log K(ow) and preferential partitioning into the particle phase. As a result, large environmental reservoirs of BDE-209 are being created in sediments, and these may present a long-term threat to biota: BDE-209 breaks down into more persistent, more bioaccumulative, more toxic, and more mobile PBDE congeners in the environment.


Parental Cancer History and Its Association With Minor Children's Unmet Food, Housing, and Transportation Economic Needs.
A cancer diagnosis is associated with substantial economic burden for patients and their families. Young adult cancer survivors with dependent children may be particularly vulnerable to financial hardship. To examine associations of parental cancer with their children's unmet economic needs. This cross-sectional study used data from the nationally representative 2013 to 2018 US National Health Interview Survey. Children aged 5 to 17 years living in families with and without parental cancer history were queried about recent 1-year experiences. Statistical analyses were conducted from January 2022 to April 2023. Parental cancer history. The main outcomes were children's unmet economic needs, including family-level food insecurity, parent's financial worry about paying for monthly bills and housing costs, and delayed child medical care owing to lack of transportation. Multivariable logistic regressions adjusted for (1) child's characteristics (ie, age group, sex, and race and ethnicity), (2) parent's characteristics (ie, age group, sex, health insurance coverage, comorbid conditions, and obesity status), and (3) family's characteristics (ie, family structure [married or cohabiting parents vs single parent families], highest educational attainment in the family, and family income). Additional analyses focused on children with a parental cancer history to identify potentially modifiable characteristics associated with unmet economic needs. In this cross-sectional study of 22 941 children with (812 children; weighted number, 860 488 children) and without (22 129 children; weighted number, 24 545 463 children) a parental cancer history, the majority of children were aged 5 to 11 years (12 022 children [52.4%]), male (11 920 children [52.0%]), and non-Hispanic White (11 863 children [51.7%]). In adjusted analyses, parental cancer history was associated with more severe family-level food insecurity, including worrying about food running out (odds ratio [OR], 1.97; 95% CI, 1.56-2.49; P < .001), food not lasting (OR, 2.01; 95% CI, 1.56-2.58; P < .001), and inability to afford balanced meals (OR, 1.38; 95% CI, 1.06-1.79; P = .02). Moreover, parental cancer history was associated with parent's worry about paying monthly bills (OR, 1.41; 95% CI, 1.15-1.74; P = .001) and housing-related costs (OR, 1.31; 95% CI, 1.07-1.60; P = .009) and delays in child medical care because of lack of transportation (OR, 2.31; 95% CI, 1.49-3.59; P < .001). Among children with parental cancer history, female children, non-Hispanic Black children, children whose parents had multiple comorbidities, and children living in low-income families were especially vulnerable to unmet economic needs. Parental cancer is associated with greater likelihood of food insecurity, unaffordability of housing and other necessities, and transportation barriers to medical care for minor children. Strategies to identify such children and address their needs are warranted.


TMBIM5 is the Ca2+ /H+ antiporter of mammalian mitochondria.
Mitochondrial Ca2+ ions are crucial regulators of bioenergetics and cell death pathways. Mitochondrial Ca2+ content and cytosolic Ca2+ homeostasis strictly depend on Ca2+ transporters. In recent decades, the major players responsible for mitochondrial Ca2+ uptake and release have been identified, except the mitochondrial Ca2+ /H+ exchanger (CHE). Originally identified as the mitochondrial K+ /H+ exchanger, LETM1 was also considered as a candidate for the mitochondrial CHE. Defining the mitochondrial interactome of LETM1, we identify TMBIM5/MICS1, the only mitochondrial member of the TMBIM family, and validate the physical interaction of TMBIM5 and LETM1. Cell-based and cell-free biochemical assays demonstrate the absence or greatly reduced Na+ -independent mitochondrial Ca2+ release in TMBIM5 knockout or pH-sensing site mutants, respectively, and pH-dependent Ca2+ transport by recombinant TMBIM5. Taken together, we demonstrate that TMBIM5, but not LETM1, is the long-sought mitochondrial CHE, involved in setting and regulating the mitochondrial proton gradient. This finding provides the final piece of the puzzle of mitochondrial Ca2+ transporters and opens the door to exploring its importance in health and disease, and to developing drugs modulating Ca2+ exchange.


Fundus-first laparoscopic cholecystectomy.
Fundus-first dissection (FFD) is an established technique to deal with difficult open cholecystectomies. Although the indications for such an approach are similar for laparoscopic cholecystectomy (LC), FFD is not widely practiced because of difficulties that arise with liver retraction, the dissection of dense adhesions, or obscured cystic pedicles, often necessitating conversion to an open procedure. The aim of this study was to evaluate the indications for FFD and the technical aspects of the procedure in cases with a difficult cystic pedicle. Prospectively collected data and video recordings of cases of fundus-first laparoscopic cholecystectomy (FFLC) were analyzed. The great majority were difficult cases, so we also reviewed the safety aspects of this approach and assessed its effect on the conversion rate. FFLC was resorted to in 35 cases (5%) of 710 consecutive LCs with difficulty grade II (two cases), III (13 cases), or IV (20 cases). There were 16 male patients (46% vs 9% males in the whole), and the mean age was 56 years (ranges, 28-87). The reasons for FFD were dense adhesions preventing the exposure of the cystic pedicle in 14 cases, large Hartmann's pouch stones in 10 cases, short dilated cystic ducts in six cases, and Mirizzi syndrome in three cases. Two cases had contracted "burn-out" gallbladders. Intraoperative cholangiography (IOC) was possible in 24 patients, failed in 10 (29%), and was not attemped in one. Seven patients had bile duct stones and required bile duct exploration. FFLC was completed in 31 patients, 28 of whom were seriously considered for conversion prior to commencing FFD. Conversion was still necessary after trial FFD in four cases (11%) two with Mirizzi abnormalities, one with bile duct stones, and one with dense adhesions. The mean operative time was 125 min, (range, 50-230). There were no operative or technique-related complications. FFLC is feasible and is a safe option for cases with a difficult cystic pedicle. Its use reduced the conversion rate of the series from a potential 5.2% to 1.2%, However, subtotal cholecystectomy or conversion must not be delayed if, after the neck of the gallbladder is reached the anatomy is still unclear.


Purification and properties of a new enzyme, NG,NG-dimethylarginine dimethylaminohydrolase, from rat kidney.
A new enzyme, NG, NG-dimethylarginine dimethylaminohydrolase which plays a role in the metabolism of NG,NG-dimethyl-L-arginine, has been purified to homogeneity from rat kidney. The enzyme consists of a single polypeptide and its molecular weight is about 33,000. The isoelectric point of the enzyme is at pH 5.2. The enzyme catalyzes the hydrolytic liberation of the dimethylamino moiety of NG,NG-dimethyl-L-arginine and forms L-citrulline and dimethylamine. It is highly specific for NG,NG-dimethyl-L-arginine and NG-monomethyl-L-arginine, and the Km values for these amino acids are 0.18 and 0.36 mM, respectively. The enzyme shows the maximum activity at pH 6.5 and requires no cofactor. The activity is strongly inhibited by SH-blocking reagents (e.g. p-chloromercuribenzoate and HgCl2) and divalent metal ions (e.g. Cd2+, Cu2+, and Zn2+).


Polymorphs and Versatile Solvates of 7-Hydroxyisoflavone.
7-hydroxyisoflavone has been crystallized, identified, and characterized as 2 solvent-free conformational polymorphs and 5 solvates, which differ from each other in the mode of packing and in molecular conformation. All the 7 crystal structures were previously unreported. The conformational polymorphs and solvates were compared by Hirshfeld surface and fingerprint plot analysis and were spectroscopically characterized by powder X-ray diffraction, differential scanning calorimetry, and thermal gravimetric analysis. Hydrogen bond played an important role in the formation of polymorphs. From this study, we can predict that more solvates could be cultivated in other polarity solvents such as isopropanol or 2-butanol at appropriate conditions.


Development and validation of polygenic risk scores for prediction of breast cancer and breast cancer subtypes in Chinese women.
Studies investigating breast cancer polygenic risk score (PRS) in Chinese women are scarce. The objectives of this study were to develop and validate PRSs that could be used to stratify risk for overall and subtype-specific breast cancer in Chinese women, and to evaluate the performance of a newly proposed Artificial Neural Network (ANN) based approach for PRS construction. The PRSs were constructed using the dataset from a genome-wide association study (GWAS) and validated in an independent case-control study. Three approaches, including repeated logistic regression (RLR), logistic ridge regression (LRR) and ANN based approach, were used to build the PRSs for overall and subtype-specific breast cancer based on 24 selected single nucleotide polymorphisms (SNPs). Predictive performance and calibration of the PRSs were evaluated unadjusted and adjusted for Gail-2 model 5-year risk or classical breast cancer risk factors. The primary PRSANN and PRSLRR both showed modest predictive ability for overall breast cancer (odds ratio per interquartile range increase of the PRS in controls [IQ-OR] 1.76 vs 1.58; area under the receiver operator characteristic curve [AUC] 0.601 vs 0.598) and remained to be predictive after adjustment. Although estrogen receptor negative (ER-) breast cancer was poorly predicted by the primary PRSs, the ER- PRSs trained solely on ER- breast cancer cases saw a substantial improvement in predictions of ER- breast cancer. The 24 SNPs based PRSs can provide additional risk information to help breast cancer risk stratification in the general population of China. The newly proposed ANN approach for PRS construction has potential to replace the traditional approaches, but more studies are needed to validate and investigate its performance.


[Detection of second tumors in 11C-choline PET/CT studies performed due to biochemical recurrence of prostate cancer].
Early localization of biochemical recurrence in patients after radical treatment of prostate cancer is a widely accepted clinical indication of (11)C-choline PET/CT. Its widespread clinical use has prompted the depiction of incidentalomas, unusual sites of metastatic lesions, as well as false positive and negative cases. Over the last 6 years, a total of 454 (11)C-choline PET/CT studies have been performed in our institution to locate biochemical recurrence of patients with prostate cancer. With these studies, a second neoplasm has been found in 7 patients (1.54%): 3 lung, 2 colorectal, 1 esophagus and 1 esophageal junction, respectively. Although the clinical usefulness of this technique for detecting cancer lesions other than prostate origin is known for those patients who undergo this technique in the accepted indication, the diagnosis of a second tumor has a significant impact on their therapeutic management.


The politics of paying for health reform: zombies, payroll taxes, and the holy grail.
This paper analyzes the politics of paying for health care reform. It surveys the political strengths and weaknesses of major options to fund universal coverage and explores obstacles to changing how the United States finances health care. Finding a politically viable means to finance universal coverage remains a central barrier to enacting health reform.


Isolation of completely inactive plasma prorenin and its activation by kallikreins. A possible new link between renin and kallikrein.
Existing views on prorenin are conflicting and its physiological activation mechanism is not clear. In an attempt to obtain clearcut views on the molecular properties of prorenin in human plasma, the renin zymogen (prorenin) was separated from active renin by two steps of affinity chromatography and it was demonstrated that prorenin is a completely inactivate zymogen contrary to the existing information. Inactive prorenin has an apparent molecular of 56,000 contrary to 46,000-43,000 of partially active prorenin. Isolated and acid-treated human prorenin was shown to be activated by kallikreins from human urine and plasma. This activation was completely blocked by Trasylol. Hog pancreatic kallikrein also activated human prorenin. The kallikrein mediated activation of prorenin indicates the existence of a new link between the vasoconstricting renin-angiotensin system and the vasodilating kallikreinkinin system.


Use of imaging to study leukocyte trafficking in the central nervous system.
The migration of leukocytes from the bloodstream into the central nervous system (CNS) is a key event in the pathogenesis of inflammatory neurological diseases and typically involves the movement of cells through the endothelium of post-capillary venules, which contains intercellular tight junctions. Leukocyte trafficking has predominantly been studied in animal models of multiple sclerosis, stroke and infection. However, recent evidence suggests that immune cells and inflammation mechanisms play an unexpected role in other neurological diseases, such as epilepsy and Parkinson's disease. Imaging leukocyte trafficking in the CNS can be achieved by epifluorescence intravital microscopy (IVM) and multiphoton microscopy. Epifluorescence IVM is ideal for the investigation of leukocyte-endothelial interactions, particularly tethering and rolling, signal transduction pathways controlling integrin activation, slow rolling, arrest and adhesion strengthening in CNS vessels. Multiphoton microscopy is more suitable for the investigation of intraluminal crawling, transmigration and motility inside CNS parenchyma. The mechanisms of leukocyte trafficking in the CNS are not well understood but the use of in vivo imaging techniques to unravel the underlying regulatory pathways will provide insight into the mechanisms of brain damage and may contribute to the development of novel therapeutic strategies. In this review, we discuss recent work in this field, highlighting the development and use of in vivo imaging to investigate leukocyte recruitment in the CNS.


Secretion of keratinocyte growth factor by cultured human endometrial stromal cells is induced through a cyclic adenosine monophosphate-dependent pathway.
To evaluate the effects of known modulators of endometrial function on the production of keratinocyte growth factor by endometrial stromal cells. The effects of dibutyryl-cyclic adenosine monophosphate (db-cAMP), 12-O-tetradecanoylphorbol 13-acetate (TPA), ethinyl estradiol-17alpha (EE), and medroxyprogesterone acetate (MPA) on the secretion of keratinocyte growth factor by endometrial stromal cells were investigated. Research laboratory at a university medical school. Eleven endometrial specimens in the late proliferative phase. Endometrial stromal cells were incubated for 24 hours with db-cAMP, TPA, EE, or MPA. The concentration of keratinocyte growth factor in the culture media was measured using an ELISA. Small amounts of keratinocyte growth factor were detected in the culture media of unstimulated endometrial stromal cells. The production of keratinocyte growth factor by endometrial stromal cells was stimulated with db-cAMP in a dose-dependent manner. The stimulatory effect of db-cAMP was inhibited by Rp-adenosine 3',5'-cyclic monophosphothioate. None of TPA, EE, nor MPA affected the keratinocyte growth factor production by these cells. These results suggest that a cAMP-dependent pathway may play an important role in the regulation of keratinocyte growth factor production by endometrial stromal cells. Keratinocyte growth factor secreted by endometrial stromal cells may be involved in the regeneration of the endometrium during the normal menstrual cycle and early pregnancy.