[Association of polymorphisms in ACE and CYP11B2 genes with antihypertensive effects of hydrochlorothiazide].
To determine whether the blood pressure (BP) response to hydrochlorothiazide (HCTZ) was associated with the angiotensin converting-enzyme (ACE) I/D and aldosterone synthase (CYP11B2)-344T/C polymorphisms. The BP response to HCTZ 12.5 mg once daily for 6 weeks was assessed in 829 subjects with mild or moderate essential hypertension, and compared across the ACE and CYP11B2 genotypes. Of the 829 enrolled subjects, 785 completed the study. The systolic BP response differed according to the ACE (DD 9.4 +/- 15.7 mm Hg, ID 4.8 +/- 16.3 mm Hg, and II 5.1 +/- 14.8 mm Hg, P < 0.01), but not the CYP11B2 genotype (P > 0.05). Subjects with the combination of ACE DD and CYP11B2 CC genotypes tended to have a more pronounced systolic BP reduction than the other genotypic combinations of these 2 genes. Multiple linear regression analyses showed that the ACE DD genotype and serum aldosterone concentration at baseline were associated with the systolic BP reduction after treatment. None of the genetic associations with changes in diastolic BP or mean arterial pressure reached statistical significance (P > 0.05). The present study suggested that the ACE DD genotype was associated with the systolic BP response to HCTZ, and that the subjects with the combination of ACE DD and CYP11B2 CC genotypes might have a better BP response to HCTZ than the other genotypic combinations of these 2 genes.


Lhermitte's sign: incidence and treatment variables influencing risk after irradiation of the cervical spinal cord.
Lhermitte's sign is a relatively infrequent sequela of irradiation of the cervical spinal cord. In this study, we sought to determine whether various treatment parameters influenced the likelihood of developing Lhermitte's sign. Between October 1964 and December 1987, 2901 patients with malignancies of the upper respiratory tract were treated at the University of Florida. The dose of radiation to the cervical spinal cord was calculated for those patients who had a minimum 1-year follow-up. A total of 1112 patients who received a minimum of 3000 cGy to at least 2 cm of cervical spinal cord were included in this analysis. Forty patients (3.6%) developed Lhermitte's sign. The mean time to development of Lhermitte's sign after irradiation was 3 months, and the mean duration of symptoms was 6 months. No patient with Lhermitte's sign developed transverse myelitis. Several variables were examined in a univariate analysis, including total dose to the cervical spinal cord, length of cervical spinal cord irradiated, dose per fraction, continuous-course compared with split-course radiotherapy, and once-daily compared with twice-daily irradiation. Only two variables proved to be significant. Six (8%) of 75 patients who received > or = 5000 cGy to the cervical spinal cord developed Lhermitte's sign compared with 34 (3.3%) of 1037 patients who received < 5000 cGy (p = .04). For patients treated with once-daily fractionation, 28 (3.4%) of 821 patients who received < 200 cGy per fraction developed Lhermitte's sign compared with 6 (10%) of 58 patients who received > or = 200 cGy (p = .02). An increased risk of developing Lhermitte's sign was demonstrated for patients who received either > or = 200 cGy per fraction (one fraction per day) or > or = 5000 cGy total dose to the cervical spinal cord.


Improved early and late continence following robot-assisted radical prostatectomy with concurrent bladder neck fascial sling (RoboSling).
To describe a novel RoboSling technique performed at the time of robot-assisted radical prostatectomy (RARP) and its utility for enhancing urinary function recovery postoperatively. The surgical technique involves harvesting a vascularised, fascial flap from the peritoneum on the posterior aspect of the bladder. Following completion of prostatectomy, the autologous flap is tunnelled underneath the bladder and incorporated into the rectourethralis and vertical longitudinal detrusor fibres at the posterior bladder neck with a modified Rocco suture. After urethra-vesical anastomosis is completed, the corners of the flap are hitched up to Cooper's ligament bilaterally with V-Loc sutures, tensioned and secured creating a bladder neck sling. A prospective, longitudinal cohort study was performed of 193 consecutive patients undergoing RARP between December 2016 and September 2019. The first 163 patients underwent standard RARP, and the last 30 patients had the RoboSling technique performed concurrently. Continence outcomes were the primary outcomes assessed using pad number and Expanded Prostate Cancer Composite (EPIC)-urinary domain questionnaire. Operative time (OT), estimated blood loss (EBL), complications and oncological outcomes were secondary outcomes. The two groups were comparable for demographics and clinicopathological variables. At 3 months, zero pad usage (p = 0.005) and continence rates, defined as EPIC score ≥ 85 (p = 0.007), were both higher in the RoboSling group. EBL, complication rate and positive surgical margin rate did not differ between the two groups. Superior zero pad usage was observed at 1 year in the RoboSling group (p = 0.029). The RoboSling technique added on average 16 min to OT. The RoboSling procedure at the time of RARP was associated with earlier return to continence without negatively impacting other postoperative outcomes. This improvement in continence outcomes was maintained long term.


Assessment of the pulmonary volume pulse in idiopathic pulmonary arterial hypertension by means of electrical impedance tomography.
Electrical impedance tomography (EIT) is a non-invasive imaging technique which can be used to measure the blood volume changes in the pulmonary vascular bed during the cardiac cycle. This study was performed to evaluate the differences in the EIT signal of the pulmonary vascular bed between healthy subjects and patients with idiopathic pulmonary arterial hypertension (IPAH), who are known to have a remodelled pulmonary vascular bed. Twenty-one patients (17 females, 4 males) with IPAH and 30 healthy controls (5 females, 25 males) were measured. EIT measurements were performed in duplicate, on the same day as right heart catheterization to obtain haemodynamic data. The maximal impedance change during systole (Delta Z(sys)) was used as a measure of the pulmonary volume pulse and expressed in arbitrary units (AU). Total lung capacity, spirometric values and diffusion capacity for carbon monoxide were measured as well. Mean Delta Z(sys) was 215 +/- 58 x 10(-2) AU (95% CI 193 x 10(-2) to 236 x 10(-2)) in the healthy subjects and 78 +/- 27 x 10(-2) AU (95% CI 66 x 10(-2) to 91 x 10(-2)) in the IPAH patient group (p < 0.0001). No significant correlation was found between Delta Z(sys) and any of the haemodynamic or lung function data. The impedance pulsation of the pulmonary vascular bed is reduced in IPAH in comparison with controls, indicating a reduced volume pulse. This might represent the reduced cross section area, as well as the reduced compliance and number of the pulmonary vessels in these patients.


Dynamic proliferation assessment in flow cytometry.
Dynamic proliferation assessment via flow cytometry is legitimately supposed to be the most powerful tool for recording cell cycle kinetics in-vitro. The preeminent feature is a single cell-based multi-informative analysis by temporal high-resolution. Flow cytometric approaches are based on labeling of proliferating cells via thymidine substitution by a base analog (e.g., 5-bromo-2'-deoxyuridine, BrdU) that is added to cell cultures either for a short period of time (pulse labeling) or continuously until cell harvesting. This unit describes the alternative use of the thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) in place of BrdU for three different applications: (1) dynamic proliferation assessment by EdU pulse cell labeling; (2) the same approach as (1) but in combination with live/dead cell discrimination; and (3) dynamic cell cycle analysis based on continuous cell labeling with EdU and Hoechst fluorochrome quenching. In contrast to the detection of BrdU incorporation, EdU-positive cells can be identified by taking advantage of click chemistry, which facilitates a simplified and fast cell preparation. Further analysis options but also limitations of the utilization of EdU are discussed.


Raman microscopy of Greek icons: identification of unusual pigments.
Five Greek icons, made between the 15th and 18th centuries and now belonging to the Victoria and Albert Museum collections, were analysed by energy-dispersive X-ray fluorescence (EDXRF), optical microscopy and Raman microscopy in order to determine the stratigraphy of the artworks and the identity of the pigments used. Together with common pigments, such as red lake, vermilion, red lead, red iron oxide, orpiment, yellow ochre, lead white, chalk, gypsum, anhydrite, Prussian blue, indigo and a copper-containing green, a few unusual materials were identified, specifically pararealgar (a yellow arsenic sulphide, As4S4), its precursor the chi-phase, and lead tin yellow type II (PbSn(1-x)SixO3). Attention is drawn to the complementarity of the techniques used for the pigment identifications.


Efficacy and safety of a novel, soluble microneedle patch for the improvement of facial wrinkle.
Various kinds of functional cosmetics are on the market, although there are a variety of opinions concerning the actual effect. Transdermal microneedle patch has been introduced as a newly developed device for drug delivery through the skin. This study was conducted to verify the face skin improvement effect and safety of a novel cosmetic microneedle patch. A total of 84 Korean females finished this prospective clinical trial. The subjects were divided into 3 groups: (1) soluble hyaluronic acid (HA) microneedle patch alone, (2) soluble HA microneedle patch plus adenosine wrinkle cream, and (3) adenosine wrinkle cream alone. The treatments were applied to the crow's feet and nasolabial fold wrinkle for 12 weeks. The test areas were measured before treatment and at 4, 8, and 12 weeks after use of the test product. At the completion of the testing period of the trial, the global assessment of efficacy and product preferences were surveyed from the subjects. Combination treatment with wrinkle cream and microneedle patch significantly improved Merz scale for crow's feet and nasolabial folds, compared to the sole application of wrinkle cream or patch. Measurement on the crow's feet showed an overall improvement in all 3 groups, yielding no significant differences among the groups. No serious adverse effects were observed during the follow-up period. Combination application of a soluble microneedle patch and wrinkle cream was an effective treatment in improving facial wrinkles, thus enhancing skin rejuvenation.


Location and speciation of gadolinium and yttrium in roots of Zea mays by LA-ICP-MS and ToF-SIMS.
Increasing production of rare earth elements (REE) might lead to future contamination of the environment. REE have been shown to accumulate in high concentrations in roots of plants. Plant experiments with Zea mays exposed to a nutrient solution containing gadolinium (Gd) or yttrium (Y) with 10 mg L(-1) Gd or Y were carried out to investigate this accumulation behaviour. Total concentrations of 3.17 g kg(-1) and 8.43 g kg(-1) of Gd and Y were measured in treated plant roots. Using a novel combination of laser ablation mass spectrometry and time-of-flight secondary ion mass spectrometry, imaging of location and concentration of Gd and Y was carried out in root thin sections of treated roots. Single spots of elevated REE concentration were found at the epidermis, while inside the cortex, weak signals of Gd(+) and Y(+) were aligning with the root cell structures. The composition of Gd-containing secondary ions proves an REE-oxide phase accumulated at the epidermis, limiting REE availability for further uptake.


What constitutes a School Readiness examination? The component parts.
The School Readiness medical examination is probably the single most important examination a child has outside of the infancy period. This examination helps to determine whether or not the child is physically, emotionally and intellectually ready to handle a more structured, academic environment. The physician should devote enough time and effort to do a thorough evaluation. The physician also, should be appropriately compensated for this medical evaluation based upon time, training, experience, and length of discussion with the family.


Diagnostic utilities of a fully automated molecular test for toxigenic Clostridium difficile.
Laboratory underdiagnosis of toxigenic Clostridium difficile can lead to inappropriate management of C. difficile infection (CDI). A fully automated molecular test (FAMT), BD MAX, and enzyme immunoassays for C. difficile glutamate dehydrogenase (GDH) and for toxin A/B antigen test were evaluated using clinical specimens. Laboratory analysis of 231 fecal specimens from patients suspected with CDI, indicated that the sensitivity (Sn), specificity (Sp), positive predictive value (PPV), and negative predictive value (NPV) of FAMT was 98.1%, 98.9%, 96.3%, and 99.4%, while that of toxin A/B antigen was 52.8%, 100.0%, 100.0%, and 87.7%, respectively, compared to toxigenic culture. Sn, Sp, PPV, and NPV of GDH test compared to toxigenic culture was 92.5%, 94.4%, 83.1%, and 97.7%, respectively. FAMT can support the accurate laboratory diagnosis of toxigenic C. difficile and be an effective tool for appropriate treatment of CDI.


Guanine nucleotide-binding protein 2, GNBP2, accelerates the progression of clear cell renal cell carcinoma via regulation of STAT3 signaling transduction pathway.
Guanine nucleotide-binding protein 2 (GNBP2) is a GTPase that has critical roles in host immunity and some types of cancer, but its function in clear cell renal cell carcinoma (ccRCC) is not fully understood. This work explored the role of GNBP2 in ccRCC progression and the underlying molecular mechanism. Two public human cancer databases TNMplot and TISIDB were employed to analyze the expression pattern of GNBP2 during ccRCC progression and the correlation between GNBP2 expression and clinical features of ccRCC patients. GNBP2 functions in ccRCC cells were determined by EdU staining, flow cytometry, scratch wound assay, transwell assay, and xenograft model. Gene expression was evaluated using qPCR, Western blot, immunofluorescence staining, and immunohistochemical staining. GNBP2 expression was significantly elevated in ccRCC tissues and increased gradually with the increasing tumor grades. Patients with higher GNBP2 expression had shorter overall survival times. Knockdown of GNBP2 suppressed tumor cell proliferation and cell cycle progression and reduced the capability of migration and invasion, while GNBP2 overexpression exhibited protumor effects. GNBP2 silencing by RNA interference significantly inhibited the tumor growth of tumor-bearing nude mice and decreased the proliferation marker Ki67. Mechanistically, GNBP2 downregulation suppressed the STAT3 signaling transduction, as it reduced the phosphorylation of STAT3 and modulated the expression of the target genes, including c-Myc, MMP2, N-cadherin, and E-cadherin. These findings reveal that GNBP2 promotes ccRCC progression by regulating STAT3 signaling transduction, indicating that GNBP2 might be a promising molecular target for ccRCC therapy.


Automatic statistical shape analysis of cerebral asymmetry in 3D T1-weighted magnetic resonance images at vertex-level: application to neuroleptic-naïve schizophrenia.
The study of the structural asymmetries in the human brain can assist the early diagnosis and progression of various neuropsychiatric disorders, and give insights into the biological bases of several cognitive deficits. The high inter-subject variability in cortical morphology complicates the detection of abnormal asymmetries especially if only small samples are available. This work introduces a novel automatic method for the local (vertex-level) statistical shape analysis of gross cerebral hemispheric surface asymmetries which is robust to the individual cortical variations. After segmentation of the cerebral hemispheric volumes from three-dimensional (3D) T1-weighted magnetic resonance images (MRI) and their spatial normalization to a common space, the right hemispheric masks were reflected to match with the left ones. Cerebral hemispheric surfaces were extracted using a deformable model-based algorithm which extracted the salient morphological features while establishing the point correspondence between the surfaces. The interhemispheric asymmetry, quantified by customized measures of asymmetry, was evaluated in a few thousands of corresponding surface vertices and tested for statistical significance. The developed method was tested on scans obtained from a small sample of healthy volunteers and first-episode neuroleptic-naïve schizophrenics. A significant main effect of the disease on the local interhemispheric asymmetry was observed, both in females and males, at the frontal and temporal lobes, the latter being often linked to the cognitive, auditory, and memory deficits in schizophrenia. The findings of this study, although need further testing in larger samples, partially replicate previous studies supporting the hypothesis of schizophrenia as a neurodevelopmental disorder.


Derivatization of phosphopeptides with mercapto- and amino-functionalized conjugate groups by phosphate elimination and subsequent Michael addition.
Kinetics of the beta-elimination of the phosphate group from H-Tyr-Ser(PO3H2)-Phe-OH and H-Tyr-Thr(PO3H2)-Phe-OH and subsequent addition of thiols and amines to the dehydroalaninyl and beta-methyldehydroalaninyl residues formed, were followed by RP HPLC under alkaline conditions in the absence and presence of Ba2+ ions. By this reaction sequence, the phosphoserinyl peptide was conjugated with mono-N-(2-mercaptoethyl)amide of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (4), a mercapto-functionalized pentapeptide, H-His-Gly-Gly-His-Gly-NH(CH2)4SH, and an amino-functionalized fluorescent dye, 5-dimethylaminonaphthalene-1-[N-(5-aminopentyl)]sulfonamide (dansyl cadaverine). The beta-methyldehydroalanine residue was, in turn, observed to be a poor Michael acceptor.


Rational design of activated graphitic carbon spheres with optimized ion and electron transfer channels for zinc-ion hybrid capacitors.
Herein, three-dimensional activated graphitic carbon spheres (AGCS) were constructed by simultaneous activation-graphitization of Fe-tannic acid coordination spheres with the assistance of KOH. Nanosheets-assembled AGCS with complex intersecting channel system can expose more active sites for charge storage. Simultaneous activation-graphitization can relieve trade-off relationship between porosity and conductivity of carbon materials. Benefiting from multiple synergistic effects of large specific surface area (2069 m2 g-1), abundant ion-accessible micropores (>0.78 nm), good electronic conductivity (IG/ID = 1.11), and moderate amount of oxygen doping, the optimized AGCS-2 has favored ion and electron transfer channels. AGCS-2 based zinc-ion hybrid capacitor (ZIHC) displays a high specific capacity of 148.6 mA h g-1 (334 F g-1) at 0.5 A g-1, a remarkable energy density of 119.0 W h kg-1 at 1440 W kg-1, and superior cycling life with 96% capacity retention after 10,000 cycles. This simultaneous activation-graphitization strategy may open up a new avenue to design novel carbon spheres linking optimal pores and graphitic carbon structure for ZIHC application.


One-year follow-up of patients of the ongoing Dutch Q fever outbreak: clinical, serological and echocardiographic findings.
In 2007, a large goat-farming-associated Q fever outbreak occurred in the Netherlands. Data on the clinical outcome of Dutch Q fever patients are lacking. The current advocated follow-up strategy includes serological follow-up to detect evolution to chronic disease and cardiac screening at baseline to identify and prophylactically treat Q fever patients in case of valvulopathy. However, serological follow-up using commercially available tests is complicated by the lack of validated cut-off values. Furthermore, cardiac screening in the setting of a large outbreak has not been implemented previously. Therefore, we report here the clinical outcome, serological follow-up and cardiac screening data of the Q fever patients of the current ongoing outbreak. The implementation of a protocol including clinical and serological follow-up at baseline and 3, 6 and 12 months after acute Q fever and screening echocardiography at baseline. Eighty-five patients with acute Q fever were identified (male 62%, female 38%). An aspecific, flu-like illness was the most common clinical presentation. Persistent symptoms after acute Q fever were reported by 59% of patients at 6 months and 30% at 12 months follow-up. We observed a typical serological response to Coxiella burnetii infection in both anti-phase I and anti-phase II IgG antibodies, with an increase in antibody titres up to 3 months and a subsequent decrease in the following 9 months. Screening echocardiography was available for 66 (78%) out of 85 Q fever patients. Cardiac valvulopathy was present in 39 (59%) patients. None of the 85 patients developed chronic Q fever. Clinical, serological and echocardiographic data of the current ongoing Dutch Q fever outbreak cohort are presented. Screening echocardiography is no longer part of the standard work-up of Q fever patients in the Netherlands.


P53 mutations in hepatocellular carcinoma patients in Egypt.
The p53 gene plays a major role in hepatocellular carcinoma (HCC). Acquired mutations may provide clues to etiology, as some carcinogenic agents are associated with specific genetic changes in p53. Our aim was to analyze the spectrum of p53 mutations in tumor tissues from subjects with HCC in Egypt, where there is a rising incidence of HCC due to hepatitis C virus (HCV). We collected tumor tissues from 41 subjects with HCC diagnosed at the National Cancer Institute of Cairo University during 2000-2003. Sequence mutations were analyzed by the Affymetrix GeneChip technique. HCV RNA was detected in the sera of 37 subjects (90%). Only one patient had a current HBV infection. A total of 17 of the 41 subjects (41%) had p53 mutations. Thirteen of these were in exon 7, of which 10 were in codon 249, but only 8 of the 10 were the R249S mutation, previously reported to be associated with aflatoxin exposure. The other three exon 7 mutations were found in codons 232, 242 and 248. A total of three mutations were detected in exon 5 codons 133, 144 and 176. One mutation was detected in exon 8 codon 275. Unlike previous studies, this population is characterized by a high prevalence of chronic HCV infection. The presence of the R249S mutation in exon 7 may indicate that these subjects with HCC have been exposed to aflatoxin (AFB1), and further investigation is in progress to measure AFB1-albumin adducts in the sera of these subjects.


Establishment and characterization of C-type RNA virus-producing cell lines from radiation-induced murine osteosarcomas.
Eight cell lines were established from murine osteosarcomas induced in vivo with the radionuclides 224Ra and 227Th. They have been compared by light and electron microscopy, by karyology, and by their growth properties. The morphology, the growth pattern, and the ability to induce tumors in mice indicate that five of them are tumor cell lines. Chromosome studies demonstrated that the five cell lines have marker chromosomes. The other cell lines only showed some criteria generally used to score for transformation of fibroblasts and they may be derived from stromal cells. All cell lines release virus particles in the culture fluid which have the typical properties of RNA tumor viruses. They possess C-type morphology, a density of 1.16--1.18 g/cm3, a 60--70 S RNA, a RNA dependent DNA polymerase and they induce syncytia in rat XC cells. The possible significance of these virus particles in radiation osteosarcomagenesis is discussed.


Contribution of diminished kidney transplant GFR to increased circulating chemokine ligand 27 level.
Inflammatory chemokine ligands (CCLs) play an important role in cardiovascular disease and allograft injury. CCLs may independently associate with diminished estimated glomerular filtration rate (eGFR) in stable renal transplant recipients (RTR). Plasma levels of 19 CCLs (1, 2, 3, 4, 5, 8, 11, 13, 15, 17, 21, 24, 26, 27, CXCL5, 8, 10, 12 and 13) were measured in a cohort of 101 RTR. The cohort was divided according to CKD-EPI equation into three groups; group 1: eGFR ≥ 60 ml/min, group 2: eGFR 30-59.9 ml/min and group 3 eGFR ≤ 29.9 ml/min. ANOVA, Krusklwallis, Mann- Whitney Spearman correlation and regression analysis tests were used to determine association between reduced eGFR and inflammatory CCLs plasma levels measured by multiplex techniques. 20 healthy subjects with eGFR above 90 ml/min were included as control. Significance was sat at < 0.05. Levels of CCLs 1, 4, 15, 27, CXCL8 and CXCL10 were significantly different among the four studied groups. Multivariate regression analysis (MVA) between eGFR and all CCLs demonstrated that CCL27 was the only ligand to remain significantly associated with diminished eGFR {P = 0.021 and r = - 0.35,(P = 0.001)}. In a second MVA between CCL 27 and patient's demographics and laboratory variables, diminished eGFR, and elevated PTH, out of the twenty one available variables remained significantly associated with elevated CCL27levels. Diminished eGFR in stable RTR is associated with elevated plasma levels of CCL27. This association may explain, at least in part, the independent contribution of reduced eGFR to enhanced inflammation in RTR.


Programmed cell death in fission yeast Schizosaccharomyces pombe.
Yeasts have proven to be invaluable, genetically tractable systems to study various fundamental biological processes including programmed cell death. Recent advances in the elucidation of the molecular pathways underlying apoptotic cell death in yeasts have revealed remarkable similarities to mammalian apoptosis at cellular, organelle and macromolecular levels, thus making a strong case for the relevance of yeast models of regulated cell death. Programmed cell death has been reported in fission yeast Schizosaccharomyces pombe, primarily in the contexts of perturbed intracellular lipid metabolism, defective DNA replication, improper mitotic entry, chronological and replicative aging. Here we review the current understanding of the programmed cell death in fission yeast, paying particular attention to lipid-induced cell death. We discuss our recent findings that fission yeast exhibits plasticity of apoptotic and non-apoptotic modes of cell death in response to different lipid stimuli and growth conditions, and that mitochondria, reactive oxygen species and novel cell death mediators including metacaspase Pca1, SpRad9 and Pck1 are involved in the lipotoxic cell death. We also present perspectives on how various aspects of the cell and molecular biology of this organism can be explored to shed light on the governing principles underlying lipid-mediated signaling and cell demise.


Incremental HMM training applied to ECG signal analysis.
This work discusses the implementation of incremental hidden Markov model (HMM) training methods for electrocardiogram (ECG) analysis. The HMMs are used to model the ECG signal as a sequence of connected elementary waveforms. Moreover, an adaptation process is implemented to adapt the HMMs to the ECG signal of a particular individual. The adaptation training strategy is based on incremental versions of the expectation-maximization, segmental k-means and Bayesian approaches. Performance of the training methods was assessed through experiments considering the QT and ST-T databases. The results obtained show that the incremental training improves beat segmentation and ischemia detection performance with the advantage of low computational effort.


Down-regulation of CD53 expression in Epinephelus coioides under LPS, poly (I:C), and cytokine stimulation.
Tetraspanins are a group of cell surface molecules involved in cell adhesion, motility, metastasis, signal transduction, and immune cell activation. Members of the tetraspanin family include CD9, CD37, CD63, CD53, and others. However, few tetraspanins have been investigated in teleosts. In this study, we obtained the open reading frame of CD53 cDNA from orange spotted grouper (Epinephelus coioices), an economically important fish. The predicted amino acid structure contains four membrane-spanning domains and a conserved CCG motif. The amino acid identity between human and grouper CD53 was only 38%; however, both CD53 proteins share the same structure. Quantitative real-time PCR revealed that mRNA is abundant in immune organs, including the head and trunk kidneys, spleen, thymus, gill, and blood. Immunochemistry and immunofluorescence analyses further revealed that CD53 was majorly expressed in the leukocytes of various organs. Finally, mRNA and protein expression for CD53 was down-regulated in fish treated with immune stimulators, including LPS, Poly (I:C), Vibrio, recombinant grouper IL-6, and CCL4. Our results indicate that the expression of CD53 may play important roles in pathogen invasion and inflammation reaction.


Serous cystadenoma causing the double duct sign.
An asymptomatic 70-year-old man was found to have elevated liver function tests on a routine screening evaluation. Abdominal ultrasound revealed a pancreatic head mass. Magnetic resonance cholangiopancreatography confirmed a heterogeneously enhancing pancreatic mass that was suspicious for malignancy due to obstruction of the common bile duct and pancreatic duct. Consequently a pylorus-sparing pancreaticoduodenectomy was performed. Histology revealed a serous cystadenoma with scattered foci of PanIN III. Serous cystadenomas are benign tumors without significant malignant potential. Unlike pancreatic adenocarcinomas, these tumors tend to be slow growing, well-demarcated, and rarely, as in this case, produce a mass effect.


Pressure modulation of cytochrome-to-cytochrome electron-transfer. Models and enzyme reactions.
The kinetics of electron-transfer involved in reactions of reduction of 2,6-dichlorophenol indophenol and Fe(CN)3-(6) by L-ascorbic acid and reduction of ferric cytochrome c by both L-ascorbic acid and reduced hydroxylamine oxidoreductase were studied as a function of three parameters: ionic strength, pressure (1-2000 bar) and temperature (4-20 degrees C) using the high-pressure stopped-flow method. From measurements, the thermodynamic parameters of activation volume (delta V++), and, when possible, activation enthalpy and entropy (delta H++ and delta S++) have been calculated. We found, for these four systems, that the pressure has revealed solvation effects involved in electron-transfer. For the reduction of ferric cytochrome c by reduced hydroxylamine oxidoreductase (a cytochrome-to-cytochrome electron-transfer), we have not obtained evidence for a conformational change.


Circulatory effects of prostaglandin endoperoxide analogues studied in the dog during left ventricular bypass.
1. Intravenous administration of both the 9alpha,11alpha-(epoxymethano) and 11alpha,9alpha-(epoxymethano) analogues of prostaglandin H2 (0.25 microgram/kg) produced a prominent rise in pulmonary arterial pressure and a moderate increase in systemic arterial pressure. 2. Direct administration of the endoperoxide analogues (1.25 microgram/kg) into the bypass reservoir produced a greater rise in systemic arterial pressure and less prominent rise in pulmonary arterial pressure. 3. An intravenous dose of prostaglandin F2alpha that was 20 times larger was needed to produce a comparable rise in pulmonary arterial pressure. 4. The pulmonary and systemic pressor responses produced by the endoperoxide analogues were due to a direct increase in the vascular resistance.


Caregiver goals and satisfaction for menstrual suppression in adolescent females with developmental disabilities: A prospective cohort study.
Adolescents with developmental disabilities and their caregivers often seek menstrual management. Caregivers frequently serve as medical decision-makers, and little is known about caregiver goals for menstrual management and satisfaction over time. Assess caregiver reasons for initiating menstrual management in adolescents with disabilities and satisfaction over 12 months. Prospective cohort study of caregivers of adolescents with developmental disabilities seeking menstrual management at a pediatric and adolescent gynecology clinic. Data derive from caregiver surveys and adolescents' electronic medical records. Ninety-two caregiver-adolescent pairs enrolled. The mean age of adolescents was 14.4 (±2.6). The most common method started was levonorgestrel intrauterine device (LNG-IUD; 52, 56.5%), followed by oral norethindrone acetate (21, 22.8%). Caregivers cited hygiene concerns (84.8%), behavioral problems (52.2%), and heavy/excessive bleeding (48.9%) as reasons for initiating menstrual suppression. Caregivers who identified hygiene or heavy/excessive bleeding as the most important reason for management were more likely to select LNG-IUD (p = 0.009). Caregivers who cited behavioral/mood or seizure concerns as the most important reason were more likely to choose other methods (p < 0.05). At 12 months, caregiver satisfaction with all methods was high (66.2-86.9 on a 100-point scale). For every additional day of bleeding, satisfaction decreased by 3.7 points (95% CI: 2.3-5.0). Caregiver satisfaction with all methods is high; however, it negatively correlates with days of bleeding. Caregiver reasons for menstrual suppression influence the method chosen. Management may reflect both patient and caregiver priorities; research is needed to better understand shared decision-making models that promote reproductive autonomy in adolescents with a developmental disability.


Acute kidney injury following peripheral angiography and endovascular therapy: A systematic review of the literature.
Radiographic contrast administration is a major cause of acute kidney injury (AKI), worldwide. Currently, contrast induced acute kidney injury (CI-AKI) is the third leading cause of hospital acquired renal failure in the United States. Over 50% of these cases are the result of contrast exposure during cardiac catheterization. The predictive risk factors for and clinical impact of AKI following coronary procedures have been extensively studied and documented in the literature. Similar data, however, are lacking for AKI following angiography or endovascular interventions for lower extremity peripheral artery disease (PAD). The present review examined the published data available for AKI in patients undergoing peripheral procedures using MEDLINE searches. Specific data on number of peripheral cases, subject characteristics, hydration strategies, and AKI incidence rates was recorded. The systematic review resulted in 50 potentially relevant studies and ultimately 15 studies were selected for detailed analysis that included AKI incidence data on patients undergoing peripheral angiography or interventions. The summated studies included 11,311 patients and 10,316 peripheral procedures. The median incidence of AKI in the studies was 10%. The retrieved publications demonstrated significant variations in patient risk factors, definitions of AKI, and specificity of description of endovascular therapies. The incidence, risk factors, and outcomes related to AKI in the context of peripheral angiography or endovascular therapy remain poorly described in the literature and warrant further study in a prospective, systematic fashion. © 2016 Wiley Periodicals, Inc.


Somatosensory evoked potentials are not a sensitive indicator of potential positioning injury in the prone patient.
This study monitored somatosensory evoked potentials (SSEP) at the median and ulnar nerves in awake volunteers placed in a simulated position for prone surgery. Neurologic symptoms were used as a surrogate endpoint for position related peripheral nerve injury; the occurrence of these symptoms was correlated with the presence or absence of SSEP changes in median and ulnar nerves. Median and ulnar nerve SSEP data was obtained from awake volunteers in the supine and prone positions. With the head midline in the prone position, SSEPs were measured as the arms were advanced in four cephalad increments. Symptoms, defined as tingling, numbness, or aching in the hand, forearm, or upper arm, were recorded at each position. SSEP changes were considered significant when amplitude decreased 60% and/or latency increased 10% compared with baseline prone measurements. Symptoms were correlated with SSEP changes using chi-squared analysis (p < 0.05), and Fisher's exact analysis (p < 0.07). Data were collected on 14 patients, mean age 34 +/- 3 years. Seven (50%) subjects reported symptoms with changes in position, while four (29%) subjects displayed SSEP changes. There was no statistically significant association between symptoms and SSEP changes. There were no false positives (no symptoms in the presence of significant SSEP changes), but there were 3 (21%) false negatives (positive symptoms without SSEP changes). While all SSEP changes were associated with symptoms, the development of symptoms in 3 of 7 patients without SSEP changes suggests that SSEPs may be an imperfect monitor for the detection of positioning injury. The limited sensitivity of SSEPs in this study may be due to the duration of the monitoring, sample size, or the validity of symptoms as a surrogate for nerve injury.


Structure-activity relationships of new 1-substitutedmethyl-4-[5-(N-methyl-N-propylamino)pentyloxy]piperidines and selected 1-[(N-substituted-N-methyl)-3-propyloxy]-5-(N-methy-l-N-propyl)-pentanediamines as H3 -antagonists.
Novel, potent non-imidazole histamine H3 receptor antagonists have been prepared and in vitro tested as H3 -receptor antagonists (the electrically evoked contraction of the guinea-pig jejunum). The present compounds contain a 4-hydroxypiperidine core, which behaves as a conformationally restricted version of the 3-amino-1-propanol moiety common to the many previously described non-imidazole H3 ligands. Detailed structure-activity studies revealed that 1-(2-benzofuranylmethyl)- 5c (pA2 = 8.47 ± 0.05) and 1-(3-benzofuranylmethyl)-4-[5-(N-methyl-N-propyl)pentyloxy]piperidine 5d (pA2 = 8.15 ± 0.07) exhibit high potency for the H3 histamine receptor. In addition, the potency of selected 1-[(N-substituted-N-methyl)-3-propyloxy]-5-(N-methyl-N-propyl)pentanediamines as antagonist of the H3 histamine receptor was also evaluated. Replacement of the 4-hydroxypiperidine of the leads 7 and 5c by a highly flexible 3-(methylamino)propyloxy chain yields compounds 6a (pA2 = 8.02) and 6b (pA2 = 6.23) with higher and lower potency than their piperidine analogues (7, pA2 = 7.79; 5c, pA2 = 8.47), respectively. The histaminergic H1 antagonism of selected compounds 5c, 5d and 6a has been established on the isolated guinea-pig ileum by conventional methods; the pA2 values have compared with the potency of pyrilamine. None of them showed any H1 -antagonistic activity (pA2 < 4; for pyrilamine pA2 = 8.5).


Ca(2+)-dependent protein kinase from the halotolerant green alga Dunaliella tertiolecta: partial purification and Ca(2+)-dependent association of the enzyme to the microsomes.
Ca(2+)-dependent protein kinase (CDPK) was purified 900-fold from the soluble fraction of Dunaliella tertiolecta cells by ammonium sulfate precipitation, DEAE-Toyopearl, phenyl-Sepharose, and hydroxylapatite column chromatography. The CDPK was activated by micromolar concentration of Ca2+ and required neither calmodulin nor phospholipids for its activation. The enzyme phosphorylated casein, myosin light chain, and histone type III-S (histone H-1), but did not phosphorylate protamine and phosvitin. The Km values for ATP and casein were 11 microM and 300 micrograms/ml, respectively. Phosphorylation of casein was inhibited by calmodulin antagonists, calmidazolium, trifluoperazine, and compound 48/80, but not affected by calmodulin. CDPK bound to phenyl-Sepharose in the presence of Ca2+ and was eluted by ethylene glycol bis(beta-aminoethyl ether) N,N'-tetraacetic acid (EGTA). This suggests that hydrophobicity of the enzyme was increased by Ca2+. CDPK was also bound to the microsomes isolated from Dunaliella cells in the presence of micromolar concentration of Ca2+ and released in the presence of EGTA, suggesting the possibility of in vivo Ca(2+)-dependent association of the enzyme. The enzyme phosphorylated many proteins in the microsomes but few in the cytosol, if at all.


Deletion of the 5'-ABL region: a recurrent anomaly detected by fluorescence in situ hybridization in about 10% of Philadelphia-positive chronic myeloid leukaemia patients.
Inclusion of the BCR-ABL ES probe in routine cytogenetics led to the identification of a subgroup of Philadelphia positive (Ph+) chronic myeloid leukaemia patients characterized by a 5'-ABL deletion. This anomaly was observed in 5/51 cases (9.8%). Cytological and clinical data suggest that the 5'-ABL deletion may be associated with dysplastic features of polymorphonuclear cells and metamyelocytes and a short chronic phase duration.


Effect of charge on microvascular permeability in early experimental sepsis in the rat.
A key feature of sepsis is hypovolemia due to increased microvascular permeability. It has been suggested that the negative charge of albumin and of the endothelial glycocalyx is important for maintenance of the normally low permeability for albumin. Here we tested the hypothesis that charge effects contribute to the increased permeability in sepsis. Transcapillary escape rate (TER) and initial distribution volume for (125)I-labeled bovine serum albumin (BSA, isoelectric point pH 4.6) and for (131)I-labeled charge modified BSA (cBSA, average isoelectric point, pH 7.1) was measured 3h after sepsis was induced by cecal ligation and incision (CLI) (n=11) and in control animals (n=12). The importance of charge for permeability in sepsis was estimated by comparing the ratio between TER for cBSA and TER for BSA during control conditions to that after CLI. Plasma concentration of the glycocalyx component glycosaminoglycans (GAGs) was measured in separate control and CLI animals. The initial distribution volume for BSA and cBSA in control animals was 38 ± 3 ml/kg and 47 ± 4 mL/kg and decreased by 17% and 19%, respectively, following CLI. TER for BSA increased from 16.7 ± 4.1% in the controls to 20.1 ± 1.9% following CLI. Corresponding values for cBSA were 26.7 ± 5.6% and 29.8 ± 3.5%, respectively. The ratio between TER for cBSA and TER for BSA was 1.62 ± 0.1 in the control group and 1.49 ± 0.1 following CLI (p<0.05). Plasma GAG concentrations were higher in CLI animals than in the control group. We conclude that CLI induce hypovolemia secondary to increased microvascular permeability. Negative charge contributes to the normally low permeability of albumin and the importance of charge is decreased in early experimental sepsis. The observed charge effects are associated with CLI-induced breakdown of the glycocalyx.


Humeral Head Morphology Influences Outcomes of Arthroscopic Interposition Glenoid Patch Allograft for Glenohumeral Arthritis.
To determine short- to midterm patient-reported outcomes of arthroscopic soft-tissue interposition arthroplasty using acellular dermal allograft with a minimum follow-up of 1 year and to assess outcomes in patients with and without flattening of the humeral head. Patients with a diagnosis of primary glenohumeral arthritis who underwent arthroscopic soft-tissue interposition arthroplasty with an acellular dermal allograft from July 2010 to November 2019 were retrospectively enrolled. Inclusion criteria were a primary diagnosis of glenohumeral arthritis and Outerbridge 4 full-thickness cartilage loss of ≥50% of the glenoid articular surface. Patients underwent arthroscopic debridement, microfracture, and biological arthroscopic soft-tissue interposition arthroplasty with an acellular dermal matrix. Postoperative outcomes included American Shoulder and Elbow Surgeon (ASES) score, Single Assessment Numeric Evaluation (SANE) score, Penn Shoulder Score (PSS), numeric rating scale (NRS) pain score, analgesic use, and conversion to total shoulder arthroplasty (TSA). Results were stratified according to humeral head morphology on preoperative radiographs. A total of 25 patients were included, with a mean age of 56.0 years (range 19.2 to 74.8) and a mean follow-up of 3.36 years (range 1.03 to 8.98). The mean postoperative ASES score was 64.1 (range 11.7 to 100.0), SANE score was 62% (range 5% to 100%), and PSS was 61.2 (range 10.6 to 97.9). Additionally, 56% of patients rated their shoulder function as improved or much improved, and 36% of patients converted to TSA at a mean of 2.35 years. Patients with and without humeral flattening had similar postoperative ASES scores (P = .44), SANE scores (P = .90), PSS (P = .73), and conversions to TSA (P = .83). Patients with humeral flattening were more likely to have shoulder pain at night (83.3% versus 28.6%, P = .02). Arthroscopic soft-tissue interposition arthroplasty with an acellular dermal allograft resulted in satisfactory short- to mid-term postoperative outcomes for younger patients with glenohumeral arthritis but demonstrated a TSA conversion rate of 36%. Patients with humeral head flattening also had satisfactory shoulder function but were more likely to experience shoulder pain at night. Level IV, therapeutic case series.


Temporal-spatial distributions of road silt loadings and fugitive road dust emissions in Beijing from 2019 to 2020.
Road silt loading (sL) is an important parameter in the fugitive road dust (FRD) emissions. In this study, the improved Testing Re-entrained Aerosol Kinetic Emissions from Roads (TRAKER) combined with the AP-42 method was firstly developed to quickly measure and estimate the sLs of paved roads in Beijing, China. The annual average sLs in Beijing was 0.59±0.31 g/m2 in 2020, and decreased by 22.4% compared with that in 2019. The seasonal variations of sLs followed the order of spring > winter > summer > autumn in the two years. The seasonal mean road sLs on the same type road in the four seasons presented a decline trend from 2019 to 2020, especially on the Express way, decreasing 47.4%-72.7%. The road sLs on the different type roads in the same season followed the order of Major arterial ∼ Minor arterial ∼ Branch road > Express road, and Township road ∼ Country highway > Provincial highway ∼ National highway. The emission intensities of PM10 and PM2.5 from FRD in Beijing in 2020 were lower than those in 2019. The PM10 and PM2.5 emission intensities at the four planning areas in the two years all presented the order of the capital functional core area > the urban functional expansion area > the urban development new area > the ecological conservation and development area. The annual emissions of PM10 and PM2.5 from FRD in Beijing in 2020 were 74,886 ton and 18,118 ton, respectively, decreasing by ∼33.3% compared with those in 2019.


Current Evidence on PET Response Assessment to Immunotherapy in Lymphomas.
Response assessment in malignant lymphoma has progressively evolved in the last 20 years, leading to continuous adaptations to clinical requirements and technology improvements. The latest challenge in treatment evaluation is represented by immunomodulatory drugs, capable of stimulating response to cancer by unleashing the immune system of the host. Despite the consolidated consensus on the use of Deauville score and Lugano criteria for the assessment of first-line therapeutic regimens, during other lines of treatment and, particularly, during the course of immunotherapy, response parameters and clinical evidence appear less clear.


Performance and microbial communities of different biofilm membrane bioreactors with pre-anoxic tanks treating mariculture wastewater.
The performance of pollutant removals, activated sludge characteristics, and microbial communities of two biofilm membrane bioreactors coupled with pre-anoxic tanks (BF-AO-MBRs) (one using fiber bundle bio-carriers (FB-MBR) and the other using suspended bio-carriers (MB-MBR)) were compared at the salinity between zero and 60 g/L. At all salinities, three bioreactors showed good COD average removal efficiencies (>94.1%), and FB-MBR showed the best TN removal efficiency (90.4% at 30 g/L salinity). Moreover, FB-MBR had the faster process start-up time and better salt shock resistance. At high salinities (30-60 g/L), more extracellular polymeric substances were produced by the BF-AO-MBRs to avoid the penetration of salt and protect the bacterial community. Because of the different attachment patterns of biofilms, the microbial community structure in the FB-MBR exposed to 30 g/L salinity had higher nitrite-oxidizing/ammonia-oxidizing bacteria ratio (6.44) with more abundance of denitrifiers, which contribute to higher TN removal efficiency and lower nitrite accumulation.


[Opsonic cooperation in the neutrophil phagocytosis system in acute myeloblastic leukemia].
Investigation of opsonic activity of the serum (OAS) and indices of neutrophil phagocytosis in 35 patients with acute myeloblastic leucosis (AML) and 20 healthy subjects indicates that reduction of OAS in AML results in disturbances of the processes of reception and absorption of microorganisms by granulocytes while insufficient activation of oxygen-dependent metabolism favours disorders of bactericidal activity of neutrophils.


Beyond the implantable cardioverter-defibrillator: are we making progress?
Sudden cardiac death due to ventricular fibrillation occurs when a dynamic interaction between triggers and substrate leads to the development of reentry, initiation of ventricular tachycardia, and its degeneration to fibrillation. To move beyond the implantable cardioverter-defibrillator as the only effective therapy for aborting sudden cardiac death, an improved understanding of trigger-substrate interaction is essential. This brief review summarizes some of the recent progress in this direction.


Identification of positive charges situated at the outer mouth of the CFTR chloride channel pore.
We have used site-directed mutagenesis and functional analysis to identify positively charged amino acid residues in the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel that interact with extracellular anions. Mutation of two positively charged arginine residues in the first extracellular loop (ECL) of CFTR, R104, and R117, as well as lysine residue K335 in the sixth transmembrane region, leads to inward rectification of the current-voltage relationship and decreased single channel conductance. These effects are dependent on the charge of the substituted side chain and on the Cl(-) concentration, suggesting that these positive charges normally act to concentrate extracellular Cl(-) ions near the outer mouth of the pore. Side chain charge-dependent effects are mimicked by manipulating charge in situ by mutating these amino acids to cysteine followed by covalent modification with charged cysteine-reactive reagents, confirming the location of these side chains within the pore outer vestibule. State-independent modification of R104C and R117C suggests that these residues are located at the outermost part of the pore. We suggest that ECL1 contributes to the CFTR pore external vestibule and that positively charged amino acid side chains in this region act to attract Cl(-) ions into the pore. In contrast, we find no evidence that fixed positive charges in other ECLs contribute to the permeation properties of the pore.


Identity confirmation of 99mTc-MAG3, 99mTc-sestamibi and 99mTc-ECD using radio-LC-MS.
Due to the low concentrations in which 99mTc-radiopharmaceuticals are obtained (4-40 ng/ml), confirmation of the identity of these tracer agents in the European Pharmacopoeia is generally performed only indirectly by assessment of their retention times on RP-HPLC. We have investigated whether it is possible to obtain more direct proof of the identity of technetium-99m labelled radiopharmaceuticals using radio-LC-MS. As representative examples, negatively charged 99mTc-MAG3, positively charged 99mTc-Sestamibi and neutral 99mTc-ECD were used. The three technetium-99m radiopharmaceuticals were prepared in several conditions to obtain variable relative amounts of radiochemical impurities and variable concentrations of the complexes (pico- to nanomolar). The preparations were analyzed on a reversed phase C18 HPLC column using a radio-LC-MS system equipped with a time of flight mass spectrometer with electrospray ionization in positive (99mTc-Sestamibi, 99mTc-ECD) or negative (99mTc-MAG3, 99mTc-ECD) mode. For each of the studied complexes, the main peak in the radiometric channel coincided with the expected molecular ion mass of the corresponding technetium complex in the mass spectrometer channel. The relative error on the measured accurate mass was in the range of 10 ppm. The identity of several radiochemical impurities of the three technetium complexes was also confirmed or established. It is concluded that radio-LC-MS can be a sensitive aid in quality control of 'no carrier added' radiopharmaceuticals.


Lissoclinolide, the first non-nitrogenous metabolite from a Lissoclinum tunicate.
The isolation and characterization of a new metabolite from the tunicate Lissoclinum patella is described. The structure of lissoclinolide [1] was solved using spectral and chemical methods, and it is shown to be the first non-nitrogenous, non-sulfur-containing compound to be isolated from the genus Lissoclinum.


Pediatric Salivary Gland Malignancies.
Pediatric malignant salivary gland tumors are extremely rare. The percentage of malignant tumors is higher than that seen in adults, although the outcomes in terms of survival are better in pediatric patients. The mainstay of treatment is surgical excision with negative margins. This article reviews current concepts in demographics, etiology, management, and outcomes of malignant salivary tumors in children.


Osteopontin gene polymorphism association with milk traits and its expression analysis in milk of riverine buffalo.
Osteopontin gene is regarded as a plausible candidate in mammary gland differentiation and development, expressed by variety of cells, tissues, and biological fluids including milk. The current study was performed in two phases. In the first phase, Osteopontin gene polymorphisms were identified and associated with milk composition such as ash, milk fat, SNF, lactose, and protein. In the second phase, milk samples from five healthy mastitis-free Nili Ravi buffaloes were analyzed for expression of Osteopontin gene at transition (day 15), mid (day 90), and end (day 250) stage of their second lactation. Briefly, blood samples were collected from Nili Ravi buffalo to isolate the genomic DNA, specific primers were designed for PCR amplification. The amplified PCR products were sequenced bi-directionally. Six polymorphisms were identified in the coding region and four in the intronic region of the gene. The results showed that SNP g.38329758 T > C causing substitution of valine to alanine (V127A) was associated with high milk protein. For mRNA expression analysis, somatic cells were separated from milk samples for RNA isolation. Analysis of differential gene expression data has permitted us to illustrate the expression pattern of osteopontin gene in lactating buffalo. The Osteopontin gene was found to be transcribed among all three lactation stages, but expression was observed with the highest value (fold change) in peak lactation and remained elevated till the end of lactation. Identified gene marker may be helpful for the prediction of superior animal for selection. The presented study also gave an insight into the genetic screening and lactation biology of riverine buffalo, offering direction for future research in lactating buffalo.


[Comparative Study on Morphological Characteristics of Root of Six Species in Sect. Cruciata].
To provide evidences for the identification of Gentianae Macrophyllae Radix by comparing the morphological characteristics of six species of Sect. Cruciata (Gentiana macrophylla, G. crassicaulis, G. straminea, G. dahurica, G. officinalis and G. siphonantha). By microscope, the tissue characteristics with freehand section of the upper, middle and lower of root and the powder characteristics with chloral hydrate were studied. The vascular cylinder of G. crassicaulis was not split. The vascular cylinder of G. macrophylla, G. dahurica and G. officinalis, were only split in the upper part. The root of G. straminea and G. siphonantha wre completely divided into several smaller roots twisting together. There were a lot of thick walled cells in the powder of G. dahurica, G. straminea and G. siphonantha, but their shapes were different. No thick walled cells were found in the other three species. There are obviously differences among the microscopic morphological characteristics of root of six species of Sect. Cruciata, which can provide the basis for identification of Gentianae macrophyllae Radix.


Simulate SubQ: The Methods and the Media.
For decades, there has been a growing interest in injectable subcutaneous formulations to improve the absorption of drugs into the systemic circulation and to prolong their release over a longer period. However, fluctuations in the blood plasma levels together with bioavailability issues often limit their clinical success. This warrants a closer look at the performance of long-acting depots, for example, and their dependence on the complex interplay between the dosage form and the physiological microenvironment. For this, biopredictive performance testing is used for a thorough understanding of the biophysical processes affecting the absorption of compounds from the injection site in vivo and their simulation in vitro. In the present work, we discuss in vitro methodologies including methods and media developed for the subcutaneous route of administration on the background of the most relevant absorption mechanisms. Also, we highlight some important knowledge gaps and shortcomings of the existing methodologies to provide the reader with a better understanding of the scientific evidence underlying these models.


Synthesis and evaluation of (17 alpha,20E)21-[125I]iodo-11-substituted-19-norpregna-1,3,5(10), 20-tetraene-3,17 beta-diols: the influence of 11-stereochemistry on tissue distribution of radioiodinated estrogens.
The 21-tri-n-butylstannyl derivatives of (17 alpha,20E)-11 alpha and beta-methoxy-19-norpregna-1,3,5(10),20-tetraene-3,17 beta-diol were synthesized and characterized. These compounds, as well as the 11-unsubstituted compound were converted via electrophilic ipso radioiododestannylation to the corresponding 21[125I]iodo analogs at the no-carrier-added level in 73-90% isolated radiochemical yields. The radiochemical 4c [IV alpha ME2, (17,20E)-21[125I]iodo-11 alpha-methoxy-19-norpregna-1,3, 5(10),20-tetraene-3,17 beta-diol] was evaluated in immature female rats and the results compared to those previously reported for 4a (IVE2) and 4b (IV beta ME2) to determine the influence of 11-substitution on the ability of the compounds to function as estrogen receptor-seeking agents in vivo. The results indicated that the uptake of 11 alpha-methoxy derivative in the target organ was substantially lower, of shorter duration, with a much smaller specific receptor binding component than the other two radioligands. The distribution profile of the three 17 alpha-iodovinyl estrogens paralleled that previously reported for the corresponding 17 alpha-ethynyl estrogens and this study suggests that the in vivo pharmacological results reported for the 17 alpha-ethynyl estrogens may be used to predict the in vivo behavior of the corresponding 17 alpha-iodovinyl analogs.


Cytokeratin polypeptide in cultured human squamous cell carcinoma as a possible marker for keratinization.
A human cancer cell line (ZK-1) has been established from a well-differentiated squamous cell carcinoma of the tongue. The cytokeratin polypeptides pattern of ZK-1 cells consists of four major polypeptides with molecular weights between 46 and 58 kilodaltons (kd). Antibodies raised against the purified 58 kd cytokeratin filament from cultured ZK-1 cells were shown to be specific by one- and two-dimensional polyacrylamide gel electrophoresis, immunofluorescence, and immunoelectron microscopy. Immunoprecipitation studies showed that the antibody reacted mainly with the 58 kd cytokeratin. The distribution of the 58 kd cytokeratin in both sparse and confluent cultures was analyzed by the indirect immunofluorescence technique. In both cases, it appeared that fibrillar arrays extended throughout the cytoplasm running over the nucleus and toward cell-to-cell boundaries. Thick bundles of filaments were seen in confluent cultures, especially in large and flattened keratinized cells. Characteristically, reactions were also seen in the intercellular boundaries, appearing as "dots." Electron microscopy using immunoperoxidase techniques indicated that the 58 kd cytokeratin was localized in tonofilaments, tonofilaments attached to desmosomes, desmosomal plaques and membrane-coating granules.


Bioenergetic failure correlates with autophagy and apoptosis in rat liver following silver nanoparticle intraperitoneal administration.
Deposition and accumulation of silver nanoparticles (Ag-nps) in the liver have been shown to induce hepatotoxicity in animal studies. The hepatotoxicity may include oxidative stress, abnormalities in energy metabolism, and cell death. Studies have indicated that autophagy is an intracellular event involving balance of energy, nutrients, and turnover of subcellular organelles. The present study was undertaken to test the hypothesis that autophagy plays a role in mediating hepatotoxicity in animal after exposure to Ag-nps. Focus was placed on interrelationship between energy metabolism, autophagy, apoptosis and hepatic dysfunction. Sprague Dawley rats were intraperitoneally injected with Ag-nps (10-30 nm in diameter) at concentration of 500 mg kg(-1). All animals were sacrificed on days 1, 4, 7, 10 and 30 after exposure and blood and liver tissues were collected for further studies. Uptake of Ag-nps was quite prompt and not proportional to the blood Ag concentration. Declination of ATP (-64% in days 1) and autophagy (determined by LC3-II protein expression and morphological evaluation) increased and peaked on the first day. The ATP content remained at low level even though the autophagy has been activated. Apoptosis (based on caspase-3 protein expression and TUNEL-positive cells staining) began to rise sigmoidally at days 1 and 4, reached a peak level at day 7, and remained at the same levels during days 7-30 post exposure. Meanwhile, autophagy exhibited a gradual decrease from days 1-10 and the decrease at day 30 was statistically significant as compared to day 0 (sham group). Inflammatory reaction (histopathological evaluation) was found at day 10 and preceded to an advanced degree at day 30 when liver function was impaired. These results indicate that following Ag-nps administration, autophagy was induced; however, failure to preserve autophagy compounded with energy reduction led to apoptosis and the eventual impairment of liver function. The study provides an in-vivo evidence of hepatotoxicity by continuous exposure of Ag-nps in rats.


[Fast-growing atypical cervical mass].
Aneurysms of the jugular vein are an uncommon condition, typical of childhood and adolescence although they can appear at any age. Clinically, they normally constitute an asymptomatic cervical mass which increases when Valsalva's manoeuvre is applied. Doppler echography is the most effective diagnostic test, with CT or NMR for doubtful situations. Treatment is conservative except for complications or major aesthetic repercussions. We present here the case of a 65-year-old female patient, and review the literature regarding aneurysms of the jugular vein.


Sports participation and preterm birth: a nationwide birth cohort in Japan.
Children born preterm may be less physically active than children born term because of neurocognitive problems, reduced lung function, and poor physical fitness. We evaluated sports participation of children and adolescents who had been born preterm (<37 weeks) and early term (37-38 weeks) in 2001. Data from a nationwide longitudinal survey (n = 47,015, including 2375 children born preterm) were analyzed. As indicators of sports participation, we used responses to questions about participation in sports clubs at 7 and 10 years old and in extracurricular school sports at 15 years old. Children born very preterm (25-31 weeks) and moderately to late preterm (32-36 weeks) were less likely to participate in sports clubs at 7, 10, and 15 years old than children born full term (39-41 weeks). Compared with children born full term, the adjusted risk ratios for participation in extracurricular school sports at 15 years old were 0.86 (95% confidence interval: 0.75-0.98) for children born very preterm, 0.92 (0.88-0.97) for children born moderately to late preterm, and 1.00 (0.98-1.02) for children born early term. Our findings suggest that preterm birth is associated with less participation in organized sports during childhood and adolescence than full-term birth. Research investigating associations between preterm birth and physical activity among children born in the 2000s is limited. This study shows that preterm birth was associated with less participation in organized sports during childhood and adolescence than full-term birth, especially in boys, and the participation in organized sports of children born preterm decreased as gestation shortened. During childhood, boys born early term were also less likely to participate in organized sports than boys born full term, suggesting a continuum with preterm births. These findings offer important additional insights into the limited evidence available for predicting future health outcomes for preterm infants.


[The autonomous quadrant-syndrome (author's transl)].
The quadrant-syndrome (Q.S.) is a functional disturbance of the vegetative system, characterized by the distribution of subjective and objective lesions to a quarter of the body (quadrant). The neurological aspect of the Q.S. varies so much in character, that it may mistakenly be considered to be psychogenic in origin. All qualities of peripheral nerve function can be affected singly or collectively and in varying degree. The duration of illness cannot be predicted; it may last from some weeks up to more than 10 years. The patients age seems to play some role in the duration of the syndrom. On account of symptomatological and pathogenetical similarities with the dystrophy of Sudeck and the causalgic state, it appears that the Q.S. would be observed more frequently, if its characteristic principle of distribution to a quarter of the body is taken into consideration. Out of the many possible causes of this syndrom the common lesions of the extremities are pointed out. These peripheral lesions act as focus, which affects a specially predisposed central vegetative nervous system. A case report is given of an 18 year-old girl suffering from a median nerve compression in the elbow region (pronator teres syndrom) and developing a Q.S. with complete anaesthesia and analgesia. Six weeks after surgical decompression of the median nerve, she was free of symptoms and remained free of symptoms after a follow up control six months later. Besides sanitation of the peripheral focus, sympathectomy is critically discussed, and the possibility of preventive treatment in common lesions of the extremities is ventilated.


Chronic keratoconjunctivitis associated with ocular adenovirus infection.
Adenovirus (types 3, 4, 5) was implicated as the etiologic agent in three cases of persistent keratoconjunctivitis. In two, virus was isolated 25 and 29 months after the onset of symptoms; the third patient has suffered intermittent flares of kerato-conjunctivitis since the initial isolation of virus 13 weeks after the onset of the disease. Such patients may represent a previously unsuspected source of adenovirus in the population. All were given corticosteroids topically. This treatment may have prolonged the adenoviral disease.


Lamotrigine and remacemide protect striatal neurons against in vitro ischemia: an electrophysiological study.
In the present study, we investigated the cellular and synaptic mechanisms underlying the neuroprotective action of lamotrigine and remacemide. Both drugs, in fact, have been reported to exert a neuroprotective action in in vivo animal models of ischemia. To address this issue, electrophysiological recordings and cell swelling measurements were performed from striatal neurons in control condition and during combined oxygen and glucose deprivation (in vitro ischemia) in a brain slice preparation. Lamotrigine, remacemide, and the active desglycinyl metabolite of remacemide, D-REMA, induced a concentration-dependent reduction of both repetitive firing discharge and excitatory postsynaptic potentials. However, while remacemide and D-REMA exerted their inhibitory action on glutamatergic transmission by blocking NMDA receptors, lamotrigine exerted a preferential presynaptic action, as indicated by its ability to increase paired-pulse facilitation. Both remacemide and lamotrigine were found to be neuroprotective against the irreversible field potential loss and cell swelling induced by in vitro ischemia, and coadministration of low concentrations of these drugs exerted an additive neuroprotective action. A combined use of lamotrigine and remacemide could be employed in clinical trials to enhance neuroprotection in neurological disorders involving an abnormal striatal glutamatergic transmission.


Three siblings with Cerebrotendinous Xanthomatosis: a novel mutation in the CYP27A1 gene.
Cerebrotendinous Xanthomatosis (CTX) is a rare autosomal recessive lipid storage disease caused by sterol 27-hydroxylase (CYP27) deficiency. We report three CTX siblings that shared a novel mutation of the CYP27A1 gene. These siblings presented with elevated cholestanol levels and typical manifestations such as tendon xanthomas, cataracts, osteopenia, mental retardation, cerebellar ataxia and peripheral neuropathy. All shared the same genetic mutation, c.1146_1151delins and c.1214G>A of CYP27A1. All were treated with 750 mg/day chenodeoxycholic acid (CDCA). In conclusion, one should consider the possibility of CTX in any individual with normocholesterolemic xanthomatosis, early-onset cataracts, mental retardation, cerebellar ataxia and peripheral neuropathy.


Exploiting isolobal relationships to create new ionic liquids: novel room-temperature ionic liquids based upon (N-alkylimidazole)(amine)BH2+"boronium" ions.
Readily prepared imidazole-based boronium ions form stable, hydrophobic, room-temperature ionic liquids (RTIL) with unique electronic and spectroscopic characteristics.


Will it hurt? Verbal interaction between child and radiographer during radiographic examination.
This study investigated the nature of verbal interactions between child, parent and radiographer and the extent to which it varied as a function of the child's age. The participants were 20 female radiographers and 32 children (3-15 years) examined for acute injuries. The verbal interactions during the examination were video recorded and analyzed using the Roter Interaction Analysis System (RIAS). Results indicated that 80% of the verbal interaction was accounted for by the radiographer, 17% by the child and 3% by the parent. The distribution of utterances varied with regard to children's age.


Rearrangement of the cytoskeletal network induced by platelet-derived growth factor in rat hepatic stellate cells: role of different intracellular signalling pathways.
Cytoskeletal reorganization plays an important role in the regulation of different cell functions, such as proliferation and migration. Since platelet-derived growth factor (PDGF) stimulates both proliferation and chemotaxis of hepatic stellate cells (HSC), we investigated the effects of this cytokine on cytoskeletal components of cultured rat HSC. Exposure of HSC to PDGF induced the formation of stress fibres and of a ruffled configuration of the plasma membrane, evaluated by both fluorescence and electron microscopy. These modifications were also induced by exposure to the protein kinase C (PKC) activator phorbol-12-myristate-13-acetate (PMA) and abolished by pretreatment with the PKC inhibitor calphostin C, with the Rho inhibitor C3 exoenzyme and with the intracellular calcium chelator MAPTAM, but not with the PI-3 kinase inhibitor wortmannin or with the mitogen-activated protein kinase kinase inhibitor PD 98059. PDGF induced a translocation of Rho from the cytosol to the membrane which was inhibited by C3 exoenzyme and by calpostin C, and which was also induced by PMA. Moreover, PDGF induced a rearrangement of vinculin which was prevented by C3 exoenzyme and calphostin C. PDGF-induced cytoskeletal reorganization in HSC is dependent on PKC and Rho, thus suggesting that these two pathways may play an important role in the response of liver to injury.


Use of tissue expanders in adult postpneumonectomy syndrome.
Mediastinal shift and rotation after pneumonectomy can lead to severe symptomatic airway compression. Historically, a variety of treatments, such as muscle-flap transposition, pericardial fixation, and plombage, have been used. In this study we retrospectively evaluated the effectiveness of intrathoracic tissue expansion in postpneumonectomy syndrome. Since 1990, our center has used tissue expanders as plombage in patients with postpneumonectomy syndrome. Between 1990 and 2005, a total of 20 patients were treated. The outcome was evaluated by using preoperative, perioperative, and postoperative bronchoscopy and imaging studies. Patient satisfaction was determined with a validated questionnaire. In 19 of the 20 patients, up to 3 tissue expanders were placed and filled within the pleural cavity. Access to the pleural cavity could not be obtained in 1 patient because of adhesions. Perioperative and postoperative bronchoscopic scans demonstrated decompression of the left main bronchus in 16 (84%) of 19 patients. On discharge, all patients reported improvement of their respiratory symptoms. Six (32%) patients required reoperation because of herniation (n = 2), luxation (n = 1), inadequate positioning (n = 2), and leakage of the tissue expander (n = 4). In 4 patients additional filling was performed in the outpatient clinic, with immediate improvement of respiratory distress. Use of tissue expanders in adults with postpneumonectomy syndrome is an effective means of decompressing the remaining bronchus, thereby leading to a significant improvement in respiratory symptoms. Although 32% of patients required reoperation for complications, each complication was readily correctable.


Transcriptomic analysis supports a role for the nervous system in regulating growth and development of Fasciola hepatica juveniles.
Fasciola spp. liver flukes have significant impacts in veterinary and human medicine. The absence of a vaccine and increasing anthelmintic resistance threaten sustainable control and underscore the need for novel flukicides. Functional genomic approaches underpinned by in vitro culture of juvenile Fasciola hepatica facilitate control target validation in the most pathogenic life stage. Comparative transcriptomics of in vitro and in vivo maintained 21 day old F. hepatica finds that 86% of genes are expressed at similar levels across maintenance treatments suggesting commonality in core biological functioning within these juveniles. Phenotypic comparisons revealed higher cell proliferation and growth rates in the in vivo juveniles compared to their in vitro counterparts. These phenotypic differences were consistent with the upregulation of neoblast-like stem cell and cell-cycle associated genes in in vivo maintained worms. The more rapid growth/development of in vivo juveniles was further evidenced by a switch in cathepsin protease expression profiles, dominated by cathepsin B in in vitro juveniles and by cathepsin L in in vivo juveniles. Coincident with more rapid growth/development was the marked downregulation of both classical and peptidergic neuronal signalling components in in vivo maintained juveniles, supporting a role for the nervous system in regulating liver fluke growth and development. Differences in the miRNA complements of in vivo and in vitro juveniles identified 31 differentially expressed miRNAs, including fhe-let-7a-5p, fhe-mir-124-3p and miRNAs predicted to target Wnt-signalling, which supports a key role for miRNAs in driving the growth/developmental differences in the in vitro and in vivo maintained juvenile liver fluke. Widespread differences in the expression of neuronal genes in juvenile fluke grown in vitro and in vivo expose significant interplay between neuronal signalling and the rate of growth/development, encouraging consideration of neuronal targets in efforts to dysregulate growth/development for parasite control.


Sulfhydryl groups in hemoglobin. A new molecular probe at the alpha1 beta 1 interface studied by Fourier transform infrared spectroscopy.
Infrared absorption bands due to sulfhydryl groups (VSH) of alpha-104(G11) and beta-112 (G14) cysteine residues of human carboxyhemoglobin (HbCO) have been observed near 2560 cm- minus 1 by use of Fourier transform infrared (FTIR) spectroscopy. The beta-93 cysteine SH groups absorb infrared radiation so weakly that they are not distinguished from background. Only single SH absorption bands due to the alpha-104 cysteines of pig and horse hemoglobin are observed. The SH absorption bands from human HbCO disappear in alkali, are broadened by detergent or guanidinium chloride, and show a complex titration curve, and an isotopic frequency shift (VSD/VSH equals to 0.7267) virtually identical with that reported for methanethiol. The integrated absorption coefficient (epsilonmM(area)) for 0.1 M ethanetiol increased with H-bond acceptor solvents in the order: CCl4 (0.07), water (0.21), acetone (0.43), and N,N-dimethyl-acetamide (1.35 mM- minus 1 cm- minus 2). Comparison of the integrated absorption coefficients for the alpha-104 cysteine SH (2.43), and the beta-112 SH (0.80), of human HbCO with those of ethanetiol solutions suggested specifically H-bonded structures with peptide carbonyl groups 4 (or 3) residues back in the G helices. This was found to agree with a molecular model of the alpha-chain G helix of horse HbO2 built to coordinates from M. F. Perutz.


Aberrantly activated Wnt/β-catenin pathway co-receptors LRP5 and LRP6 regulate osteoblast differentiation in the developing coronal sutures of an Apert syndrome (Fgfr2S252W/+ ) mouse model.
Apert syndrome is an autosomal, dominant inherited disorder characterized by craniosynostosis and syndactyly caused by gain-of-function mutations in the fibroblast growth factor receptor 2 (FGFR2) gene. Wnt/β-catenin signaling plays critical roles in regulating the skeletal development. Here, we analyzed the role of this pathway in the developing coronal sutures (CS) of a murine Apert syndrome model (Fgfr2S252W/+ ). We observed aberrantly increased mRNA expression of Lrp5 and Lrp6 in CS of Fgfr2S252W/+ mice, whereas both wild type (WT) and Fgfr2S252W/+ mice showed similar expression of other Wnt/β-catenin-related genes, such as Wnt3, Wnt3a, Fzd4, Fzd6, Axin2, and Dkk1 as evidenced by in situ hybridization. Significantly increased Lrp5 and Lrp6 mRNA expression was observed by quantitative PCR analysis of cultured cells isolated from CS of Fgfr2S252W/+ mice. Phospho-LRP5, phospho-LRP6, and non-phospho-β-catenin were upregulated in Fgfr2S252W/+ CS compared with that in WT CS. Short-interfering RNA targeting Lrp5 and Lrp6 significantly reduced runt-related transcription factor 2, collagen type 1 alpha 1, and osteocalcin mRNA expression, and alkaline phosphatase activity in cultured cells. The Wnt/β-catenin pathway was activated in the CS of Fgfr2S252W/+ mice during craniofacial development, suggesting the involvement of the Wnt/β-catenin pathway in the pathogenesis of CS synostosis in Fgfr2S252W/+ mice.


Placenta growth factor (PlGF) mRNA expression in brain tumors.
To investigate the relationship between placenta growth factor (PlGF) and brain tumor angiogenesis, we screened 36 primary and 3 metastatic brain tumors. We examined the expression of PlGF mRNA with respect to vasculature of various tumors which was determined by preoperative angiography. The expression of genes of the other angiogenic factors, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) was also tested, and compared to that of PlGF. The primary tumors consisted of 16 meningiomas, 7 gliomas, 7 schwannomas, 4 pituitary adenomas, 1 germinoma, and 1 choriocarcinoma. Using a quantitative reverse transcription-polymerase chain reaction, the mRNA for PlGF149 and PlGF170 were detected in 25 out of 39 (64.1%) brain tumors. In primary brain tumors, PIGF mRNA was expressed in all the hypervascular tumors, but only in 5 of 16 hypovascular tumors (31.3%). None of the 3 metastatic hypervascular tumors expressed PlGF mRNA. The VEGF and bFGF mRNA expression was both detected in 87.2% of the tumors examined. We conducted hypoxic experiments with cultured U-251MG human glioma cells to determine the mechanism of PIGF gene regulation. As the atmospheric oxygen concentration was decreased, the PIGF mRNA level in the U-251MG cells was markedly increased. These results suggest that PIGF may contribute to the pathogenesis of brain tumor angiogenesis.


Huge right atrial myxoma causing fixed tricuspid stenosis with constitutional symptoms.
Nonspecific constitutional symptoms are reported mostly in patients with left-atrial myxomas, which occur five times as often as its right-atrial counterpart. We present huge right-atrial myxoma, which obstructs tricuspid orifice with nonspecific constitutional symptoms without any pulmonary embolism attack.


Radiofrequency therapy in esthetic dermatology: A review of clinical evidences.
Chronological skin aging causes the modification of genetic material through enzymes and proteins changes. The process reduces cellular proliferation, along with loss of tissue elasticity, reduced ability to regulate aqueous exchanges, and inefficient tissue replication. Appearance is negatively affected by cumulative changes in coloration, texture, and elasticity over time. The increase in the population's average life expectancy boosts the search for cosmetic therapies that can delay aging, mostly for the noninvasive modalities. Among the various options, radiofrequency therapy is a technique that can help reduce the effects of skin aging. Therefore, this study aims to review clinical evidence provided by scientific literature on the benefits of using radiofrequency therapy in reducing skin aging effects. A review of the literature concerning skin aging, characteristics of radiofrequency therapy, and radiofrequency therapy in the treatment of skin laxity and mechanism of action was conducted using PubMed. The included studies have suggested that the mechanism of radiofrequency action is heating the dermis while preserving the epidermis. This heating causes immediate collagen denaturation, which is followed by the formation of new collagen, naturally providing skin tightening and greater elasticity. Even when used as single therapeutic modality, radiofrequency seems to meet the expectations in reducing the effects of skin aging.


Amelioration of excision wounds by topical application of green synthesized, formulated silver and gold nanoparticles in albino Wistar rats.
Wound healing, a complex biological process, has attained a lot of attention as dermatologists are primarily interested in stimulated wound closure without formation of scar or a faint scar. The recent upsurgence of nanotechnology has provided novel therapeutic materials in the form of silver and gold nanoparticles which accelerate the wound healing process. The effect of formulated nanoparticles using Coleus forskohlii root extract (green synthesized) has been tried out for ameliorating full thickness excision wounds in albino Wistar male rats. The evaluation of in vivo activity of nanoparticles in wound healing was carried out on open wounds made by excision on the dorsal sides of albino Wistar rats under anesthesia, and the healing of the wounds was assessed. Histological aspects of the healing process were studied by a HE (Hematoxylin and Eosin) staining method to assess various degrees of re-epithelialization and the linear alignment of the granulation tissue whereas Van Gieson's histochemical staining was performed to observe collagen fibers. The healing action shown by the formulated nanoparticles was remarkable during the early stages of wound healing, which resulted in the substantial reduction of the whole healing period. Topical application of formulated gold nanoparticles was found to be more effective in suppressing inflammation and stimulating re-epithelialization compared to silver nanoparticles during the healing process. The results throw light on the amelioration of excision wounds using nanoparticles which could be a novel therapeutic way of improving wound healing in clinical practice. The mechanism of advanced healing action of both types of nanoparticles could be due to their antimicrobial, antioxidant and anti-inflammatory properties.


Early experience with the Levitronix Centrimag device for extra-corporeal membrane oxygenation following lung transplantation.
Extra-corporeal membrane oxygenation (ECMO) is accepted as a salvage therapy to treat life-threatening complications following lung transplantation such as primary graft dysfunction, acute rejection or airway dehiscence. Levitronix Centrimag (Levitronix LLC, Waltham, MA) is a centrifugal pump that consists of a magnetically levitated bearing-less rotor designed to reduce blood friction. ECMO using the Levitronix Centrimag pump may be an ideal medium-term support for lung transplant recipients suffering these serious complications. We describe our early experience of using ECMO with Levitronix Centrimag device and the Hilite hollow fibre membrane oxygenator (Medos Hilite LT, Medos Medizintechnik AG) in three cases following lung transplantation. The device is technically easy to implant and manage with a low complication rate on minimal anticoagulation.


Ectonucleotidase CD39 and Checkpoint Signalling Receptor Programmed Death 1 are Highly Elevated in Intratumoral Immune Cells in Non-small-cell Lung Cancer.
Lung cancer is the leading cause of cancer death in both sexes worldwide and has a predicted 5-year survival rate of <20%. Immunotherapy targeting immune checkpoints such as the programmed death 1 (PD-1) signaling pathway has led to a shift of paradigm in the treatment of advanced non-small-cell lung cancer (NSCLC) but remains without effect in ∼80% of patients. Accumulating evidence suggests that several immunosuppressive mechanisms may work together in NSCLC. The contribution and cooperation between different immunosuppressive mechanisms in NSCLC remain unknown. Recently, the CD39-adenosine pathway has gained increasing attention as a crucial immunosuppressive mechanism and possible target for immunotherapy. Immune cells were extracted from lung and tumor tissue after lung resection in 12 patients by combined enzymatic and mechanical tissue disaggregation. A multiparameter flow cytometry panel was established to investigate the expression and coexpression of CD39 and PD-1 on key lymphocyte subtypes. Frequencies of CD39+, PD-1+, and CD39+/PD-1+cells were higher among both CD4+ and CD8+ T cells isolated from NSCLC tumor tissue than in T cells from normal lung tissue. Similarly, the frequency of FoxP3+ CD4+ T cells (Tregs) was highly significantly elevated in tumor tissue compared to adjacent lung tissue. The consistent upregulation of CD39 on immune cells in tumor microenvironment indicates that the CD39 signaling pathway may, in addition to the PD-1 pathway, represent another important mechanism for tumor-induced immunosuppression in NSCLC. In addition, the present study indicates that a comprehensive immune response profiling with flow cytometry may be both feasible and clinically relevant.


Clusters of branched aliphatic side chains serve as cores of stability in the native state of the HisF TIM barrel protein.
Imidazole-3-glycerol phosphate synthase is a heterodimeric allosteric enzyme that catalyzes consecutive reactions in imidazole biosynthesis through its HisF and HisH subunits. The unusually slow unfolding reaction of the isolated HisF TIM barrel domain from the thermophilic bacteria, Thermotoga maritima, enabled an NMR-based site-specific analysis of the main-chain hydrogen bonds that stabilize its native conformation. Very strong protection against exchange with solvent deuterium in the native state was found in a subset of buried positions in α-helices and pervasively in the underlying β-strands associated with a pair of large clusters of isoleucine, leucine and valine (ILV) side chains located in the α7(βα)8(βα)1-2 and α2(βα)3-6β7 segments of the (βα)8 barrel. The most densely packed region of the large cluster, α3(βα)4-6β7, correlates closely with the core of stability previously observed in computational, protein engineering and NMR dynamics studies, demonstrating a key role for this cluster in determining the thermodynamic and structural properties of the native state of HisF. When considered with the results of previous studies where ILV clusters were found to stabilize the hydrogen-bonded networks in folding intermediates for other TIM barrel proteins, it appears that clusters of branched aliphatic side chains can serve as cores of stability across the entire folding reaction coordinate of one of the most common motifs in biology.


A study of iodine concentration in drinking waters of Bryansk and Oryol regions.
The aim of the research was to study and compare iodine concentration in natural waters originating from aquifers of different ages, primarily residents use for drinking purposes. The analysis was based on the original data on the samples collected during the fieldwork in the Bryansk region (2013-2017) and in the Oryol region (2016-2017). In addition to iodine concentration, the main geochemical parameters (salinity, etc.) were determined in the selected waters. The results showed a significant variation of iodine in waters from both regions (Bryansk region-from 0.7 to 41.2 µg/l; Oryol region-from 1.12 to 36.8 µg/l), the difference being apparently due to different ages and origins of the sampled aquifers (Quaternary, Upper Devonian and Cretaceous deposits). The overall low provision of surface water with iodine was found both in the Bryansk region (median for shallow wells-5.82 µg/l, median for surface water-6.76 µg/l) and in the Oryol region (median for shallow wells-2.96 µg/l, median for surface water-7.4 µg/l). The data obtained deserve attention during organization of monitoring and implementation of measures preventing thyroid diseases.


Identification of fiber orientation in left free-wall accessory pathways: implication for radiofrequency ablation.
Previous reports on the anatomic discordance between atrial and ventricular insertion sites of left free-wall accessory pathways were limited and their findings were controversial. The purpose of this study was to explore the fiber orientation and related electrophysiologic characteristics of left free-wall accessory pathways. The study population comprised 96 consecutive patients with a single left free-wall accessory pathway (33 manifest and 63 concealed pathways), who underwent electrophysiologic study and radiofrequency catheter ablation using the retrograde ventricular approach. The atrial insertion site of the accessory pathway was defined from the cinefilms as the site with the earliest retrograde atrial activation bracketed on the coronary sinus catheter during tachycardia, and the ventricular insertion site was defined as the site where successful ablation of the pathway was achieved. Forty-two patients (44%) had their atrial insertion sites 5-20 mm (10 +/- 3 mm) distal to the ventricular insertion sites (proximal excursion), 30 (31%) patients had their atrial insertion sites 5-20 mm (12 +/- 3 mm) proximal to the ventricular insertion sites (distal excursion), and 24 (25%) patients had directly aligned atrial and ventricular insertion sites. Retrograde conduction properties, including 1:1 VA conduction and effective refractory period, were significantly poorer in the pathways with proximal excursion (302 +/- 67, 285 +/- 61 ms respectively) than in those with distal excursion (264 +/- 56, 250 +/- 48 ms respectively) or direct alignment (272 +/- 61, 258 +/- 73 ms respectively). Accessory pathways at the more posterior location had a significantly higher incidence of proximal excursion (P = 0.006), and those at the more anterior location had a higher incidence of distal excursion (P = 0.012). In conclusion, a wide variation in fiber orientations and related electrophysiologic characteristics was found in left free-wall accessory pathways. This may have important clinical implications for radiofrequency ablation.


Comparison of fresh tissue incubation assay and the in vivo localization of monoclonal antibodies to renal cell carcinoma.
Few in vitro tests currently available are able to accurately predict the in vivo localization of monoclonal antibodies (Mabs) to cancer. We report on a fresh tissue incubation assay (FTIA) and compare the results of this assay to the in vivo localization of renal cell carcinoma (RCC)-reactive Mab A6H and control Mab AFP-22 to RCC and non-RCC xenografts implanted in nude mice. Both the FTIA and in vivo localization study demonstrated highly selective uptake of A6H in RCC but not in non-RCC xenografts. Radioimmunoscintigraphy using A6H clearly visualized RCC xenografts in every attempt, while AFP-22 did not highlight any of the tumor xenografts. The results demonstrate that FTIA may be a useful in vitro assay for selecting Mabs for in vivo application, and that radioimmunoscintigraphy is a potentially useful tool in detecting cancer sites.


[Streptomyces spheroides M8-2 strain--a producer of extracellular proteolytic enzymes possessing fibrinolytic and thrombolytic action].
Strain Streptomyces spheroides M8-2 was obtained as a result of ultraviolet light action on the S. spheroides spores strain 35. The mutant is characterized by the considerable increase (in 4-5 times) of the extracellular proteolytic enzymes which are able to dissolve fibrin and thrombs. The description of the morfologic, cultural and physiologic-biochemical properties of the culture is given. The content of GC-pairs in DNK of the mutant and parent strains has been determined.


The usefulness of CHAPS as a non-cytotoxic stabilizing agent in purification of growth factors.
Among several detergents, a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), was found to be least cytotoxic for cultured mammalian cells. CHAPS improved the activity recovery and elution profile of crude and purified fibroblast growth factors (FGFs) during chromatographies. Diluted preparations of FGFs were stabilized by CHAPS against the loss during storage. Amino acid sequence analysis was not disturbed by CHAPS. CHAPS was removable by reversed-phase high-performance liquid chromatography. These results indicate that CHAPS is useful as a non-cytotoxic stabilizing agent in purification of various kinds of bioactive polypeptides.


Hemihypoglossal nerve transfer in brachial plexus repair: technique and results.
In multiple avulsions of the brachial plexus, the search for extraplexal donor nerves in the hope of achieving motor neurotization is a major goal. We explored the possibility of using the hypoglossal nerve as a transfer point to reanimate muscles in the upper limb. The hypoglossal nerve was used as a donor nerve for neurotization in seven patients with avulsive injuries of the brachial plexus. The surgical technique--an end-to-side microsuture using approximately half of the nerve fascicles--is basically the same as that used in the hypoglossal nerve-facial nerve jump graft, which is a well-known technique in facial nerve reanimation. The recipient nerves were the suprascapular (two patients), the musculocutaneous (one patient), the posterior division of the upper trunk (two patients), and the medial contribution to the median nerve (two patients). In spite of a connection documented by electromyography and selective activation in three of seven patients, the functional results in our patients were extremely disappointing: no patient had an outcome better than M1 in the reinnervated muscles. This technique was of no help to the patients and thus has been abandoned at our institution.


The immunobiology of basal cell carcinoma: an in situ monoclonal antibody study.
A semi-quantitative, immunoperoxidase monoclonal antibody technique was used to study the mononuclear cells surrounding 32 basal cell carcinoma specimens from 30 patients. Tumours were analysed in subgroups based on recurrence, size and ulceration. T cell counts were high (greater than 3 out of 4) for all groups while T helper/inducer and T suppressor/cytotoxic cell counts were equal (approx. 2). Macrophage counts were low for all groups, about I X 2, while B cell and Ia positive cell counts were high (greater than 3). T/B cell and T helper/suppressor cell ratios approached one for the tumours as a whole as well as the sub-groups. The relative importance and contribution of cell mediated vs. humoral immunity in keeping basal cell carcinomas in check is discussed.