Synthesis of mannose-containing analogues of (1-->6)-branched (1-->3)-glucohexaose (I).
alpha-D-Manp-(1-->3)-[alpha-D-Manp-(1-->6)]-alpha-D-Glcp-(1-->3)-beta-D-Glcp-(1-->3)-[alpha-D-Manp-(1-->6)]-D-Glcp and alpha-D-Manp-(1-->3)-[beta-D-Glcp-(1-->6)]-alpha-D-Glcp-(1-->3)-beta-D-Glcp-(1-->3)[-alpha-D-Manp-(1-->6)]-D-Glcp were synthesized in a regio- and stereoselective way as the mannose-containing analogues of the immunomodulating beta-D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-alpha-D-Glcp-(1-->3)-beta-D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-D-Glcp.


Integration of a dominant selectable marker into human chromosomes and transfer of marked chromosomes to mouse cells by microcell fusion.
A method for the production of stable mouse-human cell hybrids containing a single human chromosome is described. As a first step in this method, a cloned selectable marker, the E. coli xanthine-guanine phosphoribosyltransferase (Ecogpt) gene, was transferred to human cells to generate cell lines each carrying Ecogpt integrated into a different site. Human chromosomes marked with Ecogpt were transferred further into mouse cells by microcell fusion. Monochromosomal hybrids, in which the human chromosome is maintained by selection, have been produced for chromosomes 2, 5, 16, and a rearranged chromosome involving a translocation between chromosomes 1 and 2. In addition to these monochromosomal hybrids, we have also obtained monochromosomal hybrids for human chromosomes 6, 12, and 17 by selection for the loss of marked chromosome from the microcell hybrids each containing two human chromosomes. Although the human chromosome present in these hybrids cannot be maintained by selection, 80-90% of cells retained the transferred chromosome on continuous growth for 15 days. Monochromosomal hybrids would provide biological materials to construct genetic maps of human chromosomes. In addition, chromosomes marked with dominant selectable markers can be transferred further to any cell line of interest in inter- or intra-species combination.


The role of silicone ureteral stents in extracorporeal shock wave lithotripsy of large renal calculi.
Between November 1984 and December 1985 extracorporeal shock wave lithotripsy was used to treat 1,645 kidneys at our institution. A total of 646 kidneys with stone burden greater than 14 mm. was evaluated with regard to the impact of silicone ureteral stents in post-extracorporeal shock wave lithotripsy morbidity. Our results indicate that small stones were pulverized and eliminated with minimum morbidity. Larger stones frequently were associated with post-treatment ureteral obstruction by sand and fragments. Of 283 kidneys with stone burden exceeding 25 mm. pretreatment placement of silicone ureteral stents reduced complications from 26 to 7 per cent and auxiliary procedure rates from 15 to 6 per cent. Silicone ureteral stents protect the kidney from ureteral obstruction, and allow for safe and effective extracorporeal shock wave lithotripsy of large renal calculi.


[Calcium regulation of muscle contraction: the molecular regulation mechanisms of contracility].
The role of calcium in muscle activation is described with particular emphasis on interactions occurring between the regulatory proteins troponin-C, troponin-I, actin and myosin. These interactions depend on calcium, but in turn they also influence the calcium binding to troponin-C. They may be affected by short "competitive" peptides whose sequences are similar to the amino-acid sequences on the protein-interaction sites. They may be also affected by certain drugs (cardiotonic calcium sensitizers) that influence the calcium sensitivity of myofilaments.


[Investigation of the brain reactiveness depending on the type of human intrinsically present electroencephalography].
While adopting the lighting flashes 9 Hz in frequency, and while the absence of the adopting the rythm reaction during the photostimulation conduction in majority of the patients examined the alpha-oscillations amplitude according to background electroencephalography constitutes 25-50 mcV; while adopting of the lighting flashes rythm with 6 and 12 Hz frequency--50-75mcV.


Anemia - an initial manifestation of Bing-Neel syndrome: A case report.
It is very likely that we will miss Bing-Neel syndrome (BNS) when its initial sign is anemia.Patient concerns: A 59-year-old woman presented with episodic loss of consciousness, anemia, and extremity muscle strength scores (5-) and extremity tendon reflexes (++). Magnetic Resonance Imaging (MRI) showed abnormal signal in the left hippocampus, left insula, and right occipital lobe. Quantitative serum immunoglobulins showed elevated immunoglobulinm (IgM) (60.6g/L). Bone marrow biopsy showed lymphoplasmacytic lymphoma (LPL) and tested positive for the MYD88 L265P mutation suggesting Waldenström macroglobulinemia (WM). The patient underwent 3 plasma exchange treatments in the department of hematology followed by chemotherapy (cyclophosphamide for injection, bortezomib for injection). The patient's condition improved after treatment. Clinicians must remain vigilant, as BNS may be the only sign of WM progression in a patient well-controlled on treatment.


Benchmarking various green fluorescent protein variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for live cell imaging.
Green fluorescent protein (GFP) offers efficient ways of visualizing promoter activity and protein localization in vivo, and many different variants are currently available to study bacterial cell biology. Which of these variants is best suited for a certain bacterial strain, goal, or experimental condition is not clear. Here, we have designed and constructed two "superfolder" GFPs with codon adaptation specifically for Bacillus subtilis and Streptococcus pneumoniae and have benchmarked them against five other previously available variants of GFP in B. subtilis, S. pneumoniae, and Lactococcus lactis, using promoter-gfp fusions. Surprisingly, the best-performing GFP under our experimental conditions in B. subtilis was the one codon optimized for S. pneumoniae and vice versa. The data and tools described in this study will be useful for cell biology studies in low-GC-rich Gram-positive bacteria.


Lipopolysaccharide Attenuates the Cytotoxicity of Resveratrol in Transformed Mouse Macrophages.
Resveratrol and pterostilbene are natural products that are present in plants and have been incorporated into various dietary supplements. Numerous beneficial pharmacologic effects have been reported for these stilbenes; however, the mechanism by which these compounds exert a cytotoxic effect in RAW 264.7 macrophages has not been well characterized. We have previously described that resveratrol is toxic to these tumor-derived macrophages and that stimulation with lipopolysaccharide (LPS) reduces resveratrol toxicity via a mechanism that involves activation of toll like receptor 4. In the present work, we examined the cellular and molecular effects of resveratrol and the related compound pterostilbene by determining cell viability and caspase 3 activity in control and LPS-stimulated RAW 264.7 macrophages incubated with these stilbenes for 24 h. We found that LPS stimulation reduced the cytotoxicity of resveratrol but not of pterostilbene in these cells. When examined for effects on caspase 3 activation after a 24 h incubation, resveratrol and pterostilbene were each found to separately and significantly increase caspase 3 activity in these cells. LPS stimulation prevented caspase 3 activation by pterostilbene and reduced caspase 3 activation by resveratrol in RAW 264.7 macrophages. The data presented here indicate that LPS induces a phenotype switch in tumor-derived RAW 264.7 macrophages in which cells experiencing LPS in the presence of resveratrol or pterostilbene become less likely to activate the pro-apoptotic factor caspase 3.


Failure to find association between TRAR4 and schizophrenia in the Chinese Han population.
The TRAR4 gene locates in SCZD5 (MIM 603175), which a number of studies have linked with schizophrenia. One recent study suggested that three TRAR4 variants (M1: rs4305745, P=0.0014; M2: rs6903874, P=0.0026; M3: rs6937506, P=0.0052) in the 3'-UTR were associated with schizophrenia. To replicate these findings, we conducted a family-based association study within a sample of 235 Chinese Han trios. However, we didn't find significant evidence of preferential transmission of the three variants across all the trios (all P values>0.2). Thus, we conclude that TRAR4 is not a major or independent determinant in the occurrence of schizophrenia in the Chinese Han population.


Door-To-Triage Time in a Canadian Tertiary-Care Center.
The interval from patient arrival to triage is arguably the most dangerous time a patient spends in the emergency department (ED), as they are an unknown entity until assessed by a health care professional. We sought to quantify door-to-triage time (DTT), an important factor in patient safety that has not yet been quantified in Canada. Data were collected from all ambulatory patients presenting to a tertiary-care ED during a consecutive 7-day period. Demographic information, arrival time (door time), triage time, and Canadian Triage and Acuity Score (CTAS) were collected. DTT was compared across variables using Kruskal-Wallis one-way analysis of variance. Seven hundred and seventy-five patients were included in the study, representing 82.9% of ambulatory patients. DTT was variable (1-86 min) with a median of 12 min (interquartile range [IQR] 6-21 min). Patients in the 5th percentile with the longest DTT waited a median of 54 min (IQR 48-63 min). DTT varied across days of the week (p < 0.01); the longest wait was on Monday (median 22 [IQR 11-43] min) and the shortest on Sunday (median 8 [IQR 5-12] min). There was no relationship between DTT and CTAS (p = 0.12). DTT is an important variable affecting patient safety. Given site-specific factors, replication across additional centers is necessary. Additional research evaluating factors affecting DTT, different triage paradigms, and quality improvement interventions should be undertaken.


Extracellular binding of indinavir to matrix metalloproteinase-2 and the alpha-7-nicotinic acetylcholine receptor: implications for use in cancer treatment.
Results from recent studies have suggested a role for protease inhibitors in altering mechanisms involved in the initiation and proliferation of cancer cells. One such inhibitor, indinavir, may act as an anti-cancer agent by modulating the alpha-7-nicotinic acetylcholine receptor, which is a pro-carcinogenic protein that has been researched in conjunction with nicotine in lung cancer development. In our study, we compare indinavir's binding affinity towards α7-nAchR and MMP-2, another promoter of malignancy, to determine what extracellular effects the drug has before being internalized to inhibit HIV-1 protease. A computer program, PyRx, was used to compare indinavir's binding affinity with digital models for α7-nAchR, MMP-2 and HIV-1 protease, which were then compared to the results of in vitro binding assays for these targets. PyRx testing predicted the highest binding affinity values for indinavir to MMP-2 (mean = 8.77 kcal/mol, S.D. = 0.29), followed by the α7-nAchR (mean = 8.53 kcal/mol, S.D. = 0.15) and HIV-1 protease (mean = 7.5 kcal/mol, S.D. = 0.44). In vitro, indinavir's mean percent inhibition of control values were 103.2 for HIV-1 protease, 5.3 for MMP-2, and 7.7 for the α7-nAchR. Binding affinity values for indinavir to MMP-2 and α7-nAchR were not significantly different. Using PyRx to predict affinity compared with in vitro testing did not yield comparable results. However, indinavir was shown to slightly inhibit both α7-nAchR and MMP-2, which may have ramifications in the drug's delivery to the intracellularly located HIV-1 protease.


Author Correction: Engineering monocyte/macrophage-specific glucocerebrosidase expression in human hematopoietic stem cells using genome editing.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Transcutaneous bilirubin in predicting hyperbilirubinemia in term neonates.
To assess the utility of 24 and 48 hours transcutaneous bilirubin (TcB) index for predicting subsequent significant hyperbilirubinemia in healthy term neonates. TcB indices were obtained for healthy, breastfed, term AGA newborns at 24 +/- 2, 48 +/- 2 and subsequently at intervals of 24 hours. Neonates with illness, on treatment and positive Direct Coomb's test were excluded. Serum bilirubin levels were obtained whenever indicated. Neonates having serum bilirubin > or = 17 mg/dL were considered as significant hyperbilirubinemia. The 24 and 48 hour TcB indices, as risk predictors for such hyperbilirubinemia were determined. Study included 461 healthy term neonates. The mean birth weight was 2949 (+/- 390) gm and mean gestation of 38.6 (+/- 1.1) weeks. Eight one (17.6%) had significant hyperbilirubinemia. Of 461, 135 (29.3%) had TcB index. The 24 and 48 hour TcB indices are predictive for subsequent significant hyperbilirubinemia and can guide clinician in early discharge of healthy term newborns.


The inadequate treatment of pain: collateral damage from the war on drugs.
Jason Nickerson and Amir Attaran examine the vast inequities in medical pain relief around the world and argue that the global control of licit narcotics be shifted from the International Narcotic Control Board to WHO.


Experimental and simulation studies of heat flow and heterocyclic amine mutagen/carcinogen formation in pan-fried meat patties.
Heterocylic amine (HA) compounds formed in the cooking of certain foods have been shown to be bacterial mutagens and animal carcinogens, and may be a risk factor for human cancer. To help explain the variation observed in HA formation under different cooking conditions, we have performed heat-flow simulations and experiments on the pan-frying of beef patties. The simulations involve modeling the heat flow within a meat patty using empirically derived thermal transport coefficients for the meat. The predicted temperature profiles are used to integrate the Arrhenius rate equation to estimate the concentration of HAs formed in the meat. We find that our simulations accurately model experimentally determined temperature profiles, cooking times, HA spatial distributions and total HA formation in patties that are flipped once during the pan-frying process. For patties flipped every 60 s, the simulations qualitatively agree with experiment in predicting reduced cooking times and HA formation relative to the singly-flipped patties. However, the simulations overestimate the effect of rapid flipping on cooking times and underestimate the effect of flipping on total HAs formed. These results suggest that the dramatic reductions in HA formation due to rapid flipping may be due to factors other than the heating process or that there is a critical feature of the flipping process that is not captured in our model.


Characterization of a new insect cell line that is derived from the neonate larvae of Papilio xuthus (Lepidoptera: Papilionidae) and its susceptibility to AcNPV.
The cell line RIRI-PX1 was established from neonate larval tissues of Papilio xuthus by performing primary cultures in the modified Grace medium that was supplemented with 20% fetal bovine serum (FBS). The cell line primarily consisted of spindle-shaped and spherical cells which attached themselves to the flask. The population-doubling times (PDTs) at the 50th and 60th passage were 42.5 h and 42.1 h respectively. The average chromosome numbers of RIRI-PX1 cell line from passage 5 to passage 50 ranged from 103 to 199. It was confirmed that RIRI-PX1 cell line was derived from P. xuthus by comparing the mitochondrial cytochrome c oxidase subunit I gene (COI) of RIRI-PX1 cells and P. xuthus eggs. This cell line was susceptible to the Autographa californica nucleopolyhedrovirus (AcNPV) and produced high yield of polyhedral occlusion bodies (43.9OBs/cell) after 10 days of infection by AcNPV. The virus titer of AcNPV infected RIRI-PX1 cells was 3.25×10⁷ TCID₅₀/ml. We concluded that the RIRI-PX1 cell line is established from the neonate larvae tissues successfully and the cells of the cell line are sensitive to AcNPV.


INDOCYANINE GREEN ANGIOGRAPHY OF TYPE 1 MACULAR NEOVASCULARIZATION IN AGE-RELATED MACULAR DEGENERATION AND CENTRAL SEROUS CHORIORETINOPATHY REVEALS DIFFERENT DISEASE MECHANISMS.
To assess the rate of late phase hyperfluorescent plaque (LPHP) in Type 1 macular neovascularization (MNV) in central serous chorioretinopathy (CSCR) and age-related macular degeneration (AMD) and to evaluate its prognostic value. Retrospective study including Type 1 MNV in AMD and CSCR, from 2012 to 2020. Eyes with a late indocyanine green angiography image (>20 minutes) and clear visualization of MNV on optical coherence tomography angiography (OCTA) were included. Quantitative and qualitative parameters on optical coherence tomography and best-corrected visual acuity were recorded at baseline and after three monthly antivascular endothelial growth factor injections. Eighty-three eyes were included, 35 with CSCR and 48 with AMD. Patients in the CSCR group were significantly younger than in the AMD group (61.3 ± 10.4 vs. 80.2 ± 6.8 years, respectively, P < 0.001), predominantly male (68.6% CSCR vs. 35.4% AMD; P = 0.003), and with a thicker choroid (379 ± 93.3 µ m vs. 204.2 ± 93.2 µ m; P < 0.001). Type 1 MNV in CSCR showed fewer LPHP compared with AMD (31.4% vs. 77.1%; P < 0.001). The baseline visual acuity was lower in patients with LPHP (0.37 ± 0.22 vs. 0.27 ± 0.28 logarithm of the minimum angle of resolution, P = 0.03). On multivariate analysis, AMD was associated with the presence of LPHP ( P < 0.001). No significant difference in the response to antivascular endothelial growth factor was observed. Leakage of macromolecules from MNV and accumulation in the retinal pigment epithelium and/or in the stroma imaged by the LPHP is less common in eyes with Type 1 MNV in CSCR than in AMD. Late phase indocyanine green angiography imaging offers an insight into the metabolism of the dye and the environment surrounding the neovascular membrane.


Seasonal variation in catabolic enzyme activities in breast muscle of some migratory birds.
Five bird species were examined in order to ascertain if any changes in flight muscle catabolism take place between breeding season and migration. Two different patterns were discovered. The first consists of a high oxidative capacity and a low glycolytic and anaerobic capacity during migration. The converse occurs during the breeding season, i.e. low oxidative, high glycolytic and anaerobic capacity. The pattern was found in those species that deposit large amounts of fat prior to migration. The second pattern was similar to the first, but there was no change in fatty acid oxidation capacity between breeding season and migration. The pattern was found in those species that do not deposit much fat towards migration. These changes are believed to reflect differences in migration strategy and differences in locomotory activity during different seasons. Deviations from these patterns are discussed.


Lipophilic amines as potent inhibitors of N-acylethanolamine-hydrolyzing acid amidase.
N-Acylethanolamines (NAEs) including N-arachidonoylethanolamine (anandamide) and N-palmitoylethanolamine are endogenous lipid mediators. These molecules are degraded to the corresponding fatty acids and ethanolamine by fatty acid amide hydrolase (FAAH) or NAE-hydrolyzing acid amidase (NAAA). Lipophilic amines, especially pentadecylamine (2c) and tridecyl 2-aminoacetate (11b), were found to exhibit potent NAAA inhibitory activities (IC(50)=5.7 and 11.8μM), with much weaker effects on FAAH. These simple structures would provide a scaffold for further improvement in NAAA inhibitory activity.


Oncogenic transformation by BK virus and association with human tumors.
BK virus (BKV), a human polyomavirus closely related to JC virus and Simian Virus 40, is ubiquitous in human populations worldwide. After primary infection, BKV establishes a lifelong latent infection in many organs. BKV transforms rodent cells to the neoplastic phenotype and is highly oncogenic in rodents. This review considers the oncogenic potential of BKV in humans and its possible involvement in human tumors. BKV sequences and T antigen (Tag) are detected in several types of human neoplasms, although the viral load is generally low, with less than one copy of the viral genome per cell. The possible causative role of BKV in human oncogenesis rests on the ability of BKV Tag to inactivate the functions of tumor suppressor proteins p53 and pRB family as well as on its ability to induce chromosomal aberrations in human cells. A 'hit and run' mechanism and secretion of paracrine growth factors by BKV Tag-positive cells, recruiting into proliferation neighboring and distant cells, are discussed as possible BKV pathogenic elements in human oncogenesis.


The lack of effectiveness of (-)-epicatechin against alloxan induced diabetes in Wistar rats.
(-)-Epicatechin, a naturally occurring flavonoid, has been reported to protect pancreatic beta cells from alloxan-induced diabetes (1) and to stimulate beta cell regeneration when given after alloxan administration (2,3). However, in the present study, we have not been able to confirm these findings. Administration of (-)-epicatechin (100 mg/kg/day for 15 days) beginning three days after alloxan administration (140 mg/kg) had no significant effect on blood glucose levels when compared to alloxan control animals. Another study which failed to demonstrate an antidiabetic effect of (-)-epicatechin (4) has been criticized for not using fresh solutions of (-)-epicatechin because of its instability in aqueous solution (5). We have found, however, that (-)-epicatechin is stable for at least five days based on thin-layer chromatographic analysis and optical rotation measurements.


Corrigendum: Identification of an autophagy-related 12-lncRNA signature and evaluation of NFYC-AS1 as a pro-cancer factor in lung adenocarcinoma.
[This corrects the article DOI: 10.3389/fgene.2022.834935.].


Clinical Effect of Preoperative 25-OH-Vitamin D3 Level in Liver Transplant Recipients.
Vitamin D deficiency is common in patients with chronic liver disease and is associated with increased risk of infection and mortality. This study evaluated the effects of preoperative vitamin D levels on clinical outcomes after liver transplant. This single-center retrospective study included liver transplant recipients from June to November 2017 who had preoperative 25-OH-vitamin D3 (25-OH-D3) data. Severe vitamin D deficiency, insufficiency, and normal levels were defined as serum 25-OH-D3 concentrations of < 10 ng/mL, 10 to 20 ng/mL, and ≥ 20 ng/mL, respectively. The primary outcome was length of hospital stay; secondary outcomes included the duration of normalization of inflammatory markers after liver transplant, new infection rates, rejection rates, length of intensive care unit stay, and mortality according to preoperative 25-OH-D3 levels. Among 219 liver transplant recipients, 67.6% were vitamin D-deficient. The mean (standard deviation) 25-OH-D3 concentration was 17.8 (13.2) ng/mL, and 65 (29.7%) patients had levels < 10 ng/mL. Patients with lower mean 25-OH-D3 levels had significantly longer intensive care unit (13.8 [21.9] days vs 5.9 [12.3] days vs 2.7 [4.6] days, P < .001) and hospital (59.0 [66.0] days vs 42.0 [67.4] days vs 27.2 [17.1] days, P = .001) stays. The incidence of new infections was higher in the vitamin D deficiency group. (46.2% vs 28.9% vs 14.1%, P < .001). A higher Nutritional Risk Screening 2002 score (adjusted odds ratio, 1.77; 95% confidence interval [CI], 1.24-2.56; P = .002) and severe vitamin D deficiency (adjusted odds ratio, 3.43; 95% CI, 1.57-7.57; P = .002) were significant risk factors for poor outcome among patients who had been in the hospital for more than 43 days. Vitamin D deficiency before liver transplant was associated with increased intensive care unit and hospital lengths of stay. Although several factors may influence the clinical outcomes of patients with liver transplant, low vitamin D3 was an independent risk factor.


Development and Characterization of Bio-Based Composite Films for Food Packing Applications Using Boiled Rice Water and Pistacia vera Shells.
Customer demand for natural packaging materials in the food industry has increased. Biocomposite films developed using boiled rice water could be an eco-friendly and cost-effective packaging product in the future. This study reports the development of bio-based films using waste materials, such as boiled rice water (matrix) and Pistacia vera shells (reinforcement material), using an adapted solution casting method. Several film combinations were developed using various concentrations of plasticizing agent (sorbitol), thickening agent (oil and agar), and stabilizing agents (Arabic gum, corn starch, and Pistacia vera shell powder). Various packaging properties of the film were analyzed and examined to select the best bio-based film for food packaging applications. The film fabricated with Pistacia vera shell powder in the biocomposite film exhibited a reduced water solubility, swelling index, and moisture content, as compared to polyethene packaging material, whereas the biocomposite film exhibited poor antimicrobial properties, high vapor transmission rate, and high biodegradability rate. The packaging properties and characterization of the film indicated that the boiled rice water film with Pistacia vera shell powder was suitable for packaging material applications.


Concurrent Treatment of Substance Use and PTSD.
Substance use disorders (SUD) and posttraumatic stress disorder (PTSD) are chronic, debilitating conditions that frequently co-occur. Individuals with co-occurring SUD and PTSD suffer a more complicated course of treatment and less favorable treatment outcomes compared to individuals with either disorder alone. The development of effective psychosocial and pharmacological interventions for co-occurring SUD and PTSD is an active and critically important area of investigation. Several integrated psychosocial treatments for co-occurring SUD and PTSD have demonstrated promising outcomes. While recent studies examining medications to treat co-occurring SUD and PTSD have yielded encouraging findings, there remain substantial gaps in the evidence base regarding the treatment of co-occurring SUD and PTSD. This review will summarize the findings from clinical trials targeting a reduction in SUD and PTSD symptoms simultaneously. These results may improve our knowledge base and subsequently enhance our ability to develop effective interventions for this complex comorbid condition.


Developmental toxicity and estrogenic potency of zearalenone in zebrafish (Danio rerio).
Zearalenone (ZEA, F2) is one of the most common mycotoxins and the only known mycoestrogen. It enters the food and feed chain from contaminated cereals and infiltrates into sewage or natural waters posing potential threat to exposed livestock, wildlife and humans. Therefore evaluation of its biological effects is of international importance. We performed toxicological tests on zebrafish (Danio rerio) larvae and adults. Developmental toxicity was assessed by an extended (5 days) fish embryo toxicity test (FET). Effects of early ZEA exposure were concentration-dependent with LC50 and LC10 values of 893 and 335 μg/L. In larvae exposed to 500 μg/L and above, ZEA induced similar phenotype to has (heart-and soul) showing defects in heart and eye development and upward curvature of the body axis. From 250 μg/L at 72 hpf the gap in the melanophore streak at the base of the tail fin was missing and the fin fold was abnormal, suggesting disturbance in the development of the adult tail fin primordium. Estrogenic potency was measured on the basis of Vitellogenin (Vtg) protein (adults) levels and relative abundance of vitellogenin-1 mRNA (vtg-1) (larvae and adults). qRT-PCR in larvae proved to be sufficient substitute to adult tests and sensitive enough to detect ZEA in 0.1 μg/L concentrations, that is close to levels observed in wastewaters. Developmental defects reveal that besides direct estrogenic effects, zearalenone might interact with other ontogenic pathways.


Orientation and lipid-peptide interactions of gramicidin A in lipid membranes: polarized attenuated total reflection infrared spectroscopy and spin-label electron spin resonance.
Gramicidin A was incorporated at a peptide/lipid ratio of 1:10 mol/mol in aligned bilayers of dimyristoyl phosphatidylcholine (DMPC), phosphatidylserine (DMPS), phosphatidylglycerol (DMPG), and phosphatidylethanolamine (DMPE), from trifluoroethanol. Orientations of the peptide and lipid chains were determined by polarized attenuated total reflection infrared spectroscopy. Lipid-peptide interactions with gramicidin A in DMPC bilayers were studied with different spin-labeled lipid species by using electron spin resonance spectroscopy. In DMPC membranes, the orientation of the lipid chains is comparable to that in the absence of peptide, in both gel and fluid phases. In gel-phase DMPC, the effective tilt of the peptide exceeds that of the lipid chains, but in the fluid phase both are similar. For gramicidin A in DMPS, DMPG, and DMPE, the degree of orientation of the peptide and lipid chains is less than in DMPC. In the fluid phase of DMPS, DMPG, and DMPE, gramicidin A is also less well oriented than are the lipid chains. In DMPE especially, gramicidin A is largely disordered. In DMPC membranes, three to four lipids per monomer experience direct motional restriction on interaction with gramicidin A. This is approximately half the number of lipids expected to contact the intramembranous perimeter of the gramicidin A monomer. A selectivity for certain negatively charged lipids is found in the interaction with gramicidin A in DMPC. These results are discussed in terms of the integration of gramicidin A channels in lipid bilayers, and of the interactions of lipids with integral membrane proteins.


Effects of alpha-phenyl-N-tert-butyl nitrone and N-acetylcysteine on hydroxyl radical formation and dopamine depletion in the rat striatum produced by d-amphetamine.
Previous studies have shown that treatment with free radical scavengers attenuated the D-amphetamine (AMPH) neurotoxicity. But several of these agents also prevent AMPH-induced elevation of body temperature in the rat. Thus, further studies are needed to determine if blockade of the production of free radical or hypothermia are related to the neuroprotective mechanism of the free radical scavengers for AMPH neurotoxicity. In the present study, we examined the effects of the free radical scavengers alpha-phenyl-N-tert-butyl nitrone (PBN) and N-acetylcysteine (NAC) on long-term depletion of striatal dopamine (DA) and lipid peroxidation formation and on hyperthermia induced by AMPH. We also determined their effects on acute hydroxyl radical formation after direct intrastriatal infusion of AMPH. The results showed that both significantly attenuated long-term DA depletion and lipid peroxidation formation in the rat striatum at the dose range that did not block hyperthermia induced by AMPH. These agents also completely inhibited the production of hydroxyl radical after AMPH infusion into the striatum. Our results suggest that free radical scavengers such as PBN and NAC could protect against AMPH-induced oxidative stress and DAergic terminal toxicity via their free radical removing property independent of lowering the core body temperature of rats, and imply that supplement with antioxidants is a potential strategy in the treatment of AMPH neurotoxicity.


Microclimatic variation in UV perception and related disparity in tropane and quinolizidine alkaloid composition of Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger.
The aim of current research was to evaluate the physiological adjustment in three medicinal herbs viz., Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger to the winter period characterised by intense UV flux in Kashmir valley across the North Western Himalaya. Quinolizidine (QA) and tropane alkaloid (TA) concentrations were analysed in these herbs thriving at two different altitudes via GC-MS and correlated by PCA analysis. This study investigated the hypothesis that UV reflectance and absorbance at low temperatures are directly related to disparity in alkaloid accumulation. Among QAs in L. polyphyllus, ammodendrine and lupanine accumulated at higher concentration and exhibited significant variation of 186.36% and 95.91% in ammodendrine and lupanine respectively in both sites. Tetrahydrohombifoline displayed non-significant variation of about 9.60% irrespective of sites. Among tropane alkaloid (TA), hyoscyamine was recorded as the most abundant constituent irrespective of the plant and site while apotropine accumulated in lesser quantity in A. acuminata than H. niger. However, apotropine demonstrated significant variation of 175% among both sites. The final concentration of quinolizidine (QA) and tropane alkaloid (TA) reflects the interplay between reflectance and absorbance of UV radiation response field. These findings suggest that spectral response of UV light contributes directly to alkaloid biosynthesis.


Antioxidant potentials of furanodihydrobenzoxanthones from Artocarpus elasticus and their protection against oxLDL induced injury in SH-SY5Y cells.
Exposure to reactive oxygen species (ROS) leads to the oxidation of low-density lipoproteins (LDL), converting them into oxidized ones (oxLDL), which are involved in the pathogenesis of Alzheimer's disease, suggesting a potential link between lipid dysregulation and neurodegenerative processes. Phenolic metabolites derived from Artocarpus elasticus root bark were found to possess significant antioxidant properties at three different radical scavenging assays, including 2,2-diphenyl-1-picrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC), and thiobarbituric acid reactive substances (TBARS). Among them, furanodihydrobenzoxanthones (1-3) demonstrated notable protection against Cu2+ induced LDL oxidation, with IC50 values ranging from 0.9 to 2.9 μM in measurement of the malondialdehyde (MDA) production at TBARS and prolonged lag times (>180 min) in the generation of conjugated diene (CD). At a concentration of 10 μM, all three compounds (1-3) effectively protected against LDL oxidation as determined by relative electrophoretic mobility (REM). The most potent compound 1 defended human neuroblastoma SH-SY5Y cells from oxLDL-mediated dysfunction, including oxLDL-induced cytotoxicity, inhibited reactive oxygen species (ROS) formation, and enhancing mitochondrial membrane potential (ΔΨm). Individual components annotation in the ethylacetate extract was performed using LC-ESI-QTOF/MS, which serves as a chemotaxonomic marker for A. elasticus root barks.


The search-inference framework: a proposed strategy for novice clinical problem solving.
Medical education in the clinical clerkship years emphasises the systematic collection and organisation of patient information to be combined with domain-specific knowledge of disease processes. Eventually, novice clinical problem solvers will learn to recognise patterns within the patient data ('illness scripts') which suggest the main diagnostic possibilities. Before novice problem solvers develop these illness scripts, pattern recognition may not be effective for solving clinical problems. This discussion paper describes the application of a decision framework adapted from cognitive psychology (the search-inference framework) to basic medical problem solving. Emphasis is placed on problem solving by novices who have not yet developed a full compliment of illness scripts. The search-inference framework is similar to the approach taken by laypersons to diagnose their own symptoms or solve other problems. It may be especially useful for students who have not yet developed a sizeable repertoire of illness scripts, and the principles described may also be invoked by experienced clinicians confronting difficult clinical problems.


Monoclonal antibody D2-40, a new marker of lymphatic endothelium, reacts with Kaposi's sarcoma and a subset of angiosarcomas.
There is controversy over the histogenesis of Kaposi's sarcoma (KS) from lymphatic or blood vessel endothelium. D2-40 is a novel monoclonal antibody to an Mr 40,000 O-linked sialoglycoprotein that reacts with a fixation-resistant epitope on lymphatic endothelium. We sought to establish the selectivity of D2-40 for lymphatic endothelium in normal tissues and compare its reactivity with the expression of the widely used vascular endothelial marker CD31 in a series of 62 formalin-fixed and paraffin-embedded vascular lesions including KS. In normal tissues, D2-40 stained the endothelium of lymphatic channels but not of blood vessels, including arteries and capillaries defined by reactivity with the blood vessel endothelial marker PAL-E. In our series of vascular lesions, D2-40 stained lymphangiomas (10/10), benign tumors of undisputed lymphatic origin, but not benign neoplasms or tumorlike lesions of blood vessel origin, including hemangiomas (0/10), glomus tumors (0/3), angiolipomas (0/2), pyogenic granulomas (0/2), vascular malformations (0/2), hemangiopericytoma (0/1), or hemangioendothelioma (0/1). D2-40 stained all cases of cutaneous KS (24/24) at all stages of progression, including patch, plaque, and nodular stages, supporting the concept that this disease originates from a cell type capable of undergoing lymphatic differentiation. D2-40 also stained three of seven angiosarcomas, indicating that a subset of these tumors can undergo at least partial differentiation along the lymphatic endothelial lineage and could be classified as lymphangiosarcomas. In comparison, CD31 was expressed in all benign and malignant vascular lesions, except for glomus tumors (0/3) and 5/10 lymphangiomas, in which staining was absent. We conclude that D2-40 is a new selective marker of lymphatic endothelium in normal tissues and vascular lesions and is valuable for studying benign and malignant vascular disorders in routinely processed tissue specimens.


Assessment of the impact of pesticide residues on microbiological and biochemical parameters of tea garden soils in India.
The main aim of this study was to assess the impact of pesticidal residues on soil microbial and biochemical parameters of the tea garden soils. The microbial biomass carbon (MBC), basal (BSR) and substrate induced respirations (SIR), beta-glucosidase activity and fluorescein diacetate hydrolyzing activity (FDHA) of six tea garden soils, along with two adjacent forest soils (control) in West Bengal, India were measured. The biomass and its activities and biochemical parameters were generally lower in the tea garden soils than the control soils. The MBC of the soils ranged from 295.5 to 767.5 micro g g(- 1). The BSR and SIR ranged from 1.65 to 3.08 mu g CO2-C g(- 1) soil h(- 1) and 3.08 to 10.76 micro g CO2-C g(- 1)h(- 1) respectively. The beta-glucosidase and FDHA of the soils varied from 33.3 and 76.3 micro g para-nitrophenol g(- 1) soil h(- 1) and 60.5 to 173.5 micro g fluorescein g(- 1)h(- 1)respectively. The tea garden soils contained variable residues of organophosphorus and organochlorine pesticides, which negatively affected the MBC, BSR, SIR, FDHA and beta -glucosidase activity. Ethion and chlorpyriphos pesticide residues in all the tea garden soils varied from 5.00 to 527.8 ppb and 17.6 to 478.1 ppb respectively. The alpha endosulfan, beta endosulfan and endosulfan sulfate pesticide residues in the tea garden soils ranged from 7.40 to 81.40 ppb, 8.50 to 256.1 ppb and 55 to 95.9 ppb respectively. Canonical correlation analysis shows that 93% of the total variation was associated with the negative impact of chlorpyriphos, beta and alpha endosulfan and endosulfan sulfate on MBC, BSR and FDHA. At the same time ethion had negative impact on SIR and beta-glucosidase. Data demonstrated that the pesticide residues had a strong impact on the microbial and biochemical components of soil quality.


Serum tumour marker regression rate following chemotherapy for malignant teratoma.
The rate of fall of the serum tumour markers alphafetoprotein (AFP) and the beta sub-unit of human chorionic gonadotrophin (HCG) was analysed following platinum-based chemotherapy for metastatic non-seminomatous germ cell tumours. Of 90 evaluable patients 81% were alive and disease-free 1.5-4 yr (median 28 months) from the start of chemotherapy and 69 (77%) had remained continuously disease free. All three patients with an initial AFP half-life greater than 9 days relapsed; however, a further eight relapsing patients had an initial regression rate of serum AFP within the same range as patients remaining in remission (half-life 6-9 days). The HCG regression rate did not discriminate between patients remaining well or those who relapsed after chemotherapy. In 11 examples of a pattern of late slowing of the rate of marker fall (i.e. increasing half-life), five relapses were seen (45%), though this pattern was also observed in the context of large residual differentiated teratoma masses.


Coping with the consequences of a stroke.
Having a stroke constitutes a major life event which engenders some types of adaptation in order to try to resume to a "normal" life. The objectives of this study were (1) to explore the type of coping strategies used following a stroke; (2) to verify if coping strategies change over time, and (3) to verify if the coping strategies used are related to age, gender, actualization of potential, handicap level and depression. A convenience sample of 76 participants who had had a stroke was recruited upon discharge from a functional intensive rehabilitation unit. Data were collected 2 weeks later and 6 months later in their own home. Coping strategies were measured using a modified version of the Ways of Coping Questionnaire. Actualization of potential was measured using the Measure of the Actualization of Potential and handicap level was quantified with the LIFE-H. Finally, the Beck Depression Inventory was used to evaluate the presence of depressive symptoms. The results suggest that long after the end of an intensive rehabilitation programme, some coping strategies might be more effective in dealing with the consequences of a stroke whereas others might be related to some form of inadaptation. However, the results should be treated with caution. More research on coping following a stroke using mixed methodologies is needed.


White blood cell count and chronic obstructive pulmonary disease: A Mendelian Randomization study.
Blood leukocyte counts (e.g., eosinophil count) are important biomarkers for the onset, classification, and exacerbation of chronic obstructive pulmonary disease (COPD). The causal relationships between them are necessary for the development of COPD treatment strategy, but remain unclear. Here, we implement two-sample bi-directional univariable Mendelian Randomization (MR) and multivariable MR to investigate the causal relationships. Univariable MR find that elevated blood eosinophil count significantly increases the risk of COPD (odds ratio (OR) = 1.22, 95% confidence interval (CI): 1.14-1.30, P = 1.54 × 10-09) and COPD-related hospitalization (OR = 1.44, 95% CI: 1.15-1.80, P = 1.36 × 10-03). Besides, it also significantly decreases the ratio of forced expiratory volume in the first second over forced vital capacity (FEV1/FVC ratio) (OR = 0.942, 95% CI: 0.914-0.971, P = 1.02 × 10-04). These findings are fully supported by multivariate MR results. Interestingly, univariable MR reveals a weak causal relationship between elevated blood eosinophil count and COPD risk in younger people (<65 years) (OR = 1.39, 95% CI: 1.10-1.75, P = 5.52 × 10-03), but not older individuals (OR = 1.20, 95% CI: 0.926-1.55, P = 0.17). Finally, reverse univariable MR reveals the onset of COPD and the decreased FEV1/FVC ratio both lead to increased blood neutrophil count (OR = 1.03, 95% CI: 1.01-1.05, P = 3.40 × 10-03 and OR = 0.947, 95% CI: 0.91-0.986, P = 8.75 × 10-03 respectively). In summary, this MR study demonstrates that high blood eosinophil count is an independent causal mediator of COPD risk, FEV1/FVC decline, and COPD-related hospitalization. The increase in neutrophil count is induced by COPD onset or FEV1/FVC decline. This suggests eosinophil, but not neutrophil, may be used as a therapeutic target for preventing the onset and exacerbation of COPD and FEV1/FVC decline. Therefore, a non-neutrophil-targeted therapeutic strategy for neutrophilic COPD is required in the future.


Systematic molecular evolution enables robust biomolecule discovery.
Evolution occurs when selective pressures from the environment shape inherited variation over time. Within the laboratory, evolution is commonly used to engineer proteins and RNA, but experimental constraints have limited the ability to reproducibly and reliably explore factors such as population diversity, the timing of environmental changes and chance on outcomes. We developed a robotic system termed phage- and robotics-assisted near-continuous evolution (PRANCE) to comprehensively explore biomolecular evolution by performing phage-assisted continuous evolution in high-throughput. PRANCE implements an automated feedback control system that adjusts the stringency of selection in response to real-time measurements of each molecular activity. In evolving three distinct types of biomolecule, we find that evolution is reproducibly altered by both random chance and the historical pattern of environmental changes. This work improves the reliability of protein engineering and enables the systematic analysis of the historical, environmental and random factors governing biomolecular evolution.


Immunotherapy of Melanoma.
Arising from melanocytes in skin, mucosal membranes, eye, and meninges, melanoma is a tumor that has been associated with poor prognosis in advanced disease stages. Given the poor response to chemotherapy and radiation therapy, new treatment approaches with targeted therapy, immunotherapy, and adoptive T-cell therapy have revolutionized the standard of care for patients with advanced melanoma. This review provides a short overview of past, present, and future immunotherapeutic approaches and their limitations, with a focus on new combination agents in early clinical trials.


Growth control mechanisms in neuronal regeneration.
Neurons grow during development and extend long axons to make contact with their targets with the help of an intrinsic program of axonal growth as well as a range of extrinsic cues and a permissive milieu. Injury events in adulthood induce some neuron types to revert to a regenerative state in the peripheral nervous system (PNS). Neurons from the central nervous system (CNS), however, reveal a much lower capacity for regenerative growth. A number of intrinsic regeneration-promoting mechanisms have been described, including priming by calcium waves, epigenetic modifications, local mRNA translation, and dynein-driven retrograde transport of transcription factors (TFs) or signaling complexes that lead to TF activation and nuclear translocation. Differences in the availability or recruitment of these mechanisms may partially explain the limited response of CNS neurons to injury.


Recent advances of ambient ionization mass spectrometry imaging in clinical research.
The structural information and spatial distribution of molecules in biological tissues are closely related to the potential molecular mechanisms of disease origin, transfer, and classification. Ambient ionization mass spectrometry imaging is an effective tool that provides molecular images while describing in situ information of biomolecules in complex samples, in which ionization occurs at atmospheric pressure with the samples being analyzed in the native state. Ambient ionization mass spectrometry imaging can directly analyze tissue samples at a fairly high resolution to obtain molecules in situ information on the tissue surface to identify pathological features associated with a disease, resulting in the wide applications in pharmacy, food science, botanical research, and especially clinical research. Herein, novel ambient ionization techniques, such as techniques based on spray and solid-liquid extraction, techniques based on plasma desorption, techniques based on laser desorption ablation, and techniques based on acoustic desorption were introduced, and the data processing of ambient ionization mass spectrometry imaging was briefly reviewed. Besides, we also highlight recent applications of this imaging technology in clinical researches and discuss the challenges in this imaging technology and the perspectives on the future of the clinical research.


[Hemorrhagic congenital diseases: What can be the future of plasma-derived products against recombinants?].
Until 1990, congenital hemorrhagic disorders were treated by plasma-derived concentrates. The first recombinant drug, recombinant factor VIII was available after this date and few years later recombinant factor IX could also be proposed to patients. The evolution of market share in France was different between these two drugs: while recombinant factor VIII took a large place in hemophilia A treatment (85%), plasma-derived factor IX represent 50% of the French market. In the next years, the arrival of long-acting antihemophilic factors may lead to the dramatically reduce the amount of plasma-derived antihemophilic factors used to treat hemophilia. For rare bleeding coagulation disorders, plasma-derived concentrates are still widely used, while they are the only concentrates available in most diseases. This situation is unlikely to evolve significantly in the next years.


Abnormal cortical excitability with preserved brainstem and spinal reflexes in sialidosis type I.
To examine neurophysiological evidence of functional involvement of the brainstem and spinal cord and motor cortical excitability in sialidosis type I, a rare inherited neurodegenerative disorder caused by mutations in the NEU1 gene. We investigated particular pathways in the brainstem, spinal cord and motor cortex in 12 genetically proven cases of sialidosis type I by assessing blink reflex recovery cycle (BR), spinal reciprocal inhibition (RI), input-output curves (I/O), short interval intracortical inhibition (SICI), intracortical facilitation (ICF) and silent period (SP). The BR and RI were normal. The slope of I/O was significantly increased, and SICI and the duration of SP were reduced in sialidosis patients. Despite reports of pathology involving brainstem and anterior horn neurones, there were no obvious abnormalities in spinal and brainstem reflexes in the present patients, suggesting that the major clinical effects may be caused by changes at a level above the brainstem. For the first time, the integrity of certain brainstem and spinal cord reflexes in addition to motor cortical facilitatory and inhibitory circuits has been assessed in genetically proven type I sialidosis. This provides new data to aid in understanding of the pathophysiology of motor system dysfunction in this condition.


Left ventricular function after coronary artery reperfusion.
Left ventricular dysfunction and dilation after reperfusion relate to the amount of infarcted and dysfunctional myocardium and will continue to be important determinants of morbidity and mortality. There is marked heterogeneity in the anatomic and pathophysiologic presentation of patients with acute myocardial infarction prior to thrombolysis, and many of these individual settings resemble those in animal species with various degrees of collateral formation. Three major determinants of infarct size are responsible for this heterogeneity and include the risk area, the duration of the coronary occlusion, and the level of the residual coronary blood flow via collaterals or a partially patent artery. All 3 of these determinants will influence the initial and late results of reperfusion therapy on infarct size and ventricular function. However, in addition to late or unsuccessful thrombolysis, there are other important factors determining outcome: inadequate reflow, residual coronary stenosis, and coronary reocclusion, factors that can be associated with late progressive left ventricular dilation and dysfunction. The risk factors for left ventricular dysfunction and dilation after reperfusion can now be identified, and such patients should undergo coronary angiography prior to hospital discharge and, if appropriate, revascularization of the infarct-related artery (and perhaps other vessels). In other patients, if serial studies reveal progressive left ventricular failure and dilation late after reperfusion, despite therapy with an angiotensin-converting enzyme (ACE) inhibitor, and if repeat coronary angiography identifies significant coronary stenoses and areas of hibernating or stunned myocardium, revascularization may limit progression of dilation and improve left ventricular function.(ABSTRACT TRUNCATED AT 250 WORDS)


The trapezoid form of the trochlea tali.
The wedge-shaped superior surface of the trochlea tali may be seen in a proximal view as a trapezium. The angle of this trapezium was calculated by measuring the length, and the smallest and largest widths of the trochlea, and was found to be 16.04 degrees for the left talus and 12.48 degrees for the right talus, respectively. The difference between these angles is highly significant. Due to biomechanical features of the talocrural joint an increase in the angle of the trochlea tali also intensifies the incongruency of this joint in plantar-flexion. Thus, this incongruency is larger on the left side. With this diminution of the joint-surface contact, increased internal rotation or "wobbling" movements are possible. An explanation for these facts might be found in footedness, similar to handedness. When constructing a prosthesis one has to consider that the trochlea tali cannot be exactly mirrored from one side to the other but may have to be calculated separately for each side. The fact that footedness seems to correlate with the angle of the trochlea tali can be also helpful.


GAU-PED study for early diagnosis of Gaucher disease in children with splenomegaly and cytopenia.
Gaucher disease (GD) diagnosis can be delayed due to non-specific symptoms and lack of awareness, leading to unnecessary procedures and irreversible complications. GAU-PED study aims to assess GD prevalence in a high-risk pediatric population and the presence, if any, of novel clinical or biochemical markers associated with GD. DBS samples were collected and tested for β-glucocerebrosidase enzyme activity for 154 patients selected through the algorithm proposed by Di Rocco et al. Patients showing β-glucocerebrosidase activity below normal values were recalled to confirm the enzyme deficiency with the gold standard essay on cellular homogenate. Patients tested positive at the gold standard analysis were evaluated through GBA1 gene sequencing. 14 out of 154 patients were diagnosed with GD, with a prevalence of 9.09% (5.06-14.78%, CI 95%). Hepatomegaly, thrombocytopenia, anemia, growth delay/deceleration, elevated serum ferritin, elevated Lyso-Gb1 and chitotriosidase were significantly associated with GD. GD prevalence in a pediatric population at high-risk appeared to be higher compared to high-risk adults. Lyso-Gb1 was associated with GD diagnosis. The algorithm proposed by Di Rocco et al. can potentially improve the diagnostic accuracy of pediatric GD, allowing the prompt start of therapy, aiming to reduce irreversible complications.


High Periventricular T1 Relaxation Times Predict Gait Improvement After Spinal Tap in Patients with Idiopathic Normal Pressure Hydrocephalus.
The diagnosis of idiopathic normal pressure hydrocephalus (iNPH) can be challenging. Aim of this study was to use a novel T1 mapping method to enrich the diagnostic work-up of patients with suspected iNPH. Using 3T magnetic resonance imaging (MRI) we prospectively evaluated rapid high-resolution T1 mapping at 0.5 mm resolution and 4 s acquisition time in 15 patients with suspected iNPH and 8 age-matched, healthy controls. T1 mapping in axial sections of the cerebrum, clinical and neuropsychological testing were performed prior to and after cerebrospinal fluid tap test (CSF-TT). T1 relaxation times were measured in 5 predefined periventricular regions. All 15 patients with suspected iNPH showed gait impairment, 13 (86.6%) showed signs of cognitive impairment and 8 (53.3%) patients had urinary incontinence. Gait improvement was noted in 12 patients (80%) after CSF-TT. T1 relaxation times in all periventricular regions were elevated in patients with iNPH compared to controls with the most pronounced differences in the anterior (1006 ± 93 ms vs. 911 ± 77 ms; p = 0.023) and posterior horns (983 ± 103 ms vs. 893 ± 68 ms; p = 0.037) of the lateral ventricles. Montreal cognitive assessment (MoCA) scores at baseline were negatively correlated with T1 relaxation times (r < -0.5, p < 0.02). Higher T1 relaxation times were significantly correlated with an improvement of the 3‑m timed up and go test (r > 0.6 and p < 0.03) after CSF-TT. In iNPH-patients, periventricular T1 relaxation times are increased compared to age-matched controls and predict gait improvement after CSF-TT. T1 mapping might enrich iNPH work-up and might be useful to indicate permanent shunting.


A vault-prediction formula for implantable collamer lens based on preoperative parameters: a retrospective clinical study.
The aim of the present study was to predict the vault of implantable collamer lens (ICL) using a formula established based on the preoperative parameters. This retrospective study included data on 226 eyes from 114 patients(the median age and range was 27.5 ± 5.7; 18-46 years) who underwent EVO-ICL surgery between May 2021 and April 2022. Preoperative parameters, such as anterior chamber depth (ACD) horizontal angle-to-angle diameter (ATA), Crystalline lens rise (CLR), and vault (1 week after surgery) were measured by anterior segment optical coherence tomography. The axial length of eyes (AL) and white-to-white (WTW) values were measured using an IOLmaster and calipers under a slit lamp separately. The mean WTW, ACD, horizontal ATA diameter, CLR, AL, and vault 1 week after surgery were 11.42 ± 0.37 mm, 3.25 ± 0.25 mm, 11.72 ± 0.44 mm, 45.78 ± 175.42 μm, 27.61 ± 1.93 mm, and 586.51 ± 254.54 μm respectively. Multivariate regression analysis showed that the ICL-width, ACD (β = 0.293), ATA (β =-0.657), CLR (β =-0.157), and the anterior chamber angle in temporal side (T:ACA) (β=-0.277) were predictors of the vault size (adjusted-R2 = 0.39, P < 0.001). The formula built based on these preoperative parameters could be used to predict the vault before surgery. The predictors indicated that the pre-operative parameters of eye such as ACD, ATA, CLR and T:ACA play an important role in choosing optimal ICL size.


ALS-associated KIF5A mutations abolish autoinhibition resulting in a toxic gain of function.
Understanding the pathogenic mechanisms of disease mutations is critical to advancing treatments. ALS-associated mutations in the gene encoding the microtubule motor KIF5A result in skipping of exon 27 (KIF5AΔExon27) and the encoding of a protein with a novel 39 amino acid residue C-terminal sequence. Here, we report that expression of ALS-linked mutant KIF5A results in dysregulated motor activity, cellular mislocalization, altered axonal transport, and decreased neuronal survival. Single-molecule analysis revealed that the altered C terminus of mutant KIF5A results in a constitutively active state. Furthermore, mutant KIF5A possesses altered protein and RNA interactions and its expression results in altered gene expression/splicing. Taken together, our data support the hypothesis that causative ALS mutations result in a toxic gain of function in the intracellular motor KIF5A that disrupts intracellular trafficking and neuronal homeostasis.


Contrasting effects of PACAP and carbachol on [Ca2+]i and inositol phosphates in human neuroblastoma NB-OK-1 cells.
The effects of PACAPs on [Ca2+]i were compared to those of carbachol in human neuroblastoma NB-OK-1 cells. PACAP(1-27) and PACAP(1-38) increased [Ca2+]i in a biphasic manner: a transient rise and a secondary plateau. The transient phase reflected the mobilization of [Ca2+]i pool(s) via the inositol phosphate pathway. The modest sustained plateau required extracellular Ca2+. Carbachol also increased [Ca2+]i in a biphasic manner, but it mobilized intracellular Ca2+ pool(s) with a higher efficacy than PACAPs, then greatly increased Ca2+ entry, this being accompanied by a more marked and prolonged elevation of IP3 and IP4 than with PACAPs. It is likely that cAMP-mediated phosphorylations due to PACAPs facilitated desensitization at the PACAP receptor-phospholipase C level, so that there was less Ca2+ handling through PACAP receptors than with muscarinic M1 receptors.


Interpolated average CT for cardiac PET/CT attenuation correction.
Previously, we proposed interpolated averaged CT (IACT) for improved attenuation correction (AC) in thoracic PET/CT. This study aims to evaluate its feasibility and effectiveness on cardiac PET/CT. We simulated (18)F-FDG distribution using the XCAT phantom with normal and abnormal cardiac uptake. Average activity and attenuation maps represented static PET and respiration average CT (ACT), respectively, while the attenuation maps of end-inspiration/expiration represented 2 helical CTs (HCT). IACT was obtained by averaging the 2 extreme phases and the interpolated phases generated between them. Later, we recruited 4 patients who were scanned 1 hr post 315-428 MBq (18)F-FDG injection. Simulated and clinical PET sinograms were reconstructed with AC using (1) HCT, (2) IACT, and (3) ACT. Polar plots and the 17-segment plots were analyzed. Two regions-of-interest were drawn on lesion and background area to obtain the intensity ratio (IR). Polar plots of PETIACT-AC were more similar to PETACT-AC in both simulation and clinical data. Artifacts were observed in various segments in PETHCT-AC. IR differences of HCT as compared to the phantom were up to ~20%. IACT-AC reduced respiratory artifacts and improved PET/CT matching similarly to ACT-AC. It is a promising low-dose alternate of ACT for cardiac PET/CT.


A quantitative freeze-etching study on the effects of cytochalasin B, colchicine and concanavalin A on the formation of intercellular contacts in vitro.
The effects of concanavalin A (conA), cytochalasin B (CB), and colchicine acting on cell membrane-associated structures, such as surface receptors or submembranal cytoskeletal components, were studied in trypsin-isolated plexus cells reaggregated in vitro. Following reaggregation, these cells form intercellular contacts, i.e. a zonula occludens (ZO) and integrated nexus (gap junctions, GJ). In the control cultures the GJ occupied 4.7%, the tight junctions (TJ) occupied 14 microns/microns2 of the surface of the zonula occludens (ZO). All the drugs/lectins studied inhibit contact formation in a characteristic manner. ConA inhibits the formation of GJ (1% of the ZO) and causes 5 times more defective TJ strands than the controls. CB inhibits the assembly of the TJ (75% of the controls) and the GJ (1% of the ZO) when it is administered during the period of contract formation. Colchicine inhibits the formation of GJ (1% of the ZO) and produces 2.5 times more open-end strands of TJ than the controls. All effects are reversible with the exception that integrated GJ are not re-formed after colchicine administration. It is suggested that an intact interaction between surface receptors, intramembranous particles and associated cytoskeletal structures is necessary for remodelling cellular contacts.


A comparative study of meat quality and vibrational spectroscopic properties of different chicken breeds.
Chicken breed is one of the key factors that influence meat quality. The quality attributes of breast meat from commercial broiler (CB), Thai native chicken (NC, Leung Hang Khao), and the crossbred Korat chicken (KC) were investigated via synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy, Fourier transform Raman (FT-Raman) spectroscopy, and physicochemical analysis. The protein and carbonyl contents of KC and NC meats were higher than that of CB meat, but the lipid content was lower (P < 0.05). CB meat was characterized by high moisture, lightness (L*), and presence of taste-active nucleotides, namely, inosine 5'-monophosphate (IMP) and guanosine 5'-monophosphate (GMP). Moreover, NC meat had the highest insoluble collagen and inosine contents (P < 0.05). The predominant protein secondary structures of KC and NC meats were β-turns and random coils, whereas α-helices were mainly found in CB meat. Based on principal component analysis, the meat quality and spectra were clearly separated by breeds. The high moisture and lipid content of meat corresponded to O-H stretching (3,203 cm-1) and C-H stretching (2,854 cm-1) in the FT-Raman spectra, whereas PO2- stretching (1,240 cm-1), measured via SR-FTIR, was well correlated with the IMP content. In addition, the FT-Raman wavenumber of 934 cm-1, indicating C-C stretching, was correlated with high water-holding capacity (WHC) in KC meat. The quality of meat from slow- and fast-growing chickens significantly varies. Vibrational spectroscopy is a powerful technique that provides insightful molecular information correlated with various meat attributes.


Concordance of emergency department physicians' decisions on HIV post-exposure prophylaxis with national guidelines: results from a retrospective cohort study.
Post-exposure prophylaxis (PEP) is an effective tool to prevent infection with HIV. Patients seeking PEP after potential HIV exposure usually present to the emergency department (ED). Our study sought to determine the concordance of ED physicians' decisions on HIV-PEP with national guidelines (primary objective) and to assess the clinical relevance of drug-drug interactions (DDIs) between the HIV-PEP regimen and patients' concomitant medication (secondary objective). We conducted a retrospective cohort study at the ED of Hannover Medical School, Germany. Between 1 January 2018 and 31 December 2019, 113 of 11 246 screened patients presented to the ED after potential HIV exposure and were enrolled in the study. The median age of the patients (82.3% male) was 30 y (IQR 25-35.5), 85.8% of potential HIV exposures were characterised as sexual and 85.0% presented within 72 h. ED physicians' decisions on HIV-PEP were concordant with national guidelines in 93.8%. No clinically relevant DDIs were detected. ED physicians' decisions on HIV-PEP were highly concordant with national guidelines. Approximately 1% of patient presentations to the ED were related to HIV exposure; therefore, training ED physicians on HIV transmission risk assessment and indications/contraindications for HIV-PEP is paramount.


[Asthma: diagnosis and treatment in general pratice].
Based on the last international guidelines, this paper summarise the current concepts on diagnosis and treatment of asthma, adapted to the general medicine.


Chronic exposure of sheep to a zinc smelter in Peru.
Liver levels of arsenic, cadmium, copper, lead, manganese, and zinc were assessed in adult female sheep living in the vicinity of a zinc smelter in Peru. The study was conducted on a large sheep-raising cooperative divided into seven management units, operated under a standardized husbandry system. Soil samples were analyzed for the same metals at varying distances from the smelter. A gradient of concentration of heavy metals in soil was found between 1 and 56 km from the smelter. Soil concentrations for all metals except manganese decreased significantly with increasing distance from the smelter. The strongest correlations were found for copper, lead, and zinc (P less than 0.001). Topographic features and prevailing wind direction appeared to play a role in the dispersion of pollutants. Liver samples were obtained from 153 sheep on five units of the cooperative. The centers of the units sampled were between 13 and 56 km from the smelter. Hepatic concentrations of arsenic, lead, manganese, and zinc decreased significantly with increasing distance from the smelter. Liver arsenic and liver zinc were significantly related to soil concentrations. Liver levels of arsenic, cadmium, lead, and manganese in sheep from the cooperative were significantly higher than those detected in control sheep from southern Peru. Sheep populations may serve as biological monitors for heavy metal accumulation and environmental carcinogenesis.


Hypothalamic temperature and the 22 kHz vocalization of the male rat.
Following ejaculation the male rat emits a 22 kHz vocalization that has been hypothesized to play a communicative role. We found previously that this vocalization is often accompanied by rapid and selective hypothalamic cooling, and we hypothesized that the vocalization or, more precisely, the thoracic-laryngeal maneuver underlying the vocalization, is primarily a thermoregulatory behavior. Accordingly, one prediction made herein was that heating the brain of the isolated male, through the infusion of prostaglandin E2, would be accompanied by the vocalization. Another was that cooling the brain of the copulating male through the injection of sodium salicylate would significantly reduce the post-ejaculatory vocalization. Both predictions were confirmed.


Voice disorder classification using convolutional neural network based on deep transfer learning.
Voice disorders are very common in the global population. Many researchers have conducted research on the identification and classification of voice disorders based on machine learning. As a data-driven algorithm, machine learning requires a large number of samples for training. However, due to the sensitivity and particularity of medical data, it is difficult to obtain sufficient samples for model learning. To address this challenge, this paper proposes a pretrained OpenL3-SVM transfer learning framework for the automatic recognition of multi-class voice disorders. The framework combines a pre-trained convolutional neural network, OpenL3, and a support vector machine (SVM) classifier. The Mel spectrum of the given voice signal is first extracted and then input into the OpenL3 network to obtain high-level feature embedding. Considering the effects of redundant and negative high-dimensional features, model overfitting easily occurs. Therefore, linear local tangent space alignment (LLTSA) is used for feature dimension reduction. Finally, the obtained dimensionality reduction features are used to train the SVM for voice disorder classification. Fivefold cross-validation is used to verify the classification performance of the OpenL3-SVM. The experimental results show that OpenL3-SVM can effectively classify voice disorders automatically, and its performance exceeds that of the existing methods. With continuous improvements in research, it is expected to be considered as auxiliary diagnostic tool for physicians in the future.


An analysis of published trials found that current use of pragmatic trial labels is uninformative.
Randomized trials labelled as "pragmatic" are attractive to funders, patients, and clinicians as the label implies that the results are directly applicable to clinical care. We examined how authors justify use of the label (e.g., by referring to one or more PRECIS [PRagmatic Explanatory Continuum Indicator Summary]-2 domains). We reviewed primary trial reports published 2014-2019, registered in ClinicalTrials.gov and using the pragmatic label anywhere in the report. Among 415 trials, the label was justified by reference to at least one design element in 282 (68.0%); of these, 240 (85.1%) referenced trial characteristics that can be mapped to one or more of the PRECIS-2 domains, most commonly eligibility (91, 32.3%), setting (90, 31.9%), flexibility delivery (89, 31.6%), and organization (75, 26.6%); 42 (14.9%) referenced characteristics that are not PRECIS-2 domains, most commonly type of intervention/comparator (48, 17%), recruitment without consent (22, 7.8%), routinely collected data (22, 7.8%), and cluster randomization (20, 7.1%). Most reports referenced only one or two design elements. Overall, 9/415 (2.2%) provided PRECIS wheels. Current use of pragmatic labels is uninformative. Authors should clarify the decision the trial is intended to support and include a PRECIS-2 table to make the design transparent.


Early angiotensin converting enzyme inhibitor therapy after experimental myocardial infarction prevents left ventricular dilation by reducing infarct expansion: a possible mechanism of clinical benefits.
To examine the effects of early angiotensin-converting enzyme (ACE) inhibitor therapy after myocardial infarction on infarct expansion in an experimental rat model. ACE inhibitor therapy within 24 h of acute myocardial infarction (AMI) reduces mortality by unknown mechanism(s). Rats underwent permanent coronary artery occlusion. A treated group received enalapril (1.9+/-0.2 mg/kg) daily in drinking water beginning 2 h after coronary artery occlusion, a time too late to reduce infarct size. Rats were sacrificed 2 days or 2 weeks after myocardial infarction. Hearts were arrested and fixed at a constant pressure, then sectioned and photographed for morphometric analysis. Infarcts in the control group expanded between 2 days and 2 weeks after myocardial infarction (expansion index 0.7+/-0.1 versus 2.5+/-0.4, P< 0.05). However, infarct expansion remained unchanged in the enalapril group between 2 days and 2 weeks after myocardial infarction (expansion index 0.8+/-0.1 versus 1.3+/-0.1, NS). Two weeks after myocardial infarction, the enalapril group had fewer expanded infarcts than the control group (expansion index 1.3+/-0.1 versus 2.5+/-0.4, P< 0.05). While left ventricular volume increased in the control group between 2 days and 2 weeks after myocardial infarction (0.17+/-0.01 ml versus 0.36+/-0.03 ml, P< 0.05), it remained constant in the enalapril group (0.22+/-0.02 ml versus 0.25+/-0.03 ml, NS). Two weeks after myocardial infarction, the left ventricles were larger in the control group than in the enalapril group (0.36+/-0.03 ml versus 0.25+/-0.03 ml, P< 0.05). Treatment with enalapril initiated 2 h after AMI prevented left ventricular dilation by limiting infarct expansion. This may explain the mechanism by which ACE inhibitor therapy started within 24 h of an AMI improves survival 5-6 weeks after infarction.


Cellular distribution of prostacyclin synthase and specific prostacyclin binding sites in bovine corpora lutea of pregnancy.
The cellular distribution of prostacyclin (PGI2) synthase and specific PGI2 binding sites was investigated by light microscope immunocytochemistry and quantitative autoradiography. The immunostaining for the enzyme was found in small (15-18 microns) and large (18-45 microns) cells as well as in non-luteal cells, arteriole smooth muscle and endothelium. There was no consistent difference between small and large luteal cells and luteal vs. non-luteal cells in the immunostaining. Vascular smooth muscle and endothelial cells, on the other hand, contained less immunostaining than the other cells. Contrary to wide-spread distribution of PGI2 synthase, specific PGI2 binding sites were only found in the luteal cells. On a per cell basis, large luteal cells contained more sites than small luteal cells and vice versa when expressed per unit cell area. The [3H]PGI2 binding to the luteal cells was time and temperature dependent and was inhibited by excess unlabeled PGI2 but not by its metabolite, 6-keto-PGF1 alpha or other eicosanoids which bind to their own receptors. In conclusion, while PGI2 synthase is widely distributed among different cell types in bovine corpora lutea, specific binding sites which may mediate luteotropic actions of PGI2 are only present in small and large luteal cells.


Structural and functional characteristics of activated human factor IX after chemical modification of gamma-carboxyglutamic acid residues.
Activated human factor IX (factor IXa) was treated under mildly acidic conditions with a mixture of formaldehyde and morpholine. This reagent has been shown to react preferentially with gamma-carboxyglutamyl (Gla) residues and to convert these residues to gamma-methyleneglutamyl residues (Wright, S.F., Bourne, C.D., Hoke, R.A., Koehler, K.A., and Hiskey, R.G. (1984) Anal. Biochem. 139, 82-90). The modified enzyme was evaluated for coagulant activity and calcium-dependent fluorescence quenching. [14C]Formaldehyde was employed to allow quantitation of the modification and to facilitate localization of the modified residues in the primary structure of factor IXa. In the presence of the [14C]formaldehyde/morpholine reagent, factor IXa rapidly lost coagulant activity, which corresponded to incorporation of radiolabel. Examination of the relationship between protein modification (radiolabel incorporation) and the loss of coagulant activity suggested that modification of 1 mol of Gla/mol of factor IXa results in complete loss of factor IXa coagulant activity. Primary structure analysis of the radioactivity labeled factor IXa suggested that modification of any one of 11 Gla residues was responsible for the loss of coagulant activity. In the presence of calcium, modified factor IXa exhibited a smaller Gla-dependent decrease in protein fluorescence than native factor IXa, but the Gla-independent fluorescence change was the same for both proteins. It therefore appears that the Gla domain of factor IXa must be completely intact for the enzyme to undergo a functionally important calcium-dependent conformational change necessary for coagulant activity.


Relapse-free survival in Sudanese women with non-metastatic breast cancer.
Breast cancer (BC) is the most frequently diagnosed cancer and a major cause of cancer mortality in Sudan. However, there is lack of data related to BC relapse. Therefore, this study was undertaken to estimate the 5-year relapse free survival (RFS) rate and factors related to BC relapse in Sudanese women with non-metastatic BC. Data of BC women with BC diagnosed and treated at the National Cancer Institute-University of Gezira during 2012 were retrieved from medical records. The cases were followed-up through hospital records and telephone contact. Survival functions were calculated using Kaplan-Meier method and compared by log-rank test. The prognostic factors were tested using univariate and multivariable Cox regression analyses. We included 168 women with median age of 45 years (range, 22-83 years). 53.5%of women had stage III at time of diagnosis, whereas 4.2% and 42.3% of women presented with stage I and stage II, respectively. At the end of 5 years follow-up, with median follow-up period of 64 months, 94 (56.0%) women were alive in remission, 11 (6.5%) were alive with BC relapse, 49 (29.2%) were dead, and survival status was unknown in 14 (8.3%) women. Most of the occurred relapses were distant relapses. The 5-year RFS was 59%. The independent predictors of relapse were: larger primary tumor size (HR:1.84, 95% CI: 1.54-5.48, p=0.018); involved axillary lymph nodes with tumour (HR: 2.91, 95% CI: 1.53-7.91, p=0.001); not receiving adjuvant radiotherapy (HR: 2.2, 95% CI: 1.22-3.95, p=0.009); and not receiving hormone therapy (HR: 1.67, 95% CI: 1.01-2.76, p= 0.046). We found a high risk of BC relapse in our resource-constrained settings. Advanced stages, not receiving adjuvant radiotherapy, and not receiving adjuvant hormone therapy were independent predictors associated with worse 5-year RFS. Therefore, enhancing the early diagnosis of BC and improving timely access to appropriate treatments represent key approaches to achieving better treatment outcomes.


C32-O-imidazol-2-yl-methyl ether derivatives of the immunosuppressant ascomycin with improved therapeutic potential.
A series of C32-O-aralkyl ether derivatives of the FK-506 related macrolide ascomycin have been prepared based on an earlier reported C32-O-cinnamyl ether design. In the present study, the nature of the aryl tethering group was varied in an attempt to improve oral activity. An imidazol-2-yl-methyl tether was found to be superior among those investigated and has resulted in an ascomycin analog, L-733,725, with in vivo immunosuppressive activity comparable to FK-506 but with an improved therapeutic index.


Auditory brainstem responses in ground squirrels arousing from hibernation.
Auditory brainstem responses (ABRs) were recorded in ground squirrels (Citellus lateralis) arousing from hibernation. Squirrels implanted with recording screws to record ABRs, and a thermistor to record brain temperature, were placed in a cold room at 9 degrees C on a 2L:22D light-dark cycle. Hibernating animals were moved from the cold room and ABRs recorded during arousal. The responses showed a gradual development of all brainstem peaks. At low temperatures there were very long latencies to the peaks. The amplitudes of the peaks increased (with fluctuations) as brain temperature increased. The data indicate that neural generators on the brainstem auditory pathway were all activated early in arousal. These results do not support the hypothesis that successive peaks appear and grow in amplitude only after previous peaks are fully developed.


Plasma clonidine levels in hypertension.
Ten patients with essential hypertension were treated orally with clonidine, and the dose was titrated until the diastolic pressure fell below 90 mm Hg or at least 10 mm Hg below baseline, or until a maximum dose of clonidine of 1.2 mg/day was reached. After at least 2 wk on a constant dose of clonidine, the patients were hospitalized and blood pressure and plasma clonidine concentrations were measured by a highly specific and sensitive gas chromatographic, electron-capture method (sensitivity as low as 0.1 ng clonidine/ml). There was good correlation between daily clonidine dose and plasma clonidine concentration. The average blood pressure reduction during 12 hr after clonidine correlated poorly with steady-state plasma levels of clonidine, but when the individual maximum decrease in mean blood pressure was paired with the corresponding peak plasma concentration of clonidine, the correlation was strong.


Cardiac Magnetic Resonance Feature Tracking Global Longitudinal Strain and Prognosis After Heart Transplantation.
This study determined the long-term prognostic significance of GLS assessed using CMR-FT in a large cohort of heart transplant recipients. In heart transplant recipients, global longitudinal strain (GLS) assessed using echocardiography has shown promise in the prediction of clinical outcomes. We hypothesized that CMR feature tracking (CMR-FT) GLS is independently associated with long-term outcomes in heart transplant recipients. In a cohort of consecutive heart transplant recipients who underwent routine CMR for clinical surveillance, CMR-FT GLS was calculated from 3 long-axis cine CMR images. Associations between GLS and a composite endpoint of death or major adverse cardiac events (MACE), including retransplantation, nonfatal myocardial infarction, coronary revascularization, and heart failure hospitalization, were investigated. A total of 152 heart transplant recipients (age 54 ± 15 years; 29% women; 5.0 ± 5.4 years after heart transplantation) were included. The median GLS was -11.6% (interquartile range: -13.6% to -9.2%). Over a median follow-up of 2.6 years, 59 recipients reached the composite endpoint. On Kaplan-Meier analyses, recipients with GLS worse than the median had a higher estimated cumulative incidence of the composite endpoint compared with recipients with GLS better than the median (log rank p = 0.004). On multivariate Cox proportional hazards regression, GLS was independently associated with the composite endpoint after adjustment for cardiac allograft vasculopathy, history of rejection, left ventricular ejection fraction (LVEF), right ventricular EF, and presence of myocardial fibrosis, with a hazard ratio of 1.15 for every 1% worsening in GLS (95% confidence interval: 1.06 to 1.24; p < 0.001). Similar results were seen in subgroups of recipients with LVEF >50% and with no myocardial fibrosis. GLS provided incremental prognostic value over other variables in the multivariate model as determined by the log-likelihood chi-squared test. In a large cohort of heart transplant recipients, CMR-FT GLS was independently associated with the long-term risk of death or MACE.


Antitumor potential of SLPI promoter controlled recombinant caspase-3 expression in laryngeal carcinoma.
The purpose of this study is to develop a specific and efficient targeted gene therapy candidate approach for laryngeal carcinomas. Several promoters of human squamous cell carcinoma antigen 2(SCCA2), secretory leukocyte protease inhibitor (SLPI) and Survivin genes were cloned from human genomic DNA and evaluated for tumor-specific transcription potential in human laryngeal carcinoma Hep-2 cells by dual luciferase assays. One SLPI promoter fragment (677 bp) showed the highest efficiency and specificity, and was used to control the expression of a recombinant active caspases-3 (revCasp3), which could trigger apoptosis without activation of its upstream cascade elements once expressed in a cell, in an adenoviral vector (Ad-SLPI-revCasp3), and its antitumor efficacy was assessed. In vitro infection with Ad-SLPI-revCasp3 showed revCasp3 could be specifically expressed in Hep-2 cells, resulting in efficient activation of endogenous Caspase-3 and subsequent apoptosis of Hep-2 cells. In Hep-2 nude mice xenograft model, intratumoral administration of Ad-SLPI-revCasp3 significantly inhibited tumor growth without obvious loss of body weight and obvious hepatic toxicity. In summary, our study showed the specific and efficient apoptosis-inducing potential of Ad-SLPI-revCasp3, and this makes it a new candidate approach of targeted gene therapy for laryngeal squamous cell carcinoma, which needs further systematic investigation.


Transplacental transfer of anthracyclines, vinblastine, and 4-hydroxy-cyclophosphamide in a baboon model.
The paucity of data on the fetal effects of prenatal exposure to chemotherapy prompted us to study transplacental transport of chemotherapeutic agents. Fluorouracil-epirubicin-cyclophosphamide (FEC) and doxorubicin-bleomycin-vinblastine-dacarbazine (ABVD) were administered to pregnant baboons. At predefined time points over the first 25 h after drug administration, fetal and maternal blood samples, amniotic fluid (AF), urine, fetal and maternal tissues, and cerebrospinal fluid (CSF) were collected. High-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) were used for bioanalysis of doxorubicin, epirubicin, vinblastine, and cyclophosphamide. In nine baboons, at a median gestational age of 139 days (range, 93-169), FEC 100% (n = 2), FEC 200% (n=1), ABVD 100% (n = 5), and ABVD 200% (n = 1) were administered. The obtained ratios of fetal/maternal drug concentration in the different simultaneously collected samples were used as a measure for transplacental transfer. Fetal plasma concentrations of doxorubicin and epirubicin averaged 7.5 ± 3.2% (n = 6) and 4.0 ± 1.6% (n = 8) of maternal concentrations, respectively. Fetal tissues contained 6.3 ± 7.9% and 8.7 ± 8.1% of maternal tissue concentrations for doxorubicin and epirubicin, respectively. Vinblastine concentrations in fetal plasma averaged 18.5 ± 15.5% (n=9) of maternal concentrations. Anthracyclines and vinblastine were neither detectable in maternal nor in fetal brain/CSF. 4-Hydroxy-cyclophosphamide concentrations in fetal plasma and CSF averaged 25.1 ± 6.3% (n = 3) and 63.0% (n = 1) of the maternal concentrations, respectively. This study shows limited fetal exposure after maternal administration of doxorubicin, epirubicin, vinblastine, and 4-hydroxy-cyclophosphamide.


A comparison of the lipolytic activities in liver perfusates and liver plasma membranes from rats.
We have undertaken a study to resolve the conflicting reports on the substrate specificity of the lipolytic enzyme(s) released by heparin from liver and report the following: (1) Heparin perfusates from liver contain an enzyme(s) capable of degrading triacylglycerol, diacylglycerophosphorylethanolamine and monoacylglycerol, whereas a heparin-solubilized fraction from liver plasma membranes hydrolyzes diacylglycerophosphorylethanolamine and monoacylglycerol only; (2) The lipolytic activities for the two sources behave differently on gel filtration but have the same behavior on heparin-Sepharose affinity chromatography; (3) Treatment of the preparation from the plasma membrane with Triton X-100 followed by heparin-Sepharose affinity chromatography produces forms of the enzyme(s) that now have activity on triacylglycerol This study suggests that the enzyme(s) from the two sources may be the same and that some change occurs when the enzyme is released from the intact liver.


Safety Validation of Repeated Blood-Brain Barrier Disruption Using Focused Ultrasound.
The purpose of this study was to investigate the effects on the brain of multiple sessions of blood-brain barrier (BBB) disruption using focused ultrasound (FUS) in combination with micro-bubbles over a range of acoustic exposure levels. Six weekly sessions of FUS, using acoustical pressures between 0.66 and 0.80 MPa, were performed under magnetic resonance guidance. The success and degree of BBB disruption was estimated by signal enhancement of post-contrast T1-weighted imaging of the treated area. Histopathological analysis was performed after the last treatment. The consequences of repeated BBB disruption varied from no indications of vascular damage to signs of micro-hemorrhages, macrophage infiltration, micro-scar formations and cystic cavities. The signal enhancement on the contrast-enhanced T1-weighted imaging had limited value for predicting small-vessel damage. T2-weighted imaging corresponded well with the effects on histopathology and could be used to study treatment effects over time. This study demonstrates that repeated BBB disruption by FUS can be performed with no or limited damage to the brain tissue.


FGL2 regulates IKK/NF-κB signaling in intestinal epithelial cells and lamina propria dendritic cells to attenuate dextran sulfate sodium-induced colitis.
Inflammatory bowel disease (IBD) is an autoimmune disease characterized by an abnormal immune response. Fibrinogen-like protein 2 (FGL2) is known to have immunoregulatory and anti-inflammatory activity. The level of FGL2 is elevated in patients with IBD; however, its comprehensive function in IBD is almost unknown. In our study, we explored the effect of FGL2 on dextran sulfate sodium (DSS)-induced colitis in mice and on NF-κB signaling in intestinal epithelial cells (IECs) and lamina propria dendritic cells (LPDCs). We founded that FGL2-/- mice in the colitis model showed more severe colitis manifestations than WT mice did, including weight loss, disease activity index (DAI), and colon histological scores. FGL2-/- mice treated with DSS produced more proinflammatory cytokines (IL-1β, IL-6, TNF-α) in serum than WT mice did and demonstrated upregulated expression of TNF-α and inflammatory marker enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (Cox-2) in the colon tissue. Our data suggested that DSS-treated FGL2-/- mice showed stronger activation of NF-κB signaling, especially in IECs. Next, we demonstrated that recombinant FGL2 (rFGL2) inhibited the production of proinflammatory cytokines and the expression of inflammatory marker enzymes by downregulating the NF-κB signaling in HT-29 cells. Finally, we discovered that LPDCs from the colon of DSS-treated FGL2-/- mice showed significantly upregulated expression of surface maturation co-stimulatory molecules, including CD80, CD86, CD40, and MHC class II molecules compared with that in WT mice. In addition, LPDCs in FGL2-/- treated with DSS exhibited excessive NF-κB activity and the administration of rFGL2 to FGL2-/- mice could rescue the aggravated results of FGL2-/- mice. Taken together, our findings demonstrated that FGL2 might be a target for further therapy of IBD.


18F-FDG-PET/CT predicts survival in hypopharyngeal squamous cell carcinoma.
We investigated the relationship between overall survival of patients and pretreatment [(18)F]-2-fluorodeoxyglucose ((18)F-FDG) uptake, assessed by positron emission tomography combined with computed tomography (PET/CT) in hypopharyngeal squamous cell carcinoma. Thirty-one patients who were newly diagnosed as resectable hypopharyngeal squamous cell carcinoma underwent pretreatment (18)F-FDG-PET/CT. We used the maximum standardized uptake value (SUVmax) as (18)F-FDG uptake. Overall survival rate was calculated by the Kaplan-Meier method. The median SUVmax was 11.53 (range 2.49-22.33). Patients with SUVmax ≥ 13 significantly exhibited shorter overall survival in univariate analysis (p < 0.01). Moreover, by Cox proportional hazards model of multivariate analysis, SUVmax ≥ 13 was a significant prognostic factor independent of clinical T and N classification, and treatment group (p < 0.02). These results suggested that SUVmax obtained by pretreatment (18)F-FDG PET/CT assessment is an important prognostic factor in patients with hypopharyngeal squamous cell carcinoma.


Short-term individual housing induced social deficits in female Mongolian gerbils: attenuation by chronic but not acute imipramine.
Mongolian gerbils are highly sensitive to manipulations of their social environments. Housing females individually for short periods (in the order of 7-21 days) has been shown to produce robust and reliable impairments of their subsequent social behaviour. These effects are typified by a marked reduction in the social investigation of an unfamiliar male in a neutral arena and/or a marked increases in levels of freezing whilst and only whilst they are being socially investigated (Immobile in contact). These responses demonstrate housing induced impaired motivation to socially interact. These effects have also been shown to be sensitive to treatment with chronic (but not acute) administration of the selective serotonin reuptake inhibitor (SSRI) fluoxetine. It was therefore of interest to know if similar effects would be produced by treatment with the tricyclic antidepressant Imipramine. This mixed NA/5-HT reuptake inhibitor first developed in the 1950's is a commonly used standard in animal models of depression and remains in clinical use today. Female gerbils were individually housed for 7 days or maintained in single-sex groups of 4 for the same period. All animals were then randomly allocated to be administered with either 0, 10 or 20 mg/kg imipramine. Acute administration did not reverse the social impairments produced by the individual housing but did produce non-specific stimulant effects on locomotion in both housing conditions. These social impairments were however reduced after a further 14 days chronic treatment with 10 or 20 mg/kg imipramine and stimulant effects were no longer seen. Following chronic administration in group-housed animals locomotor stimulation was replaced with sedation, which resulted in a reduction in social behaviour. That is, opposite to the effect seen in Individual housed animals. It is therefore concluded that chronic treatment with imipramine serves to increase social behaviour but only in those animals with a pre-existing social deficit.


Changes in Iris Stiffness and Permeability in Primary Angle Closure Glaucoma.
To evaluate the biomechanical properties of the iris by evaluating iris movement during pupil constriction and to compare such properties between healthy and primary angle-closure glaucoma (PACG) subjects. A total of 140 subjects were recruited for this study. In a dark room, the anterior segments of one eye per subject were scanned using anterior segment optical coherence tomography imaging during induced pupil constriction with an external white light source of 1700 lux. Using a custom segmentation code, we automatically isolated the iris segments from the AS-OCT images, which were then discretized and transformed into a three-dimensional point cloud. For each iris, a finite element (FE) mesh was constructed from the point cloud, and an inverse FE simulation was performed to match the clinically observed iris constriction in the AS-OCT images. Through this optimization process, we were able to identify the elastic modulus and permeability of each iris. For all 140 subjects (95 healthy and 45 PACG of Indian/Chinese ethnicity; age 60.2 ± 8.7 for PACG subjects and 57.7 ± 10.1 for healthy subjects), the simulated deformation pattern of the iris during pupil constriction matched well with OCT images. We found that the iris stiffness was higher in PACG than in healthy controls (24.5 ± 8.4 kPa vs. 17.1 ± 6.6 kPa with 40 kPa of active stress specified in the sphincter region; P < 0.001), whereas iris permeability was lower (0.41 ± 0.2 mm2/kPa s vs. 0.55 ± 0.2 mm2/kPa s; p = 0.142). This study suggests that the biomechanical properties of the iris in PACG are different from those in healthy controls. An improved understanding of the biomechanical behavior of the iris may have implications for the understanding and management of angle-closure glaucoma.