Biosynthesis of dolichyl pentasaccharide diphosphate in calf pancreas microsomes.
Incubation of synthetic dolichyl pyrophosphate tetrasaccharide and GDP-[14C]mannose with calf pancreas microsomes gave three lipid-linked oligosaccharides, which could be extracted with chloroform/methanol (2:1) and separated on silica gel plates. The fastest migrating product was characterized as dolichyl pyrophosphate pentasaccharide based on gel filtration and high pressure liquid chromatography. The formation of the pentasaccharide-lipid was greatly stimulated by addition of synthetic tetrasaccharide-lipid and required the presence of Triton X-100. Dolichyl phosphate mannose could not replace GDP-mannose as a sugar donor. The structure of the pentasaccharide was determined by degradation with endo-beta-N-acetylglucosaminidase D, acetolysis, alpha-D-mannosidase, and concanavalin A-Sepharose chromatography, showing that the following reaction was taking place: alpha-D-Manp-(1 leads to 3)-beta-D-Manp-(1 leads to 4)-beta-D-GlcpNAc-(1 leads to 4)-alpha-D-GlcpNAcPPDol + GDPMan leads to GDP + alpha-D-Manp-(1 leads to 3)-[alpha-D-Manp-(1 leads to 6)]-beta-D-Manp-(1 leads to 4)-beta-D-GlcpNAc-(1 leads to 4)-alpha-D-GlcpNAcPPDol. The mannosyltransferase was partially characterized.


[3H]AF-DX 116 labels subsets of muscarinic cholinergic receptors in rat brain and heart.
The in vitro binding properties of the novel muscarinic antagonist [3H]AF-DX 116 were studied using a rapid filtration technique. Association and dissociation rates of [3H]AF-DX 116 binding were rapid at 25 degrees C (2.74 and 2.70 X 10(7) min-1 M-1 for K+1; 0.87 and 0.93 min-1 for k-1) but 20-40 times slower at 0-4 degrees C (0.13 and 0.096 X 10(7) min-1 M-1 for k+1; 0.031 and 0.022 min-1 for k-1 in cerebral cortical and cardiac membranes, respectively). Kinetic dissociation constants (Kds) were estimated to be 31.8 nM and 30.9 nM at 25 degrees C; 23.1 nM and 0-4 degrees C for the cerebral cortex and heart, respectively. In saturation studies, [3H]AF-DX 116 labeled 29 percent of the total [3H](-)QNB binding sites in the cerebral cortical membranes and 87 percent in the cardiac membranes, with Kd values of 28.9 nM and 17.9 nM, respectively. Muscarinic antagonists inhibited [3H]AF-DX 116 binding in a rank order of potency of atropine greater than dexetimide greater than AF-DX 116 greater than PZ greater than levetimide in both tissues. Except for PZ/[3H]AF-DX 116 and AF-DX 116/[3H]AF-DX 116 in the cerebral cortex, all the antagonist competition curves had Hill coefficients close to one. Carbachol and oxotremorine produced shallow inhibition curves against [3H]AF-DX 116 binding in both tissues. Regional distribution studies with [3H](-)QNB, [3H]PZ and [3H]AF-DX 116 showed that most of the muscarinic receptors in the cerebral cortex, hippocampus, nucleus accumbens and corpus striatum are of the M1 subtype while those in the brainstem, cerebellum and other lower brain regions are of the M2 subtype. These results indicate that [3H]AF-DX 116 is a useful probe for the study of heterogeneity of muscarinic cholinergic receptors.


Beta2-ADR haplotypes/polymorphisms associate with bronchodilator response and total IgE in grass allergy.
Association and linkage studies of beta2-adrenergic receptor (beta2-ADR) polymorphisms in relation to the expression of asthmatic phenotypes and immune regulatory mechanisms have shown inconsistent results. In order to analyse the relevance of particular combinations of single nucleotide polymorphisms (SNPs) or haplotypes of beta2-ADR gene to bronchial asthma, bronchodilator response and total immunoglobulin E (IgE) we determined by direct DNA sequencing five SNPs (in positions: -47, -20, 46, 79, 252) in a group of 180 Caucasian subjects (110 patients with grass allergy and 70 nonatopic controls). The eight different beta2-ADR haplotypes were identified, with three the most common of them representing 92% of the studied cohort. Significantly higher (pcor = 0.0045) bronchodilator response was observed in patients with homozygotic genotype 46A/A in comparison with respective homo- and hetero-zygotes. There was no significant difference in bronchodilator response when beta2-ADR haplotypes were analysed. Significantly higher (pcor = 0.0005) total IgE levels were found in patients with beta2-ADR haplotype -47T/-20T/46A/79C/252G and homozygotic carriers of 46A (pcor = 0.0015) and 79C (pcor = 0.003) genotypes. No significant associations were found in regards to asthmatic phenotype and atopy. These results indicate that depending on phenotype studied, either an individual beta2-ADR SNP or beta2-ADR haplotype might affect disease manifestation.


Contributions of magnetic resonance imaging in the evaluation of optic gliomas.
Eleven patients with gliomas in the optic chiasm were studied using a 0.5 Tesla superconducting system and third generation computed tomography scanners. All tumors were identified with both modalities, but in the majority of cases, the posterior extension of the tumor and its relationship to adjacent brain, important to both prognosis and management, were shown better by magnetic resonance imaging.


Organocatalysts wrapped around by poly(ethylene glycol)s (PEGs): a unique host-guest system for asymmetric Michael addition reactions.
Asymmetric Michael addition reactions of unmodified ketones to nitroalkenes were performed in PEGs catalyzed by novel pyrrolidinyl-thioimidazolium salts to give products in up to 97% yield and 99% enantioselectivity; ESI mass spectrometric detection for the first time gave evidence of the presence of the PEG-organocatalyst host-guest complex.


Extracellular matrix alterations in the ketamine model of schizophrenia.
The neural extracellular matrix (ECM) plays an important role in regulation of perisomatic GABAergic inhibition and synaptic plasticity in the hippocampus and cortex. Decreased labeling of perineuronal nets, a form of ECM predominantly associated with parvalbumin-expressing interneurons in the brain, has been observed in post-mortem studies of schizophrenia patients, specifically, in brain areas such as prefrontal cortex, entorhinal cortex, and amygdala. Moreover, glial ECM in the form of dandelion clock-like structures was reported to be altered in schizophrenia patients. Here, we verified whether similar abnormalities in neural ECM can be reproduced in a rat model of schizophrenia, in which animals received sub-chronic administration of ketamine to reproduce the aspects of disease related to disrupted signaling through N-methyl-D-aspartate receptors. Our study focused on two schizophrenia-related brain areas, namely the medial prefrontal cortex (mPFC) and hippocampus. Semi-quantitative immunohistochemistry was performed to evaluate investigate ECM expression using Wisteria floribunda agglutinin (WFA) and CS56 antibody, both labeling distinct chondroitin sulfate epitopes enriched in perineuronal nets and glial ECM, respectively. Our analysis revealed that ketamine-treated rats exhibit reduced number of WFA-labeled perineuronal nets, and a decreased intensity of parvalbumin fluorescence in mPFC interneurons somata. Moreover, we found an increased expression of CS56 immunoreactive form of ECM. Importantly, the loss of perineuronal nets was revealed in the mPFC, and was not detected in the hippocampus, suggesting regional specificity of ECM alterations. These data open an avenue for further investigations of functional importance of ECM abnormalities in schizophrenia as well as for search of treatments for their compensation.


Assessment of dual-energy computed tomography derived virtual monoenergetic imaging for target volume delineation of brain metastases.
Objective and subjective assessment of image quality of brain metastases on dual-energy computed tomography (DECT) virtual monoenergetic imaging (VMI) and its impact on target volume delineation. 26 patients with 37 brain metastases receiving Magnetic Resonance Imaging (MRI) and DECT for stereotactic radiotherapy planning were included in this retrospective analysis. Lesion contrast (LC), contrast-to-noise ratio (CNR) and signal-to-noise ratio (SNR) were assessed for reconstructed VMI at 63 keV and artificial 120 kV Computed Tomography (CT). Image contrast and demarcation of metastases between 120 kV CT, VMI and MRI were subjectively assessed. Brain metastases were delineated by four radiation oncologists on VMI with a fixed or free brain window and contours were compared to solely MRI-based delineation using the Dice similarity coefficient. LC, CNR and SNR were significantly higher in VMI than in 120 kV CT (p < 0.0001). Image contrast and lesion demarcation were significantly better on VMI compared to 120 kV CT (p < 0.0001). Mean gross tumor volume (GTV)/planning target volume (PTV) Dice similarity coefficients were 0.87/0.9 for metastases without imaging uncertainties (no artifacts, calcification or impaired visibility with MRI) but worse for metastases with imaging uncertainties (0.71/0.74). Target volumes delineated on VMI were around 5-10% smaller compared to MRI. Image quality of VMI is objectively and subjectively superior to conventional CT. VMI provides significant advantages in stereotactic radiotherapy planning with improved visibility of brain metastases and geometrically distortion-free representation of brain metastases. Beside a plausibility check of MRI-based target volume delineation, VMI might improve reliability and accuracy in target volume definition particularly in cases with imaging uncertainties with MRI.


Astrocytic proliferation and mitochondrial dysfunction induced by accumulated glutaric acidemia I (GAI) metabolites: possible implications for GAI pathogenesis.
Glutaric (GA) and 3-hydroxyglutaric (OHGA) acids accumulate in glutaric acidemia I (GAI), a neurometabolic disease characterized by acute striatal degeneration and chronic progressive cortical atrophy. To explore the hypothesis that astrocytes are involved in GAI pathogenesis and targets of accumulating metabolites, we determined the effects of GA and OHGA on cultured rat cortical astrocytes. Remarkably, both acids induced mitochondria depolarization and stimulated proliferation in confluent cultures without apparent cell toxicity. Newborn rats injected with GA systemically also showed increased cell proliferation in different brain regions. Most of the proliferating cells displayed markers of immature astrocytes. Antioxidant iron porphyrins prevented both mitochondria dysfunction and increased in vitro and in vivo proliferation, suggesting a role of oxidative stress in inducing astrocytosis. Taken together, the data suggest that mitochondrial dysfunction induced by GA metabolites causes astrocytes to adopt a proliferative phenotype, which may underlie neuronal loss, white matter abnormalities and macrocephalia characteristics of GAI.


Studies on the potential protective effect of cinnamon against bisphenol A- and octylphenol-induced oxidative stress in male albino rats.
Among the numerous chemicals discharged into the surrounding environment, bisphenol A (BPA) and octylphenol (OP) have been shown to increase oxidative stress in body by disturbing the prooxidant/antioxidant balance of cells. Cinnamon aqueous extract (CAE) is a natural product rich in polyphenolic compounds that have antioxidant activity. This study was designed to investigate the protective efficacy of CAE against oxidative disorders induced by BPA and OP in male albino rats. Animals were divided into 6 groups (10 rats each) and treated orally, 3 times weekly for 50 days. Group 1: control vehicle (olive oil); group 2 (25 mg BPA/kg b.wt./day); group 3 (25 mg OP/kg b.wt./day); group 4 (200 mg CAE/kg b.wt./day); group 5 (CAE 2 h before BPA administration); and group 6 (CAE 2 h before OP administration). BPA- and OP-exposed groups showed insignificant elevation in the final body weight; weight gains and significant reduction only in the relative kidneys weight. Also, BPA and OP exposure resulted in significant increase in serum urea, creatinine and kidney, brain, testicular malondialdehyde (MDA) levels. Significant reduction in tissues reduced glutathione (GSH) contents; catalase (CAT) and superoxide dismutase (SOD) activities were also recorded in BPA and OP exposed animals compared to the control vehicle group. Pretreatment with CAE 2 h either before BPA or OP administration ameliorated the BPA- and OP-induced body weight; weight gains and relative organs weight changes and biochemical adverse effects. CAE pretreatment also protected against the recorded pathological changes in kidney, brain and testis. In conclusion, CAE could ameliorate the oxidative toxic effects of BPA and OP indicating its protective antioxidant effect.


On the origin of the Helmholtz's square illusion: An attentional account.
A square filled with parallel horizontal or vertical lines appears perceptually extended in the direction orthogonal to the lines. Here, we suggest that this Helmholtz illusion arises due to changes in spatial attention that entail changes at very early stages of perceptual processing. Three experiments are reported which tested this assumption. In Experiment1 and Experiment2, transient attentional cues were flashed in such a way that they either promoted (congruent condition) or hindered (incongruent condition) the attentional state presumably induced by the target objects. We predicted a decline of the illusion in the incongruent condition compared with the congruent condition. This prediction was confirmed in both experiments. However, the influence of (in)congruent attention cuing on the Helmholtz illusion depended on more sustained distributions of attention as well. An influence of sustained attention on the illusion was confirmed in Experiment 3, in which changes of attentional focus were induced by a secondary task. Overall, the results were consistent with our claim that the origin of the Helmholtz illusion is closely linked to the distribution of spatial attention.


Characterization of MHC class I alleles in sooty mangabeys as a tool for evaluating cellular immunity in natural hosts of SIV infection.
Although immune pressure exerted by MHC class I-restricted cytotoxic T lymphocytes (CTL) are an important determinant of outcome in pathogenic HIV and SIV infection, lack of data on MHC class I genes has hampered study of its role in natural hosts with nonpathogenic SIV infection. In this study, we cloned and characterized full-length MHC class I genes derived from the cDNA library of two unrelated naturally infected sooty mangabeys (Cercocebus atys) in whom SIV-specific CTL epitopes were previously mapped. Twenty one full-length MHC class I alleles consisting of five MHC-A (Ceat-A), 13 MHC-B (Ceat-B), and three MHC-E (Ceat-E) alleles were identified. Sequence-specific primers (SSP) for high-throughput screening of genomic DNA by PCR were developed for 16 of the 18 Ceat-A and Ceat-B alleles. Screening of 62 SIV-negative and 123 SIV-infected sooty mangabeys at the Yerkes National Primate Research Center (YNPRC) revealed the presence of up to four MHC-A and eight MHC-B alleles in individual mangabeys, indicating that similar to macaque species, mangabeys have at least two duplications of the MHC-A locus and four duplications of the MHC-B locus in the absence of an MHC-C locus. Using stable transfectants of Ceat MHC Class I alleles in the MHC-null 721.221 cell line, we identified Ceat-B*12:01 as the restricting allele of a previously reported Nef20-28 CTL epitope. Ceat-B*1201/Nef20-28 tetramers identified tetramer-positive CD8+ T lymphocytes in Ceat-B*1201-positive SIV-infected mangabeys. This study has laid the groundwork for comprehensive analysis of CTL and SIV evolution in a natural host of SIV infection.


Bioactive Properties of Maillard Reaction Products Generated From Food Protein-derived Peptides.
Food protein-derived peptides are promising food ingredients for developing functional foods, since various bioactive peptides are released from food proteins. The Maillard reaction, which plays an important role in most processed foods, generates various chemical components during processing. Although changes of amino acids or proteins and reduced sugars by the Maillard reaction have been studied extensively, such changes of peptides by the Maillard reaction are still not resolved enough. Since food protein-derived peptides are widely utilized in many processed foods, it deserves concern and research on the changes of peptides by the Maillard reaction in foods during processing or storage. This chapter initially overviewed food protein-derived bioactive peptides. Then, Maillard reaction products generated from peptides are discussed. We focused particularly on their bioactivities.


The effects of tenotomy upon the contraction characteristics of motor units in rabbit soleus muscle.
1. Isometric contractions of tenotomized rabbit solei have been compared with a group of unoperated control muscles. A large decrease in the tension developed by the tenotomized muscle was accompanied by a slight shortening of the twitch contraction time, and a larger, progressive, reduction in the time to half relaxation.2. A comparison of motor units obtained from muscles which had been tenotomized for six weeks with those of the control muscles showed a large reduction in the range of contraction and half relaxation times of the units from the tenotomized muscles.3. The mean motor unit tension (expressed as a percentage of the whole muscle tension) was similar for both those units from the control muscles and those from muscles which had been tenotomized for six weeks, indicating a uniform atrophy of motor units within the tenotomized muscles.4. It was concluded that the change in the pattern of motor unit contraction times was not the result of a process of differential atrophy favouring the preservation of the faster contracting motor units.5. A correlation between axon conduction velocity and both the speed of contraction and the size (tension) of the motor units was demonstrated in the control muscles. Following tenotomy the relationship between axon conduction velocity and motor unit tension was lost.


Electro-optical measurements on aqueous suspension of purple membrane from Halobacterium halobium.
The permanent dipole moment, polarizability, and the retinal angle of Halobacterium halobium purple membranes were determined at different pH values. All of the parameters have a maximum between pH 5 and 6. There is a reversal in the direction of the permanent dipole moment near pH 5. The value of permanent dipole moment was determined to be 60 D/protein at pH 6.6, and the value obtained for polarizability was 3 X 10(-28) Fm2/membrane fragment. The retinal angle of all-trans retinal was 0.8 degrees smaller than that of the 13-cis conformation.


Detection of Enteroviruses and SARS-CoV-2 in Tunisian Wastewater.
Monitoring the circulation of enteric viruses in environmental wastewater is a valuable tool for preventing the emergence of waterborne and food-borne diseases in humans. The detection of viruses was performed in five Tunisian wastewater treatment plants, three located in the Grand Tunis City (WWTP 1, WWTP 2, WWTP 3) and two in the Sahel of Tunisia (WWTP 4, WWTP 4), known as very developed and crowded zones, to assess the effectiveness of three biological wastewater treatment procedures namely natural oxidizing lagoons, rotating biodisks procedure, activated sludge procedure, and one tertiary sewage treatment using UV-C254 reactor for this enteric viruses' removal. Thus, 242 sewage samples were collected between June 2019 and May 2020 from different lines of wastewater treatment procedures implemented in the five wastewater treatment plants investigated. SARS-CoV-2 was analyzed using real-time multiplex reverse-transcription polymerase chain reaction (multiplex real-time RT-PCR) and enteroviruses using reverse-transcription polymerase chain reaction (RT-PCR). The enteroviruses detection showed 93% and 73% respective high frequencies only in the two WWTPs of the Grand Tunis (WWTP 1 and WWTP 2). SARS-CoV-2 was detected in 58% of the all wastewater samples collected from the five studied WWTPs with a respective dominance of N gene (47%), S gene (42%), RdRp gene (42%) and at last E gene (20%). These enteroviruses and SARS-CoV-2 detection were revealed in all steps of the wastewater treatment procedures, so poor virological quality is found at the exit of each biological and tertiary step of treatment investigated. For the first time in Tunisia, these results highlighted the enterovirus and SARS-CoV-2 detection with high rates, and the ineffectiveness of the biological and UV-C254 treatment implemented to remove these viruses. The preliminary results of SARS-CoV-2 circulation in Tunisian wastewater confirmed the wide positivity rate underlined by other works worldwide and allowed showing a move towards integrating wastewater as a way for this virus to spread in different areas and environments. So, this last result about SARS-CoV-2 circulation allowed us to caution about the strong probability of diffusion of this hazardous virus through water and sewage; despite its enveloped character and nature, as a labile and sensitive virus in these environments. Thus, establishing a national surveillance strategy is needed to improve the sanitary quality of treated wastewater and prevent public health problems related to these viruses in treated wastewater.


High-throughput in-silico prediction of ionization equilibria for pharmacokinetic modeling.
Chemical ionization plays an important role in many aspects of pharmacokinetic (PK) processes such as protein binding, tissue partitioning, and apparent volume of distribution at steady state (Vdss). Here, estimates of ionization equilibrium constants (i.e., pKa) were analyzed for 8132 pharmaceuticals and 24,281 other compounds to which humans might be exposed in the environment. Results revealed broad differences in the ionization of pharmaceutical chemicals and chemicals with either near-field (in the home) or far-field sources. The utility of these high-throughput ionization predictions was evaluated via a case-study of predicted PK Vdss for 22 compounds monitored in the blood and serum of the U.S. population by the U.S. Centers for Disease Control and Prevention National Health and Nutrition Examination Survey (NHANES). The chemical distribution ratio between water and tissue was estimated using predicted ionization states characterized by pKa. Probability distributions corresponding to ionizable atom types (IATs) were then used to analyze the sensitivity of predicted Vdss on predicted pKa using Monte Carlo methods. 8 of the 22 compounds were predicted to be ionizable. For 5 of the 8 the predictions based upon ionization are significantly different from what would be predicted for a neutral compound. For all but one (foramsulfuron), the probability distribution of predicted Vdss generated by IAT sensitivity analysis spans both the neutral prediction and the prediction using ionization. As new data sets of chemical-specific information on metabolism and excretion for hundreds of chemicals are being made available (e.g., Wetmore et al., 2015), high-throughput methods for calculating Vdss and tissue-specific PK distribution coefficients will allow the rapid construction of PK models to provide context for both biomonitoring data and high-throughput toxicity screening studies such as Tox21 and ToxCast.


[The significance of topic differences and cadaveric changes of histamine and serotonin levels for the determination of the age of wounds in animal experiments (author's transl)].
Even when the extraction method of Fazekas for the determination of the so-called free histamine and serotonin is used, different dorsal skin areas of laboratory animals show certain differences in the amine content, which reach higher values for histamine than for 5-HT. 1) Related to the animal collective, no differences in the mean values of the 5-HT and histamine content between the right and left side of damaged dorsal skin of guinea pigs resulted, although in single cases remarkable differences (up to 1.26 microgram/g) were found. Descending from nuchal to caudal a decrease of the amine content--less in case of 5-HT, more in case of histamine--is noticed. The observed topical differences possibly depend on the continous moulting of the animals. 2) From these analytical data a method for statistical prediction is derived, which allows to state, when and with which probability a single difference value exceeds the normal variance. Applied to the comparative measurements wounded skin/control skin it is thus made possible to test the validity of the findings obtained in practical cases. 3) Postmortal changes of the amine content of the wounded as well as the normal skin are not detectable for the 5-HT, while in the case of histamine solely the values of the damaged skin first show a decrease (24 h p.m.), followed by an increase (48 h p.m., bacterial neogenesis?).


The recurrence and progression risk after simultaneous endoscopic surgery of urothelial bladder tumour and benign prostatic hyperplasia: a systematic review and meta-analysis.
To evaluate recurrence and progression risk after simultaneous endoscopic surgery of bladder cancer and benign prostatic hyperplasia (BPH), as simultaneous surgery is not an unusual scenario and theoretically simultaneous transurethral resection of bladder tumour (TURBT) and transurethral resection of the prostate (TURP) can lead to an increased risk of recurrence in the bladder neck and prostatic urethra (BN/PU). We conducted a systematic review and meta-analysis to assess the risk of recurrence (i.e. whole bladder and/or BN/PU) and tumour progression as outcomes after a simultaneous endoscopic surgery of bladder tumour and BPH, as compared to TURBT alone. We queried PubMed and Web of Science database on 1 January 2020. We used random- and/or fixed-effects meta-analytic models in the presence or absence of heterogeneity according to the I2 statistic, respectively. Nine retrospective and three clinical trial studies were selected after considering inclusion and exclusion criteria. We conducted the meta-analysis on retrospective and randomised controlled trials (RCTs) separately. Eight retrospective and three RCT studies were included to assess the BN/PU recurrence risk and the summarised risk ratio (RR) was 1.02 (95% confidence interval [CI] 0.74-1.41) and 0.93 (95% CI 0.47-1.84), respectively. Five retrospective and two RCT studies were included to assess the progression risk and the summarised RR was 0.91 (95% CI 0.56-1.48) and 1.16 (95% CI 0.30-4.51), respectively. Eight retrospective and three RCT studies were included to assess the whole bladder recurrence risk and the summarised RR was 0.87 (95% CI 0.78-0.97) and 0.89 (95% CI 0.65-1.21), respectively. We did not observe any increased risk of total bladder recurrence, BN/PU recurrence, or progression after a simultaneous endoscopic surgery of bladder tumour and BPH, as compared to TURBT alone.


Specific therapy in severe fetal intrauterine growth retardation: failure of prostacyclin.
Two case histories of severe, early onset intrauterine growth retardation are presented. Intravenous prostacyclin was administered in an attempt to promote fetal growth and thus prolong the pregnancy, but was unsuccessful and resulted in intrauterine death in each case. The possible reasons for the failure of this treatment and alternative therapeutic options are discussed.


Surface-enhanced Raman scattering detection of silver nanoparticles in environmental and biological samples.
Growing concerns over the potential release and threat of silver nanoparticles (AgNPs) to environmental and biological systems urge researchers to investigate their fate and behavior. However, current analytical techniques cannot meet the requirements for rapidly, sensitively and reliably probing AgNPs in complex matrices. Surface-enhanced Raman spectroscopy (SERS) has shown great capability for rapid detection of AgNPs based on an indicator molecule that can bind on the AgNP surface. The objective of this study was to exploit SERS to detect AgNPs in environmental and biological samples through optimizing the Raman indicator for SERS. Seven indicator molecules were selected and determined to obtain their SERS signals at optimal concentrations. Among them, 1,2-di(4-pyridyl)ethylene (BPE), crystal violet and ferric dimethyl-dithiocarbamate (ferbam) produced the highest SERS intensities. Further experiments on binding competition between each two of the three candidates showed that ferbam had the highest AgNPs-binding ability. The underlying mechanism lies in the strong binding affinity of ferbam with AgNPs via multiple sulfur atoms. We further validated ferbam to be an effective indicator for SERS detection of as low as 0.1mg/L AgNPs in genuine surface water and 0.57 mg/L in spinach juice. Moreover, limited interference on SERS detection of AgNPs was found from environmentally relevant inorganic ions, organic matter, inorganic particles, as well as biologically relevant components, demonstrating the ferbam-assisted SERS is an effective and sensitive method to detect AgNPs in complex environmental and biological samples.


Localization of methylene blue paramolybdate in vitally stained nerves.
Methylene blue taken up by living neurons can be preserved for electron microscopy in a fixative containing osmium tetroxide and ammonium paramolybdate at pH 5.2. Paramolybdate is the buffer, precipitating agent and main osmotic ingredient; it does not function as an electron stain unless methylene blue is present. The low pH keeps the dye/paramolybdate complex from dissolving. Neither the low pH nor drastic dehydration from water to absolute ethanol harm the tissue. The staining mechanism involves cationic methylene blue associating with anionic structures such as microtubules and neurofilaments in the living cell; during fixation paramolybdate forms a precipitate with the dye at the staining sites. This fixative does not preserve microtubules unless they are first vitally stained.


Trauma of occult hydronephrotic kidney.
The occult unilateral hydronephrotic kidney is often discovered during the genitourinary evaluation of patients sustaining blunt abdominal trauma. Few cases have been reported documenting the angiographic, computerized tomography (CT), and ultrasound appearances. Two cases are described which demonstrate that relatively minor trauma can precipitate hematuria and hypovolemic shock. Angiography demonstrated the bleeding site in both cases and was utilized in conjunction with other parameters of clinical assessment to plan initial management. CT and ultrasound proved to be useful noninvasive diagnostic parameters for baseline and follow-up studies in patients undergoing conservative management. They accurately demonstrated the degree of hydronephrosis, residual renal parenchymal, and resolving hematoma.


Phase I/II trial of combined kyphoplasty and intraoperative radiotherapy in spinal metastases.
Spinal metastases occur in 30%-50% of patients with systemic cancer. The primary goals of palliation are pain control and prevention of local recurrence. This study aimed to test the safety and efficacy of a combined modality approach consisting of kyphoplasty and intraoperative radiotherapy (Kypho-IORT). Kyphoplasty and intraoperative radiotherapy was a prospective, single-center phase I/II trial. Patients were enrolled in a classical 3+3 scheme within the initial phase I, where Kypho-IORT was applied using a needle-shaped 50 kV X-ray source at three radiation dose levels (8 Gy in 8-mm, 8 Gy in 11-mm, and 8 Gy in 13-mm depth). Thereafter, cohort expansion was performed as phase II of the trial. The trial is registered with clinicaltrials.gov, number NCT01280032. Patients aged 50 years and older with a Karnofsky Performance Status of at least 60% and with one to three painful vertebral metastases confined to the vertebral body were eligible to participate. The primary end point was safety as per the occurrence of dose-limiting toxicities. The secondary end points were pain reduction, local progression-free survival (L-PFS), and overall survival (OS). Pain was measured using the visual analog scale (VAS) and local control was assessed in serial computed tomography or magnetic resonance imaging scans. None of the nine patients enrolled in the phase I showed dose-limiting toxicities at any level and thus, 52 patients were subsequently enrolled into a phase II, where Kypho-IORT was performed at various dose levels. The median pain score significantly dropped from 5 preoperatively to 2 at the first postoperative day (p<.001). Of 43 patients who reported a pre-interventional pain level of 3 or more, 30 (69.8%) reported a reduction of ≥3 points on the first postoperative day. A persistent pain reduction beyond the first postoperative day of ≥3 points was seen in 34 (79.1%) patients. The 3, 6, and 12 month L-PFS was excellent with 97.5%, 93.8%, and 93.8%. The 3, 6, and 12 months OS was 76.9%, 64.0%, and 48.4%. Kyphoplasty and intraoperative radiotherapy is safe and immediately provided sustained pain relief with excellent local control rates in patients with painful vertebral metastases.


Uveitis in patients with oligoarticular juvenile idiopathic arthritis and juvenile spondyloarthritis/enthesitis related arthritis: is there any difference?
Juvenile idiopathic arthritis (JIA) is the most common systemic disease causing uveitis in childhood and adolescence.


Identification of a Recognizable Progressive Skeletal Dysplasia Caused by RSPRY1 Mutations.
Skeletal dysplasias are highly variable Mendelian phenotypes. Molecular diagnosis of skeletal dysplasias is complicated by their extreme clinical and genetic heterogeneity. We describe a clinically recognizable autosomal-recessive disorder in four affected siblings from a consanguineous Saudi family, comprising progressive spondyloepimetaphyseal dysplasia, short stature, facial dysmorphism, short fourth metatarsals, and intellectual disability. Combined autozygome/exome analysis identified a homozygous frameshift mutation in RSPRY1 with resulting nonsense-mediated decay. Using a gene-centric "matchmaking" system, we were able to identify a Peruvian simplex case subject whose phenotype is strikingly similar to the original Saudi family and whose exome sequencing had revealed a likely pathogenic homozygous missense variant in the same gene. RSPRY1 encodes a hypothetical RING and SPRY domain-containing protein of unknown physiological function. However, we detect strong RSPRY1 protein localization in murine embryonic osteoblasts and periosteal cells during primary endochondral ossification, consistent with a role in bone development. This study highlights the role of gene-centric matchmaking tools to establish causal links to genes, especially for rare or previously undescribed clinical entities.


A simple technique for neoumbilical reconstruction for congenital hernia associated proboscis deformity.
Umbilical reconstruction in pediatric patients who have developed a large proboscoid redundancy can be challenging after standard umbilical repair. We present a simple and unique surgical technique that results in a cosmetically appealing reconstruction. The operation is initiated with circumferential redundant skin excision and isolation of the hernia sac. Primary fascial repair is performed. Reconstruction utilizes the cut dermal/epidermal edge by approximating it to the exposed fascia just below the skin edge outside of the fascial repair in a purse string fashion for the creation of a neoumbilicus. This simple technique is unique from any currently published methods and results in a cosmetically pleasing reconstruction without evidence of any incision. Level V: Expert Opinion.


The Stabilization of Central Sympathetic Nerve Activation by Renal Denervation Prevents Cerebral Vasospasm after Subarachnoid Hemorrhage in Rats.
The poor prognosis of subarachnoid hemorrhage (SAH) might be associated with sympathetic nerve activation (catecholamine surge) initiated by hypothalamic injury. As renal denervation (RD) has been shown to exert protective effects on cardiovascular dysfunction by suppressing increased central sympathetic nerve activation, we examined whether RD improved the experimental SAH prognosis in this study. Two hundred thirty-eight male Sprague-Dawley rats were divided into sham-operated and SAH-operated groups, and then each rat was further separated into Sham-operated and RD-operated groups. Bilateral RD was performed approximately 45 min after SAH induction. We examined the effect of RD on early brain injury (EBI) and delayed cerebral ischemia (DCI) as a primary endpoint, and also explored the effect on cerebral vasospasm (CVS) as a secondary endpoint. Although RD did not exert significant effects on primary endpoint, RD significantly prevented CVS and reduced SAH-induced increases in the number of phosphorylated extracellular signal-regulated kinase (ERK)-positive endothelial cells, cyclooxygenase-2 expression, and macrophage infiltration in major cerebral arteries. Moreover, RD significantly decreased the areas displaying dopamine β-hydroxylase and glial fibrillary acidic protein immunopositivity in the paraventricular nucleus of the hypothalamus and serum angiotensin II levels, all of which were increased by SAH. Although RD decreased systolic blood pressure, significant changes in cerebral blood flow were not observed compared with SAH + Sham group. Based on the findings, RD improved CVS by reducing endothelial cell damage and the effects were associated with the stabilization of central sympathetic nerve activation in a SAH model.


Expression and identification of a thermostable malate dehydrogenase from multicellular prokaryote Streptomyces avermitilis MA-4680.
A malate dehydrogenase (MDH) from Streptomyces avermitilis MA-4680 (SaMDH) has been expressed and purified as a fusion protein. The molecular mass of SaMDH is about 35 kDa determined by SDS-PAGE. The recombinant SaMDH has a maximum activity at pH 8.0. The enzyme shows the optimal temperature around 42 °C and displays a half-life (t(1/2)) of 160 min at 50°C which is more thermostable than reported MDHs from most bacteria and fungi. The k(cat) value of SaMDH is about 240-fold of that for malate oxidation. In addition, the k(cat)/K(m) ratio shows that SaMDH has about 1,246-fold preference for oxaloacetate (OAA) reduction over L-malate oxidation. The recombinant SaMDH may also use NADPH as a cofactor although it is a highly NAD(H)-specific enzyme. There was no activity detected when malate and NADP(+) were used as substrates. Substrate inhibition studies show that SaMDH activity is strongly inhibited by excess OAA with NADH, but is not sensitive to excess L-malate. Enzymatic activity is enhanced by the addition of Na(+), NH(4)(+), Ca(2+), Cu(2+) and Mg(2+) and inhibited by addition of Hg(2+) and Zn(2+). MDH is widely used in coenzyme regeneration, antigen immunoassays and bioreactors. The enzymatic analysis could provide the important basic knowledge for its utilizations.


Epigenetics: monoallelic expression in the immune system.
Epigenetic modifications to DNA and chromatin programme important genome functions including gene expression, chromosomal architecture and stability, and the maintenance of developmental states. Recent findings further implicate epigenetic modifications in the control of allelic choice in the immune system.


An in vivo evaluation of a novel spiral cut flexible ureteral stent.
The flow characteristics, ureteral conformance, and histopathologic changes of a novel spiral cut flexible ureteral stent (Percuflex Helical, Boston Scientific, Boston, MA) were evaluated in vivo in an acute and chronic porcine model. Flow characteristics and ureteral conformance of the novel stent were determined in 6 acute and 6 chronic swine models and compared with a control ureteral stent (Percuflex Plus, Boston Scientific). The flow characteristics were determined in vivo after ligating the renal vessels and via a nephrostomy tube delivering a standard rate of 0.9% saline at 35 cm H(2)O. Flows in the unobstructed ureter, normal stent, intraluminally obstructed stent, extraluminal obstructed stent, and both intraluminally and extraluminally obstructed conditions were determined. In the chronic animals, flow was determined at day 10, with the stent in place and immediately after stent removal. Conformance and hydronephrosis was assessed on pyelograms. Histopathologic changes were also evaluated in the chronic animals. The acute and chronic flow characteristics in the novel stent were equivalent to the control stent. Size and weight of the kidney, degree of hydronephrosis, stent migration, and presence of urinary tract infection were also similar between the test and control stents. There were no differences seen in histopathologic grading or degree of encrustation in either stent. The novel stent appeared to conform better to the shape of the ureter in both acute and chronic animals. The novel helical stent appears to drain as well as a standard stent and causes no increased degree of histopathologic changes in the ureter.


An integrative review on the uses of plant-derived bioactives formulated in conventional and innovative dosage forms for the local treatment of damaged nasal cavity.
Plants and their derivates have been used as medicines for centuries and today is being re-discovered their usefulness for the human health. The therapeutic properties of phytochemicals are re-evaluated under the light of medical and pharmacological research, pushed by a constantly growing market demand, where consumers trust more natural products than synthetic drugs. New studies are enlightening the effectiveness of phytochemicals against a wide range of ailments, nevertheless very few evaluate the efficacy of topical formulations based on natural bioactive molecules in the treatment of nasal mucosal diseases. This review aims at exploring this little covered topic. An overview on the properties and functionality of the nasal mucosa and the different diseases affecting it has been provided. We summarized various nasal dosage forms containing natural bioactive and explored how innovative delivery systems loading phytochemicals can improve the treatment results. Finally, the potential use of novel nanocarriers for the treatment of nasal ailments has been covered as well.


Distribution pharmacokinetics of warfarin in the rat, a non-linear multicompartment model.
Preliminary analysis and linear two-compartment solutions of warfarin plasma concentrations recorded in the rat after intravenous bolus injections of 1, 2, 8 and 40 mg/kg of sodium warfarin revealed marked non-linearities. The half-life of total warfarin concentration in the plasma from 1-12h remained unchanged with all the doses used, but that of free warfarin was shorter with 40 mg/kg, possibly as the result of an increase in the binding of the drug to plasma proteins as the high total warfarin concentration decreased. The apparent volume of distribution generally increased with increasing dose, and differed according to the method used for its calculation. Liver warfarin data could be solved with Langmuir type saturation kinetics, but the saturation phenomena were slight in the concentration range studied. A non-linear multicompartment model was constructed, the physiological spaces of which were plasma, interstitial fluid and tissue. The binding of free warfarin to plasma proteins, interstitial fluid proteins and tissue structures was assumed to occur instantaneously, with saturable binding to plasma and interstitial fluid proteins, and a constant binding to tissues. The fluxes between the free warfarin pools of plasma and interstitial fluid as well as elimination were assumed to be linear. Following parameters were simulated simultaneously, using an analog hybrid computer: two for the above-mentioned fluxes, four for zero time drug mass distribution between plasma and interstitial fluid, and one for tissue binding. According to the best fits, warfarin is preferentially distributed into plasma, interstitial fluid and highly perfused tissues. The solution suggests that non-linearities in the pharmacokinetics of warfarin, a highly plasma protein-bound drug, first occur in plasma and interstitial fluid. Therefore, it is believed that the quantitative non-linear multicompartment approach presented in this paper might be useful in studying the kinetic behaviour of other highly plasma protein-bound drugs, too.


Fluorescent body distribution in spermatozoa in the male with exclusively female offspring.
Fluorescent (F) body distribution was determined in a group of men who did not have a fertility problem, but rather had fathered exclusively female offspring. The study was designed to analyze spermatozoa for the frequency of zero F-body (X-bearing) and one F-body (Y-bearing) spermatozoa. Semen samples were separated (processed) for Y-bearing spermatozoa enrichment and reanalyzed for fluorescent body distribution. The study consisted of 50 control (10 males) samples (unprocessed), 35 preseparation (35 patients) samples (unprocessed), and 18 postseparation (18 patients) samples (processed). A significantly higher frequency (P less than 0.05) of zero F-body spermatozoa were observed in the preseparation samples when compared with the control samples. The presence of more spermatozoa without fluorescent body correlates with the occurrence of more female births.


The relationship between cultural sensitivity and perceived stress among nurses working with foreign patients.
To determine the relationship between cultural sensitivity and perceived stress of nurses working at Kilis State Hospital. As foreigners live in and visit Turkey for various reasons, it is essential to provide culturally appropriate healthcare. Descriptive and cross-sectional design. This study was implemented at the State Hospital in Kilis on the southeast border of Turkey, between June-July 2014. The study sample consisted of 120 nurses. Data collection tools included a questionnaire about the socio-demographic and professional characteristics of participants, the Chen and Starosta's Intercultural Sensitivity Scale, and Perceived Stress Scale. The average score for nurse cultural sensitivity was 84·32 (SD, 11·40) and the average score for perceived stress was 27·97 (SD, 7·32), corresponding to a medium level. We identified negative correlation between cultural sensitivity and perceived stress. Nurses working at Kilis State Hospital have a medium level of cultural sensitivity and perceived stress, and cultural sensitivity tended to be affected by perceived stress. This study suggests that training programmes for cultural sensitivity and stress management should be available for nurses. Furthermore, patient care plans should be adapted to consider different cultural backgrounds of patients. These findings should be considered when designing nurses' education and continuing education programmes.


The relation between carotid plaque echogenicity and oxidative stress marker 8-iso-prostaglandin F2α.
Echolucent plaques are associated with high risk of ischemic cerebrovascular events. Oxidative stress has been implicated in the process of atherosclerotic plaque development from initiation to progression. We assessed the relation between carotid plaque echogenicity and urinary 8-iso-prostaglandin F2α, as an index of oxidative stress. This cross-sectional study was conducted prospectively on 290 consecutive outpatients. Each patient was evaluated for carotid plaque echogenicity using the gray-scale median at the maximal thickness plaque and urinary 8-iso-prostaglandin F2α using enzyme linked immunosorbent assay. By Pearson correlation analysis, we found significant negative linear relation between gray-scale median values and the urinary 8-iso-prostaglandin F2α levels (r = -0.133, p = 0.023). This correlation remained significant after adjustment for atherosclerotic risk factors, thickness of the maximal plaque and medication use (β = -0.137, p = 0.031). We herein show that higher levels of urinary 8-iso-prostaglandin F2α is associated with lower plaque echogenicity.


[A comparative analysis of the effect of nootropic preparations with different chemical structures on the failure of the escape reaction in rats].
The nootropic agents pyroglutamyl-D-alanine amide (TGS-20), mechlofenoxate, and mexidole versus piracetam were tested for their effects on the reversible impairments of cognitive brain functions, on the demolition of active avoidance responses in rats. TGS-20 (1 mg/kg/day) and mexidole (10 mg/kg/day) injected intraperitoneally 30 minutes prior to the experiment showed a normalizing effect in smaller doses than did piracetam (300 mg/kg/day). Mechlofenoxate (50 mg/kg/day) displayed a lower protective effect.


Solution conformation of lactate dehydrogenase as studied by saturation transfer ESR spectroscopy.
Several binary and ternary inhibitor and 'dead end' complexes of pig heart lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 22.214.171.124) were studied by saturation transfer ESR spectroscopy by means of an active NAD analog, spin-labeled at N6. The mobility of the spin-label depends on the nature of small molecules bound at the remote catalytic end of the coenzyme. The spin-label was found to serve as a reporter group monitoring the conformation of the peptide loop that is folded down over the active cleft in crystals of ternary complexes. The data suggest a fluctuation of the loop between open and closed forms in solution. The structure of the inhibitor molecules has been correlated with their ability to stabilize a more closed conformation of the loop.


Characteristics of gadolinium-based contrast media cancellation at the point of care: a 15-month assessment of FDA-inspired medication guides on gadolinium retention.
To assess the effect of United States Food and Drug Administration (FDA)-inspired patient handouts, regarding gadolinium retention, on the frequency of patient-initiated cancellations of gadolinium-based contrast media (GBCM) at the point of care. This is a single center retrospective cohort review of MRI examinations protocolled to receive GBCM but performed without GBCM from October 2017 to January 2019. The study period spans an FDA-inspired safety intervention (December 2017 to May 2018) to provide patients with a handout on gadolinium retention. Examinations were coded according to rationale for GBCM cancellation. Process control charts were created to statistically evaluate trends in GBCM cancellations over time. From October 2017 to January 2019, GBCM was canceled at the point of care for 0.22% (131/58,837; 95% CI 0.19-0.26) of examinations in which GBCM was indicated. This did not significantly vary during the study period. No cancellations (0/131) were due to stated concern for gadolinium retention. However, 38% (50/131; 95% CI 30-47) were canceled due to kidney impairment and 21% (28/131; 95% CI 13-30) were canceled for an unknown reason. FDA-inspired handouts informing patients about gadolinium retention did not significantly change the frequency of GBCM cancellation. GBCM cancellations at the point of care remain uncommon.


Precipitation regulates the responses of xylem phenology of two dominant tree species to temperature in arid and semi-arid forest of the southern Altai Mountains.
Arid and semi-arid forests are important carbon sinks, with implications for the global carbon balance. However, the impacts of climate warming on the growth of arid and semi-arid forest tree species and ecosystem carbon sink dynamics remain uncertain because the effects of the complex interactions between precipitation and temperature on xylem phenology are not clearly understood. Here, we monitored xylem formation over two years in two dominant tree species (Siberian larch, Larix sibirica Ledeb.; Siberian spruce, Picea obovata Ledeb.) along the arid and semi-arid southern Altai Mountains of Central Asia. We determined that temperature interaction with precipitation plays a key role in regulating xylem phenology of these two species, with differences between species. Under rising mean annual temperatures, the growth of L. sibirica advanced as the onset of xylem formation was not limited by early season water availability. However, the earlier cessation of cell enlargement, likely due to legacy effects, compensated for such advancement. In contrast, water stress constrained the advancement of xylem formation under rising temperatures in P. obovata. Nevertheless, water stress was seemingly relieved later in the growing season and consequently did not lead to the earlier cessation of xylem formation. Our results demonstrate that precipitation drives species-specific response to rising temperatures and thus is a key driver of growing season length and carbon sink dynamics in arid and semi-arid forests under climate warming. Integrating the effects of temperature and precipitation on xylem phenology in climate models may improve estimates of climate-carbon feedback in arid and semi-arid forests under future warming scenarios.


[Seasonal dynamics of the development and reproduction of parthenogenetic generations of Xiphidiocercaria sp. VII Odening 1962 (Trematoda: Plagiorchiidae) in the mollusk, Coretus corneus].
Intensity of reproduction of parthenogenetic generations of Xiphidiocercaria sp. VII Odening 1962 was shown to exhibit a seasonal dependence. Comparative studies of generative and somatic cell nuclei size, amount of divisional cells, quantity of cercariae and embryos were conducted in summer and winter.


Indole alkaloids from Ophiorrhiza cantoniensis with immunosuppressive activity.
Three new indole alkaloids, ophiorrhines C-D (1-3), together with one known analogue, have been isolated from the plant of Ophiorrhiza cantoniensis Hace. The structures of the new alkaloids with the absolute configurations were elucidated by means of spectroscopic methods, electronic circular dichroism (ECD) and calculated nuclear magnetic resonance (NMR) with DP4+ analysis. Compounds 1 and 2 exhibited certain activity to Con-A induced T cell proliferation, and 1 exhibited good inhibition on LPS-induced B cell proliferation with an IC50 value of 8.7 μM.


Strategies for Improving the Efficacy of CAR T Cells in Solid Cancers.
Therapy with T cells equipped with chimeric antigen receptors (CARs) shows strong efficacy against leukaemia and lymphoma, but not yet against solid cancers. This has been attributed to insufficient T cell persistence, tumour heterogeneity and an immunosuppressive tumour microenvironment. The present article provides an overview of key strategies that are currently investigated to overcome these hurdles. Basic aspects of CAR design are revisited, relevant for tuning the stimulatory signal to the requirements of solid tumours. Novel approaches for enhancing T cell persistence are highlighted, based on epigenetic or post-translational modifications. Further, the article describes CAR T strategies that are being developed for overcoming tumour heterogeneity and the escape of cancer stem cells, as well as for countering prevalent mechanisms of immune suppression in solid cancers. In general, personalised medicine is faced with a lack of drugs matching the patient's profile. The advances and flexibility of modern gene engineering may allow for the filling of some of these gaps with tailored CAR T approaches addressing mechanisms identified as important in the individual patient. At this point, however, CAR T cell therapy remains unproved in solid cancers. The further progress of the field will depend on bringing novel strategies into clinical evaluation, while maintaining safety.


Mesenteric vein thrombosis: is it one disease?
to compare the clinical course and outcome of mesenteric vein thrombosis due to known causes and mesenteric vein thrombosis of unknown aetiology. a retrospective study. two major general hospitals in Kuwait. 12 patients who had mesenteric vein thrombosis due to known causes and seven patients who had mesenteric vein thrombosis of unknown aetiology were treated over eight years. severity of clinical findings and outcome. There was no statistical difference between the two groups in the clinical presentation or in outcome. 36% of mesenteric vein thrombosis occurred without an obvious cause although the disease appeared to be the same. More work on the aetiology of this condition is required.


Delivering Transgenic DNA Exceeding the Carrying Capacity of AAV Vectors.
Gene delivery using recombinant adeno-associated virus (rAAV) has emerged to the forefront demonstrating safe and effective phenotypic correction of diverse diseases including hemophilia B and Leber's congenital amaurosis. In addition to rAAV's high efficiency of transduction and the capacity for long-term transgene expression, the safety profile of rAAV remains unsoiled in humans with no deleterious vector-related consequences observed thus far. Despite these favorable attributes, rAAV vectors have a major disadvantage preventing widespread therapeutic applications; as the AAV capsid is the smallest described to date, it cannot package "large" genomes. Currently, the packaging capacity of rAAV has yet to be definitively defined but is approximately 5 kb, which has served as a limitation for large gene transfer. There are two main approaches that have been developed to overcome this limitation, split AAV vectors, and fragment AAV (fAAV) genome reassembly (Hirsch et al., Mol Ther 18(1):6-8, 2010). Split rAAV vector applications were developed based upon the finding that rAAV genomes naturally concatemerize in the cell post-transduction and are substrates for enhanced homologous recombination (HR) (Hirsch et al., Mol Ther 18(1):6-8, 2010; Duan et al., J Virol 73(1):161-169, 1999; Duan et al., J Virol 72(11):8568-8577, 1998; Duan et al., Mol Ther 4(4):383-391, 2001; Halbert et al., Nat Biotechnol 20(7):697-701, 2002). This method involves "splitting" the large transgene into two separate vectors and upon co-transduction, intracellular large gene reconstruction via vector genome concatemerization occurs via HR or nonhomologous end joining (NHEJ). Within the split rAAV approaches there currently exist three strategies: overlapping, trans-splicing, and hybrid trans-splicing (Duan et al., Mol Ther 4(4):383-391, 2001; Halbert et al., Nat Biotechnol 20(7):697-701, 2002; Ghosh et al., Mol Ther 16(1):124-130, 2008; Ghosh et al., Mol Ther 15(4):750-755, 2007). The other major strategy for AAV-mediated large gene delivery is the use of fragment AAV (fAAV) (Dong et al., Mol Ther 18(1):87-92, 2010; Hirsch et al., Mol Ther 21(12):2205-2216, 2013; Lai et al., Mol Ther 18(1):75-79, 2010; Wu et al., Mol Ther 18(1):80-86, 2010). This strategy developed following the observation that the attempted encapsidation of transgenic cassettes exceeding the packaging capacity of the AAV capsid results in the packaging of heterogeneous single-strand genome fragments (<5 kb) of both polarities (Dong et al., Mol Ther 18(1):87-92, 2010; Hirsch et al., Mol Ther 21(12):2205-2216, 2013; Lai et al., Mol Ther 18(1):75-79, 2010; Wu et al., Mol Ther 18(1):80-86, 2010). After transduction by multiple fAAV particles, the genome fragments can undergo opposite strand annealing, followed by host-mediated DNA synthesis to reconstruct the intended oversized genome within the cell. Although, there appears to be growing debate as to the most efficient method of rAAV-mediated large gene delivery, it remains possible that additional factors including the target tissue and the transgenomic sequence factor into the selection of a particular approach for a specific application (Duan et al., Mol Ther 4(4):383-391, 2001; Ghosh et al., Mol Ther 16(1):124-130, 2008; Hirsch et al., Mol Ther 21(12):2205-2216, 2013; Trapani et al., EMBO Mol Med 6(2):194-211, 2014; Ghosh et al., Hum Gene Ther 22(1):77-83, 2011). Herein we discuss the design, production, and verification of the leading rAAV large gene delivery strategies.


[Placental mechanisms underlying the effects of maternal stress on the fetal development].
Placenta is the only link between the pregnant woman and fetus, and the basis for maintaining the normal pregnancy process and fetal development. Maternal stress is the maternal physiological and psychological changes caused by various factors, characterized by the increased level of glucocorticoid, which affects the hypothalamic-pituitary-target gland axis and regulates the expression of target genes. Maternal stress also changes the weight, metabolism and nutrient transportation of the placenta, which will substantially influence the development of fetus. This paper will firstly summarize the characteristics of maternal stress and its influence on offspring. Then, the changes in the body under maternal stress will be described. Finally, we will clarify the proven mechanisms underlying maternal stress and raise some important problems that have not been clarified in this area. The study of maternal stress on fetus and its underlying mechanisms will serve as theoretical basis for the diagnosis and treatment of the stress-related pregnant diseases and disorders.


Homeobox gene expression in cancer: insights from developmental regulation and deregulation.
Homeobox genes encode transcription factors that play essential roles in controlling cell growth and differentiation during embryonic development. Many homeobox genes are aberrantly expressed in a wide variety of solid tumours, and their deregulation appears to enhance cell survival and proliferation and to inhibit differentiation. In hematologic malignancies, deregulated homeobox genes profoundly perturb self-renewal and proliferation of hematopoietic stem cells and progenitors. It is increasingly recognised that solid tumours, like hematologic malignancies, could arise from cancer stem cells, and that targeting these cells could be the most effective means of inhibiting tumour progression and disease recurrence. Studying the biological effects and mechanisms of homeobox genes in cancers could provide valuable insights into identifying cancer stem cells and targeting the self-renewal pathways in these cell populations.


Characterization of a murine model of endotoxin-induced acute lung injury.
Endotoxin-induced microvascular lung injury in mice is a commonly used experimental model of the acute respiratory distress syndrome (ARDS). The present paper aimed to characterize this popular model in a comprehensive and systematic fashion. Male C57bl/6 mice (n = 5) were administered an LD55 dose of E. coli endotoxin (15 mg/kg, i.p.), and lungs were harvested at several time points and evaluated for injury as well as for expression of a variety of inflammatory mediators. Endotoxin induced many features characteristic of acute microvascular lung injury. These included early (1-2 h) expression of inflammatory mediators (IL-1alpha, IL-1beta, IL-4, IL-6, IL-10, TNF-alpha, interferon-alpha, interferon gamma, and MCP-1) and leukocyte accumulation in lung tissue (lung myeloperoxidase activity 18.5 +/- 7.8 U/g tissue, P < 0.05), followed by pulmonary edema (lung water content index 17.4% +/- 2.5%, P < 0.05) and mortality. Histopathological evaluation of lung tissue was compatible with these findings. The characterization of this murine model of endotoxin-induced microvascular injury will facilitate its utilization in ARDS research.


Percutaneous endourologic approach for transitional cell carcinoma of the renal pelvis.
A small mass was detected in the renal pelvis during investigation for hypertension and diminished renal function. Its presence was confirmed by computed tomography. Since the patient was not a candidate for nephrectomy, interventional percutaneous techniques were used to confirm the diagnosis of transitional cell carcinoma and ultimately permit endourologic access for resection and cure.


Uptake and accumulation of imidazolium ionic liquids in rice seedlings: Impacts of alkyl chain length.
The uptake and accumulation of three imidazolium ionic liquids with different alkyl chain lengths ([C2min]Br, [C4min]Br, [C8min]Br) in rice seedlings were investigated. All three different ILs were primarily accumulated in roots, while only a little amount of ILs were translocated and accumulated in stems and leaves. Accumulation and transportation of ILs in rice depend on the concentration and the alkyl chain length of ILs. ILs contents in the roots, stems and leaves decreased as ILs alkyl chain length increased. Growth inhibition results showed that the toxic effects of ILs on rice growth depends on the alkyl chain length: [C8min]Br >[C4min]Br >[C2min]Br. As markers of defense and phytotoxicity, the plant antioxidant enzymes and biochemical stress responses were also assessed. All different ILs significantly increased malondialdehyde (MDA), catalase (CAT), peroxidase (POD) and dismutase (SOD) activities in rice tissue. Compared to the control group, the contents of chlorophyll a reduced by 59.56%, 62.28% and 69.74% after addition of [C2min]Br, [C4min]Br, and [C8min]Br, respectively. This study provides important information for a better understanding on the uptake and accumulation of imidazolium ILs by agricultural plants.


Effects of ai-assisted colonoscopy on adenoma miss rate/adenoma detection rate: A protocol for systematic review and meta-analysis.
Colonoscopy can detect colorectal adenomas and reduce the incidence of colorectal cancer, but there are still many missing diagnoses. Artificial intelligence-assisted colonoscopy (AIAC) can effectively reduce the rate of missed diagnosis and improve the detection rate of adenoma, but its clinical application is still unclear. This systematic review and meta-analysis assessed the adenoma missed detection rate (AMR) and the adenoma detection rate (ADR) by artificial colonoscopy. Conduct a comprehensive literature search using the PubMed, Medline database, Embase, and the Cochrane Library. This meta-analysis followed the direction of the preferred reporting items for systematic reviews and meta-analyses, the preferred reporting item for systematic review and meta-analysis. The random effect model was used for meta-analysis. A total of 12 articles were eventually included in the study. Computer aided detection (CADe) significantly decreased AMR compared with the control group (137/1039, 13.2% vs 304/1054, 28.8%; OR,0.39; 95% CI, 0.26-0.59; P < .05). Similarly, there was statistically significant difference in ADR between the CADe group and control group, respectively (1835/5041, 36.4% vs 1309/4553, 28.7%; OR, 1.54; 95% CI, 1.39-1.71; P < .05). The advanced adenomas missed rate and detection rate in CADe group was not statistically significant when compared with the control group. AIAC can effectively reduce AMR and improve ADR, especially small adenomas. Therefore, this method is worthy of clinical application. However, due to the limitations of the number and quality of the included studies, more in-depth studies are needed in the field of AIAC in the future.


Ahaptoglobinaemia in Melanesia: DNA and malarial antibody studies.
To assess the relative contributions of genetic and acquired factors, particularly malaria, to the high frequencies of ahaptoglobinaemia found in Melanesia we have performed DNA and malarial antibody studies in a population from Vanuatu. No gene deletion or rearrangement was found on gene mapping in any ahaptoglobinaemic individual and the frequencies of the Hp1 and Hp2 alleles in the ahaptoglobinaemic group were similar to controls. However, antibodies to Plasmodium falciparum were significantly elevated in the ahaptoglobinaemics. These data suggest that malaria rather than genetic factors is the major cause of ahaptoglobinaemia in Melanesia.


Ultrastructural demonstration of oxytocin and vasopressin release sites in the neural lobe and median eminence of the rat by tannic acid and immunogold methods.
We have investigated the localization of oxytocin (OXT) and vasopressin (AVP) release sites in the neural lobe and median eminence (ME) of the rat. Ultrastructural and post-embedding immunocytochemical studies with protein A-gold as a marker were used for the identification of OXT and AVP neurosecretory granules (NSG). Release of the contents of these OXT and AVP NSG by exocytosis has been clearly demonstrated with the tannic acid-Ringer incubation method in axon terminals of the neural lobe and in varicose fibres with OXT and AVP NSG in the internal zone of the ME. As these fibers lack synaptic specializations, and are not located in the direct vicinity of capillaries of the primary portal plexus, these observations suggest that OXT and AVP are released in a paracrine way in the ME of the rat.


Methane mass balance at three landfill sites: what is the efficiency of capture by gas collection systems?
Many developed countries have targeted landfill methane recovery among greenhouse gas mitigation strategies, since methane is the second most important greenhouse gas after carbon dioxide. Major questions remain with respect to actual methane production rates in field settings and the relative mass of methane that is recovered, emitted, oxidized by methanotrophic bacteria, laterally migrated, or temporarily stored within the landfill volume. This paper presents the results of extensive field campaigns at three landfill sites to elucidate the total methane balance and provide field measurements to quantify these pathways. We assessed the overall methane mass balance in field cells with a variety of designs, cover materials, and gas management strategies. Sites included different cell configurations, including temporary clay cover, final clay cover, geosynthetic clay liners, and geomembrane composite covers, and cells with and without gas collection systems. Methane emission rates ranged from -2.2 to >10,000 mg CH(4) m(-2) d(-1). Total methane oxidation rates ranged from 4% to 50% of the methane flux through the cover at sites with positive emissions. Oxidation of atmospheric methane was occurring in vegetated soils above a geomembrane. The results of these studies were used as the basis for guidelines by the French environment agency (ADEME) for default values for percent recovery: 35% for an operating cell with an active landfill gas (LFG) recovery system, 65% for a temporary covered cell with an active LFG recovery system, 85% for a cell with clay final cover and active LFG recovery, and 90% for a cell with a geomembrane final cover and active LFG recovery.


Roles Of EAAT1, DHFR, And Fetuin-A In The Pathogenesis, Progression And Prognosis Of Chondrosarcoma.
Chondrosarcoma (CS) is a high-morbidity, relatively common bone malignancy without well-established biomarkers. The proteins EAAT1, DHFR, and fetuin-A have been investigated in many cancers, but their specific relationship to CS has not been reported. The present study examined EAAT1, DHFR, and fetuin-A expression in CS and the clinicopathological significance of these proteins in CS pathogenesis, progression, and prognosis. EAAT1, DHFR, and fetuin-A protein levels in 80 CS and 25 chondroma specimens were measured by immunohistochemistry and related to histological and clinical factors with chi-squared tests. Following univariate survival analysis, ROC curves calculation, and multivariate analysis. EAAT1, DHFR, and fetuin-A expression levels were significantly higher in the CS group than in the chondroma group (p < 0.05). Their immunopositivity rates were significantly greater in tissues with moderate or poor tumor differentiation, AJCC stage III or IV, Enneking stage II or III, and metastasis (p<0.05 or p<0.01 or p<0.001). Kaplan-Meier survival analysis showed significantly shorter survival in patients with moderately or poorly differentiated tumors, AJCC stage III or IV CS, Enneking stage II or III CS, metastasis, invasion, or EAAT1, DHFR, and fetuin-A immunopositivity (p < 0.05 or p < 0.001). Cox regression analysis showed that moderate or poor tumor differentiation, AJCC stage III or IV, Enneking stage II or III, metastasis, invasion, and EAAT1, DHFR, or fetuin-A immunopositivity correlated negatively with postoperative survival and positively with mortality (p < 0.05). The AUCs for EAAT1, DHFR, and fetuin-A were 0.654 (95% CI: 0.532-0.776, p = 0.025), 0.638 (95% CI: 0.519-0.756, p = 0.039), and 0.670 (95% CI: 0.556-0.784, p = 0.011), respectively. EAAT1, DHFR, and fetuin-A may be important biomarkers of the pathogenesis and progression of CS and predictors of its prognosis.


Family association studies of markers on chromosome 2q and Type 1 diabetes in subjects from South India.
Several Type 1 diabetes susceptibility loci have been located to chromosome 2q12-21. However, results have not always been consistent and this may reflect study design and the population analysed. We have used a family-based design to look for an association between Type 1 diabetes and markers located to 2q12-21. Ninety-one South Indian families consisting of subjects with Type 1 diabetes and their parents were genotyped for eight polymorphic markers localised to 2q12-21, which includes the interleukin-1 gene cluster. Radiation hybrid mapping was used to localise the map position of D2S308 and D2S363 on 2q12-21. The extended transmission disequilibrium test was used for statistical analysis. No associations were found between Type 1 diabetes and markers located in and around the interleukin-1 gene cluster or the interleukin-1 Type 1 receptor. In contrast, a suggestive association was found between Type 1 diabetes and two closely-linked markers telomeric of the interleukin-1 gene cluster (D2S308 and D2S363, separated by 3.3 cR) (p=0.004 and p=0.002, respectively). This preliminary study suggests that a locus close to D2S308 and D2S363 is involved in the aetiology of Type 1 diabetes in the South Indian population.


Linking Essential Tremor to the Cerebellum-Neuroimaging Evidence.
Essential tremor (ET) is the most common pathological tremor disorder in the world, and post-mortem evidence has shown that the cerebellum is the most consistent area of pathology in ET. In the last few years, advanced neuroimaging has tried to confirm this evidence. The aim of the present review is to discuss to what extent the evidence provided by this field of study may be generalised. We performed a systematic literature search combining the terms ET with the following keywords: MRI, VBM, MRS, DTI, fMRI, PET and SPECT. We summarised and discussed each study and placed the results in the context of existing knowledge regarding the cerebellar involvement in ET. A total of 51 neuroimaging studies met our search criteria, roughly divided into 19 structural and 32 functional studies. Despite clinical and methodological differences, both functional and structural imaging studies showed similar findings but without defining a clear topography of neurodegeneration. Indeed, the vast majority of studies found functional and structural abnormalities in several parts of the anterior and posterior cerebellar lobules, but it remains to be established to what degree these neural changes contribute to clinical symptoms of ET. Currently, advanced neuroimaging has confirmed the involvement of the cerebellum in pathophysiological processes of ET, although a high variability in results persists. For this reason, the translation of this knowledge into daily clinical practice is again partially limited, although new advanced multivariate neuroimaging approaches (machine-learning) are proving interesting changes of perspective.


One-step synthesis of phosphorus/oxygen co-doped g-C3N4/anatase TiO2 Z-scheme photocatalyst for significantly enhanced visible-light photocatalysis degradation of enrofloxacin.
Anatase TiO2 nanoparticles coated with P and O co-doped g-C3N4 were prepared via a single-step procedure. The resulting POCN/anatase TiO2 demonstrated remarkable performance in the degradation of enrofloxacin (ENFX). The photocatalytic activity of this heterojunction was 28.9 and 3.71 times better than that of the CN and anatase TiO2, respectively. The microtopography of the POCN/anatase TiO2 was revealed in this study. Co-doping with P and O increased the visible light adsorption capacity of the g-C3N4, whereas the anatase TiO2 nanoparticles enhanced the adsorption properties of the ENFX and the separation of the photoinduced carriers of the POCN/anatase TiO2. The O2·- and h+ were the main reactive oxidative species in the photocatalytic degradation of ENFX. The results of the detection of H2O2 and ESR confirmed that POCN/anatase TiO2 was a type Z-scheme photocatalyst. Finally, the ENFX degradation pathways were estimated through the detection of by-products.


Health-Related Quality of Life in Community-Dwelling Older People with Cognitive Impairment: EQ-5D-3L Measurement Properties.
Assessing quality of life (QoL) in older people with cognitive impairment is a challenge. There is no consensus on the best tool, but a short, user-friendly scale is advised. This study aimed to assess the psychometric properties of the self-reported and generic EQ-5D (including the EQ index and EQ visual analog scale [VAS]) in community-dwelling older adults with cognitive impairment. Cross-sectional study analyzing the feasibility, acceptability, reliability, and validity of the EQ-5D based on 188 self-administered questionnaires in a sample of community-dwelling older adults with Mini-Mental State Examination (MMSE) scores of 10 to 24 points. The EQ index was 0.69 (±0.27) and the EQ VAS was 63.8 (±28.54). Adequate measurement properties were found in acceptability and feasibility. Cronbach's alpha was 0.69. Good validity was observed in the correlation of each dimension of the EQ-5D with geriatric assessment scales. Higher validity was observed for the EQ index compared to the EQ VAS. The EQ-5D scale could be a good tool for assessing health-related QoL in community-dwelling older adults with cognitive impairment, though it is necessary to assess the dimensions and the EQ index.


[Regulations for the commercialization of new disinfectants: efficacy studies required for products to be used in foodstuffs, and in industrial, domestic and institutional areas].
Regulations for the commercialization of new disinfectants in Europe are currently going through a transition phase. Up to the present time these were set down by each member State of the European Union while in May 2010 a single European set of regulations (Directive 98/8/CE) will become effective in all Member States, the main aspects of which are discussed in this paper. In particular, we discuss the main points regarding studies and other information to be submitted for obtaining approval of new active substances and for reviewing products already in the market.


Chromosomal breakage in Crohn's disease: anticlastogenic effect of D-penicillamine and L-cysteine.
The incidence of chromosome breakage was found to be elevated in 42 patients with Crohn's disease. This phenomenon was much more striking in cultures set up with TCM 199 than in cultures set up with RPMI 1629 rich in L-cysteine. The drug D-penicillamine, a close analog of L-cysteine, gave an apparent therapeutic response in several patients and reduced the chromosome breakage frequency in the lymphocytes of these patients in vitro and in vivo.


The significant value of predicting prognosis in patients with colorectal cancer using 18F-FDG PET metabolic parameters of primary tumors and hematological parameters.
The purpose was to evaluate the correlation of the pre-treatment hematological parameters with metabolic parameters of primary tumor in baseline 18F-FDG PET/CT in patients with colorectal cancer (CRC) and estimate the prognostic value of both. We retrospectively investigated 231 patients with CRC who underwent baseline 18F-FDG PET/CT. Routine blood sampling was tested in the same term. PET parameters in term of hematological parameters and pathological characteristics of primary tumor were compared. Kaplan-Meier survival analysis was performed in the patients without distant metastasis. The differences of disease-free survival between groups were compared by log-rank tests. Neutrophil-to-lymphocyte ratio (NLR) and lymphocyte-to-monocyte ratio (LMR) were significantly correlated with all the metabolic parameters including maximum standardized uptake value (SUVmax), metabolic tumor volume (MTV) and tumor lesion glycolysis (TLG). The patients with NLR > 3 had higher MTV (24.82 ± 18.16 vs 19.06 ± 13.30, P = 0.039) and TLG (219.04 ± 186.94 vs 166.45 ± 146.39, P = 0.047) than those whose NLR ≤ 3. NLR in those patients with distant metastasis was significantly higher than those without distant metastasis (P = 0.018) while LMR in those patients with distant metastasis was significantly lower than those without distant metastasis (P = 0.032). Survival analysis showed that those patients with low MTV (P = 0.015), low NLR (P = 0.008) and high LMR (P = 0.027) revealed significant survival benefit. There was a significant association between the pre-treatment hematological parameters and metabolic parameters of baseline 18F-FDG PET/CT in the patients with CRC. It might be helpful in those patients with high NLR and low LMR to undergo 18F-FDG PET/CT to detect distant metastasis and predict prognosis.


Hereditary Predisposition to Hematopoietic Neoplasms: When Bloodline Matters for Blood Cancers.
With the advent of precision genomics, hereditary predisposition to hematopoietic neoplasms- collectively known as hereditary predisposition syndromes (HPS)-are being increasingly recognized in clinical practice. Familial clustering was first observed in patients with leukemia, which led to the identification of several germline variants, such as RUNX1, CEBPA, GATA2, ANKRD26, DDX41, and ETV6, among others, now established as HPS, with tendency to develop myeloid neoplasms. However, evidence for hereditary predisposition is also apparent in lymphoid and plasma--cell neoplasms, with recent discoveries of germline variants in genes such as IKZF1, SH2B3, PAX5 (familial acute lymphoblastic leukemia), and KDM1A/LSD1 (familial multiple myeloma). Specific inherited bone marrow failure syndromes-such as GATA2 haploinsufficiency syndromes, short telomere syndromes, Shwachman-Diamond syndrome, Diamond-Blackfan anemia, severe congenital neutropenia, and familial thrombocytopenias-also have an increased predisposition to develop myeloid neoplasms, whereas inherited immune deficiency syndromes, such as ataxia-telangiectasia, Bloom syndrome, Wiskott Aldrich syndrome, and Bruton agammaglobulinemia, are associated with an increased risk for lymphoid neoplasms. Timely recognition of HPS is critical to ensure safe choice of donors and/or conditioning-regimen intensity for allogeneic hematopoietic stem-cell transplantation and to enable direction of appropriate genomics-driven personalized therapies. The purpose of this review is to provide a comprehensive overview of HPS and serve as a useful reference for clinicians to recognize relevant signs and symptoms among patients to enable timely screening and referrals to pursue germline assessment. In addition, we also discuss our institutional approach toward identification of HPS and offer a stepwise diagnostic and management algorithm.


UDP-glucuronosyltransferase 1A6 overexpression in breast cancer cells resistant to methotrexate.
Methotrexate is a chemotherapeutic agent used in breast cancer treatment, but the occurrence of resistance limits its therapeutic use. A microarrays analysis between sensitive and methotrexate resistant MCF7 and MDA-MB-468 breast cancer cells pointed out the UDP-glucuronosyltransferase 1A (UGT1A) family as a common deregulated node in both cell lines. This family of genes is involved in Phase II metabolism. UGT1A6 was the main isoform responsible for UGT1A family overexpression in these cells. Its overexpression was not due to gene amplification. Transfection of a vector encoding for UGT1A6 in sensitive cells counteracted the cytotoxicity caused by methotrexate. Methotrexate increased the transcriptional activity from a luciferase reporter driven by the UGT1A6 promoter and induced UGT1A6 mRNA and enzymatic activity. Promoter analysis suggested that UGT1A6 induction by methotrexate could be driven by the transcription factors ARNT (HIF-1) and AhR/ARNT. Cells incubated with anticancer drugs susceptible to glucuronidation, such as tamoxifen or irinotecan, together with methotrexate, showed a lesser degree of cytotoxicity, due to UGT1A6 induction. The pharmacological effect of this induction should be taken into account when combining methotrexate with other drugs that are glucuronidated.


Stowaways in the cargo: Contaminating nucleic acids in rAAV preparations for gene therapy.
Recombinant AAV (rAAV) is the most used delivery vector for clinical gene therapy. However, many issues must be addressed before safer and more widespread implementation can be achieved. At present, efficacies are highly variable across trials and patients, and immune responses after treatment are widely reported. Although rAAV is capable of directly delivering gene-encoded therapeutic sequences, increased scrutiny of viral preparations for translational use have revealed contaminating nucleic acid species packaged within rAAV preparations. The introduction of non-therapeutic nucleic acids into a recipient patient adds to the risk burden, immunogenic or otherwise, of rAAV therapies. DNA from incomplete expression cassettes, portions of plasmids or vectors used to facilitate viral replication, and production cell line genomes all have the potential to be packaged within rAAV. Here, we review what is currently known about the profile, abundance, and post-treatment consequences of nucleic acid impurities within rAAV and cover strategies that have been developed to improve rAAV purity. Furthering our understanding of these aberrantly packaged DNA species will help to ensure the continued safe implementation of rAAV therapies as the number of patients treated with this modality increases.


Estramustine phosphate sodium in heavily pretreated patients with metastatic breast cancer: a retrospective consecutive case series.
Estramustine phosphate sodium (EMP) is an oral agent poorly developed--although active--in patients with metastatic breast cancer (MBC). To resume interest in EMP in MBC, we analyzed a retrospective series of consecutive patients with estrogen receptor-positive disease. EMP was given orally at a dose of 140 mg daily. Treatment discontinuation rates due to progressive disease/toxicity and response rates were assessed. Twenty postmenopausal patients with mainly visceral disease were treated with EMP, in five cases in combination with other anticancer drugs. Median numbers of previous chemotherapies and hormonal treatments were six and four, respectively. From the entire cohort, one complete response and four partial responses were observed. The proportions of patients free of progression at 6 and 12 months were 39 and 8 %, respectively. Six patients discontinued EMP, three each for toxicity and adverse events. Good disease control was obtained in heavily pretreated MBC patients receiving EMP. Toxicity was manageable and reversible although treatment discontinuation has to be considered. A prospective study should be encouraged to identify the optimal use of the drug.


Integrated, multi-scale, spatial-temporal cell biology--A next step in the post genomic era.
New microscopic approaches, high-throughput imaging, and gene editing promise major new insights into cellular behaviors. When coupled with genomic and other 'omic information and "mined" for correlations and associations, a new breed of powerful and useful cellular models should emerge. These top down, coarse-grained, and statistical models, in turn, can be used to form hypotheses merging with fine-grained, bottom up mechanistic studies and models that are the back bone of cell biology. The goal of the Allen Institute for Cell Science is to develop the top down approach by developing a high throughput microscopy pipeline that is integrated with modeling, using gene edited hiPS cell lines in various physiological and pathological contexts. The output of these experiments and models will be an "animated" cell, capable of integrating and analyzing image data generated from experiments and models.


Preformed oligomeric epidermal growth factor receptors undergo an ectodomain structure change during signaling.
Fluorescence resonance energy transfer (FRET) was used to reveal aspects of the mechanism of signal transduction by epidermal growth factor receptors (EGFR). The superpositions of epidermal growth factor (EGF), transforming growth factor-alpha (TGFalpha) and an antibody fragment (29.1) to the carbohydrate extremity of the receptor's ectodomain as measured by FRET, show that 14% of EGFRs in A431 cells are oligomerized before growth factor binding. After binding growth factor and signaling, these oligomers dissociate before releasing growth factor. Time courses of the FRET-derived distances between constitutively oligomerized EGFRs during signal transduction show a transient structural change in the extracellular domain, which occurs simultaneously with the production of intracellular Ca2+ signals. The FRET measurements also show a slow increase in oligomerization of EGFR monomers after growth factor binding. The structural change found in the extracellular domain of oligomeric EGFRs is similar to that shown by others for EPO, Neu, Fas, and tumor necrosis factor receptors, and may therefore be a common property of the transduction of the receptor-mediated signals.


Conformational change in coronary artery structure assessed by optical coherence tomography in patients with vasospastic angina.
The aim of this study was to investigate the conformational change of arterial structure in the vasospastic lesion with optical coherence tomography. Coronary artery spasm plays an important role in the pathogenesis of ischemic heart diseases. The conformational change of each arterial layer during vasospasm has not been studied in detail. We assessed 19 coronary arteries (10 spasm and 9 nonspasm lesions) with optical coherence tomography during the provocation test for coronary spasm. An intimal bump was defined as 1 or more intimal projections into the lumen that disappeared after the administration of nitroglycerine (NTG). Intimal gathering was defined as a folding/gathering of the intima, resulting in multiple kinks in the luminal contour that resolved after the administration of NTG. The spasm lesion more frequently showed an intimal bump at baseline and intimal gathering during spasm compared with the nonspasm lesion (spasm 80% vs. nonspasm 0%, p < 0.01, spasm 100% vs. nonspasm 0%, p < 0.01, respectively). The spasm lesion demonstrated a thicker maximum media thickness (spasm 0.24 ± 0.04 mm vs. nonspasm 0.12 ± 0.03 mm, p < 0.01) at baseline, whereas no differences were observed after the administration of NTG (spasm 0.13 ± 0.03 mm vs. nonspasm 0.13 ± 0.02 mm, p = 0.65). Our results suggest that medial contraction occurs even in an asymptomatic state and facilitates the formation of an intimal bump in patients with vasospastic angina. Luminal narrowing during spasm is associated with intimal gathering without alteration of intimal area.


Pretreatment hemoglobin level as a predictor to evaluate the efficacy of immune checkpoint inhibitors in patients with advanced non-small cell lung cancer.
Targeting immune checkpoints represents an immense breakthrough in cancer therapeutics. The prognostic value of hemoglobin (Hb) has been investigated in many malignancies including non-small cell lung cancer (NSCLC). However, the prognostic impact of pretreatment Hb count for immune checkpoint inhibitors (ICIs) in advanced NSCLC patients remains unclear. A total of 310 late-stage NSCLC patients who received ICI therapies between January 2015 and March 2019 were prospectively enrolled. We used a propensity score-matched cohort analysis for this study. Patients' clinicopathological characteristics and pretreatment Hb concentration were assessed against the progression-free survival (PFS) and overall survival (OS) using the Kaplan-Meier method and Cox proportional hazards regression. A propensity score (PS)-matched cohort analysis was applied to adjust for potential bias and to create two comparable groups according to patients' clinicopathological characteristics. The patients with normal baseline Hb levels (⩾110 g/L) had significantly longer PFS [median: 10.0 versus 4.0 months, hazard ratio (HR): 0.63, 95% confidence interval (CI): 0.46-0.86; p = 0.001] and OS [median: 17.6 versus 10.5 months, HR (95% CI): 0.56 (0.40-0.79); p < 0.001] than those with decreased Hb count (<110 g/L) in a PS-matched cohort (n = 255). For patients with normal pretreatment Hb levels, ICI combination therapy was significantly associated with better PFS [median: 11.1 versus 8.0 months, HR (95% CI): 0.74 (0.50-1.06); p = 0.09] and OS [median: 26.0 versus 12.9 months, HR (95% CI): 0.56 (0.37-0.86); p = 0.008] than monotherapy, but there was no such trend for patients with decreased baseline Hb levels. Our findings showed that normal pretreatment Hb count served as a favorable prognostic marker in advanced NSCLC patients treated with ICIs, representing an economical biomarker with readily measuring performance among all reported ones.


[Preconceived ideas, paradoxes, and new concepts in retinal vein occlusion].
Retinal vein occlusion is the second cause of vascular retinopathy after diabetic retinopathy and often leads to poor visual outcome. In a simplified form, the authors attempt to rectify a number of preconceived ideas about retinal vein occlusion and introduce new concepts in order to assist in the understanding of their mechanisms. Various paradoxes are explained, such as the presence of arterial risk factors in this venous disorder, the atypical combination of retinal ischemia and the absence of vascular non-perfusion on fluorescein angiography, the fact that macular edema is larger in the ischemic form than in the well-perfused form, the aggravating role of contraceptive hormonal therapy, the protective role of substitutive hormonal therapy, etc. The current controversies are underlined: the seasonal onset of the disease, the role of vasodilator treatment, the place of etiologic screening, and the significance of prepapillary loops. Lastly, the authors summarize important basics about treatment modalities, even if there is not yet consensus on the treatment of retinal vein occlusion.


Extraordinarily conserved chromosomal synteny of Citrus species revealed by chromosome-specific painting.
Reliable identification of individual chromosomes in eukaryotic species is the foundation for comparative chromosome synteny and evolutionary studies. Unfortunately, chromosome identification has been a major challenge for plants with small chromosomes, such as the Citrus species. We developed oligonucleotide-based chromosome painting probes for all nine chromosomes in Citrus maxima (Pummelo). We were able to identify all C. maxima chromosomes in the same metaphase cells using multiple rounds of sequential fluorescence in situ hybridization with the painting probes. We conducted comparative chromosome painting analysis in six different Citrus and related species. We found that each painting probe hybridized to only a single chromosome in all other five species, suggesting that the six species have maintained a complete chromosomal synteny after more than 9 million years of divergence. No interchromosomal rearrangement was identified in any species. These results support the hypothesis that karyotypes of woody species are more stable than herbaceous plants because woody plants need a longer period to fix chromosome structural variants in natural populations.


Thymic hormones in radiation-induced immunodeficiency. I. Induction of mature interleukin 1 responsive cell in the thymus by thymosin fraction 5.
The restorative effect of thymosin fraction 5 (TF5) on the thymus of gamma-irradiated mice was examined. Four different mouse strains were used in this study since earlier work determined that the degree of response to TF5 is strain dependent. The responsiveness to comitogenic effect of interleukin 1 (IL-1) was used to measure the rate of recovery of immunocompetent cells in the thymus, since only more mature PNA-, Lyt-1+-2- medullary cells respond to this monokine. Contrary to several earlier reports that radioresistant cells repopulating the thymus within the first 10 days after irradiation are mature, corticosteroid resistant, immunocompetent cells, the thymic cells from irradiated mice in all strains used had greatly reduced responses to IL-1. Daily intraperitoneal injections of TF5 increased significantly the responses of thymic cells to IL-1 in 10- to 13-weeks-old C57Bl/KsJ, C57Bl/6, C3H/HeJ, and DBA/1 mice. Older mice, 5 months or more in age, of DBA/1 strain did not respond to treatment with TF5. However, C3H/HeJ mice of the same age were highly responsive. In conclusion, (1) cells repopulating the thymus within 12 days after irradiation contain lower than normal fraction of mature IL-1 responsive cells, (2) thymic hormones increase the rate of recovery of immunocompetent cells in the thymus, and (3) the effect of thymic hormones is strain and age dependent.


Mechanisms of Complete Dissociation of CO2 on Iron Clusters.
Dissociation of CO2 on iron clusters was studied by using semilocal density functional theory and basis sets of triple-zeta quality. Fe2 , Fe4 , and Fe16 clusters were selected as the representative host clusters. When searching for isomers of Fen CO2 , n=2, 4 and 16 corresponding to carbon dioxide attachment to the host clusters, its reduction to O and CO, and to the complete dissociation, it was found that the total spin magnetic moments of the lowest energy states of the isomers are often quenched with respect to those of initial reagents Fen +CO2 . Dissociation pathways of the Fe2 +CO2 , Fe4 +CO2 , and Fe16 +CO2 reactions contain several transition states separated by the local minima states; therefore, a natural question is where do the spin flips occur? Since lifetimes of magnetically excited states were shown to be of the order of 100 fs, the search for the CO2 dissociation pathways was performed under the assumption that magnetic deexcitation may occur at the intermediate local minima. Two dissociation pathways were obtained for each Fen +CO2 reaction using the gradient-based methods. It was found that the Fe2 +CO2 reaction is endothermic with respect to both reduction and complete dissociation of CO2 , whereas the Fe4 +CO2 and Fe16 +CO2 reactions are exothermic to both reduction and complete dissociation of carbon dioxide. The CO2 reduction was found to be more favorable than its complete dissociation in the Fe4 case.


Identification of the novel allele, HLA-B*15:388, in a Chinese bone marrow donor.
The novel allele, HLA-B*15:388, was identified in a Chinese bone marrow donor by sequence-based typing.