Presentation and Diagnosis of Hypersensitivity to Platinum Drugs.
Hypersensitivity reactions (HSRs) to platinum drugs are increasing due to their extensive use in a wide variety of malignancies and the repeated exposures in patients with increased life expectancy. Understanding the incidence of HSR to platinum drugs and associated risk factors can help with the diagnosis and may provide protection against severe HSRs. A thorough clinical history with identification of the typical and atypical symptoms, the relationship with the platin administration, and the number of previous exposures are the key to the diagnosis. An elevated serum tryptase at the time of the HSR indicates that IgE and/or mast cells/basophils were involved in the HSR. Skin testing to platinum drugs is a highly sensitive and specific diagnostic tool, which helps provide risk stratification and management recommendations. Platinum specific IgE measurement and basophil activation test (BAT) are emerging as new diagnostic tools and in combination with skin testing can help support the diagnosis and the cross-reactivity between the three most commonly used platinum drugs, namely carboplatin, cisplatin, and oxaliplatin.


Quantitative determination of paclitaxel in human plasma using semi-automated liquid-liquid extraction in conjunction with liquid chromatography/tandem mass spectrometry.
This paper describes a high-throughput sample preparation procedure combined with LC-MS/MS analysis to measure paclitaxel in human plasma. Paclitaxel and an internal standard were extracted from plasma by a semi-automated robotic method using liquid-liquid extraction. Thereafter compounds were separated on a RP C18 column. Detection was by a PE Sciex API 3000 mass spectrometer equipped with a TurboIonSpray interface. The compounds were detected in positive ion mode using the mass transition m/z 854.6-->286.2 and m/z 831.6-->263.2 for paclitaxel and the internal standard, respectively. The limit of quantitation for paclitaxel was 1 ng/ml with an imprecision of 5.2% following extraction of 0.1 ml of plasma. Linearity was confirmed over the whole calibration range (1-1000 ng/ml) with correlation coefficients higher than 0.99 indicating good fits of the regression models. The inter and intra-day precision was better than 9.5% and the accuracy ranged from 90.3 to 104.4%. The assay was simple, fast, specific and exhibited excellent ruggedness.


Assessment of the willingness of Nigerian Orthodontists to offer face-to-face orthodontic services to patients infected with Covid-19.
Efforts to contain the spread of the COVID-19 disease led to suspension of many services which caused orthodontists to resort to providing only emergency services. The aims of this study were to assess the willingness of Nigerian Orthodontists and Orthodontic resident doctors to treat patients infected with Covid-19 as well as to ascertain advice given regarding traumatic arch wires and aligners during the pandemic. This was a descriptive cross-sectional study. We included dentists who were Orthodontists and orthodontic resident doctors practicing in Nigeria. Data was collected via an online questionnaire (Google form) which was sent three times weekly throughout May/June, 2020 and analysed using SPSS version 23. Descriptive statistics were used to analyse the data. Forty-eight out of the 90 members of the WhatsApp group responded within the allotted time, giving a response rate of 53.3%. Out of a total of 48 respondents, 39 (81.3%) were unwilling to treat patients infected with COVID-19 during the pandemic. The major reason was fear of infection with the virus (12, 25.0%). Most of the respondents (41, 55.4%) suggested the use of wax for relief of trauma from arch wires. Majority of Orthodontists and orthodontic residents practicing in Nigeria expressed unwillingness to treat patients infected with COVID-19. Aligner therapy was proffered as an alternative to bracket therapy.


Cellular origin of human lymphotoxin and its purification.
The ability of various subsets of human mononuclear cells to produce human lymphotoxin (LT) was examined. Peripheral blood mononuclear cells were separated into OKT4+, OKT8+, and Leu-11a+ subpopulations by flow cytometry. Both OKT4+ and OKT8+ cells produced LT upon phytohemagglutinin (PHA) stimulation, but the OKT4+ T cells were the major source of LT. In contrast, Leu-11a+ cells failed to produce LT. The LT production and the proliferative response to PHA, of OKT4+ and OKT8+ cells, were inhibited by prostaglandin E2 and histamine. The LT, derived from PHA-stimulated peripheral blood lymphocytes, was purified by a procedure involving Blue-agarose, Con A-Sepharose chromatography, preparative gradient polyacrylamide gel electrophoresis and preparative sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The LT activity was recovered from SDS gel with major activity peak in the Mr 76,000 region and the minor activity peak in the Mr 24,000 region. The LT derived from 1788 lymphoblastoid cell line also showed heterogeneity on SDS gel. The activity was recovered from two peaks in the region of Mr 70,000 and 20,000.


Characterization of a neutral pectin lyase produced by Oidiodendron echinulatum MTCC 1356 in solid state fermentation.
A neutral pectin lyase produced by a new fungal strain Oidiodendron echinulatum MTCC 1356 under solid state fermentation using wheat bran as agro waste has been studied. The enzyme was purified by ammonium sulphate precipitation (30-60%), DEAE anion exchange and Sephadex G-100 column chromatographies. The SDS-PAGE and native PAGE revealed two bands of sizes 42 and 47 kDa. The enzyme was purified 37 fold with specific activity of 4.5 U/mg and 2.25% yield. The K(m) and V(max) values determined using citrus pectin were 1.2 mg/ml and 0.36 IU/min respectively. The pH and temperature optima were pH 7.0 and 50 °C, respectively. The pH stability was around 5.0 for 24 h at 20 °C. The purified enzyme retained maximum activity for 30 min upto 50 °C. The activation energy for thermal denaturation of the purified enzyme was found to be 60.0 kJ/Mol. The effects of various metal ions and protein inhibitors on enzyme activity have revealed total inhibition of the enzyme activity in the presence of Ag(+) and Cu(+) and KMnO(4) at 1 mM. The neutral pectin lyase showed retting of Crotalaria juncea fibre in the presence of EDTA.


Sustained clinical and structural benefit after joint distraction in the treatment of severe knee osteoarthritis.
Treatment of severe osteoarthritis (OA) in relatively young patients is challenging. Although successful, total knee prosthesis has a limited lifespan, with the risk of revision surgery, especially in active young patients. Knee joint distraction (KJD) provides clinical benefit and tissue structure modification at 1-year follow-up. The present study evaluates whether this benefit is preserved during the second year of follow-up. Patients included in this study presented with end-stage knee OA and an indication for total knee replacement (TKR); they were less than 60 years old with a VAS pain ≥60 mm (n = 20). KJD was applied for 2 months (range 54-64 days) and clinical parameters assessed using the WOMAC questionnaire and VAS pain score. Changes in cartilage structure were measured using quantitative MRI, radiography, and biochemical analyses of collagen type II turnover (ELISA). Average follow-up was 24 (range 23-25) months. Clinical improvement compared with baseline (BL) was observed at 2-year follow-up: WOMAC improved by 74% (P < 0.001) and VAS pain decreased by 61% (P < 0.001). Cartilage thickness observed by MRI (2.35 mm (95%CI, 2.06-2.65) at BL) was significantly greater at 2-year follow-up (2.78 mm (2.50-3.09); P = 0.03). Radiographic minimum joint space width (JSW) (1.1 mm (0.5-1.7) at BL) was significantly increased at 2-year follow-up as well (1.7 mm (1.1-2.3); P = 0.03). The denuded area of subchondral bone visualized by MRI (22% (95%CI, 12.5-31.5) at BL) was significantly decreased at 2-year follow-up (8% (3.6-12.2); P = 0.004). The ratio of collagen type II synthesis over breakdown was increased at 2-year follow-up (P = 0.07). Clinical improvement by KJD treatment is sustained for at least 2 years. Cartilage repair is still present after 2 years (MRI) and the newly formed tissue continues to be mechanically resilient as shown by an increased JSW under weight-bearing conditions.


Resin infiltration of enamel white spot lesions: An ultramorphological analysis.
There is not a clear understanding of the ultramorphology of enamel white spot lesions (WSLs). The purpose of this study is to characterize resin infiltration of enamel WSLs using electron microscopy. Enamel sections with sound enamel and WSLs were sectioned from extracted teeth and assigned to three groups: (a) left untreated; (b) etched with 15% hydrochloric acid (Icon-Etch); (c) restored with the resin infiltration sequence (Icon-Etch, Icon-Dry, and Icon-Infiltrant). Restored specimens were demineralized to obtain replicas. Observations were carried out under a field-emission scanning electron microscope. Icon-Etch resulted in an array of pits and funneled holes on the WSL. Replicas of WSLs depicted 0.5-6.0-μm-thick shaggy resin tags up to a depth of 465 μm. Enamel crystallites were enveloped with resin at the bottom of the WSL forming a hybrid layer. The resin infiltrant filled the spaces between the crystallites and resulted in an enamel hybrid layer. In addition to masking enamel WSLs, resin infiltration is able envelop residual enamel crystallites forming an enamel hybrid layer. This hybridization makes resin-embedded enamel more resistant to acid attack than sound enamel.


Engineering Cupriavidus necator H16 for enhanced lithoautotrophic poly(3-hydroxybutyrate) production from CO2.
A representative hydrogen-oxidizing bacterium Cupriavidus necator H16 has attracted much attention as hosts to recycle carbon dioxide (CO2) into a biodegradable polymer, poly(R)-3-hydroxybutyrate (PHB). Although C. necator H16 has been used as a model PHB producer, the PHB production rate from CO2 is still too low for commercialization. Here, we engineer the carbon fixation metabolism to improve CO2 utilization and increase PHB production. We explore the possibilities to enhance the lithoautotrophic cell growth and PHB production by introducing additional copies of transcriptional regulators involved in Calvin Benson Bassham (CBB) cycle. Both cbbR and regA-overexpressing strains showed the positive phenotypes for 11% increased biomass accumulation and 28% increased PHB production. The transcriptional changes of key genes involved in CO2-fixing metabolism and PHB production were investigated. The global transcriptional regulator RegA plays an important role in the regulation of carbon fixation and shows the possibility to improve autotrophic cell growth and PHB accumulation by increasing its expression level. This work represents another step forward in better understanding and improving the lithoautotrophic PHB production by C. necator H16.


Intragenic Variations in BTLA Gene Influence mRNA Expression of BTLA Gene in Chronic Lymphocytic Leukemia Patients and Confer Susceptibility to Chronic Lymphocytic Leukemia.
The aim of this study was to determine the association between polymorphisms in gene encoding B- and T-lymphocyte attenuator (BTLA) and susceptibility to chronic lymphocytic leukemia (CLL) and their influence on mRNA expression of BTLA gene in T and B cells from CLL patients (pts.). The following BTLA single-nucleotide polymorphisms (SNPs): rs2705511, rs1982809, rs9288952, rs76844316, rs16859633, rs9288953, rs2705535, rs1844089, rs2705565, rs2633580 were genotyped with use of TaqMan probes in 321 CLL pts. and in 470 controls. The mRNA levels of human BTLA were determined in subpopulations of T and B cells from 37 CLL patients with use of Applied Biosystems assays. Three SNPs: rs1982809, rs2705511 and rs9288953 were associated with susceptibility to CLL. The frequency of rs1982809[G] allele and rs2705511[C] allele carriers was higher in patients compared to the controls (0.51 vs. 0.41, OR 1.51, 95% CI 1.14-2.02, p = 0.004 and 0.56 vs. 0.44, OR 1.62, 95% CI 1.22-2.16, p = 0.0009, respectively). Furthermore, rs9288953[TT] genotype was overrepresented in CLL pts. compared to the controls (0.22 vs. 0.14, OR 1.74, 95% CI 1.20-2.53, p = 0.004). The evaluation of the influence of BTLA SNPs on BTLA mRNA expression in CLL pts. showed that the presence of rs1982809[G] allele was associated with lower median (±SD) BTLA mRNA expression in T cells (expressed as 2-delta Ct) in CLL pts. as compared to [AA] homozygotes (0.009 ± 0.013 vs. 0.026 ± 0.012, p = 0.03). Our results indicate that rs1982809 BTLA gene polymorphism is associated with mRNA expression level and that variations in the BTLA gene might be considered as potentially low-penetrating CLL risk factor.


Decision-making in home care package spending.
To understand the motivations and drivers of spending decisions made by home care package recipients, in the context of high levels of unspent funds. Qualitative study using 30 semi-structured interviews with 38 home care package clients and/or client representatives in rural and urban Victoria conducted between December 2018 and March 2019. Interview transcripts were analysed to determine themes. Spending decisions were influenced by assessment and entry experiences, knowledge and understanding of package funds, availability and acceptability of services, and attitudes to spending. Clients need confidence in their individual funds and their use in order to maximise the potential benefits. Clear, consistent communication at all stages, and additional supports to build consumer capability, will enable clients to better understand packages and their application. This in turn will optimise the capacity of home care package recipients to maximise their health and well-being.


Towards a More Connected Field of Community Development Education: Encouraging Community Well-Being Through an Engaged Symposium.
As society continues to shift and progress, the field of community development becomes more salient to the well-being and sustainability of rural and urban communities. Globalization, global pandemics, technological change, and devolution are among several issues impacting the well-being of rural townships and urban neighborhoods nationally and globally. Post-secondary community development degree programs may be vital in developing individuals to address these unique issues. However, due to the multidisciplinary nature of the field, there has been a lack of communication and consensus among educators regarding the necessary skills, theories, and practices to teach in these programs. In 2017, a foundational initiative was implemented to examine community development education higher education programs across the United States. A unique component of this initiative was the inaugural Community Development Education Symposium. This innovative symposium engaged many community development educators from a variety of fields in an intense, purposeful three day event. This article captures the synergy of bringing together community development educators in a uniquely structured symposium, to network, share expertise, and ultimately build a Community of Practice (CoP) towards a more connected field of community development education and ultimately enhancing the well-being of communities through their student graduates.


Spontaneous closure of esophageal tear in Boerhaave's syndrome.
An unusual case of spontaneous rupture of the esophagus is reported in which the patient survived without surgical closure or drainage procedure. The site of perforation and subsequent closure are documented radiologically.


Two-dimensional benzyldimethyl-n-hexadecylammonium chloride/SDS-PAGE for membrane proteomics.
Despite the importance of membranes in any living system, the global analysis of membrane subproteomes is still a common obstacle. In particular, the widely used 2-DE technique consisting of IEF in the first dimension and SDS-PAGE in the second dimension has some major drawbacks regarding the separation of hydrophobic proteins. Therefore, we applied an alternative electrophoretic technique for separating membrane proteins: two-dimensional BAC/SDS electrophoresis (2-DB) using the cationic detergent benzyldimethyl-n-hexadecylammonium chloride in the first and the anionic detergent SDS in the second dimension. The use of 2-DB resulted in an improved separation of hydrophobic proteins. Thus, extremely hydrophobic proteins such as cytochrome-c oxidase subunit I with a grand average hydrophobicity (GRAVY) index of 0.74 and a total of 12 known transmembrane domains (TMD) or Sec61alpha with a GRAVY index of 0.56 and a total of ten known TMD could be identified by MS/MS analyses of protein spots derived from 2-DB gels.


Acute and subacute oral administration of mycotoxin deoxynivalenol exacerbates the pro-inflammatory and pro-pruritic responses in a mouse model of allergic dermatitis.
Deoxynivalenol (DON) contamination in food is a public health concern; however, the effect of DON exposure on immune disorders including allergies remains unclear. The aim of this study is to elucidate the effect of oral exposure to DON on pro-inflammatory and pro-pruritic responses in a mouse model of allergic dermatitis, which was generated by topical application of toluene-2,4-diisocyanate (TDI), a hapten that induces type-2 helper T cells. To evaluate acute exposure to DON, the mice were orally administered vehicle alone, 0.1 mg/kg DON, or 0.3 mg/kg DON 48, 24, and 1 h before the final challenge with TDI. To study subacute exposure, the mice were fed DON-contaminated rodent diet (0.3 ppm) during the experimental period. After the itch behavior and ear-swelling response were monitored, the serum, auricular lymph node, and skin tissue were collected for analyzing immunocyte differentiation, cytokine determination, and histological changes. Acute oral administration of DON significantly enhanced pro-inflammatory responses including ear-swelling response, immunocyte infiltration, and cytokine productions. Histological evaluation supported the occurrence of pro-inflammatory responses. In contrast, acute DON exposure only slightly increased itch behavior. Subacute oral exposure to DON significantly up-regulated the inflammatory responses, but showed almost no effect on pruritic response. In vitro evaluation in dendritic cells and keratinocytes indicated that DON pre-exposure induced a dose-dependent significant increase in cytokine production. Our results imply that both acute and subacute exposures to DON are associated with pro-inflammatory responses in cutaneous allergy.


TIMP-2 fusion protein with human serum albumin potentiates anti-angiogenesis-mediated inhibition of tumor growth by suppressing MMP-2 expression.
TIMP-2 protein has been intensively studied as a promising anticancer candidate agent, but the in vivo mechanism underlying its anticancer effect has not been clearly elucidated by previous works. In this study, we investigated the mechanism underlying the anti-tumor effects of a TIMP-2 fusion protein conjugated with human serum albumin (HSA/TIMP-2). Systemic administration of HSA/TIMP-2 effectively inhibited tumor growth at a minimum effective dose of 60 mg/kg. The suppressive effect of HSA/TIMP-2 was accompanied by a marked reduction of in vivo vascularization. The anti-angiogenic activity of HSA/TIMP-2 was directly confirmed by CAM assays. In HSA/TIMP-2-treated tumor tissues, MMP-2 expression was profoundly decreased without a change in MT1-MMP expression of PECAM-1-positive cells. MMP-2 mRNA was also decreased by HSA/TIMP-2 treatment of human umbilical vein endothelial cells. Zymographic analysis showed that HSA/TIMP-2 substantially decreased extracellular pro-MMP-2 activity (94-99% reduction) and moderately decreased active MMP-2 activity (10-24% reduction), suggesting MT1-MMP-independent MMP-2 modulation. Furthermore, HSA/TIMP-2 had no effect on in vitro active MMP-2 activity and in vivo MMP-2 activity. These studies show that HSA/TIMP-2 potentiates anti-angiogenic activity by modulating MMP-2 expression, but not MMP-2 activity, to subsequently suppress tumor growth, suggesting an important role for MMP-2 expression rather than MMP-2 activity in anti-angiogenesis.


Safety of tropicamide and phenylephrine in pupillary mydriasis for cataract surgery.
A prospective study to evaluate the adverse cardiovascular effects of topical phenylephrine and tropicamide used for pupillary mydriasis before cataract surgery. A total of 517 consecutive eyes in 517 patients subjected to routine 0.8% tropicamide and 5% phenylephrine eye drops before undergoing cataract surgery (phacoemulsification and manual small-incision surgery) under local or topical anesthesia in a medical college hospital were analyzed. No untoward cardiovascular effects were seen. The increase in blood pressure after 0.8% tropicamide and 5% phenylephrine eye drops was statistically significant, but it was not relevant as it was within clinically permissible limits. The combination of 0.8% tropicamide and 5% phenylephrine eye drops is a safe and effective option for pupillary mydriasis before cataract surgery.


Acute intraoperative choroidal effusion.
We studied five cases of acute choroidal effusion that occurred during intraocular surgery and one case of a localized choroidal hemorrhage. Acute intraoperative choroidal effusion is a poorly recognized surgical complication and may precede expulsive hemorrhage in many cases. The treatment is immediate closure of the wound, after which the intraocular pressure may rise to 80 mm Hg or more. However, the pressure will return to normal after 15 to 30 minutes and the area of effusion will remain localized. Scleral puncture to drain fluid is not indicated if the wound can be closed before there is a prolapse of the iris or other intraocular tissues.


Determination of the optical purity and absolute configuration of threo-methylphenidate by proton nuclear magnetic resonance spectroscopy with chiral solvating agent.
The direct determination of both the optical purity and absolute configuration of threo-methylphenidate has been accomplished in a simple, specific, and accurate manner by 1H-NMR spectroscopy. The enantiomeric resonances of threo-methylphenidate were effectively resolved in CDCl3 solution by the addition of the chiral solvating agents (R)-(-)- or (S)-(+)-2,2,2,-trifluoro-1-(9-anthryl)ethanol. Optical purities were determined on the basis of the intensities of the enantiomeric ester methyl proton resonances; the assignment of enantiomeric configurations was based on the relative field positions of these resonances and the examination of molecular models. The analysis of synthetic enantiomeric mixtures of threo-methylphenidate by the proposed NMR method resulted in assay values that agreed closely with the known quantities of each enantiomer in the mixtures tested. The mean +/- SD recovery value for the (2S,2'S)-(-)-threo-enantiomer, amounting to 99.9 +/- 0.6% of added (n = 10), correlated well with that previously found by 1H-NMR spectroscopy with a chiral Eu(III) shift reagent. However, the present approach is simpler, shows less reliance on reagents and solvents of a high purity, and does not require strict anaerobic working conditions.


Biology of the protein kinase C family.
Protein kinase C (PKC) is composed of a family of isozymes that transduce signals of certain hormones, growth factors, lectins, and neurotransmitters. This review addresses the role of PKC in the regulation of cellular proliferation and its disorders. PKC is directly activated in vivo by the second messenger diacylglycerol, a lipid produced by phospholipase C-catalyzed hydrolysis of phosphatidylinositol and polyphosphoinositides. Diacylglycerol activates PKC by reducing the enzyme's requirement for Ca2+. Phorbol ester tumor promoters and related agents potently activate PKC by a mechanism analogous to that of diacylglycerol, providing evidence that PKC activation is a critical event in tumor promotion. However, the role of PKC activation in tumor promotion is not entirely clear. For example, bryostatin is a potent PKC activator that antagonizes phorbol ester-mediated tumor promotion, and mezerein is a second-stage tumor promoter that potently activates PKC. In addition to studies concerned with tumor promotion, studies of oncogene action also indicate a role for PKC in carcinogenesis. A number of plasma membrane-associated oncogene products and related proteins are PKC substrates, and PKC activation leads to induction of the expression of oncogenes that code for nuclear proteins. PKC is implicated in human breast and colon carcinogenesis. Tumor-promoting bile acids activate PKC, and PKC expression studies in rat colonic epithelial cells and human breast cancer cells indicate a positive role for PKC in the proliferation of the cells. Altered expression of PKC in human colon and breast tumors indicates that PKC isozymes may be useful markers for these diseases.


Attitudes and Perception of the REFLECT Communication Curriculum for Clinical Oncology Graduate Medical Education.
Communication and interpersonal skills are essential components of oncology patient care. The REFLECT (Respect, Empathy, Facilitate Effective Communication, Listen, Elicit Information, Compassion, and Teach Others) curriculum is a novel framework to improve and refine physician/patient interactions for oncology graduate medical trainees. We seek to evaluate the attitudes and perceptions of the REFLECT communication curriculum among oncology trainees. Seven-question and 8-question Likert scale surveys (1 = not beneficial and 5 = beneficial) were distributed to resident/fellow participants and faculty mentors, respectively. Questions asked trainees and faculty about their perceptions of improvement in communication, handling of stressful situations, the value of the curriculum, and overall impression of the curriculum. Descriptive statistics determined the survey's baseline characteristics and response rates. Kruskal-Wallis rank sum tests were used to compare the distribution of continuous variables. Thirteen resident/fellow participants completed the participant survey. Six (43.6%) Radiation Oncology trainees and 7 (58.3%) Hematology/Oncology fellows completed the trainee survey. Eight (88.9%) Radiation Oncologists and 1 (11.1%) Medical Oncologist completed the observer survey. Faculty and trainees generally felt that the curriculum increased communication skills. Faculty responded more favorably to the program's impact on communication skills (median 5.0 vs. 4.0, p = 0.008). Faculty were more assertive about the curriculum's capabilities to improve a learner's ability to handle stressful situations (median 5.0 vs. 4.0, p = 0.003). Additionally, faculty had a more favorable overall impression of the REFLECT curriculum than the residents/fellows (median 5.0 vs. 4.0, p < 0.001). Radiation Oncology residents felt more strongly that the curriculum enhanced their ability to handle stressful topics, compared to Heme/Onc fellows (median 4.5 vs. 3.0, range 1-5, p = 0.379). Radiation Oncology trainees felt more consistently that the workshops improved their communication skills, compared to Heme/Onc fellows (median 4.5 vs. 3.5, range 1-5, p = 0.410). The overall impression between Rad Onc resident and Heme/Onc fellows was similar (median 4.0, p = 0.586). Conclusions: Overall, the REFLECT curriculum enhanced communication skills of trainees. Oncology trainees and faculty physicians feel that the curriculum was beneficial. As interactive skills and communication is critical to build positive interactions, further work is needed to improve the REFLECT curriculum.


Photophysical properties of Pr(III) and Er(III) complexes of poly(pyrazolyl)borates.
The complexes [M(L(1))(2)(NO(3))] and [M(L(2))(NO(3))(2)](M = Pr, Er; L(1)= the tetradentate ligand dihydrobis-[3-(2-pyridyl)pyrazolyl]borate; L(2)= the hexadentate ligand hydrotris-[3-(2-pyridyl)pyrazolyl]borate) were prepared and their structural and photophysical properties studied. All complexes are 10-coordinate. Crystallographic analysis of [M(L(1))(2)(NO(3))](M = Pr, Er) showed that for the smaller Er(iii) ions steric congestion at the metal centre results in two of the Er-N(pyridyl) distances being particularly long, which does not occur with the larger Pr(iii) ion that is better able to accommodate 10-fold coordination. On UV irradiation, both Pr(iii) complexes show, in the visible region of their luminescence spectra, transitions originating from both the (3)P(0) level (at ca. 21,000 cm(-1)) and the (1)D(2) level (at ca. 17,000 cm(-1)), a consequence of the fact that the lowest triplet state of the coordinated pyrazolylborate ligands lies at ca. 24,000 cm(-1) in each case so is high enough in energy to populate both levels. This contrasts with Pr(iii) complexes based on diketonate ligands in which the lower triplet energies of the ligands result in emission from the (1)D(2) level only. At longer wavelengths, near-infrared luminescence arising from the (1)D(2) emissive level is observed with lifetimes (in both the solid state and solution) being in the range 50-110 ns. For both Er(iii) complexes, luminescence at 1530 nm occurs following UV excitation of ligand-centred transitions. In CH(2)Cl(2) both complexes gave dual-exponential luminescence, with the major component having a lifetime characteristic of an intact Er(iii) complex (approximately 1.5 micros) and the minor component being much shorter lived (0.2-0.5 micros), suggestive of a species in which a ligand is partially detached and the metal is solvated, with the two forms interconverting slowly. This behaviour is consistent with the steric congestion and long M-N(pyridyl) bonds that were observed in [Er(L(1))(2)(NO(3))]. In the solid state both Er(iii) complexes gave very weak luminescence, which could be fitted to a single exponential decay with a lifetime similar to the longer-lived of the solution components.


Metal work-function-dependent barrier height of Ni contacts with metal-embedded nanoparticles to 4H-SiC.
Metal, typically gold [Au], nanoparticles [NPs] embedded in a capping metal contact layer onto silicon carbide [SiC] are considered to have practical applications in changing the barrier height of the original contacts. Here, we demonstrate the use of silver [Ag] NPs to effectively lower the barrier height of the electrical contacts to 4H-SiC. It has been shown that the barrier height of the fabricated SiC diode structures (Ni with embedded Ag-NPs) has significantly reduced by 0.11 eV and 0.18 eV with respect to the samples with Au-NPs and the reference samples, respectively. The experimental results have also been compared with both an analytic model based on Tung's theory and physics-based two-dimensional numerical simulations.


Sublingual grass and ragweed immunotherapy: Clinical considerations-a PRACTALL consensus report.
Sublingual allergen immunotherapy provides a new option for patients with allergic rhinitis in the United States. The efficacy of these sublingual immunotherapy tablets in the treatment of allergic rhinitis has been firmly established in large multicenter clinical trials. In addition, the clinical benefits of sublingual immunotherapy might persist after treatment is discontinued. Local reactions, such as gastrointestinal or oropharyngeal symptoms, are common. However, severe anaphylaxis is rare, and therefore the immunotherapy tablets can be administered at home. Sublingual immunotherapy for allergic rhinitis has been used successfully for years in Europe, and these products might be appropriate for patients who do not do well with standard drug therapy or for those who prefer a disease-modifying approach.


Ginsenoside Rg3 inhibits endothelial progenitor cell differentiation through attenuation of VEGF-dependent Akt/eNOS signaling.
Endothelial progenitor cells (EPCs) play a critical role both in vascular repair after cell transplantation for ischemic diseases and in the growth of early tumors by intervening with the angiogenic switch during tumor progression. This paper reports on the effect of ginsenoside Rg3 in EPCs as a candidate angiogenesis inhibitor for in vitro functional assays. CD34⁺ cells were isolated from human cord blood and the study investigated whether or not ginsenoside Rg3 regulated EPC bioactivities including cell proliferation, differentiation, migration and tube formation. Although ginsenoside Rg3 did not affect the ex vivo expansion of CD34 and/or KDR (VEGFR2) stem/progenitor cells, treatment with ginsenoside Rg3 led to a significant decrease in CD34-expressing cells, specifically the absolute number of expanded CD34⁺ cells. Importantly, a significantly decreased number of EPC colony-forming units among human cord blood-derived CD34⁺ cells was observed, implying that ginsenoside Rg3 inhibited EPC differentiation, in particular, the commitment to primitive EPC colonies (the early stage of EPC differentiation). Moreover, treatment of CD34-derived EPCs with ginsenoside Rg3 resulted in the attenuation of VEGF-dependent Akt/eNOS signaling as well as the inhibition of migration and tube formation. In conclusion, this study provides in vitro evidence for ginsenoside Rg3 as a potential therapeutic molecule, specifically as an angiogenesis inhibitor that functions by attenuating EPC bioactivities.


Augmentation of SIV DNA vaccine-induced cellular immunity by targeting the 4-1BB costimulatory molecule.
DNA vaccines are effective at inducing antigen-specific cellular immune responses. Approaches to improve these responses, however, are needed. We examined the effect of stimulating 4-1BB, an activation-inducible T-cell costimulatory receptor, by intravenously co-administering anti-human 4-1BB monoclonal antibody (mAb) in DNA-immunized cynomolgus macaques. Three groups of six cynomolgus macaques were immunized intramuscularly with a DNA vaccine encoding SIV Gag antigen (pSIVgag) at weeks 0, 4 and 8. At days 12, 15, and 19, six macaques received anti-4-1BB 4E9 mAb and six macaques received anti-4-1BB 10C7 mAb. Treatment with 10C7 mAb led to a significant augmentation of SIV Gag-specific IFN-gamma, granzyme B and perforin responses. Treatment with humanized 4E9 mAb also resulted in an enhancement of SIV Gag-specific cellular responses but the magnitude was lower compared to animals receiving 10C7 mAb. These responses persisted up to week 40 and were mostly mediated by CD8(+) T cells. Treatment with anti-4-1BB mAb was more effective in driving the CD8(+) T cells toward a more differentiated CCR7(-)/CD45RA(+) effector state. This study demonstrates that targeting the 4-1BB molecule in vivo results in an enhanced and long-lasting cellular immune response. 4-1BB stimulation may be a promising approach to enhance the effectiveness of DNA vaccines.


Do In-Hospital Rothman Index Scores Predict Postdischarge Adverse Events and Discharge Location After Total Knee Arthroplasty?
There have been efforts to reduce adverse events and unplanned readmissions after total joint arthroplasty. The Rothman Index (RI) is a real-time, composite measure of medical acuity for hospitalized patients. We aimed to examine the association among in-hospital RI scores and complications, readmissions, and discharge location after total knee arthroplasty (TKA). We hypothesized that RI scores could be used to predict the outcomes of interest. This is a retrospective study of an institutional database of elective, primary TKA from July 2018 until December 2019. Complications and readmissions were defined per Centers for Medicare and Medicaid Services. Analysis included multivariate regression, computation of the area under the curve (AUC), and the Youden Index to set RI thresholds. The study cohort's (n = 957) complications (2.4%), readmissions (3.6%), and nonhome discharge (13.7%) were reported. All RI metrics (minimum, maximum, last, mean, range, 25th%, and 75th%) were significantly associated with increased odds of readmission and home discharge (all P < .05). RI scores were not significantly associated with complications. The optimal RI thresholds for increased risk of readmission were last ≤ 71 (AUC = 0.65), mean ≤ 67 (AUC = 0.66), or maximum ≤ 80 (AUC = 0.63). The optimal RI thresholds for increased risk of home discharge were minimum ≥ 53 (AUC = 0.65), mean ≥ 69 (AUC = 0.65), or maximum ≥ 81 (AUC = 0.60). RI values may be used to predict readmission or home discharge after TKA.


Inhibitors for the hepatitis C virus RNA polymerase explored by SAR with advanced machine learning methods.
Hepatitis C virus (HCV) is a global health challenge, affecting approximately 200 million people worldwide. In this study we developed SAR models with advanced machine learning classifiers Random Forest and k Nearest Neighbor Simulated Annealing for 679 small molecules with measured inhibition activity for NS5B genotype 1b. The activity was expressed as a binary value (active/inactive), where actives were considered molecules with IC50 ≤0.95 μM. We applied our SAR models to various drug-like databases and identified novel chemical scaffolds for NS5B inhibitors. Subsequent in vitro antiviral assays suggested a new activity for an existing prodrug, Candesartan cilexetil, which is currently used to treat hypertension and heart failure but has not been previously tested for anti-HCV activity. We also identified NS5B inhibitors with two novel non-nucleoside chemical motifs.


Temporomandibular Joint Fibrocartilage Contains CD105 Positive Mouse Mesenchymal Stem/Progenitor Cells with Increased Chondrogenic Potential.
A specific type of mesenchymal stem/progenitor cells (MSPCs), CD105+ is reported to aid in cartilage regeneration through TGF-β/Smad2-signalling. The purpose of this study was to identify and characterize CD105+ MSPCs in temporomandibular joint (TMJ) cartilage. MSPCs were isolated from mouse TMJ condyle explants and evaluated for their clonogenicity and pluripotential abilities. MSPC were examined for CD105 antigen using immunohistochemistry and flow cytometry. Immunohistochemistry revealed presence of CD105+ MSPCs in the proliferative zone of condyle's cartilage. Only 0.2% of isolated MSPCs exhibited CD105, along with the stem cell surface markers CD44 and Sca-1. In CD105+ MSPCs, intracellular immunostaining revealed significantly higher (p < 0.05) protein levels of collagen type 1, 2, proteoglycan 4. Ability for chondrogenic differentiation was found to be significantly higher (p < 0.05) after 4 weeks compared to CD105- cells, using alcian blue staining. CD105+ cells were found to resemble an early MSPC subgroup with significantly higher gene expression of biglycan, proteoglycan 4, collagen type 2, Gli2, Sox5 (p < 0.001) and Sox9 (p < 0.05). In contrast, significantly lower levels of Runx2 (p < 0.05), Osterix, Trps1, Col10a1 (p < 0.01), Ihh (p < 0.001) related to chondrocyte senescence and commitment to osteogenic lineage, were observed compared to CD105- cells. The study showed the existence of a CD105+ MSPC subgroup within TMJ fibrocartilage that may be activated to aid in fibrocartilage repair.


Effect of Pilates Matwork exercises on posterior chain flexibility and trunk mobility in school children: A randomized clinical trial.
To investigate the effects of an exercise program, based on the Pilates Matwork method, on posterior chain flexibility and trunk mobility in healthy school age children. The study was a parallel-group randomized clinical trial. The participants were randomly assigned to groups: Pilates Group (PG) and Control Group (CG). The program was developed at the Early Childhood Education Institute, Londrina-PR. 43 children with age between eight to 12 years, no prior knowledge of the Pilates method, and no exercise training in the last six months. Four months of twice a week 50 min Pilates Matwork exercises were administered. Flexibility and mobility, assessed using the sit-and-reach test, fingertip-to-floor test and photogrammetry. The assessors were blinded to the allocation of participants. Three children were excluded before randomization and 40 were randomized (PG n = 20; CG n = 20).12 children were excluded during the protocol (PG n = 7; CG n = 5) and included in the intention to treat analysis. No significant difference between groups was observed for flexibility measures. There was a significant difference in the following outcomes for the PG: distance reached in the sit-and-reach test between pre-test (median 14.25[11.25-28.38]) and post-test (median 20.25[12.00-29.63]) (ES = 0.29, SRM = 0.73); Posterior angle of the knee in the fingertip-to-floor test between pre-test (median 191.60[187.20-191.60]) and post-test (median 189.00[185.90-191.50]) (ES = 0.56, SRM = 0.54). There were no differences in posterior chain flexibility and trunk mobility between school age children who underwent Pilates Matwork exercises and the control. However, children who participated in the exercise program showed improvement in some results of flexibility. Brazilian Registry of Clinical Trials (REBEC) (N° RBR-8t5p7d).


Context-Oriented Model Development in Psychotherapy Planning ('COMEPP'): a useful adjunct to diagnosis and therapy of severe personality disorders.
Pathogenous interpersonal (e.g. interfamilial) relationships and reference styles can compromise treatment efforts in severely disturbed (i.e. psychotic or borderline) patients. The integration of family- and individual-centred starting points may be useful in establishing interdisciplinary treatment concepts in these patients. Context-Oriented Model Development in Psychotherapy Planning (COMEPP) represents a diagnostic and therapy planning process, integrating both systemic and psychoanalytic conceptualizations. COMEPP is exemplified by the case of a young man with psychotic personality disorder who had previously been unresponsive to pharmacological and psychological treatment. After psycho-dynamical conflicts (i.e. primitive projective processes from the patient's mother to her son) had been elucidated during the COMEPP process, a sufficient treatment setting could be established. COMEPP provides a psychotherapeutical approach to treatment planning on case-specific premises and may serve as an adjunct to concomitant pharmacological and psychological treatment strategies in so-called 'therapy refractory' patients.


MicroRNA-503 inhibits the G1/S transition by downregulating cyclin D3 and E2F3 in hepatocellular carcinoma.
Increasing evidence indicates that deregulation of microRNAs (miRNAs) is involved in tumorigenesis. Downregulation of microRNA-503 has been observed in various types of diseases, including cancer. However, the biological function of miR-503 in hepatocellular carcinoma (HCC) is still largely unknown. In this study we aimed to elucidate the prognostic implications of miR-503 in HCC and its pathophysiologic role. Quantitative reverse transcriptase polymerase chain reaction was used to evaluate miR-503 expression in HCC tissues and cell lines. Western blotting was performed to evaluate the expression of the miR-503 target genes. In vivo and in vitro assays were performed to evaluate the function of miR-503 in HCC. Luciferase reporter assay was employed to validate the miR-503 target genes. miR-503 was frequently downregulated in HCC cell lines and tissues. Low expression levels of miR-503 were associated with enhanced malignant potential such as portal vein tumor thrombi, histologic grade, TNM stage, AFP level and poor prognosis. Multivariate analysis indicated that miR-503 downregulation was significantly associated with worse overall survival of HCC patients. Functional studies showed miR-503 suppressed the proliferation of HCC cells by induction of G1 phase arrest through Rb-E2F signaling pathways, and thus may function as a tumor suppressor. Further investigation characterized two cell cycle-related molecules, cyclin D3 and E2F3, as the direct miR-503 targets. Our data highlight an important role for miR-503 in cell cycle regulation and in the molecular etiology of HCC, and implicate the potential application of miR-503 in prognosis prediction and miRNA-based HCC therapy.


Primary standardisation of technetium-99m by liquid scintillation coincidence counting.
Technetium-99m was standardised by the 4π(LS)ce-γ coincidence extrapolation method. Sensitivity of the 4π(LS) channel to two types of radiation, namely conversion electrons and γ-rays, resulted in incorrect activity values being obtained when this was not adequately accounted for. Measurements were more robust when the LS detection efficiency was optimised, and when a γ-window setting was used that monitored the combined LS efficiency for conversion electrons and γ-rays. The primary standard was internationally compared through participation in the BIPM.RI(II)-K4.Tc99m key comparison.


Glu496 to Ala polymorphism in the P2X7 receptor impairs ATP-induced IL-1 beta release from human monocytes.
Priming of monocytes with LPS produces large quantities of intracellular, biologically inactive IL-1beta that can be processed and released by subsequent activation of the P2X7 receptor by extracellular ATP. We examined whether a loss-of-function polymorphism of the human P2X7 receptor (Glu496Ala) impairs this process. Both ATP-induced ethidium+ uptake and ATP-induced shedding of L-selectin (CD62L) were nearly absent in monocytes from four subjects homozygous for Glu496Ala confirming that this polymorphism impairs P2X7 function. The level of ATP-induced IL-1beta released in 2 h from LPS-activated whole blood from homozygous subjects was 50% of that from wild-type samples. A more marked defect in IL-1beta release was observed from LPS-activated monocytes of homozygous subjects which was only 22% of that released from wild-type monocytes after a 30-min incubation with ATP. However, after a 60-min incubation with ATP, the amount of IL-1beta released from homozygous monocytes was 70% of that released from wild-type monocytes. Incubation of monocytes of either genotype with nigericin resulted in a similar release of IL-1beta. Western blotting demonstrated that ATP induced the release of mature 17-kDa IL-1beta from monocytes, and confirmed that this process was impaired in homozygous monocytes. Finally, ATP-induced 86Rb+ efflux was 9-fold lower from homozygous monocytes than from wild-type monocytes. The results indicate that ATP-induced release of IL-1beta is slower in monocytes from subjects homozygous for the Glu496Ala polymorphism in the P2X7 receptor and that this reduced rate of IL-1beta release is associated with a lower ATP-induced K+ efflux.


Adherence of uropathogenic E. coli to differentiated human uroepithelial cells grown in vitro.
A quantitative in vitro model to measure E. coli adherence to differentiated human uroepithelial cells has been developed. Primary cultures of uroepithelial cells were initiated from normal ureteral epithelium. Adherence of uropathogenic 3H-labelled Escherichia coli to postconfluent human uroepithelial cells was directly related to the bacteria:epithelial cell ratio during incubation. Bacterial attachment was inhibited either by mannose or by urine containing anti-E. coli antibodies. Transmission electron microscopy showed that epithelial cells differentiated in vitro to resemble normal uroepithelium in vivo. Furthermore, electron microscopy showed specific adherence of bacteria to the glycocalyx of microvilli of the superficial uroepithelial cells in vitro in a manner which closely mimics the in vivo interaction. This model of bacterial adherence permits in vitro analysis of adhesin-receptor interactions between uropathogenic E. coli and a layer of viable uroepithelial cells similar to those lining the bladder.


Combination of epigallocatechin 3 gallate and curcumin improves D-galactose and normal-aging associated memory impairment in mice.
Previously, we observed curcumin improves aging-associated memory impairment in D-galactose (D-gal) and normal-aged (NA) mice. Evidence showed that multiple agents can be used in managing aging-induced memory dysfunction, drawn by the contribution of several pathways. Curcumin and Epigallocatechin 3 gallate (EGCG) combination substantially reduced the oxidative stress that commonly mediates aging. This study examined the combined effect of EGCG and curcumin on memory improvement in two recognized models, D-gal and normal-aged (NA) mice. The co-administration of EGCG and curcumin significantly (p < 0.05) increased retention time detected by passive avoidance (PA) and freezing response determined in contextual fear conditioning (CFC) compared to the discrete administration of EGCG or curcumin. Biochemical studies revealed that the combination of EGCG and curcumin remarkably ameliorated the levels (p < 0.05) of glutathione, superoxide dismutase, catalase, advanced oxidation protein products, nitric oxide, and lipid peroxidation compared to the monotherapy of EGCG or curcumin in mice hippocampi. The behavioral and biochemical studies revealed that the combination of EGCG and curcumin showed better improvement in rescuing aging-associated memory disorders in mice. EGCG and curcumin combination could serve as a better choice in managing aging-related memory disorders.


Metal-Free Synthesis of 4-Chloroisocoumarins by TMSCl-Catalyzed NCS-Induced Chlorinative Annulation of 2-Alkynylaryloate Esters.
4-Chloroisocoumarins can be conveniently prepared from 2-alkynylaryloate esters via the activation of alkynes by electrophilic chlorine, generated in situ from N-chlorosuccinimide (NCS) in the presence of 10 mol % trimethylsilyl chloride (TMSCl), which leads to 6-endo-dig-selective chlorinative annulation to give the desired products in moderate to quantitative yields. The procedure employs readily available reagents and can be conveniently carried out on a wide scope of substrates under mild conditions (0 °C to rt). Furthermore, the reaction is scalable for gram-scale preparation of 4-chloroisocoumarins. Additionally, 4-bromo- and 4-iodoisocoumarins can be prepared in moderate to good yields by replacing NCS with N-bromosuccinimide (NBS) and N-iodosuccinimide (NIS), respectively.


The role of stromal immune microenvironment in the progression of ductal carcinoma in situ (DCIS) to invasive breast cancer.
The first aim of the study was to compare the scores and types of stromal immune cells in 30 patients with primary DCIS and in the same patients after invasive breast recurrence in order to assess possible differences in both during tumor progression. The second aim was to evaluate possible differences in stromal cells of 30 patients with primary DCIS before progression and in the control group of 11 DCIS patients without recurrence during long-term follow-up. Evaluation of tumor-infiltrating lymphocytes (TILs) and immunohistochemical stains for immune cell markers CD4, CD8, CD20, CD138, FOXP3, CD163 and TGF beta was performed on the stroma of primary DCIS before progression, invasive breast cancer of the same patients after progression and DCIS without progression. The comparison of stromal cells in 30 patients with initial DCIS and its invasive recurrence revealed an increased level of CD20 + immune cells (median score 5% vs. 17%, respectively, p < 0.001) and CD163 + cells (median score 1% vs. 5%, respectively, p < 0.001) in invasive breast cancer. The comparison of stromal cells in 30 patients with initial DCIS before recurrence and the control group of 11 patients with DCIS without recurrence showed statistically significant difference for CD138 + cells, which were more prevalent in patients with worse prognosis (median score 0 vs. 2%, respectively, p < 0.001). No similar relationship was found for the other tested cells as well as for TGF-beta. CD138 + immune cells that were more prevalent in patients with a worse prognosis should be explored in further studies to confirm or exclude their role as a potential biological marker of DCIS invasive recurrence.


The Stockholm classification of stillbirth.
To design and validate a classification system for audit groups working with stillbirth. The classification includes well-defined primary and associated conditions related to fetal death. Descriptive. All delivery wards in Stockholm. Stillbirths from 22 completed weeks in Stockholm, Sweden. Parallel to audit work, the Stockholm stillbirth group has developed a classification of conditions related to stillbirth. The classification has been validated. The classification and the results of the validation are presented. The classification with 17 groups identifying underlying conditions related to stillbirth (primary diagnoses) and associated factors which may have contributed to the death (associated diagnoses) is described. The conditions are subdivided into definite, probable and possible relation to the death. An evaluation of 382 cases of stillbirth during 2002-2005 resulted in 382 primary diagnoses and 132 associated diagnoses. The most common conditions identified were intrauterine growth restriction/placental insufficiency (23%), infection (19%), malformations/chromosomal abnormalities (12%). The 'unexplained' group together with the 'unknown' group comprised 18%. Validation was done by reclassification of 95 cases from 2005 by six investigators. The overall agreement regarding primary diagnosis was substantial (kappa=0.70). The Stockholm classification of stillbirth consists of 17 diagnostic groups allowing one primary diagnosis and if needed, associated diagnoses. Diagnoses are subdivided according to definite, probable and possible relation to stillbirth. Validation showed high degree of agreement regarding primary diagnosis. The classification can provide a useful tool for clinicians and audit groups when discussing cause and underlying conditions of fetal death.


MRI Features as Prognostic Factors in Myxofibrosarcoma: Proposal of MRI Grading System.
Myxofibrosarcoma (MFS) is a common soft tissue sarcoma in the elderly patients with both clinical and magnetic resonance imaging (MRI) peculiar features: very high recurrence rate, relatively low risk of distant metastases. On MRI it shows an infiltrative pattern ("tail sign") and high myxoid matrix content with water-like appearance on fluid-sensitive sequences. Due to these unusual characteristics, we propose a specific MRI grading system to stratify the risk of local recurrence (LR) and offer other prognostic information. Two expert radiologists retrospectively and blindly reviewed preoperative MRI of 150 patients affected by MFS of the extremities treated at a single Institution. Myxoid matrix component and contrast enhancement of the tumor were evaluated and graded with a semiquantitative method. The presence of an infiltrative pattern, the depth of the tumor (deep and/or superficial) and tumor sizes were also recorded. MRI features were analyzed separately and correlated to LR risk, sarcoma specific survival and distant metastases rate. Then, according to the statistical significance of the correlation between MRI features and prognosis a 3-grade scoring system was proposed and evaluated to assess the risk of LR. Mean age was 66.1 ± 14.4 years; mean follow-up was 16 ± 28.3 months. The MRI features most associated with higher risk of LR resulted to be: lesion sizes (both volume and maximum diameter with a cut-off of 20 cm - p = 0.01), the "tail sign" (p = 0.045), and high myxoid matrix content with MRI water-like appearance (p = 0.0493). Ninety-four patients (94 of 150- 62.7%) were grade 1, 33 (22.0%) grade 2, and 23 (15.3%) grade 3. Interobserver agreement was substantial with K= 0.779 (95%CI 0.685-0.874). Higher grades of MRI grading system proposed were significantly associated with an increased LR risk, hazard ratio = 2.031 (95%CI 1.366-3.019; p < 0.001). This is the largest series evaluating MRI features as prognostic factors for MFS. The MRI grading system proposed is significantly able to stratify the risk of LR in MFS of the extremities. The system is applicable to all the standard MRI studies protocols, might help in surgical planning, and may offer prognostic information.


Energy coupling to K+ uptake via the Trk system in Escherichia coli: the role of ATP.
The dual energy requirement (protonmotive force and ATP) of the Escherichia coli Trk potassium transport system has been investigated. Using inhibitors and unc mutants we show that Trk is not an ATPase but may be regulated by ATP. Possible mechanisms of energy coupling to Trk are discussed.


Synthesis and evaluation of 18F labeled alanine derivatives as potential tumor imaging agents.
This paper reports the synthesis and labeling of (18)F alanine derivatives. We also investigate their biological characteristics as potential tumor imaging agents mediated by alanine-serine-cysteine preferring (ASC) transporter system. Three new (18)F alanine derivatives were prepared from corresponding tosylate-precursors through a two-step labeling reaction. In vitro uptake studies to evaluate and to compare these three analogs were carried out in 9L glioma and PC-3 prostate cancer cell lines. Potential transport mechanisms, protein incorporation and stability of 3-(1-[(18)F]fluoromethyl)-L-alanine (L-[(18)F]FMA) were investigated in 9L glioma cells. Its biodistribution was determined in a rat-bearing 9L tumor model. PET imaging studies were performed on rat bearing 9L glioma tumors and transgenic mouse carrying spontaneous generated M/tomND tumor (mammary gland adenocarcinoma). New (18)F alanine derivatives were prepared with 7%-34% uncorrected radiochemical yields, excellent enantiomeric purity (>99%) and good radiochemical purity (>99%). In vitro uptake of the L-[(18)F]FMA in 9L glioma and PC-3 prostate cancer cells was higher than that observed for the other two alanine derivatives and [(18)F]FDG in the first 1h. Inhibition of cell uptake studies suggested that L-[(18)F]FMA uptake in 9L glioma was predominantly via transport system ASC. After entering into cells, L-[(18)F]FMA remained stable and was not incorporated into protein within 2h. In vivo biodistribution studies demonstrated that L-[(18)F]FMA had relatively high uptake in liver and kidney. Tumor uptake was fast, reaching a maximum within 30 min. The tumor-to-muscle, tumor-to-blood and tumor-to-brain ratios at 60 min post injection were 2.2, 1.9 and 3.0, respectively. In PET imaging studies, tumors were visualized with L-[(18)F]FMA in both 9L rat and transgenic mouse. L-[(18)F]FMA showed promising properties as a PET imaging agent for up-regulated ASC transporter associated with tumor proliferation.


Organic copper complexes as a new class of proteasome inhibitors and apoptosis inducers in human cancer cells.
Here we report that organic copper complexes can potently and selectively inhibit the chymotrypsin-like activity of the proteasome in vitro and in vivo. Several copper compounds, such as NCI-109268 and bis-8-hydroxyquinoline copper(II) [Cu(8-OHQ)(2)], can inhibit the chymotrypsin-like activity of purified 20S proteasome. In human leukemia cells, proteasome inhibition occurs within 15min after treatment, followed by apoptosis. Neither proteasome inhibition nor apoptosis occurs in non-transformed, immortalized human natural killer cells under the same treatment. Furthermore, proteasome inhibition and apoptosis induction were detected in prostate cancer cells treated with the ligand 8-OHQ alone following pre-treatment with copper(II) chloride. None of these events occurred in cells treated with copper(II) chloride alone, 8-OHQ alone (without growth in copper-enriched media), or nickel(II) chloride pre-treatment followed by 8-OHQ. Furthermore, we found that copper-mediated inhibition of purified 20S proteasome cannot be blocked by a reducing agent and that organic copper compounds do not generate hydrogen peroxide in the cells, suggesting that proteasome inhibition and apoptosis induction are not due to copper-mediated oxidative damage of proteins. Our results suggest that certain types of organic ligands could bind to tumor cellular copper, forming potent proteasome inhibitors and apoptosis inducers at copper concentrations found in tumor tissues.


Cell surface expression of a human IgG Fc chimera activates macrophages through Fc receptors.
Antibody-dependent cell-mediated cytotoxicity plays an important role in the macrophage-mediated destruction of target cells. While the selectivity is based on antibody specificity, the lytic attack is triggered by Fc receptor-mediated respiratory burst. To mimic IgG opsonization, a chimeric antibody-like molecule, containing human IgG1 Fc, was expressed on the surface of mammalian cells. The transmembrane domain of the human transferrin receptor was fused in-frame to the N-terminus of the second and third domains of human IgG1 heavy-chain constant region. This fusion molecule was designed to take advantage of the type II membrane anchor property of the transferrin receptor to express the Fc portion of the molecule in a reverse orientation, such that the Fc portion projected away from the cell surface. This is in contrast to the conventional cell surface IgG, which is anchored by a C-terminal type I transmembrane domain. The cell surface expressed reverse Fc no longer activated complement, but retained Fc receptor-binding capability and activated superoxide production by macrophages. This activity was completely blocked by an FcgammaR I-specific monoclonal antibody.


Probing the Behaviour of Cas1-Cas2 upon Protospacer Binding in CRISPR-Cas Systems using Molecular Dynamics Simulations.
Adaptation in CRISPR-Cas systems enables the generation of an immunological memory to defend against invading viruses. This process is driven by foreign DNA spacer (termed protospacer) selection and integration mediated by Cas1-Cas2 protein. Recently, different states of Cas1-Cas2, in its free form and in complex with protospacer DNAs, were solved by X-ray crystallography. In this paper, molecular dynamics (MD) simulations are employed to study crystal structures of one free and two protospacer-bound Cas1-Cas2 complexes. The simulated results indicate that the protospacer binding markedly increases the system stability, in particular when the protospacer containing the PAM-complementary sequence. The hydrogen bond and binding free energy calculations explain that PAM recognition introduces more specific interactions to increase the cleavage activity of Cas1. By using principal component analysis (PCA) and intramolecular angle calculation, this study observes two dominant slow motions associated with the binding of Ca1-Cas2 to the protospacer and potential target DNAs respectively. The comparison of DNA structural deformation further implies a cooperative conformational change of Cas1-Cas2 and protospacer for the target DNA capture. We propose that this cooperativity is the intrinsic requirement of the CRISPR integration complex formation. This study provides some new insights into the understanding of CRISPR-Cas adaptation.


Molecular etiology of defective nuclear and mitochondrial ribosome biogenesis: Clinical phenotypes and therapy.
Ribosomopathies are rare congenital disorders associated with defective ribosome biogenesis due to pathogenic variations in genes that encode proteins related to ribosome function and biogenesis. Defects in ribosome biogenesis result in a nucleolar stress response involving the TP53 tumor suppressor protein and impaired protein synthesis leading to a deregulated translational output. Despite the accepted notion that ribosomes are omnipresent and essential for all cells, most ribosomopathies show tissue-specific phenotypes affecting blood cells, hair, spleen, or skin. On the other hand, defects in mitochondrial ribosome biogenesis are associated with a range of clinical manifestations affecting more than one organ. Intriguingly, the deregulated ribosomal function is also a feature in several human malignancies with a selective upregulation or downregulation of specific ribosome components. Here, we highlight the clinical conditions associated with defective ribosome biogenesis in the nucleus and mitochondria with a description of the affected genes and the implicated pathways, along with a note on the treatment strategies currently available for these disorders.


Mutations in the S4-H2 loop of eIF4E which increase the affinity for m7GTP.
Eukaryotic initiation factor 4E (eIF4E) binds the 5'-cap of eukaryotic mRNAs and overexpression of eIF4E in epithelial cell cancers correlates with the metastases/tissue invasion phenotype. Photolabeling of eIF4E with [gamma-32P]8-azidoguanosine 5'-triphosphate (8-N3GTP) demonstrated cross-linking at Lys-119 in the S4-H2 loop which is distant from the m7GTP binding site [Marcotrigiano et al. (1997) Cell 89, 951-961; Friedland et al. (1997) Protein Sci. 6, 125-131]. Modeling studies indicate that 8-N3GTP cross-linked with Lys-119 because it binds a site that is occupied by the second nucleotide of a bound mRNA. Mutagenesis of the S4-H2 loop produced proteins with a 5-10-fold higher affinity for m7GTP than wild-type eIF4E. These mutants of eIF4E may have uses in selectively purifying mRNAs with intact 5'-ends or in determining how the promyelocytic leukemia protein decreases the affinity of eIF4E for mRNA caps.


Genetic and molecular characterization of embryonic mutants identified following seed transformation in Arabidopsis.
Over 5000 transgenic families of Arabidopsis thaliana produced following seed transformation with Agrobacterium tumefaciens were screened for embryonic lethals, defectives, and pattern mutants. One hundred and seventy-eight mutants with a wide range of developmental abnormalities were identified. Forty-one mutants appear from genetic studies to be tagged (36% of the 115 mutants examined in detail). Mapping with visible markers demonstrated that mutant genes were randomly distributed throughout the genome. Seven mutant families appeared to contain chromosomal translocations because the mutant genes exhibited linkage to visible markers on two different chromosomes. Chromosomal rearrangements may therefore be widespread following seed transformation. DNA gel blot hybridizations with 34 tagged mutants and three T-DNA probes revealed a wide range of insertion patterns. Models of T-DNA structure at each mutant locus were constructed to facilitate gene isolation. The value of such models was demonstrated by using plasmid rescue to clone flanking plant DNA from four tagged mutants. Further analysis of genes isolated from these insertional mutants should help to elucidate the relationship between gene function and plant embryogenesis.


Targeting cancer cells by exploiting their resistance.
Cancer cells are intrinsically resistant to growth arrest and can further acquire multidrug resistance. Current approaches to this problem are intended to reverse, overcome or prevent the drug resistance. However, the resistance of cancer cells can be exploited to kill resistant cells selectively, while sparing sensitive normal cells. As the simplest example, multidrug-resistant cells pump out protectors, such as pharmacological inhibitors of apoptosis. A sequence of at least two agents must include an exclusive protector, which is ineffective in resistant cancer cells, and an inclusive cytotoxic drug, which kills unprotected cells. By abolishing several dose-limiting side effects of chemotherapy, this strategy provides a means to treat selectively most deranged, aggressive and resistant cancers.


Laparoscopic thoracic duct clipping for persistent chylothorax after extrapleural pneumonectomy.
We describe a 68-year-old man who was treated by laparoscopic thoracic duct clipping for persistent chylothorax after an extrapleural pneumonectomy for malignant pleural mesothelioma. Initial conservative treatment did not resolve the postoperative chylothorax. A second surgery through the thoracic approach was considered invasive and difficult after extrapleural pneumonectomy. A laparoscopic approach proved effective and resolved the chylothorax. Thus, laparoscopic thoracic duct clipping is considered very useful for treating chylothorax.


Osteoblast low-molecular-weight proteinase inhibitor. I. Isolation and characterization of activity from osteoblastic cells and bone.
Isolated mouse calvarial cells having phenotypic characteristics of osteoblasts, mouse parietal bone segments, mouse serum, and control mouse lung fibroblasts were extracted in NaCl and ultrafiltered to recover final concentrates having nominal molecular weights between 50,000 and 1000 daltons. Final concentrates of osteoblasts and bone but not of serum or control fibroblasts were positive for the inhibition of trypsin degradation of fibrin. Osteoblast final concentrates inhibited trypsin hydrolysis of the synthetic substrate p-tosyl-L-arginine methyl ester. Osteoblast and bone final concentrates comigrated with Trasylol but were electrophoretically distinct from alpha 1-antiproteinase. Final concentrates of osteoblast and bone extracts did not inhibit tadpole collagenase using the [3H]glycine-labeled diffuse chick collagen fibril assay. High-performance liquid chromatography (HPLC) of osteoblast final concentrates after purification using immobilized trypsin affinity chromatography revealed the presence of a major peak that was positive for the inhibition of trypsin. Molecular weight determination by HPLC indicated that the inhibitor(s) range in nominal molecular weight from 4300 to 5100 daltons. The presence of low-molecular-weight serine proteinase inhibitory activity in bone suggests its participation in the regulation of bone resorption through the regulation of enzyme activation of collagenase, and possibly its role in defense against bone matrix enzymatic degradation during tumor cell invasion.


Permutationally Invariant Polynomial Basis for Molecular Energy Surface Fitting via Monomial Symmetrization.
We describe a procedure to develop a fitting basis for molecular potential energy surfaces (PESs) that is invariant with respect to permutation of like atoms. The method is based on a straightforward symmetrization of a primitive monomial basis and illustrated for several classes of molecules. A numerically efficient method to evaluate the resulting expression for the PES is also described. The fitting basis is used to obtain a new PES for H3O(+) based on roughly 62 000 ab initio energies.


Biological activity of pectic polysaccharides investigated through biomembrane models formed at the air-water interface.
Pectin, a polysaccharide with potential bioactivity, was inserted in the aqueous subphase of monolayers of the selected lipids DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), representing mammalian and bacterial membranes, respectively. Pectin condensed both monolayers but made the DPPC monolayer more fluid, while for DPPE, it made its monolayer more rigid, as detected with dynamic interfacial rheology. Complementary data using surface potential, infrared spectroscopy, and Brewster angle microscopy also showed distinctive effects of pectin on DPPE and DPPC. We believe these data can be correlated with the action of this polysaccharide with biological lipidic surfaces with different polar heads, which may be relevant, generally speaking, to understanding the molecular mechanism of this bioactive compound for pharmaceutical purposes.


Structural insights into the gating mechanism of human SLC26A9 mediated by its C-terminal sequence.
The human SLC26 transporter family exhibits various transport characteristics, and family member SLC26A9 performs multiple roles, including acting as Cl-/HCO3- exchangers, Cl- channels, and Na+ transporters. Some mutations of SLC26A9 are correlated with abnormalities in respiration and digestion systems. As a potential target colocalizing with CFTR in cystic fibrosis patients, SLC26A9 is of great value in drug development. Here, we present a cryo-EM structure of the human SLC26A9 dimer at 2.6 Å resolution. A segment at the C-terminal end is bound to the entry of the intracellular vestibule of the putative transport pathway, which has been proven by electrophysiological experiments to be a gating modulator. Multiple chloride and sodium ions are resolved in the high-resolution structure, identifying novel ion-binding pockets for the first time. Together, our structure takes important steps in elucidating the structural features and regulatory mechanism of SLC26A9, with potential significance in the treatment of cystic fibrosis.


Prevalence of metabolic syndrome in Iran: A 2011 update.
The aim of the present study was to determine the prevalence of metabolic syndrome and its individual components among the Iranian adult population in 2011 and to investigate changes between 2007 and 2011. Data from two rounds of the Surveillance of Risk Factors of Non-communicable Diseases national surveys conducted in 2007 and 2011 were pooled. Metabolic syndrome was defined according to International Diabetes Federation criteria. In 2007, the prevalence of metabolic syndrome among adults aged 25-64 years was 35.95 (95% confidence interval [CI] 34.27-37.63), which decreased to 32.96 (95% CI 30.73-35.18) in 2011 (P = 0.0108). Despite this overall decline, the prevalence of central obesity (P = 0.1383), raised triglycerides (P = 0.3058), and reduced high-density lipoprotein cholesterol (HDL-C; P = 0.5595) remained constant. There was a trend towards a decline in the proportion of individuals with increased blood pressure (P = 0.0978), and the proportion of adults with increased fasting plasma glucose (FPG) increased (P < 0.0001). In 2011, the prevalence of central obesity, raised triglycerides, reduced HDL-C, increased blood pressure and increased FPG was 51.88 (95% CI 48.97-54.79), 36.99 (95% CI 34.52-39.45), 54.72 (95% CI 50.87-58.57), 38.92 (95% CI 36.19-41.64), and 24.97 (95% CI 22.02-27.93) respectively. Over the period 2007-11, the prevalence of metabolic syndrome has decreased slightly in Iran, although prevalence of increased FPG has increased significantly. One-third of the Iranian adult population is diagnosed with metabolic syndrome.


Cell fusion in osteoclasts plays a critical role in controlling bone mass and osteoblastic activity.
The balance between osteoclast and osteoblast activity is central for maintaining the integrity of bone homeostasis. Here we show that mice lacking dendritic cell specific transmembrane protein (DC-STAMP), an essential molecule for osteoclast cell-cell fusion, exhibited impaired bone resorption and upregulation of bone formation by osteoblasts, which do not express DC-STAMP, which led to increased bone mass. On the contrary, DC-STAMP over-expressing transgenic (DC-STAMP-Tg) mice under the control of an actin promoter showed significantly accelerated cell-cell fusion of osteoclasts and bone resorption, with decreased osteoblastic activity and bone mass. Bone resorption and formation are known to be regulated in a coupled manner, whereas DC-STAMP regulates bone homeostasis in an un-coupled manner. Thus our results indicate that inhibition of a single molecule provides both decreased osteoclast activity and increased bone formation by osteoblasts, thereby increasing bone mass in an un-coupled and a tissue specific manner.


Macrophage (monocyte)-derived mucous secretagougue (MMS) is released into the fluid of the middle ear of patients with otitis media with effusion.
Macrophage (monocyte)-derived secretagogue (MMS) is a low molecular weight peptide released by activated macrophages. This substance can enhance incorporation of carbohydrates into protein and result in the production of glycoproteins. It is believed that increased glycoproteins lead to the increased tenaciousness of mucoid effusions. Sixteen of 28 middle ear effusions demonstrated significant amounts of MMS. It is suggested that MMS is produced by activated macrophages in MEE and may lead to the development of glycoprotein by secretory epithelial cells. This substance may lead to the development of the mucoid effusion in otitis media with effusion in children.


INTRAVITREAL CORTICOSTEROIDS IN DIABETIC MACULAR EDEMA: PHARMACOKINETIC CONSIDERATIONS.
To review the relationship between kinetics, efficacy, and safety of several corticosteroid formulations for the treatment of diabetic macular edema. Reports of corticosteroid use for the treatment of diabetic macular edema were identified by a literature search, which focused on the pharmacokinetics, efficacy, and safety of these agents in preclinical animal models and clinical trials. Available corticosteroids for diabetic macular edema treatment include intravitreal triamcinolone acetonide, dexamethasone, and fluocinolone acetonide. Because of differences in solubility and bioavailability, various delivery mechanisms are used. Bioerodible delivery systems achieve higher maximum concentrations than nonbioerodible formulations. There is a relationship between visual gains and drug persistence in the intravitreal compartment. Safety effects were more complex; level of intravitreal triamcinolone acetonide exposure is related to development of elevated intraocular pressure and cataract; this does not seem to be the case for dexamethasone, where two different doses showed similar mean intraocular pressure and incidence of cataract surgery. With fluocinolone acetonide, rates of intraocular pressure elevations requiring surgery seem to be dose related; rates of cataract extraction were similar regardless of dose. Available corticosteroids for diabetic macular edema exhibit different pharmacokinetic profiles that impact efficacy and adverse events and should be taken into account when developing individualized treatment plans.


Analysis of Daily and Diurnal O3-NOx Relationships and Assessment of Local/Regional Oxidant (OX = O3 + NO2) Levels and Associated Human Health Risk at a Coastal Suburban Site of Sfax (Tunisia).
The present study investigated the temporal variability of surface ozone and its nitrogen precursors at the proximity of a traffic crossroad (≈ 22,580 vehicles per day) located in a coastal suburban site of Sfax city (Tunisia). It was performed during January-October, 2010. The study results show that the surface ozone is characterized by a slightly modulated regime between day and night. At traffic-peak hours, the decrease of ozone concentration levels is due to the oxidation reaction of NO into NO2. Complementary statistical approaches (inter-variable correlation matrix, cluster analysis, representation quality of variables and multiple regression analysis) reveal that the excess of O3 is mainly affected by the wind speed, temperature, solar radiation and NO2 with contribution rates of 127, 21, 22 and 12%, respectively. The decrease of O3 is, however, controlled by NO, relative humidity and boundary layer height with contribution rates of 25, 21 and 16%, respectively. The regional daytime and night-time contributions to O3 are very different. The daytime intercept which is greater than that of night-time indicates there was a large NOx independent regional contribution. This could be attributed to the biogenic VOCs effect interfering in the photochemical cycle. It, therefore, implies that the study site is VOC-sensitive. The investigation of the air quality index (AQI) for O3 and NO2 showed that more than 86% of the total studied period has a good quality level. Only about 14% of total days are characterized by an acceptable air quality level, however, for a very small number of people are unusually sensitive to air pollution.


Safety of Intravenous Push Levetiracetam Compared to Intravenous Piggyback at a Tertiary Academic Medical Center: A Retrospective Analysis.
Medication administration via intravenous push presents multiple potential advantages; however, there may be an increased risk of adverse drug reactions. In 2020, Brigham and Women's Hospital changed levetiracetam intravenous administration to intravenous push (IVP). The purpose of this analysis was to compare the safety profile of IVP to intravenous piggyback (IVPB) levetiracetam administration. This institutional review board-approved, single-center, pre-post analysis was performed between 1 November, 2019 and 30 May, 2020. The electronic health record was used to identify all administrations of intravenous levetiracetam greater than 1000 mg in patients ≥ 18 years old. The major safety outcomes included hypotension, bradycardia, drug-induced sedation, and intravenous site reactions such as phlebitis and infiltration. The major efficiency outcome was the time from pharmacy order verification to first-dose administration. A total of 498 administrations in 162 patients were included in the analysis: 252 administrations in 84 patients in the IVP group and 246 administrations in 78 patients in the IVPB group. The incidence of bradycardia was 7 vs 3 (3.2% vs 1.5%, p = 0.34); hypotension 10 vs 6 (5.2% vs 3.5%, p = 0.44); sedation 21 vs 36 (19.3% vs 27.9%, p = 0.12); and peripheral IV site reactions 0 vs 1 (0% vs 0.6%, p = 0.39) in the IVP vs IVPB groups, respectively. The median time between order verification and first-dose administration was significantly reduced in the IVP vs IVPB group (23.5 vs 55 min, p < 0.001). Intravenous push levetiracetam administration of doses up to 4000 mg was associated with a similar incidence of cardiovascular, sedation, and infusion site-related adverse events compared to IVPB and resulted in a significant reduction in time to first-dose administration. Intravenous push levetiracetam in doses as high as 4000 mg may be considered safe with appropriate monitoring.


A seven-degree-of-freedom, time-dependent quantum dynamics study on the energy efficiency in surmounting the central energy barrier of the OH + CH3 → O + CH4 reaction.
A time-dependent, quantum reaction dynamics calculation with seven degrees of freedom was carried out to study the energy efficiency in surmounting the approximate center energy barrier of OH + CH3. The calculation shows the OH vibration excitations greatly enhance the reactivity, whereas the vibrational excitations of CH3 and the rotational excitations hinder the reactivity. On the basis of equal amount of total energy, although this reaction has a slight early barrier, it is the OH vibrational energy that is the dominate force in promoting the reactivity, not the translational energy. The studies on both the forward O + CH4 and reverse OH + CH3 reactions demonstrate, for these central barrier reactions, a small change of the barrier location can significantly change the energy efficacy roles on the reactivity. The calculated rate constants agree with the experimental data.


No association of CpG SNP rs9357140 with onset age in Belgian C9orf72 repeat expansion carriers.
We investigated the impact of the recently described chromosome 6 open reading frame 10 (C6orf10)/LOC101929163 locus as age-at-onset modifier in an extended cohort of Belgian chromosome 9 open reading frame 72 (C9orf72) G4C2 repeat expansion carriers. We genotyped the tagging CpG single-nucleotide polymorphism rs9357140 in 224 confirmed C9orf72 repeat expansion carriers, 102 index cases and 122 relatives, and tested association with onset age. The C9orf72 repeat expansion cohort consisted of 131 symptomatic carriers, that is, 78 with dementia only, 13 with frontotemporal dementia (FTD)-amyotrophic lateral sclerosis (ALS), and 40 ALS only, and 93 presymptomatic carriers. Cox proportional hazard regression analysis failed to identify significant association (adjusted hazard ratio = 1.15, p = 0.3). We further extended our analysis to a Belgian cohort of unrelated, mutation-negative FTD index patients (n = 230), but also found no association (adjusted hazard ratio = 0.96, p = 0.3). Overall, our findings suggest that in the Belgian cohort, the C6orf10/LOC101929163 locus cannot explain the marked variability in age at onset, and other genetic or environmental modifiers must drive the clinical heterogeneity observed among C9orf72 repeat expansion carriers.


Repeated Surgery in a Case of Epithelial-Myoepithelial Carcinoma of the Parotid Gland that was Difficult to Distinguish from Pleomorphic Adenoma.
Epithelial-myoepithelial carcinoma (EMC) of the parotid gland is a comparatively rare tumor that accounts for less than 1% of all salivary gland tumors. A patient with EMC of the parotid gland that was initially diagnosed as pleomorphic adenoma and that recurred locally during the watchful waiting period but was controlled by surgery under local anesthesia is reported. An 80-year-old man had noticed a swelling in the left infra aural region. A left parotid gland tumor was suspected, and he was referred to our department. Magnetic resonance imaging (MRI) and fine-needle aspiration cytology findings were suggestive of pleomorphic adenoma of the superficial lobe of the parotid gland, and this was therefore resected under general anesthesia. Postoperative histopathological examination, immunostaining, and genetic tests resulted in a diagnosis of EMC. Postoperative pathological review showed that part of the resection margin was positive. The possibility of recurrence was explained to the patient, and additional treatment was recommended, but since the patient did not desire this, a policy of watchful waiting was adopted. Signs of cutaneous metastasis in the left infra aural region were detected at 13 months postoperatively, and this metastasis was excised under local anesthesia. The resection margin was negative, and the patient's course remains uneventful. EMC is classified as a low-grade malignant tumor, but it requires stringent monitoring because of its frequent local recurrence. Since local control can usually be achieved by surgical treatment alone, and postoperative adjuvant therapy may not necessarily warranted.


Obscure gastrointestinal bleeding: single centre experience of capsule endoscopy.
The advent of capsule endoscopy (CE) has resulted in a paradigm shift in the approach to the diagnosis and management of patients with obscure gastrointestinal bleeding (OGIB). With increasing global availability of this diagnostic tool, it has now become an integral part of the diagnostic algorithm for OGIB in most parts of the world. However, there is scant data on optimum timing of CE for maximizing diagnostic yield. OGIB continues to be a challenge because of delay in diagnosis and consequent morbidity and mortality. We evaluated the diagnostic yield of CE in identifying the source of bleeding in patients with OGIB. We identified patients who underwent CE at our institution from May 2006 to May 2011. The patients' medical records were reviewed to determine the type of OGIB (occult, overt), CE results and complications, and timing of CE with respect to onset of bleeding. Out of 346 patients investigated for OGIB, 246 (71.1%) had some lesion detected by CE. In 206 patients (59.5%), definite lesions were detected that could unequivocally explain the OGIB. Small bowel angiodysplasia, ulcer/erosions secondary to Crohn's disease, non-steroidal anti-inflammatory agent use, and neoplasms were the commonest lesions detected. Visualization of the entire small bowel was achieved in 311 (89.9%) of cases. Capsule retention was noted in five patients (1.4%). In this study, CE was proven to be a safe, comfortable, and effective, with a high rate of accuracy for diagnosing OGIB.


Inflammatory response to mucosal barrier injury after myeloablative therapy in allogeneic stem cell transplant recipients.
We noted a significant increase of interleukin-8 (IL-8), LBP and CRP mirroring the pattern of mucosal barrier injury as measured by gut integrity (lactulose/rhamnose ratio), daily mucositis score (DMS) and serum citrulline concentrations of 32 haematopoietic stem cell transplant (HSCT) recipients following intensive myeloablative therapy. Concentrations of IL-8, LBP and CRP were already significantly elevated before the onset of fever or bacteraemia due to oral viridans streptococci (OVS) in the first week after transplant during profound neutropenia. These markers reached their peak when citrulline concentrations reached their nadir, the highest scores of DMS were attained and when there was significantly decreased gut integrity. This suggests that the degree of mucosal barrier injury rather than bacteraemia due to OVS determines the intensity of the inflammatory response.


The surgical repair of membranous urethral strictures in children.
Eighteen patients have been treated for membranous urethral strictures following rupture of the prostatomembranous urethra. In 16, satisfactory results have been attained, 1 patient has been diverted permanently, and the remaining patient has not yet undergone complete repair.


An intercomparison of the accuracy of computer planning systems for brachytherapy.
The accuracy of brachytherapy computer planning systems has been investigated by means of an intercomparison of the dose distributions as calculated by these systems for five specific test cases. In this intercomparison, 13 radiotherapy institutions in The Netherlands participated. The planning systems evaluated comprise all commercial systems in clinical use in The Netherlands (different versions of in total four manufacturers) as well as in-house developed systems. The five test cases were designed in order to quantify the errors introduced by: (a) the calculation algorithm including the method of source specification used; (b) the localisation procedure via orthogonal radiographs; and (c) the clinical dose specification system in the case of a geometrically irregular implantation for which no established, rigid dose specification method is available. The results of this comparison can be summarized as follows: The accuracy of most calculation algorithms appears to be satisfactory except for the region close to the longitudinal axis of linear sources. Source strength specification in terms of activity or milligram radium equivalent can result in dosimetric errors which may be clinically relevant and therefore specification of the source strength in terms of "output", i.e. air-kerma rate at a specified distance, is to be preferred. This concurs with recent national and international recommendations regarding source strength specification. Localisation via orthogonal radiographs, as commonly used, appears to cause noticeable errors. This aspect could be improved in a quality assurance program. In case of "irregular" implantations, the uncertainty in clinical dose specification clearly dominates the overall accuracy. This illustrates the need for a consistent dose specification system, which is capable of including interstitial implants with a source geometry deviating from the rules given by the existing specification systems. It can be concluded that in brachytherapy, whenever possible, a well-defined, rigid clinical dose specification system should be applied, but that even in such cases the delivered dose may be in error by more than 5% (1 S.D.).


Mutation induction in Escherichia coli incubated in the reaction mixture of NADPH-dependent lipid peroxidation of rat-liver microsomes.
Experiments were carried out to examine mutation induction in E. coli cells incubated in the reaction mixture of NADPH-dependent lipid peroxidation of microsomes isolated from rat liver. The results obtained were as follows: (1) Lipid peroxidation of microsomes occurred extensively on incubation with NADPH and Fe2+. In the E. coli WP2uvrA(pKM101) system, the mutation frequency to streptomycin resistance increased markedly when the cells were incubated in the reaction mixture of microsomal lipid peroxidation. The induced mutation frequencies were dependent on the extent of the lipid peroxidation. (2) It was also found that the mutations were induced at the same rate as in the case of (1) when the cells were added to the microsomal suspensions after the reactions due to the short-lived free radicals had terminated. (3) The cytotoxicity of the lipid peroxidation products was larger in the DNA repair-defective mutant, E. coli SR18 (uvrArecA) than the wild-type strain, SR749. From these results it is concluded that some DNA-damaging and mutagenic substances are indeed produced in the degradation process of peroxidized polyunsaturated fatty acids in liver microsomal lipids.


A putative flowering-time-related Dof transcription factor gene, JcDof3, is controlled by the circadian clock in Jatropha curcas.
Plant-specific DNA-binding transcription factors with one finger (Dof) perform important roles in several biological processes. A yeast one-hybrid cDNA library of Jatropha curcas was used to identify Dof-type transcription factors. JcDof3, isolated from the library as a full-length cDNA, encoded a protein of 518 amino acids and contained a highly conserved Dof domain. Yeast one-hybrid systems and subcellular localization assays confirmed that JcDof3 was a typical transcription factor. In contrast to arrhythmic expression at basal level in etiolated cotyledons under continuous dark conditions, the circadian oscillations of JcDof3 transcripts were observed under long day, short day or continuous light regimes. A phylogenetic analysis showed that JcDof3 was clustered into the same clade with CYCLING DOF FACTOR (CDF), which interacts with F-box protein to regulate photoperiodic flowering. Moreover, a yeast two-hybrid assay showed that JcDof3 also interacted with F-box proteins. Our results suggest that JcDof3 is a circadian clock regulated gene, and might be involved in the flowering time regulation of J. curcas.


High dose vitamin D may improve lower urinary tract symptoms in postmenopausal women.
Lower urinary tract symptoms (LUTS) are common in postmenopausal women, and have been reported inversely associated with vitamin D intake and serum 25-hydroxyvitamin D (25(OH)D) levels. The aim of this study was to investigate if high dose vitamin D supplementation would affect LUTS in comparison to standard dose. In a randomized controlled study including 297 postmenopausal women with low bone mineral density, the participants were allocated to receive capsules of 20 000IU of vitamin D3 twice a week (high dose group) or similar looking placebo (standard dose group). In addition, all the participants received 1g of calcium and 800IU of vitamin D daily. A validated questionnaire regarding LUTS was filled in at baseline and after 12 months. At baseline, 76 women in the high dose group and 82 in the standard dose group reported any LUTS. Levels of serum 25(OH)D increased significantly more in the high dose group (from 64.7 to 164.1nmol/l compared to from 64.1 to 81.8nmol/l, p<0.01). No differences between the groups were seen regarding change in LUTS except for a statistically significant reduction in the reported severity of urine incontinence in the high dose group as compared to the standard dose group after one year (p<0.05). The results need confirmation in a study specifically designed for this purpose.


Asymptomatic interventricular septal dissection and giant coronary artery aneurysms simulating cardiac cysts: Multi-modality imaging.
we report an exceptionnel case of asymptomatic interventricular septal dissection and giant coronary artery aneurysms simulating cardiac cysts with a focus in the interst of Multi-modality Imaging.


Synthesis, characterization, and structural studies of mixed-ligand diorganotin esters, [R2Sn(OP(O)(OH)Ph)(OS(O)2R1)]n [R = n-Bu, R1 = Me (1), n-Pr(2); R = Et, R1 = Me (3)] with 1D and 3D coordination polymeric motifs.
Mixed-ligand diorganotin esters, [R 2Sn(OP(O)(OH)Ph)(OS(O) 2R (1))] n [R = n-Bu, R (1) = Me ( 1), n-Pr ( 2); R = Et, R (1) = Me ( 3)], have been synthesized by reacting the tin precursors, R 2Sn(OR (1))OS(O) 2R with an equimolar amount of phenylphosphonic acid under mild conditions (room temperature, 6-8 h, CH 2Cl 2). These have been characterized by IR, multinuclear ( (1)H, (13)C{ (1)H}, (31)P, and (119)Sn) NMR, and single crystal X-ray diffraction studies. The asymmetric unit of 1 is comprised of a tetramer with four crystallographically unique tin atoms. The structure reveals a central eight-membered (Sn-O-S-O) 2 cyclic ring with two exocyclic tin atoms, which results from micro 3-binding of the two methanesulfonate groups. The remaining two sulfonates are monodentate and contribute in O...HO(P) hydrogen bonding. The molecular structure is extended into a 3D coordination polymer with the aid of hydrogenphenylphosphonate group on each tin atom, acting in a micro 2-O 2P mode and forms a series of eight-membered (Sn-O-P-O) 2 rings in the structural framework. 2 and 3 are isostructural and represent linear 1D coordination polymers via micro 2-binding mode of both alkanesulfonate and hydrogenphenylphosphonate groups.


Structure of E. coli ketopantoate hydroxymethyl transferase complexed with ketopantoate and Mg2+, solved by locating 160 selenomethionine sites.
We report the crystal structure of E. coli ketopantoate hydroxymethyltransferase (KPHMT) at 1.9 A resolution, in complex with its product, ketopantoate. KPHMT catalyzes the first step in the biosynthesis of pantothenate (vitamin B(5)), the precursor of coenzyme A and the acyl carrier protein cofactor. The structure of the decameric enzyme was solved by multiwavelength anomalous dispersion to locate 160 selenomethionine sites and phase 560 kDa of protein, making it the largest structure solved by this approach. KPHMT adopts the (betaalpha)(8) barrel fold and is a member of the phosphoenolpyruvate/pyruvate superfamily. The active site contains a ketopantoate bidentately coordinated to Mg(2+). Similar binding is likely for the substrate, alpha-ketoisovalerate, orienting the C3 for deprotonation.


[Immunological status and somatotropic activity of the pituitary in combined abdominal injuries complicated by peritonitis].
Associated injuries of the abdomen result in peritonitis twice more often than isolated ones, the peritonitis having more severe course and being the cause of lethal outcomes in 57% of the victims. The immune deficient state is thought to be one of the causes responsible for the reduced resistance to infection and the development of peritonitis in the postshock period. Patients with associated traumas show a considerable inhibition of the immune system within several hours and an impairment of its regulation which grows during the next day. So, even at the early terms the operative methods must be similar to those used in operations for the developed peritonitis.