Cyclodextrin-modified monolithic columns for resolving dansyl amino acid enantiomers and positional isomers by capillary electrochromatography.
We describe beta- and gamma-cyclodextrins (beta- and gamma-CD)-modified monolithic columns prepared by sol-gel process and chemical modifications. The monolithic silica column was fabricated inside a fused-silica capillary with 100 microm inner diameter by sol-gel process. The monolithic silica matrix was chemically modified chiral selectors of beta- or gamma-CDs with a spacer of 3-glycidoxypropyltrimethoxysilane by on-column reactions. Gamma-CD-modified monolithic column has successfully been applied for the separation of dansyl amino acid enantiomers. Beta-CD-modified monolithic column has been used for the separation of the positional isomers of o-, m-, and p-cresols and the enantioseparation of racemates of benzoin and several dansyl amino acids by capillary electrochromatography, respectively. For the separation of neutral positional isomers, a positive electric field was applied. However, for the separation of negatively charged analytes, a negative electric field was applied at the inlet of column. The separation efficiency of 5.0 x 10(4) theoretical plates/m for dansyl-L-threonine was obtained at electric field strength of -300 V/cm in the mobile phase of 50 mM 2-(N-morpholino)ethanesulfonic acid (MES)-Tris/methanol (70/30) buffer at pH 7.0. L-enantiomers were eluted as the first peak. Scanning electron micrograph showed that monolithic columns have the morphology of continuous skeleton and large through-pores.


Prospective Randomized Comparison of Knee Stability and Proprioception for Posterior Cruciate Ligament Reconstruction With Autograft, Hybrid Graft, and γ-Irradiated Allograft.
To evaluate posterior stability and proprioception after arthroscopic posterior cruciate ligament reconstruction with autograft, hybrid graft, and γ-irradiated allograft. This prospective, randomized controlled trial (with 5 years' follow-up) included 90 patients who underwent posterior cruciate ligament reconstruction with autograft (gracilis and semitendinosus tendons), hybrid graft (γ-irradiated tibialis anterior tendon allograft and semitendinosus tendon autograft), or γ-irradiated allograft (tibialis anterior tendons). All the γ-irradiated allografts (tibialis anterior tendons) received an irradiation dose of 2.5 Mrad before distribution. Patients in this study had undergone a preoperative magnetic resonance imaging scan and physical examination. Patients were excluded from the study if they had a concomitant injury to other knee ligaments, previous surgery on the injured knee, or articular cartilage lesions greater than Outerbridge grade II. Clinical and proprioceptive evaluation results were collected preoperatively and at 3, 6, 12, 24, 36, and 60 months after surgery. The autograft, allograft, and hybrid graft groups were followed up for 5.5 ± 0.2 months (range, 5 to 6 years), 5.7 ± 0.3 months (range, 5 to 6 years), and 5.6 ± 0.5 months (range, 5 to 6 years), respectively. No statistically significant differences were found among the 3 groups regarding preoperative demographic factors (P > .05). The differences in physical examination findings and subjective evaluations among the 3 groups were not significant (P > .05). However, a significant difference was detected in instrumented anteroposterior measurements, with more laxity shown in the γ-irradiated allograft group than in the other 2 groups (P = .006). The differences in proprioceptive and functional outcomes among the 3 groups were not significant. In contrast, a significant difference was detected in instrumented anteroposterior measurements, which showed more laxity in the γ-irradiated allograft group than in the other 2 groups. However, this may not be clinically significant. Level II, prospective comparative study.


Protective effect of Shenmai injection on doxorubicin-induced cardiotoxicity via regulation of inflammatory mediators.
Doxorubicin (DOX) is a chemotherapy drug for malignant tumors. The clinical application of DOX is limited due to its dosage relative cardiotoxicity. Oxidative damage and cardiac inflammation appear to be involved in DOX-related cardiotoxicity. Shenmai injection (SMI), which mainly consists of Panax ginsengC.A.Mey.and Ophiopogon japonicus (Thunb.) Ker Gawl, is widely used for the treatment of atherosclerotic coronary heart disease and viral myocarditis in China. In this study, we investigated the protective effect of Shenmai injection on doxorubicin-induced acute cardiac injury via the regulation of inflammatory mediators. Male ICR mice were randomly divided into seven groups: control, DOX (10 mg/kg), SMI (5 g/kg), DOX with pretreatment with SMI (0.5 g/kg, 1.5 g/kg or 5 g/kg) and DOX with post-treatment with SMI (5 g/kg). Forty-eight hours after the last DOX administration, all mice were anesthetized for ultrasound echocardiography. Then, serum was collected for biochemical and inflammatory cytokine detection, and heart tissue was collected for histological and Western blot detection. A cumulative dose of DOX (10 mg/kg) induced acute cardiotoxicity in mice manifested by altered echocardiographic outcome, and increased tumor necrosis factor, interleukin 6 (IL-6), monocyte chemotactic protein 1, interferon-γ, and serum AST and LDH levels, as well as cardiac cytoplasmic vacuolation and myofibrillar disarrangement. DOX also caused the increase in the expression of IKK-α and iNOS and produced a large amount of NO, resulting in the accumulation of nitrotyrosine in the heart tissue. Pretreatment with SMI elicited a dose-dependent cardioprotective effect in DOX-dosed mice as evidenced by the normalization of serum inflammatory mediators, as well as improve dcardiac function and myofibril disarrangement. SMI could recover inflammatory cytokine levels and suppress the expression of IKK-α and iNOS in vivo, which was increased by DOX. Overall, there was evidence that SMI could ameliorate DOX-induced cardiotoxicity by inhibiting inflammation and recovering heart dysfunction.


Time-resolved 3D Rotational Angiography (4D DSA) of the Lenticulostriate Arteries: Display of Normal Anatomic Variants and Collaterals in Cases with Chronic Obstruction of the MCA.
The lenticulostriate arteries (LSA) and other perforators may play a role for collateral supply in cases with ischemia due to stenosis or occlusions of the middle cerebral artery (MCA). Purpose of this case series was to evaluate the potential of time-resolved 3D rotational angiography data sets (4D DSA) for detailed visualization of anatomic variants of LSA feeders and for display of local collaterals involving the LSA in cases with chronic MCA obstruction. Multiplanar and volume rendering reconstructions of 4D DSA data were computed in addition to standard postprocessing in 24 patients who had indications for 3D rotational angiography (3DRA) of the internal carotid artery (ICA) without pathologies of the ICA, middle cerebral artery (MCA) and anterior cerebral artery (ACA) main stems (n = 18) or with stenosis or chronic occlusion of the MCA (n = 6). For acquisition of 3DRA, we used a modified digital subtraction angiography (DSA) image acquisition protocol with an extended rotation angle of 260° and a prolonged scan time of 12 s on a Siemens Axiom Artis Zee biplane neuroangiography equipment. The 4D reconstructions of existing 3DRA data were computed on a dedicated workstation. Origin and course of LSA and other perforators were analyzed according to coronal multiplanar reconstructions (MPRs) with slice thicknesses between 6 and 28 mm. In all cases 4D reconstructions of the LSA were technically feasible and evaluable. As expected, origin and course of LSA showed a wide range of variations: The most common pattern was a common trunk dividing into multiple ascending branches originating from the proximal M1 (n = 5) or the proximal A1 segment (n = 4). Alternatively, 8 patients showed several individual branches that directly originated from the proximal M1 segment of the MCA and occasionally from the A1 segment of the ACA. In patients with M1 stenosis or occlusion, 4 out of 6 cases had local collaterals with involvement of proximal LSA trunks and a network parallel to the obstructed vessel segment. The 4D reconstructions were found to be equivalent (n = 16) or superior to 3D reconstructions (n = 8). The 4D DSA reconstructions provide a reliable display of normal LSA variants and connections to local collateral networks in cases with chronic MCA obstruction. The possibility to select a correct angiographic phase is advantageous compared to 3D DSA.


Exploring the Effect of Size Variability on Efficiency of Upconversion Nanoparticles as Optical Contrast Agents.
The current works explores the optical contrast property and nanaotheranostic capabilities of the rare earth based Upconversion nanoparticles. Optical coherence tomography (OCT) has been used to explore the particles ability to increase contrast and improve signal intensity. Photo thermal OCT, a function extension of OCT has been used to gauge the photo thermal potential of the nanoparticles. The nanoparticles successfully improved contrast and information from the deeper layers of the sample. The particles also showed excellent capability for use as photo thermal agents.


Reactive oxygen intermediates as mediators of programmed cell death in plants and animals.
Programmed cell death (PCD) is a physiological process occurring during development and in pathological conditions of animals and plants. The cell death program can be subdivided into three functionally different phases: a stimulus-dependent induction phase, an effector phase during which the wide range of death-stimuli are translated to a central coordinator, and a degradation phase during which the alterations commonly considered to define PCD (apoptotic morphology of the nucleus and chromatin fragmentation) become apparent. Recent studies suggest that mitochondrial permeability transition is the central coordinator of PCD and deciding whether or not a cell will die. There is increasing evidence that reactive oxygen intermediates (ROI) serve as direct and indirect mediators of PCD in mammalian and plant cells. Overexpression of genes encoding pro- and antioxidant enzymes in transgenic animals and plants has been informative regarding the function of ROI. Recent data imply a dual role of ROI in the apoptotic process: first, as a facultative signal during the induction phase, and, second, as a common consequence of mitochondrial permeability transition leading to the final destruction of the cell. The present review discusses and compares new insights into the function of ROI during PCD in mammalian cells and in human and plant diseases.


Hypericin and its anticancer effects: From mechanism of action to potential therapeutic application.
Emerging studies indicate that hypericin has diverse pharmacological actions and exhibits potential for treatment of various types of cancer. The current review evaluates the pharmacological activity, associated molecular mechanism, and therapeutic application of hypericin as an anticancer agent according to the most recent state of knowledge with special emphasis on clinical trials and safety profile. This review follows The Preferred Reporting Items for Systematic Reviews criteria. Various databases, including PubMed, Scopus and Science Direct, were used to search and collect relevant literature. The major keywords used included the following: cancer, distribution, property, signaling pathway, pharmacological effect, treatment, prevention, in vitro and in vivo studies, toxicity, bioavailability, and clinical trials. One hundred three articles met the established inclusion and exclusion criteria. Hypericin has shown anticancer activity against the expansion of several cell types including breast cancer, cervical cancer, colorectal cancer, colon cancer, hepatocellular carcinoma, stomach carcinoma, leukemia, lung cancer, melanoma, and glioblastoma cancer. Hypericin exerts its anticancer activity by inhibiting pro-inflammatory mediators, endothelial growth factor, fibroblast growth factor, cell adhesion, angiogenesis, and mitochondrial thioredoxin. It has also been shown to cause an increase in the levels of caspase-3 and caspase-4, arrest the cell cycle at metaphase leading to cancer cell apoptosis, and affect various protein and gene expression patterns. Hypericin exhibits significant inhibitory activity against various types of in vitro and in vivo cancer models. However, well-designed, high quality, large-scale and multi-center randomized clinical studies are required to establish the safety and clinical utility of hypericin in cancer patients.


Creating three-dimensional virtual patients by superimposing intraoral and facial digital scans guided with an aligner system: A dental technique.
A technique for creating 3-dimensional virtual patients (3DVPs) by superimposing intraoral and facial digital scans guided with a novel aligner system is described. This aligner system supports design modifications to adapt to different facial scanning methods (FSMs) and reduce the impact of FSMs on the accuracy of 3DVPs. Two different designs of the aligner system are described: one for use with less-accurate FSMs and another for use with more-accurate FSMs. These virtual designs are available for download and use.


Application of PCR for Mycoplasma pneumoniae detection in children with community-acquired pneumonia.
Between April 2002 and March 2003, to detect Mycoplasma pneumoniae by polymerase chain reaction (PCR), a primer set designed for the 16S rRNA gene was used to examine clinical samples from 369 children with community-acquired pneumonia. Samples were collected from 12 Japanese institutions participating in a study group concerning acute respiratory infectious diseases. The sensitivity of primers--2 CFU per reaction tube, using M. pneumoniae M129, a standard strain--was calculated to represent 1.1 x 10(3) M. pneumoniae organisms adherent to the tip of the swab used to collect clinical samples. Results for PCR were obtained within 2.6 h. Cases identified by PCR, cultures, and serologic tests were 68 (18.4%), 53 (14.4%), and 76 (20.6%) respectively. Among 57 PCR-positive patients tested serologically, 56 showed a significant elevation or rise in antibody titer. PCR positivity was high among patients prescribed beta-lactam antibiotics (86.7%) or no antibiotic (87.0%) before PCR analysis, but was low among patients receiving macrolides, new quinolones, or tetracyclines (37.5%). We concluded that the PCR constructed by us had a high probability for confirming a diagnosis of M. pneumoniae pneumonia and for guiding antibiotic choice for patients not yet treated.


[Blood serum biochemistry of ponies (author's transl)].
A wide blood serum biochemical profile, concerning especially blood lipids has been performed in 31 apparently healthy Shetland and Potiock ponies. These blood standard values will be useful for studies on ponies' hyperlipemia.


Retinal detachment after choroidal ischemia.
Injection of a 15-micron microsphere suspension through one or two vortex veins of nine monkey eyes caused various degrees of sectorial choroidal ischemia, which were documented by fluorescein angiography and electron microscopy. The severity of the lesions to the fundus depended on the volume of microspheres injected (0.4 to 1.6 ml of a suspension of 600,000 microspheres/ml). Three hours after embolization white patches appeared in the retinal pigment epithelium as well as a posterior pole serous retinal detachment in five eyes. Delayed choroidal filling was noted in the quadrant involved, but a few choriocapillaris units slowly perfused, leading to fluorescein leakage in the serous retinal detachments. Histologic examination showed various types of damage to the retinal pigment epithelium, including vacuolization and cell membrane rupture.


Functional assembly of the foreign carotenoid lycopene into the photosynthetic apparatus of Rhodobacter sphaeroides, achieved by replacement of the native 3-step phytoene desaturase with its 4-step counterpart from Erwinia herbicola.
Photosynthetic organisms synthesize a diverse range of carotenoids. These pigments are important for the assembly, function and stability of photosynthetic pigment-protein complexes, and they are used to quench harmful radicals. The photosynthetic bacterium Rhodobacter sphaeroides was used as a model system to explore the origin of carotenoid diversity. Replacing the native 3-step phytoene desaturase (CrtI) with the 4-step enzyme from Erwinia herbicola results in significant flux down the spirilloxanthin pathway for the first time in Rb. sphaeroides. In Rb. sphaeroides, the completion of four desaturations to lycopene by the Erwinia CrtI appears to require the absence of CrtC and, in a crtC background, even the native 3-step enzyme can synthesize a significant amount (13%) of lycopene, in addition to the expected neurosporene. We suggest that the CrtC hydroxylase can intervene in the sequence of reactions catalyzed by phytoene desaturase. We investigated the properties of the lycopene-synthesizing strain of Rb. sphaeroides. In the LH2 light-harvesting complex, lycopene transfers absorbed light energy to the bacteriochlorophylls with an efficiency of 54%, which compares favourably with other LH2 complexes that contain carotenoids with 11 conjugated double bonds. Thus, lycopene can join the assembly pathway for photosynthetic complexes in Rb. sphaeroides, and can perform its role as an energy donor to bacteriochlorophylls.


Endothelium-derived relaxing factor and the effects of acetylcholine and histamine on resistance blood vessels.
1. The role of endothelium-derived relaxing factor (EDRF) in the action of vasodilator (acetylcholine, histamine, nitroprusside) and vasoconstrictor (noradrenaline, vasopressin) drugs on vascular resistance in the isolated perfused kidney and mesentery of the rat was studied. 2. Acetylcholine (EC50 = 0.18 +/- 0.05 nmol and 3.1 +/- 0.06 nmol, n = 8) and histamine (EC50 = 31.2 +/- 4.9 nmol and 46.2 +/- 3.9 nmol, n = 8) produced dose-related vasodilatation in noradrenaline-preconstricted (i.e. 'high tone') rat renal and mesenteric blood vessels. The response to both vasodilators (but not nitroprusside) was abolished by infusion of CHAPS (4.7 mg ml-1, 30 s). By use of an immunocytochemical staining procedure CHAPS was demonstrated to remove vascular endothelial cells lining intrarenal blood vessels. 3. Gossypol (3 microM), metyrapone (10 microM) and nordihydroguaiaretic acid, (NDGA, 30 microM), presumed inhibitors of EDRF biosynthesis, reduced or abolished the response to acetylcholine and histamine in perfused kidney and mesentery of the rat without affecting vasodilatation due to nitroprusside. Mepacrine (10 microM) similarly abolished the response to acetylcholine and histamine but in addition, reduced the response to nitroprusside in both preparations. 4. Methylene blue (100 microM), a presumed antagonist of the effect of EDRF, abolished vasodilatation due to acetylcholine and histamine and reduced the response to nitroprusside in perfused rat kidney and mesentery. Superoxide dismutase, SOD (15 u ml-1), was without effect. 5. While CHAPS treatment significantly augmented the vasoconstrictor response to both noradrenaline and vasopressin in perfused renal and mesenteric vessels this effect was not mimicked by metyrapone or gossypol suggesting that the enhanced effect of vasopressor agents in CHAPSperfused rat organs is due to the removal of a permeability barrier rather than impaired EDRF formation. 6. Responses to vasoconstrictor and vasodilator drugs in the perfused kidney and mesentery were obtained in the presence of indomethacin (8 microM) which produced in excess of 90% inhibition of prostacyclin (PGI2) release as measured by radioimmunoassay of 6-oxo-prostaglandin F1 alpha,. (6-oxo- PGF1 alpha) in the Krebs effluent. 7. We provide evidence that EDRF mediates the vasodilator response to acetylcholine and histamine in resistance blood vessels in perfused rat kidney and mesentery. The possibility that EDRF has a physiological role to play in regulating the calibre of resistance blood vessels is discussed.


The transfer and stable integration of the HSV thymidine kinase gene into mouse cells.
Treatment of mutant mouse cells (Ltk-) deficient in thymidine kinase with Bam I restriction endonuclease-cleaved HSV-1 DNA results in the appearance of numerous surviving colonies which stably express thte tk+ phenotype. Through a series of electrophoretic fractionations in concert with transfection assays, we isolated a 3.4 kb fragment which contains the thymidine kinase gene and which alone is competent in the biochemical transformation of Ltk- cells. In this report, we have examined the distribution of tk sequences in the DNA of several transformed clones following stable gene transfer. A series of complementary experiments involving reassociation kinetics in solution and annealings with tk DNA to restriction-cleaved cellular DNA following electrophoresis and transfer to filters allow us to make the following general conclusions concerning the fate of the tk gene in all clones examined: the tk gene is present in all cells at a frequency of one copy per chromosomal complement; the tk gene is stably integrated in the DNA of all transformants; and integration is not site-specific and occurs at different loci in the DNA of all transformants examined. The existence of a single active tk gene in tk+ transformants now facilitates an analysis of the sequence organization of tk- mutant cells and provides a useful model system for studies on the transfer of cellular genes.


Occurrence of Pneumocystis jiroveci pneumonia after allogeneic stem cell transplantation: a 6-year retrospective study.
Pneumocystis jiroveci pneumonia (PCP) has become a rare opportunistic infection due to the efficacy of prophylactic regimens. We conducted a 6-year retrospective study at our institution. A total of 13 cases of PCP were diagnosed among 519 patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) (2.5%). In three patients, PCP occurred within the first 5 months following HSCT. These severely immunocompromised patients were receiving prophylaxis and had concomitant aspergillosis that caused rapid death in two of them. In 10 other patients, PCP occurred a median of 14.5 months after HSCT. In all these patients, PCP prophylaxis had been discontinued, mainly because of the suspected bone-marrow toxicity of the prophylactic regimen. Median CD4+ T cell count was 131/microl at diagnosis. Seven of these 10 patients were receiving immunosuppressive therapy for chronic graft versus host disease and three had a relapse of their hematological malignancy. One patient died from PCP despite high doses of cotrimoxazole. We conclude that PCP is still occurring after allogeneic HSCT, mainly as a late complication in patients in whom PCP prophylaxis had been prematurely discontinued. Long-term PCP prophylaxis should be maintained in patients receiving immunosuppressive drugs, and in those with low CD4+ T cell counts or a relapse of their hematological malignancy.


Low-background electrochemical biosensor for one-step detection of base excision repair enzyme.
We develop a low-background electrochemical biosensor for one-step detection of uracil DNA glycosylase (UDG) based on the host-guest interaction and iron-embedded nitrogen-rich carbon nanotube (Fe-N-C) that mimics enzyme-mediated electrocatalysis to achieve signal amplification. In this work, Fe-N-C is initially immobilized on a glassy carbon electrode, followed by the immobilization of β-cyclodextrin (β-CD). We construct the signal probes by assembling the methylene blue (MB)-labeled hairpin DNAs onto the surface of Au nanoparticles (AuNPs) to form the MB-hairpin/AuNP probes. Due to the steric effect of AuNPs and the stem-loop structure of hairpin DNA, MB is prevented from entering the cavity of β-CD on the electrode. In contrast, UDG enables the removal of uracil from the U•A pairs in the stem of hairpin DNA probe to generate apurinic/apyrimidinic (AP) sites, leading to the assembly of MB-hairpin/AuNP probes on the electrode based on host-guest reaction between β-CD and MB. Meanwhile, L-cysteine (RSH) is oxidized by O2 to disulfide L-cystine (RSSR) and H2O2. In the presence of H2O2, Fe-N-C catalyzes the oxidation of MB to generate an amplified electrochemical signal. Notably, the Fe-N-C-catalyzed oxidation of MB is mediated by the oxidation of RSH by O2 instead of external H2O2, greatly simplifying the experimental procedures and improving the electrochemical signal. Due to the introduction of host-guest recognition, this electrochemical biosensor displays a low-background signal and high signal-to-noise ratio, enabling the one-step sensitive measurement of UDG with a detection limit of 7.4 × 10-5 U mL-1. Moreover, this biosensor can measure UDG in crude cell extracts and screen the inhibitors, providing a new platform for biomedical research.


Roles of vascular endothelial growth factor and endothelial nitric oxide synthase during revascularization and regeneration after partial hepatectomy in a rat model.
Angiogenesis is an essential process in liver regeneration. Nitric oxide (NO) and vascular endothelial growth factor (VEGF) are the main regulators of normal and pathological angiogenesis. This study aimed to determine the roles of NO derived from endothelial nitric oxide synthase (eNOS) and VEGF in sinusoidal endothelial cell (SEC) proliferation during liver regeneration. Sprague-Dawley rats underwent a 70% partial hepatectomy (PHx), and were euthanized 0, 24, 48, 72, or 168 h later. Liver regeneration and SEC proliferation were evaluated. The protein expression of VEGF and eNOS was examined by a Western blot analysis. The rats were also treated with the NO synthase inhibitor N (G)-nitro-L-arginine-methyl ester (L-NAME) to examine its effects on liver regeneration and SEC proliferation. The proliferating cell nuclear antigen (PCNA) labeling index of hepatocytes was significantly increased at 24 h after PHx. The eNOS protein expression and NO production were significantly increased from 72 to 168 h. The expression of VEGF protein was significantly increased at 72 h. L-NAME significantly inhibited the increases in the liver mass and decreased the PCNA labeling index of hepatocytes at 24 h. L-NAME also inhibited the induction of VEGF protein at 72 h. Endothelial NOS and VEGF coordinately regulate SEC proliferation during liver regeneration. Sinusoidal endothelial cell proliferation is necessary and is an important step in liver regeneration.


RNA isolation from human skin tissues for colorimetric differential display.
High-quality RNA is essential when analyzing expression patterns of tissues by the differential display technique. However, the isolation of intact RNA can be very difficult when tissues are used that contain many RNAses or that are hard to homogenize (e.g. skin samples). We describe an improved protocol for the extraction of high-quality RNA of snap-frozen biopsies from skin tissues that works in a reproducible and reliable manner. In addition, we developed a simplified non-radioactive differential display technique using RNA from small amounts of skin samples.


Effects of diisopropyl phosphorofluoridate (DFP) on CA3 and CA1 responses in rat hippocampus.
Diisopropyl phosphorofluoridate (DFP), an insecticide, is a potent anticholinesterase that binds essentially irreversibly to acetylcholinesterase, resulting in severe, acute neurologic pathology, and less severe, but longer-lasting, delayed neuropathy. We report here on the short-term effects of bath-applied DFP on extracellularly recorded responses from CA3 and CA1 of rat hippocampus. Exposure to 10 microM DFP evokes low amplitude, spontaneous bursts in CA3 generally within 10 minutes, and the bursting does not reverse with washing. The CA1 neuronal population usually bursts synchronously with CA3, but the population events are of low amplitude and sometimes not detectable, implying a differential sensitivity to DFP. These effects were partially blocked by the muscarinic antagonist atropine, while the cholinergic antagonist gallamine had little effect. Also, the reversible anticholinesterase physostigmine could, within temporal limits, protect slices from DFP's effects, implicating the cholinergic system as the probable mediator in the first stages of DFP-induced epileptogenesis.


Expression of a gene encoding a rabbit sperm membrane protein in mammalian cells.
A general mammalian expression vector designated pSV2-EP was reconstructed by inserting an oligonucleotide fragment into pSV2-dhfr. This vector allowed insertion of cDNAs with EcoRI cohesive ends. The pSV2-EP contains a simian virus 40 (SV40) early promoter, origin for DNA replication, SV40 poly-A site, splicing site, an initiator ATG downstream from the promoter and an EcoRI site for the insertion of cDNA fragment screened from lambda gt11 expression libraries. A recombinant plasmid (pS-VRS-1) was constructed by inserting RSD-1, a cDNA encoding a rabbit sperm tail protein, into the EcoRI site of the pSV2-EP vector. Chinese hamster ovarian (CHO) dhfr-negative cells were cotransformed with pSV2-dhfr and pSVRS-1 by the calcium phosphate method. In selective culture medium without thymidine and hypoxanthine, several cell lines were obtained containing mRNA and DNA that hybridized with RSD-1. One of these transformed cell lines stained intensely with anti-rSMP-B antibodies, demonstrating that the RSD-1 was expressed in the transformed CHO cells.


Randomized trial of single-dose intramuscular dexamethasone compared with prednisolone for children with acute asthma.
To compare the clinical efficacy of single-dose intramuscular (IM) dexamethasone phosphate to a 5-day course of oral prednisolone for the treatment of moderate asthma exacerbations in young children discharged from the emergency department (ED). We performed a prospective, randomized trial in children aged 18 months to less than 7 years with asthma who presented to the ED with a clinical asthma score of 3 to 7 on a 9-point scale. Children were randomized to 1 dose of IM dexamethasone (0.6 mg/kg, maximum 15 mg) or oral prednisolone (2 mg/kg, maximum 50 mg) once daily for 5 days. The primary outcome, measured only for patients discharged from the ED, was change in asthma score from ED presentation to 4-day follow-up examination, as assessed by a physician masked to group assignment. We assessed secondary clinical course outcomes by a structured interview at 4 and 14 days. Eighty-eight patients were appropriately randomized to dexamethasone and 93 to prednisolone. Group characteristics were similar at baseline. Among those discharged from the ED, 62 (90%) of 69 and 64 (90%) of 74 patients in the dexamethasone and prednisolone groups, respectively, were reassessed at 4 days for the primary outcome. The mean change in total asthma score at 4-day follow-up was 3.6 in the dexamethasone group and 3.4 in the prednisolone group (difference, 0.2; 95% confidence interval, -0.4 to 0.7). Of patients initially discharged, 5.9% of dexamethasone patients and 4.1% of prednisolone patients were admitted before the 2-week follow-up (difference, 1.8%; 95% confidence interval, -5.4% to 9.0%). A single dose of IM dexamethasone showed no clinically meaningful difference in outcomes compared with a 5-day course of oral prednisolone for the treatment of moderate acute asthma exacerbations in young children who are discharged from the ED.


Steady state analysis of modern industrial variable speed drive systems using controllers adjusted via grey wolf algorithm & particle swarm optimization.
This paper presents the reconfiguration of control circuit designed to control four-quadrant chopper placed in the variable speed drive system (VSDS)'s DC-link. The purpose of this design is to reduce the overall total harmonic distortion THD% of input current, and the ripple factor (RF) of the DC-link current in this system. Both of Grey Wolf Algorithm (GWO) & Particle Swarm Optimization (PSO) have been used to get the optimal parameters of proportional integral PI and proportional integral differential with filter PIDN controllers. The variable speed drive system and the proposed filter have been modeled in integration with the suggested algorithms to determine the optimal values of the controllers' parameters. The grey wolf algorithm GWO outperformed the PSO algorithm in term of reaching the optimum parameters in less number of iterations in both dynamic and static work conditions. Also, the time response of the system with GWO is better than with PSO.


Characterization of size-differentiated airborne particulate matter collected from indoor environments of childcare facilities.
Inhalation of particulate matter (PM) present in indoor atmospheres has been associated with poor health and wellbeing of occupants. Here we report the characteristics of airborne PM collected from twenty-two air-conditioned childcare centres in Singapore. Airborne PM were collected using cascade impactors and characterized for morphology, elemental composition, endotoxin levels, ability to generate abiotic reactive oxygen species, and oxidative stress-dependent cytotoxicity in BEAS-2B cell lines. The mass concentrations of ultrafine particles (PM0.06-1) were more abundant than that of larger particles (PM1-4, PM4-20, and PM20-35 particles). PM20-35 and PM4-20 were irregularly shaped particles, PM1-4 particles had membranous flaky structures and PM0.06-1 particles were pseudo-spherical with the occasional presence of crystalline structures. Carbonaceous matter dominated PM20-35 particles, and the abundance of inorganic salts, iron and sulfur increased with decreasing PM size. Measured endotoxin levels were especially higher in PM4-20 particles. Compared to other particle size fractions, PM0.06-1 particles generated the highest ROS and were also the most potent in generating intracellular ROS in BEAS-2B cell lines. However, total mass concentrations, elemental compositions, abiotic responses, and PM collected from centres with split air-conditioning systems and no active outdoor air supply (SAC) were not statistically different compared with PM collected from centres with air conditioning with mechanical ventilation (ACMV). In conclusion, our study showed obvious distinctions in mass concentrations, morphology, elemental compositions, and cytotoxic potential of different sized particles collected from childcare centres, where the smallest particles (PM0.06-1) exhibited higher hazard potential.


Decreased numbers of regulatory T cells are associated with human atherosclerotic lesion vulnerability and inversely correlate with infiltrated mature dendritic cells.
Mature dendritic cells (DCs) play a crucial role in the inflammatory process within atherosclerotic lesions by stimulation of effector T cells, which can contribute to plaque instability. Interactions between DCs and regulatory T cells (Treg), which regulate immune response by attenuating acute inflammation, are postulated to be involved in the pathogenesis of autoimmune diseases. We investigated a possible correlation between infiltrated DCs and Treg in human atherosclerotic plaques. Cross-sections of 40 human carotid endarterectomy specimens were classified into groups of stable and vulnerable plaques using Trichrome staining. Immunohistochemical staining of plaques was used to detect infiltrated total (S100) and mature DCs (fascin, DC-LAMP, CD83), Treg (CD3, Foxp3), and to analyze the inflammatory state of the plaques (CD3, COX-2, CD68). In addition, RNA was isolated from plaque specimens and quantitative real-time PCR was performed to analyze transcription rates of DC markers (CD11c, CD209, HLA-DR), maturation markers (CD80, CD83, CD86), Treg-associated genes (CTLA-4, Foxp3) and of pro- and anti-inflammatory cytokines (TGFβ-family, IL-10, IFN-γ, IL-17α, IL-6). Migration assays and adhesion experiments were performed, to investigate the effects of Treg on mature DCs in vitro. As compared with stable plaques, vulnerable lesions were characterized by increased numbers of COX-2-expressing cells and T lymphocytes, indicating an enhanced inflammatory process. In vulnerable plaques, numbers of total and mature DCs were significantly higher in the inflammatory plaque shoulder, whereas the numbers of Treg were decreased compared to stable plaques. This inverse correlation and the association of the observed infiltration rates with plaque stability, were confirmed by PCR analyses, showing increased transcription levels of DC-specific markers, decreased mRNA expression of Treg-associated genes and decreased anti-inflammatory cytokines in vulnerable atherosclerotic plaques. In vitro, pre-incubation of mature DCs with Treg resulted in decreased DC migration and inhibited the adhesion of DCs to endothelial cells under non-uniform shear stress. The results of our study provide novel insights in the direct interaction of mature DCs and Treg in plaque inflammation and stability.


Branched chain amino acids in the treatment of latent portosystemic encephalopathy. A double-blind placebo-controlled crossover study.
Branched chain amino acids have been recommended for the treatment of portosystemic encephalopathy based on the false neurotransmitter hypothesis. This hypothesis implies that by correction of the deranged amino acid pattern in the blood of cirrhotics, false neurotransmission and then portosystemic encephalopathy is improved. We conducted a double-blind crossover placebo-controlled trial in 22 inpatients with liver cirrhosis and obtained evidence of latent (subclinical) portosystemic encephalopathy using an extensive psychometric test program. Patients received a defined diet of 35 cal/kg X day containing 1 g of protein. In addition, branched chain amino acids or casein in a dosage of 0.25 g/kg X day was administered in a crossover fashion, each for 1 wk. Semiquantitative nitrogen balance increased during both treatments, with a tendency of a larger increase during branched chain amino acid treatment. At the same time ammonia concentration tended to decrease during branched chain amino acid treatment. Taking into account the crossover design, significant improvements attributable to branched chain amino acid treatment could be demonstrated in psychomotor functions (line tracing, tapping, steadiness, auditory reaction time), attention (digit table), and practical intelligence (digit symbol, number connection test).


IP3 production in A10 cells, an established aortic smooth muscle cultured cell line: effects of agonist administration procedure and culture conditions.
1. After normal culture (10% FBS), medium exchange itself evoked marked increases in IP3 levels in A10 cells that masked the net change in IP3 in response to vasopressin (100 nM). 2. Low-serum (0.3% FBS) culture for 10 days abolished the increase in IP3 by medium exchange, and increased expression of cGMP kinase. 3. Stimulation of cells with vasopressin (100 nM) by a pipetting procedure, increased IP3 levels irrespective of culture conditions. 4. This cell line therefore could be used in a contractile form (after low-serum culture) with pipetting procedure to observe IP3 response, especially with regard to interactions with the cGMP signaling system.


Dopamine regulates spine density in striatal projection neurons in a concentration-dependent manner.
Dopaminergic afferents innervate spiny projection neurons (SPNs) in the striatum, maintaining basal ganglia activity. The loss of striatal innervation is the hallmark of Parkinson's disease (PD), which is characterized by dopaminergic denervation. A lack of dopamine in the dorsal striatum induces plasticity changes in SPNs. However, PD-associated denervation is progressive, and how plasticity is modified in partially innervated areas is poorly understood. The most studied models of PD are based on the use of neurotoxins that induce an almost complete striatal denervation. To investigate the impact of partial dopamine (DA) innervation in striatal plasticity, we use a genetic model of PD, Aphakia (Ak) mice, whose striatum presents an increasing dorso-ventral gradient of dopamine innervation. We studied SPNs in three different areas (dorsal, middle and ventral, with low, moderate and high innervation by tyrosine hydroxylase TH-positive axons, respectively) using fast scan cyclic voltammetry, microiontophoresis, immunohistochemistry and patch clamp techniques. Our data show an increasing dorso-ventral gradient of extracellular DA levels, overlapping with the gradient of TH innervation. Interestingly, spine loss in both direct (d-SPN) and indirect SPNs (i-SPN) decreases from dorsal to ventral in the parkinsonian striatum of Ak mice, following the decrease in DA levels. However, their dendritic trees and the number of nodes are only reduced in the poorly innervated dorsal areas and remain unaltered in moderate and highly innervated areas. The firing rate of direct SPNs does not change in either moderate or highly innervated areas, but increases in poorly innervated areas. In contrast, action potential frequency of indirect SPNs does not change along the dorso-ventral innervation gradient. Our findings indicate that spine density in d-SPNs and i-SPNs varies in a dopamine concentration-dependent manner, indicating that both d- and i-SPN are similarly innervated by DA.


[Clinical efficacy of Gengxueting and its effect on estrogen receptor and progesterone receptor in patients with hysteromyoma].
To explore the clinical efficacy of Gengxueting (GXT) in treating hysteromyoma and its effects on estrogen receptor (ER) and progesterone receptor (PR). Sixty-four hysteromyoma patients with surgical indication were equally assigned to the treated group and the control group. Patients in the treated group were treated with GXT one capsule every day for 90 consecutive days before surgical operation, while those in the control group were treated with surgery alone. Serum levels of reproductive hormones were determined in the follicular phase before medication and one day before operation by RIA, and colored Doppler ultrasound examination was conducted for measuring the size of uterus and myoma. Moreover the protein expressions of ER and PR in tumor and uterine muscular tissues were detected by immunohistochemistry assay with streptomycin avidin-biotin peroxidase complex method. In the treated group after medication, the serum level of estradiol was (167.0 +/- 85.9) pmol/L, progesterone (1.9 +/- 1.0) nmol/L, follicle-stimulating hormone (10.4 +/- 2.1) IU/L, and luteinizing (12.0 +/- 9. 8) IU/L, all reached the levels of early follicular phase, with the maximal size of myoma significantly decreased from (380.4 +/- 21.0) cm3 to (162.3 +/- 14. 8) cm3 (P < 0.01); and the ER and PR expressions in tumor tissue were significantly lower than those in the control group respectively (P < 0.01). Expressions of ER and PR in hysteromyoma tissue could be significantly reduced by medication of GXT, which leads to significant shrinkage of tumor size and improvement of clinical symptoms.


Absence of correlations among three putative in vivo probes of human cytochrome P4503A activity in young healthy men.
In vitro studies with human liver microsomes have shown that erythromycin N-demethylation, dapsone N-hydroxylation, and the 6 beta-hydroxylation of cortisol are all primarily mediated by P4503A4. Trait measurements to assess the in vivo level of activity of these separate oxidations have also been developed previously. This study investigated the relationships among the three phenotypic trait measurements in 30 young healthy white men. The frequency distributions of the trait values were all unimodal, with a twofold range for the erythromycin breath test and the urinary dapsone recovery ratio; the urinary 6 beta-hydroxycortisol/cortisol ratio was more variable, with a 17-fold range of values. No statistically significant correlations were observed among any of the trait measurements (dapsone recovery ratio versus erythromycin breath test: r = -0.07, p = 0.7; urinary 6 beta-hydroxycortisol/cortisol ratio versus erythromycin breath test: r = -0.12, p = 0.6; urinary 6 beta-hydroxycortisol/cortisol ratio versus dapsone recovery ratio: r = 0.13, p = 0.5. This lack of any relationship was unexpected and the reason(s) is unknown; however, it is possible that factors such as route of administration and extrahepatic metabolism in the intestinal epithelium and kidney are involved. Further studies are required to identify and validate the use of an appropriate in vivo probe of P4503A4 in humans.


Oxidation of gas phase trichloroethylene and toluene using composite sol-gel TiO2 photocatalytic coatings.
The previously developed composite sol-gel (CSG) process is proposed for the deposition of thick (10-50 microm) porous films of photocatalytic TiO2. The CSG titania was developed by binding pre-calcined TiO2 particles with TiO2 sol. It had relatively high surface area (15-35 m2/g) and good resistance against mechanical stress and abrasion. Photocatalytic activity tests were carried out on trichloroethylene (TCE) and toluene, and compared with those of standard Degussa P-25 titania. The CSG photocatalyst provided good photo-efficiency in removing both pollutants from contaminated air streams. When compared with P-25 titania, the CSG photocatalyst showed a similar photo-efficiency with first-order kinetic rate constants not significantly different from that of P-25. For both photocatalysts the rate of photocatalytic oxidation of TCE was significantly greater than that obtained for toluene. Overall, the combination of better mechanical integrity, resistance against abrasion, and comparable photocatalytic efficiency of the CSG titania versus that of P-25 titania, make the composite sol-gel (CSG) photocatalyst a viable alternative for industrial applications where long term stability, superior mechanical properties, and good photo-efficiency are of critical value.


Synaptogenic effect of estrogen on the hypothalamic arcuate nucleus of the adult female rat.
In order to examine the effect of estrogen on the synaptic structures in the hypothalamic arcuate nucleus (ARCN), semi-quantitative studies were performed by counting synapses in an 18,000 micron 2 area in the middle part of the ARCN in each brain. In ovariectomized female rats injected with 2 micrograms of estradiol benzoate (EB) for three weeks, the mean numbers of axodendritic and axosomatic synapses were not significantly different from those in the intact and ovariectomized controls. When the medial basal hypothalamus (MBH) including the ARCN was isolated by use of a Halász knife (MBH island), the mean number of axodendritic synapses was decreased to about half of the controls. However, EB treatment for three weeks from the day of surgery effectively restored the axodendritic synaptic population of the deafferented ARCN. This may suggest that estrogen has a facilitatory effect on axodendritic synapse formation in the deafferented ARCN, presumably by stimulating axonal sprouting and synaptic regeneration of intact axons in the MBH island.


Characterization of microbial intolerances and ruminal dysbiosis towards different dietary carbohydrate sources using an in vitro model.
This study aimed to characterize the critical points for determining the development of dysbiosis associated with feed intolerances and ruminal acidosis. A metabologenomics approach was used to characterize dynamic microbial and metabolomics shifts using the rumen simulation technique (RUSITEC) by feeding native cornstarch (ST), chemically modified cornstarch (CMS), or sucrose (SU). SU and CMS elicited the most drastic changes as rapidly as 4 h after feeding. This was accompanied by a swift accumulation of d-lactate, and the decline of benzoic and malonic acid. A consistent increase in Bifidobacterium and Lactobacillus as well as a decrease in fibrolytic bacteria was observed for both CMS and ST after 24 h, indicating intolerances within the fibre degrading populations. However, an increase in Lactobacillus was already evident in SU after 8 h. An inverse relationship between Fibrobacter and Bifidobacterium was observed in ST. In fact, Fibrobacter was positively correlated with several short-chain fatty acids, while Lactobacillus was positively correlated with lactic acid, hexoses, hexose-phosphates, pentose phosphate pathway (PENTOSE-P-PWY), and heterolactic fermentation (P122-PWY). The feeding of sucrose and modified starches, followed by native cornstarch, had a strong disruptive effect in the ruminal microbial community. Feed intolerances were shown to develop at different rates based on the availability of glucose for ruminal microorganisms. These results can be used to establish patterns of early dysbiosis (biomarkers) and develop strategies for preventing undesirable shifts in the ruminal microbial ecosystem.


Albumin effect on hemorheological parameters in patients with liver transplant.
Liver transplantation is a life-saving treatment in end-stage liver failure. Hemorheological features as blood fluidity and red blood cell aggregation may alter effective tissue perfusion, graft function and hemodynamic variables. The aim of the study is to investigate effect of albumin infusion on red blood cell deformability and aggregation, blood viscosity and hemodynamics in liver transplant patients. Seventeen live or cadaveric donors were included in this prospective study. Hemorheological and hemodynamic measurements were performed in order to evaluate the effects of albumin infusion in perioperative period. Erythrocyte aggregation was significantly reduced 90 minutes after albumin infusion (p < 0.01). Mean blood viscosity revealed significant decrease at 20 rpm and 50 rpm after 90 minutes of albumin infusion (p < 0.05). Plasma viscosity decreased significantly compared to the value before albumin infusion at 20 rpm (p < 0.05). Albumin replacement improved hemodynamic variables in patients with low blood pressure and cardiac index measurements (p > 0.05). Human albumin infusion led to decrease in whole blood and plasma viscosities, red blood cell aggregation and induced blood pressure and cardiac index elevation in perioperative liver transplant patients. Determination of hemodynamic and hemorheological effects of human albumin replacement in various patient populations may serve beneficial clinical data.


Differential activity of two oncogenes from chromosome #7 in human glioblastoma cell lines.
Human glioblastoma cell lines are known to develop polysomy of cytogenetically intact chromosomes #7 and overexpression of the erbB oncogene (7p12-p14) at a level even higher than is to be expected from the number of #7 chromosomes. The met oncogene, however, which is also located on chromosome #7 (7q31-q32), was shown not to be overexpressed in a panel of 7-polysomic glioblastoma cell lines overexpressing erbB. Molecular analysis of the cell line HeRo gave proof that there is no detectable amplification or rearrangement of the erbB gene which could be responsible for its overexpression. These findings favor the assumption of differential regulation of the met and erbB oncogenes, e.g. by means of insufficient activity of a trans-acting erbB suppressor gene possibly located on a chromosome with a low copy number.


Germline pathogenic variants associated with ovarian cancer: A historical overview.
The risk of ovarian, tubal, and peritoneal cancer is related to germline pathogenic variants, and over time, the number of known disease-associated genes has increased significantly. This study reviews the literature regarding the topic from a historical perspective. The aim is to present a timeline of the knowledge gained from the early 1900s until today. The findings are put into perspective by looking at the current gene panel used for screening for suspected hereditary ovarian cancer in Denmark compared to what is known internationally. In 1929, the first familial ovarian cancer incidents were registered, and in 1950, the involvement of a genetic component was suggested for the first time. During the 1970s, several studies reported an accumulation of ovarian cancer in certain families, and during this time, it was discovered that ovarian cancer was linked to both breast cancer and colorectal cancer. The inheritance of cancer disposition has been thoroughly investigated, leading to the discovery of the BRCA genes in the 1990s. Furthermore, new studies based on new genetic technologies have revealed several genes with germline pathogenic variants that increase the risk of ovarian cancer. The identification of these pathogenic variants has led to preventive measures and specific treatment of women with genetic disposition to ovarian cancer. In Denmark, consensus is to include at least ten genes in the screening panel for hereditary ovarian cancer, and in the future additional genes will probably be added.


Mammary analog secretory carcinoma of the thyroid gland: A primary thyroid adenocarcinoma harboring ETV6-NTRK3 fusion.
ETV6-NTRK3 fusion was identified in several cancers including the recently described mammary analog secretory carcinoma (MASC) of the salivary glands and a minority of papillary thyroid carcinomas. We describe three cases of primary MASC of the thyroid gland and provide a detailed clinical and pathological characterization of the tumor morphology, immunoprofile, and genetic background. Immunohistochemistry for PAX8, TTF-1, thyroglobulin, mammaglobin, GCDFP-15, S-100 protein, and p63 was used to define the tumor immunophenotype. Fluorescence in situ hybridization for ETV6 rearrangement was performed in three, and the next-generation sequencing assay MSK-IMPACT™ (Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets) was performed in two cases. Primary MASC of the thyroid occurred in two women and one man, age 47-72 years. All patients presented with high T stage, infiltrative, locally aggressive tumors with extrathyroidal extension. Two cases were associated with well-differentiated papillary thyroid carcinoma. Histologically, they appeared as low-grade tumors, resembling MASC of the salivary glands and labeled positive for mammaglobin, GCDFP-15, S-100 protein, p63, weakly positive for PAX8, and negative for TTF-1 and thyroglobulin. Fluorescence in situ hybridization revealed ETV6 rearrangement in all cases. In two tested cases MSK-IMPACT™ confirmed the presence of ETV6-NTRK3 gene fusion. Two patients had at least two local recurrences, one was alive with disease, and one was alive and free of disease after 14 and 17 years, respectively. The third patient was alive and free of disease after 2 years. MASC of the thyroid is histologically, immunophenotypically, and genetically similar to its salivary gland counterpart. Thyroid MASC can be associated with a well-differentiated papillary thyroid carcinoma component, supporting follicular cell origin. Clinically, these carcinomas may show frequent recurrences but are associated with long-term survival. Patients with MASC of the thyroid may potentially benefit from Trk molecular-targeted therapy.


Epidermal growth factor receptor blockade mediates smooth muscle cell apoptosis and improves survival in rats with pulmonary hypertension.
We previously reported that administration of elastase inhibitors reverses fatal pulmonary arterial hypertension (PAH) in rats by inducing smooth muscle cell (SMC) apoptosis. We showed in pulmonary artery (PA) organ culture that the mechanism by which elastase inhibitors induce SMC apoptosis involves repression of matrix metalloproteinase (MMP) activity and subsequent signaling through alphavbeta3-integrins and epidermal growth factor receptors (EGFRs). This suggests that blockade of these downstream effectors may also induce regression of PAH. In this study, we first showed in PA organ culture that MMP inhibition or alphavbeta3-integrin blockade with agents in clinical and preclinical use (SC-080 and cilengitide, respectively) mediates SMC apoptosis and regression of medial hypertrophy. We also documented similar results with an EGFR tyrosine kinase inhibitor. We then induced PAH in rats by injection of monocrotaline and, at day 21, began a 2-week treatment with SC-080, cilengitide, or the EGFR inhibitor PKI166. No vehicle- or cilengitide-treated animal survived beyond 2 weeks. Administration of SC-080 resulted in 44% survival at 2 weeks, and PKI166 therapy resulted in 78% and 54% survival in daily or 3-times-weekly treated animals, respectively. Four weeks after cessation of PKI166, we documented survivals of 50% and 23% in the 2 treatment groups, associated with reductions in pulmonary pressure, right ventricular hypertrophy, and abnormally muscularized distal arteries. We propose that selective blockade of EGFR signaling may be a novel strategy to reverse progressive, fatal PAH.


Monitoring white blood cell count in adult patients with schizophrenia who are taking clozapine: a cost-effectiveness analysis.
Long-term monitoring of white blood cell count is compulsory in patients taking clozapine, although the incidence of drug-induced agranulocytosis is lower than previously expected. The cost-effectiveness of this monitoring is unknown. We aimed to assess the cost-effectiveness of various strategies to monitor white blood cell count in adult patients with schizophrenia taking clozapine. We assessed the cost-effectiveness of four strategies for monitoring white blood cell count (national strategies used in the UK, USA, and European countries, and a hypothetical 8-week strategy) compared with that of no monitoring. We used a semi-Markov model to do the cost-utility analysis from a health-care perspective with a 3-year time horizon, assuming a probability of 0·7% that a patient would develop agranulocytosis. Clinical and resource parameters were based on data from national registries of patients treated with clozapine, study cohorts, and a pharmacovigilance database; we derived estimates of health-related quality of life and mortality from the scientific literature. We assessed model uncertainty, including time horizon, with one-way and probabilistic sensitivity analyses. Compared with no monitoring, all four monitoring strategies increased quality-adjusted survival by less than 1 day per patient; more than 5000 patients would need to be monitored to avoid one death. The incremental cost-effectiveness ratios (ICERs) were at least US$970 000 per quality-adjusted life-year gained for all four strategies compared with no monitoring. The ICERs were highest in the strategies with highest frequencies and longest durations of monitoring. The results remained robust in the one-way and probabilistic sensitivity analyses, suggesting that no monitoring had the highest probability of being cost effective. Existing strategies for monitoring white blood cell count in patients taking clozapine, based on divergent national requirements, do not seem to be cost effective. This finding should be taken into account by public health authorities and policy makers in the revision of guidance for clozapine prescription. University Hospitals of Geneva.


'I am not who I was': Old age and masculinity in Maximianus Etruscus' elegies.
The Elegies of Maximianus Etruscus (sixth century CE) are first-person narratives in which the poet writes about the life cycle, making love and sex the building blocks of the story. Through his failed relationships with different women, Maximianus talks about the perception of his aging, especially the changes in his body and his eroticism, and the progressive loss of masculinity that to him comes with old age. The aim of this paper is to analyze the relationship between old age and gender in Maximianus' elegies from a double perspective: the perception of old age itself as a unique and personal fact, and age and masculinity as interacting social constructions that are historically constructed. We will address issues such as the emotions linked to aging, the representation of old age as a living death, the relationship of masculinity with sexuality and the importance of erection. Finally, we will place Maximianus' poems within their historical context, analyzing how they can be interpreted as a metaphor for the period in which its author lived, in which old age and loss of power symbolized the disappearance of the Ostrogothic kingdom of Italy and, in general, the end of the Roman Empire. A literary exercise but also a life story, this set of six elegies allows us to approach the social construction of gender and age in Late Antiquity.


Program for couples improves adherence. Researchers study impact of bringing in partner.
A couples adherence program called SMART (Sharing Medical Adherence Responsibilities Together) significantly improves medication adherence, a recent study shows. SMART encourages HIV sero-discordant couples to support each other with both HIV prevention and the infected partner's medication adherence.


Activation and release of enzymes and major basic protein from guinea pig eosinophil granulocytes induced by different inflammatory stimuli and other substances. A histochemical, biochemical, and electron microscopic study.
In order to investigate the availability and release of enzymes from eosinophilic granulocytes in response to a variety of stimuli, guinea pig peritoneal eosinophils were obtained after repeated intraperitoneal injections of freeze-dried Trichinella spiralis larvae. The activities of the enzymes peroxidase, arylsulfatase B, beta-glucuronidase, aminopeptidase, histaminase, cytochrome c oxidase, acid phosphatase, adenosine triphosphatase and glucose 6-phosphatase, and the major basic protein (MBP) were studied histochemically and, in part, also biochemically. Eosinophils were incubated with the following substances: histamine, platelet activating factor, calcium ionophore, compound 48/80, leukotriene B4, prostaglandins E1, and E2, heparin, and eosinophil-chemotactic factors from neutrophils and lymphocytes. Eosinophils displayed a selective and stimulus-dependent enzyme and MBP reaction. Calcium ionophore and compound 48/80 provoked a release of cytotoxic major basic protein, partly associated with peroxidase release, while leukotriene B4 and eosinophil chemotactic factors caused histaminase and peroxidase release and activated leucinaminopeptidase. Heparin and calcium ionophore induced release of both MBP and histaminase. These data support the concept that eosinophils exhibit either inflammatory or cytotoxic, or antiinflammatory properties upon stimulation by various agents.


Bone disorders after organ transplantation.
Three main bone disorders may occur after organ transplantation: epiphyseal impactions, avascular bone necrosis, and osteoporosis. Their main clinical characteristics, causes, diagnostic approaches, and therapies are discussed in this review.


Localization of ACTH in the human hypothalamus.
Using an antiserum to porcine ACTH and the unlabeled antibody preoxidase-antiperoxidase technique, we have found that ACTH is present in neuronal cell bodies located exclusively in the arcuate nucleus of the human hypothalamus. ACTH-containing fibers are distributed extensively throughout the hypothalamus with a greatest density in the periventricular nucleus. No concentration of ACTH fibers could be observed in the neurovascular zone of the pituitary stalk.


Geriatric assessment in hematology scale predicts treatment tolerability in older patients diagnosed with hematological malignancies: The RETROGAH study.
The GAH (Geriatric Assessment in Hematology) scale is a psychometrically valid tool aimed at identifying older patients with hematological malignancies at higher risk of treatment-related toxicity. Our objective in this study was to determine the weights for each dimension of the GAH scale and the cut-off point to reliably predict treatment tolerability in this population, estimated by a weighted receiver operating characteristic (ROC) analysis and quantified by the area under the curve (AUC). The RETROGAH was a retrospective cohort study including 126 patients who had previously participated in the GAH study. Patients were ≥ 65 years old with newly diagnosed myelodysplastic syndrome (MDS)/acute myeloid leukemia (AML), multiple myeloma (MM), or chronic lymphoid leukemia (CLL) and treated with standard front-line therapy within three months after having completed the GAH scale. The optimal cut-off value of the GAH total score to discriminate patients at higher risk of treatment toxicity was 42, with 68.5% sensitivity and 55.8% specificity. Using this value, 66.1% of patients evaluated were found to develop some type of toxicity. The AUC was 0.6259 (95% CI: 0.512-0.739; p = 0.035). The GAH scale not only would enable clinicians to individualize therapy based on individual risk of toxicity but also discriminate patients that will benefit most from intensive treatments from those requiring an adapted approach. While futures studies in clinical practice may improve the model and overcome its limitations, the GAH scale should not be used alone when making treatment decisions.


An Open-Label, Randomized, Parallel, Phase II Trial to Evaluate the Efficacy and Safety of a Cremophor-Free Polymeric Micelle Formulation of Paclitaxel as First-Line Treatment for Ovarian Cancer: A Korean Gynecologic Oncology Group Study (KGOG-3021).
Genexol-PM is a biodegradable cremophor EL-free polymeric micelle formulation of paclitaxel. Here,we compared efficacy and safety of Genexol-PM plus carboplatin versus Genexol plus carboplatin for ovarian cancer treatment. In this multicenter, randomized, phase II study, patients with International Federation of Gynecology and Obstetrics IC-IV epithelial ovarian cancer were randomly assigned (1:1) to receive Genexol-PM 260 mg/m2 or Genexol 175 mg/m2 with 5 area under the curve carboplatin every 3weeks (6 cycles). The primary endpointwas the carbohydrate antigen 125 and Response Evaluation Criteria In Solid Tumor composite overall response rate (ORR). Of 131 enrolled patients, 98 were included in intention-to-treat analysis. Mean dosages were 260.00±0.00 mg/m2 Genexol-PM or 174.24±3.81 mg/m2 Genexol. Median followup was 18.0 months (range, 6.1 to 33.8 months). ORR was 88.0% (95% confidence interval [CI], 80.4 to 95.6) with Genexol-PM, and 77.1% (95% CI, 67.1 to 87.1) with Genexol (noninferiority threshold, 16.3%). Median time to progression was 14.8 months (95% CI, 11.3 to 20.2) with Genexol-PM and 15.4 months (95% CI, 13.2 to 29.6) with Genexol (p=0.550). Overall, six patients died. Neutropenia was the most common toxicity (incidences of 86.0% vs. 77.1%, p=0.120). Peripheral neuropathy incidences were 84.0% versus 64.6% (p= 0.148). Peripheral neuropathy of ≥ grade 3 occurred in one patient receiving Genexol. All toxicities were manageable. Genexol-PM plus carboplatin as first-line treatment in patients with epithelial ovarian cancer demonstrated non-inferior efficacy and well-tolerated toxicities compared with the standard paclitaxel regimen. Further studies are warranted to optimize the dose and schedule, and to investigate long-term outcomes.


Connected vehicle real-time traveler information messages for freeway speed harmonization under adverse weather conditions: Trajectory level analysis using driving simulator.
This paper employed a high-fidelity driving simulator to investigate the impacts of the Wyoming Department of Transportation (WYDOT) Connected Vehicle (CV) Pilot's Traveler Information Messages (TIMs) on drivers' speed selection and the safety benefits of their speed harmonization. Three driving simulator experiment scenarios were developed to simulate the typical traffic and weather conditions on the rural Interstate 80 (I-80) in Wyoming. A total of 25 professional drivers from the WYDOT and trucking industry were recruited to participate in the driving simulator experiment. Participants' instantaneous speeds at various locations were collected to reveal the effects of CV TIMs on their speed selection. The results showed that average speed profiles under CV scenarios were generally lower than under baseline scenarios, particularly for winter conditions (snowy and severe weather). The variance of speed under CV scenarios was found to be significantly lower than the baseline scenarios, indicating that CV TIMs have the potential to harmonize the variations in speed. In addition, for the work zone driving simulator experiment, this research revealed that the mean time-to-collision (TTC) under baseline scenario is approximately 40 % lower than CV scenario, and the mean deceleration to avoid a crash (DRAC) under baseline scenario is approximately 19.3 % higher than CV scenario. These findings suggest that CV TIMs can reduce the risk of crashes. Research findings would provide the WYDOT with early insights into the effectiveness of CV TIMs, which could assist with developing more efficient transportation management strategies under adverse weather conditions.


Prediction of radiosensitivity of oral cancers by serial cytological assay of nuclear changes.
To identify the relationship between the radiosensitivity of oral cancers and the induction of micronucleation, nuclear budding and multinucleation (polynucleation) evaluated by serial cytology during fractionated radiotherapy. Forty-four patients with epidermoid cancer of the oral cavity receiving radiotherapy (60 Gy in 25 fractions over 5 weeks) were studied. Serial scrape smears were taken from the tumour before and during radiotherapy and stained by Giemsa and the frequency of micronucleated cells (MNC), nuclear budded cells (NBC) and multinucleated cells (PNC) was evaluated by light microscopy. After a minimum follow-up period of 30 months the patients were classified as having resistant or sensitive tumours, depending on whether the primary tumour had recurred or not within that time. Within-group and between-group analysis on the induction of the above individual parameters and two combined parameters, the micro- or multinucleated cell (MPC) count and the abnormally nucleated cell (ANC) count, was done. The counts were expressed per 1000 uni-nucleated cells. In both groups each parameter showed a statistically significant increase with dose, the increase being higher in the sensitive group. The ANC count showed the greatest increase, the mean counts before treatment and after 28.8 Gy being 24.3 and 157.8 (P < 0.0005), respectively, in the sensitive group and 21.0 and 65.2 (P < 0.0005), respectively, in the resistant group. After 28.8 Gy the sensitive tumours had significantly higher ANC (P = 0.01), MPC (P < 0.05) and PNC (P < 0.05) counts. The study shows that serial cytological assay of nuclear changes (SCANCing) during radiotherapy is a potentially useful test to predict radiosensitivity. The fact that multinucleation showed the greatest relation with radiosensitivity suggests that injury to the cytokinetic apparatus is important in determining tumour radiosensitivity.


Synthesis and characterization of gold-chitosan nanocomposite and application of resultant nanocomposite in sensors.
Chitosan gold nanocomposite (CGNC) was synthesized in a single step process and the pH dependent properties of the composite were investigated. The structure of the polymer was pH dependent and gelation of the polymer observed at pH 4-5. Transmission electron microscopy (TEM) analysis showed that the distribution of gold nanoparticles within unit area varied with the gelation of the polymer, without affecting the size of the nanoparticles. The resulting CGNC system was explored for its use as a sensor for the detection of lead. It was found that lead could be detected at a concentration of 1 μM by absorption spectroscopy. The CGNC was electrodeposited on a gold electrode and the deposited film was used for the detection of lead using cyclic voltammetry. Lead could be detected with a detection limit of 10 μM. The fabrication reproducibility for four sensor electrodes showed a relative standard deviation (RSD) of 2.09% for the determination of 1 mM lead nitrate. The developed sensor system showed high reproducibility, stability and satisfactory selectivity.


Determination of the Re-188 Calibration Number for the Capintec CRC- 25PET Dose Calibrator.
During the development of novel Re-188 radiopharmaceuticals, it was discovered that no calibration settings were published to calibrate Re-188 on the Capintec CRC-25PET dose calibrator. Sodium [188Re]perrhenate was eluted from an OncoBeta 188W/188Re generator to measure activity on a Capintec CRC-25R dose calibrator using established dose calibrator settings provided by the manufacturer. The eluent was then used to tune the calibra on settings on a Capintec CRC-25PET dose calibrator, accounting for geometry. Radionuclidic purity of the [188Re]perrhenate source was verified via gamma spectroscopy. The calibrator number for Re-188 was determined to be 469 x 10 for the Capintec CRC-25PET dose calibrator, which differed from the manufacturer provided calibra on number of 496 x 10 for the Capintec CRC-25R dose calibra on model. W-188 breakthrough was characterised as < 0.01%. This previously unreported calibration number can be used to determine the activity of Re- 188 labelled radiopharmaceuticals using the Capintec CRC-25PET dose calibrator model.


Nucleation time: a key factor in the pathogenesis of cholesterol gallstone disease.
In earlier studies, we concluded that biliary cholesterol supersaturation may be a necessary but not sufficient cause for gallstone formation. In the present studies, we calculated cholesterol saturation indices (CSI) for 120 bile specimens of cholesterol gallstone patients and controls, using the solubility boundaries of Carey and Small (J Clin Invest 61:998-1026, 1978) for artificial biles. The mean CSI + SD was 1.42 +/- 0.68 for controls and 1.80 +/- 1.02 for cholesterol gallstone patients. Of the control bile samples, 68% were supersaturated. Since the two groups could not be sharply distinguished by saturation index, we studied another property of bile samples: nucleation time for cholesterol crystal formation. The mean nucleation time for 16 control bile samples was 15 days, and a strong correlation (r = 0.84) was found between CSI and nucleation time. Twelve bile samples of cholesterol gallstone patients had a mean nucleation time of 3 days and no correlation between CSI and nucleation time. A discriminant analysis of the data, which took into account both the CSI and the nucleation time for each sample, allowed a sharp distinction between bile samples of cholesterol gallstone patients and controls. This distinction could not have been drawn from studies with artificial biles, since these differ only in their CSI.


The effect of graded haemorrhage on erythropoietin production in the immature ovine foetus.
1. Basal and haemorrhage-stimulated erythropoietin (Epo) and ACTH levels were measured in the chronically cannulated immature ovine foetus (less than 125 days) by radio immunoassay (RIA). 2. Basal erythropoietin levels were found to be higher than those previously reported in the late gestation (greater than 130 days) ovine foetus, but were lower than those observed in the neonatal lamb. 3. In control foetuses (Protocol 1) the small degree of haemorrhage associated with the sampling procedure increased the plasma Epo values from 11.4 +/- 3.0 (n = 5) mU/mL to 23.8 +/- 4.3 mU/mL at 24 h (mean +/- s.e.m.). There was a significant monotonic increase with time (F = 16.4; d.f. 1,19; P = 0.001). An initial haemorrhage of approximately 10% blood volume (Protocol 2) increased plasma Epo values from 7.3 +/- 2.3 to 24.2 +/- 7.1 mU/mL (n = 3). 4. Haemorrhage of 20% fetal blood volume (Protocol 3) produced an increase in plasma Epo from 9.3 +/- 1.7 to 54.7 +/- 15.5 mU/mL at 6 h and to 57.6 +/- 7.3 mU/mL at 24 h (n = 5). By repeated measures ANOVA, the effect of the 20% haemorrhage was significant when compared with the control group (F = 7.32, d.f. 2,16, P = 0.006). There was a significantly greater decrease in haematocrit (F = 6.7, d.f. 2,20, P = 0.004) and haemoglobin (F = 5.0, d.f. 2,20, P = 0.013) in animals of Protocol 3 than in those of Protocol 1. 5. Fetal blood gases and plasma adrenocorticotropic hormone (ACTH) did not alter with haemorrhage, indicating the tolerance of the foetus to this degree of haemorrhage.


A soluble mannose receptor immunoadhesin enhances phagocytosis of Pneumocystis carinii by human polymorphonuclear leukocytes in vitro.
Pneumocystis carinii is an opportunistic pathogen that causes pneumonia in immunocompromised hosts. In the normal host, P. carinii is susceptible to an array of first line host defense mechanisms that are operative in the lung. Alveolar macrophages play a central role in the clearance of inhaled organisms. The macrophage mannose receptor (MR) appears to be sufficient for P. carinii phagocytosis. In individuals infected with the human immunodeficiency virus, MR expression on alveolar macrophages and P. carinii phagocytosis are decreased, however, Fc-receptor mediated phagocytosis remains intact. In this study, we demonstrate that a recombinant soluble MR immunoadhesin, consisting of the essential carbohydrate binding MR ectodomain and the Fc-region of human immunoglobulin (Ig)G1, binds P. carinii and leads to an 8.2-fold increased uptake of P. carinii by phagocytic cells. Our results suggest that the soluble MR immunoadhesin may have therapeutic potential in the treatment of P. carinii infections.


Direct radioimmunoassay for rat T-kininogen.
Antibodies raised in rabbits against pure rat T-kininogen did not cross-react with Ile-Ser-Bradykinin, bradykinin, nor with kininogens from other mammalian species. They presented a 1 to 15% cross-reaction with pure rat HMW kininogen, depending on the quantity of HMW kininogen. A direct radioimmunoassay for rat T-kininogen in plasma was developed and it enabled 89 fmol of the protein to be detected. A good correspondence was obtained between the direct RIA and the T-kinin generating assay. By the direct assay, it was found that T-kininogen is increased about ten fold in rats subcutaneously injected with turpentine. These data were confirmed by HPLC analysis of the plasma kinins released by trypsin which demonstrated that only T-kinins are increased, bradykinin being unchanged. It was possible according to the results obtained by the direct RIA and HPLC analysis to estimate that in the normal rat, HMW and LMW kininogen represent about 35% and T-kininogen 65%. In the turpentine-treated rat, T-kininogen reached 95%. This RIA will allow the study of the regulation of T-kininogen in the rat and the synthesis of this protein in cells in culture.


Intrathecal complement activation in neurological diseases evaluated by analysis of the terminal complement complex.
The terminal complement complex (TCC) was determined in plasma and cerebrospinal fluid (CSF) from 208 neurological patients. Elevated CSF TCC levels were observed in higher frequencies in patients with infectious diseases (80%), radiculoneuritis (62%), multiple sclerosis (30%), and miscellaneous autoimmune diseases (27%) than in patients with miscellaneous non-inflammatory diseases (2-13%). The plasma level of TCC was significantly increased only in the infectious group. No positive correlation was observed between the plasma and the CSF TCC concentration in the whole patient population nor in subgroups divided according to blood-brain barrier function. Furthermore, the CSF TCC concentration did not correlate with the serum/CSF albumin ratio or with CSF total protein concentration when this was below 1.0 g/l. It is concluded that an elevated TCC concentration in CSF reflects intrathecal complement activation and that quantification of TCC in CSF may be a valuable supplement in the examination of neurological diseases.


Successful treatment of tachycardia-induced cardiomyopathy with LVAD in a 12-year-old boy.
Tachycardia-induced cardiomyopathy is an unusual cardiac disease that is life-threatening if tachycardia is not controlled. We report a 12-year-old boy who suffered from ectopic left atrial tachyarrhythmia that was refractory to medications and caused tachycardia-induced cardiomyopathy with severe heart failure. The patient required a left ventricular assist device (ABIOMED BVS5000 [ABIOMED Inc, Danvers, MA]) as a bridge to recovery. Tachycardia was finally controlled with flecainide while the patient was on left ventricular assist device support. The device was successfully explanted after 28 days of support. The temporary use of a left ventricular assist device was necessary to maintain a good hemodynamic status during the treatment of pharmacological refractory tachycardia, and it allowed a successful bridge to recovery.


The effect of interlaboratory variability on antimicrobial susceptibility determination.
In the minimum inhibitory concentration (MIC) test literature, discussion concerning the effect of laboratory-to-laboratory variation is lacking. We present 2 sets of drug dilution test quality control data that illustrate considerable laboratory differences in measured MIC. In both isolates (Escherichia coli, ATCC 25922; Staphylococcus aureus, ATCC 29213) the laboratory-to-laboratory variability accounts for approximately half of the total variability. We illustrate the impact of this variability on the probability of correctly classifying the susceptibility level of an isolate and on the estimation of resistance prevalence. For example, we show that laboratory differences in the probability of correctly classifying the isolate (specifically near the lower breakpoint) can vary up to 80%.


The effect of previous defoliation of pole-stage lodgepole pine on plant chemistry, and on the growth and survival of pine beauty moth (Panolis flammea) larvae.
A study of the effects of defoliation by insects on the chemistry of lodgepole pine (Pinus contorta), and on the performance of Panolis flammea (Lepidoptera; Noctuidae) larvae, was carried out in a forest in northwest Scotland I year after a severe outbreak of P. flammea had caused extensive defoliation. Larval weight and survival were not significantly different on trees that had experienced different levels of defoliation in 1986. The nitrogen and tannin content of current and previous years' pine needles was not significantly affected by defoliation (although both were slightly greater in the foliage of defoliated trees). Phosphorus content of young pine foliage was lower (but not significantly lower except on one occasion) on heavily defoliated trees. On all sampling occasions, however, the nitrogen: phosphorus ratio was significantly higher on heavily defoliated trees. There were large differences in monoterpene composition of the previous year's shoots associated with defoliation intensity, but these differences had largely disappeared in the new growth. The results are discussed in relation to other studies on the effects of insect damage on plant chemistry and insect performance and in relation to the abundance of P. flammea in Scotland.


Pattern-evoked responses and luminance-evoked responses in the human electroretinogram.
Electrical potentials were recorded from the cornea of the human eye in response to the onset and offset of square-wave stripe patterns varying in spatial frequency, luminance and contrast. Under high retinal illumination (50119 photopic td) the response to pattern onset was a positive wave (pattern-onset wave) followed by a negative after-potential. As long as the pattern was presented, a steady potential of positive polarity (plateau potential) was observed. The response to pattern offset was a biphasic negative-positive wave followed by a negative after-potential. A comparison between wave forms obtained with pattern onset-offset stimuli and luminance increase-decrease stimuli suggests: (a) the pattern-onset wave is the result of an interaction between a luminance-increase and a luminance-decrease response; (b) the plateau potential is mainly an additive superposition of two receptor processes originating from retinal areas that receive increases and decreases in local luminance; (c) the negative-positive potential is an a-wave and a b-wave originating from retinal areas that receive increases in local luminance. The amplitude of the pattern-onset wave was greatest at a spatial frequency around 3-4 c/deg. This behaviour was closely correlated with contrast sensitivity determined psychophysically by previous investigators. Therefore, the pattern-onset wave seems to be a pattern-evoked response. The amplitude of the b-wave following pattern offset showed a monotonic decrease with increasing spatial frequency. It is mainly a luminance-evoked response. Under low retinal illumination (457 photopic td) the pattern-onset wave and the b-wave at pattern offset were smaller. However, the pattern-onset wave had its maximum amplitude at a lower spatial frequency. When increases or decreases in space-average luminance were combined with pattern onset or offset, transitions between pattern- and luminance-evoked responses could be observed. The results suggest that a decrease in local luminance is the essential stimulus for evoking a pattern-related response while increases in local light intensity generate luminance-evoked responses.


Allogeneic whole-cell vaccine: a phase I/II study in men with hormone-refractory prostate cancer.
To establish the safety and toxicity of an allogeneic human tumour cell vaccine in patients with hormone-refractory prostate cancer, and to determine any biochemical, immunological or clinical response to vaccination. Sixty patients with hormone-refractory prostate cancer were recruited and randomly allocated into four equal groups. Three cell lines (from a bank of four) were administered initially every 2 weeks and then monthly, in conjunction with the immunostimulant Mycobacterium vaccae (SRL-172), each group receiving a different combination of the four cell lines. The patients' serum prostate-specific antigen (PSA) levels were monitored regularly, and the immune response to the vaccine measured using nonspecific intracellular cytokines and specific humoral and cell-mediated assays. The vaccine was safe and well tolerated with no major side-effects. Whilst several patients had a decline in PSA from the entry level, there was no significant decrease that could be attributed solely to the vaccine. However, the immunological data were more encouraging, with several patients from each arm of the trial having an increase in cytokine production, increases in specific antibodies and evidence of T-cell proliferation in response to the vaccinations. The failure of the vaccine to produce a PSA response in the patients in the trial is not surprising considering the stage of the disease. The high PSA levels on entry indicate that the burden of disease was probably high and thus this was an extremely challenging group of patients in which to try and elicit a response through immunotherapy. However, the immunological evidence of a response to the vaccine was encouraging and suggests that further exploration of immunotherapy in less advanced disease may yield more encouraging clinical responses.


GERD and acid reduction medication use following gastric bypass and sleeve gastrectomy.
Gastroesophageal reflux disease is a common comorbid medical condition of obesity. Laparoscopic sleeve gastrectomy has been associated with de novo and worsening GERD following surgery. For this reason, patients who suffer from GERD and are considering bariatric surgery are often counseled to undergo gastric bypass. Given this practice, we sought to determine acid reduction medication (ARM) utilization in bariatric surgical patients who undergo one of these procedures prior to surgery and at 1 year following surgery. A retrospective review of prospectively maintained data on patients to undergo gastric bypass or sleeve gastrectomy between November 2012 and December 2014 was conducted after IRB approval. ARM utilization and Gastroesophageal Reflux Disease Health-Related Quality of Life (GERD-HRQL) scores [range 0 (no symptoms)-50 (severe GERD)] were compared prior to surgery and at 1 year postoperatively. 334 patients underwent an eligible procedure in the study interval. 147 patients (44 %) had data on both preoperative and 1 year postoperative ARM use (93 gastric bypass and 54 sleeve gastrectomy). ARM utilization prior to surgery in gastric bypass patients did not reach statistical significance when compared to sleeve gastrectomy (40.9 vs. 26 %, p = 0.07). GERD-HRQL scores were greater prior to surgery in gastric bypass patients (GERD-HRQL 8.2 vs. 1.9; p < 0.01). At 12 months postoperatively, sleeve gastrectomy patients had a significantly higher rate of overall ARM use (48.1 vs. 16.1 %, p < 0.01), new ARM use (35 vs. 7.3 %, p < 0.01), and persistent ARM use (78.6 vs. 21.9 %, p < 0.01) when compared to gastric bypass patients. GERD-HRQL scores were similar overall at 12 months postoperatively (4.4 bypass vs. 4.8 sleeve; p = 0.72). Laparoscopic sleeve gastrectomy is associated with a significantly increased likelihood that acid reduction medications will be necessary for GERD symptom control 12 months postoperatively when compared to gastric bypass.


New dimension of glucocorticoids in cancer treatment.
Glucocorticoids have been used in clinical oncology for over half a century. The clinical applications of glucocorticoids in oncology are mainly dependent on their pro-apoptotic action to treat lymphoproliferative disorders, and also on alleviating side effects induced by chemotherapy or radiotherapy in non-hematologic cancer types. Researches in the past few years have begun to unveil the profound complexity of glucocorticoids signaling and have contributed remarkably on therapeutic strategies. However, it remains striking and puzzling how glucocorticoids use different mechanisms in different cancer types and different targets to promote or inhibit tumor progression. In this review, we provide an update on glucocorticoids and its receptor, GR-mediated signaling and highlight some of the latest findings on the actions of glucocorticoids signaling during tumor progression and metastasis.


Progenitor translatome changes coordinated by Tsc1 increase perception of Wnt signals to end nephrogenesis.
Mammalian nephron endowment is determined by the coordinated cessation of nephrogenesis in independent niches. Here we report that translatome analysis in Tsc1+/- nephron progenitor cells from mice with elevated nephron numbers reveals how differential translation of Wnt antagonists over agonists tips the balance between self-renewal and differentiation. Wnt agonists are poorly translated in young niches, resulting in an environment with low R-spondin and high Fgf20 promoting self-renewal. In older niches we find increased translation of Wnt agonists, including R-spondin and the signalosome-promoting Tmem59, and low Fgf20, promoting differentiation. This suggests that the tipping point for nephron progenitor exit from the niche is controlled by the gradual increase in stability and possibly clustering of Wnt/Fzd complexes in individual cells, enhancing the response to ureteric bud-derived Wnt9b inputs and driving synchronized differentiation. As predicted by these findings, removing one Rspo3 allele in nephron progenitors delays cessation and increases nephron numbers in vivo.


An inflammatory myofibroblastic tumor of the small intestine presenting as jejunojejunal intussusception; A case report.
An inflammatory myofibroblastic tumor (IMT) is an uncommon solid neoplasm of mesenchymal origin. They are usually seen in children and adolescents and commonly affect the lung, but they can nearly arise from every organ. The prevalence of IMT in the small bowel is very rare. The tumors have generally a benign clinical course, with some risk of local recurrence or distant metastasis. This is a 55-year-old male patient who presented with intermittent abdominal pain, vomiting of ingested matter and loss of appetite for 2 weeks duration. On physical examination, he had stable vital signs and the abdominal examination was non-revealing. Abdominal CT scan with contrast showed a long segment jejunojejunal intussusception. He underwent en-bloc resection of the mass and end-to-end anastomosis of the jejunum. IMTs have a mesenchymal origin and are grouped into a mixture of fibroinflammatory disorders. They show a variable mix of inflammatory cells with spindle cells. The diagnosis of IMT preoperatively is challenging often mimicking malignant lesions. The diagnosis is often confirmed by histopathology after surgery. Complete excision with a negative margin is the preferred treatment. We report a rare case of jejunal IMT presenting with intussusception. An intestinal IMT is a rare and an underdiagnosed entity, and should be considered in the differential diagnosis of small bowel intussusception. Surgery is still the most favored and effective treatment for intestinal IMT. Complete surgical excision with a negative margin has the least chance of disease recurrence.


Intercellular genetic tracing of cardiac endothelium in the developing heart.
Cardiac resident macrophages play vital roles in heart development, homeostasis, repair, and regeneration. Recent studies documented the hematopoietic potential of cardiac endothelium that supports the generation of cardiac macrophages and peripheral blood cells in mice. However, the conclusion was not strongly supported by previous genetic tracing studies, given the non-specific nature of conventional Cre-loxP tracing tools. Here, we develop an intercellular genetic labeling system that can permanently trace heart-specific endothelial cells based on cell-cell interaction in mice. Results from cell-cell contact-mediated genetic fate mapping demonstrate that cardiac endothelial cells do not exhibit hemogenic potential and do not contribute to cardiac macrophages or other circulating blood cells. This Matters Arising paper is in response to Shigeta et al. (2019), published in Developmental Cell. See also the response by Liu and Nakano (2023), published in this issue.


Safety and early efficacy of endoscopic sleeve gastroplasty (ESG) for obesity in a multi-ethnic Asian population in Singapore.
Endoscopic sleeve gastroplasty (ESG) is an alternative nonsurgical treatment option for obesity. However, most studies on the utility and efficacy of ESG are derived from the Western population. It is unknown if ESG elicits similar results in Asians with different fat distribution, sociocultural customs, and dietary practices. Our study aims to assess the safety and efficacy of ESG among a multi-ethnic Asian population. We reviewed 35 patient records who underwent primary ESG for obesity at our unit. We followed a U-shaped suture pattern. Our primary outcome was to assess technical feasibility and safety. The secondary outcome was to determine the percentage total body weight loss (TBWL) at the last follow-up. The mean ± SD age and body mass index were 43.6 ± 11.3 years and 34 ± 4.9 kg/m2, respectively. The majority were female (57%) and of Chinese ethnicity (51%). The procedure was technically successful in all patients. We used an average of five sutures (range, 4-7), and the mean ± SD procedure time was 65 ± 10 min. No major complications occurred, and the average length of stay was 1 day. Twenty-one patients completed 3 months of follow-up, and 10 patients 6 months. The mean ± SD TBWL at 3 and 6 months were 14.5 ± 4.8% and 16.2 ± 4.9%, respectively. We observed improvement in diabetes mellitus (87%), fatty liver (86%), and hypertension (58%) during the follow-up. ESG is a safe and effective option for promoting weight loss in a multi-ethnic Asian population. ESG-induced weight loss may improve obesity-related comorbidities.


Salmonella infection and the genitourinary system.
Six cases of severe salmonella genitourinary infection and a case of salmonella septicemia after prostatectomy are reported. The mechanism whereby the urinary tract is involved in salmonellosis is described, as are the different manifestations of genitourinary salmonellosis. Host factors predisposing to the development of genitourinary salmonella infection include other urologic pathology, malignant neoplasms and states of depressed immunity. Recommendations are made with regard to the diagnosis and management of this sometimes lethal condition.


MexTAg mice exposed to asbestos develop cancer that faithfully replicates key features of the pathogenesis of human mesothelioma.
Animal models provide an important tool for investigating the biology of cancer and for testing the efficacy of novel treatments. Here we describe several aspects of the transgenic MexTAg mouse that develops asbestos-induced mesothelioma. Targeted expression of the TAg transgene causes mesothelioma to develop more rapidly after asbestos exposure in wild-type mice with 100% incidence compared to 30% incidence in wild-type mice. MexTAg mice do not develop spontaneous mesothelioma and exhibit a very low incidence of other tumours. Here we show that TAg does not affect the aggressiveness or rate of progression of the mesotheliomas, suggesting that the oncogene alters only the rate at which disease is initiated. The instillation of an alternative inflammatory agent, thioglycollate, did not induce mesotheliomas, demonstrating acute inflammation is not sufficient for tumour development in MexTAg mice. We found that neither the age of a mouse at the time of exposure nor its gender were prognostic factors. MexTAg mice with mesotheliomas respond to treatment with a cytotoxic drug in a similar way to that of patients with mesothelioma, demonstrating the validity of the model. We also show that long-term treatment with a COX-2 inhibitor prior to the development of disease does not have a survival benefit, suggesting that this is not a useful cancer prevention therapy for asbestos-exposed individuals. The model is robust and suitable for testing a wide variety of protocols and a range of translational studies.


Hemolysin chrysolysin from Penicillium chrysogenum promotes inflammatory response.
Some strains of Penicillium chrysogenum produce a proteinaceous hemolysin, chrysolysinTM, when incubated on sheep's blood agar at 37 degrees C but not at 23 degrees C. However, 92% (11/12) of the indoor air isolates produced hemolysis but only 43% (3/7) of the non-indoor air isolates did so. Chrysolysin is an aggregating protein composed of approximately 2kDa monomers, contains one cysteine amino acid, and has an isoelectric point of 4.85. Treatment of murine macrophage cell line RAW 264.7 with purified chrysolysin caused statistically significant (T-test, p < 0.05) increased production of macrophage inflammatory protein-2 (MIP-2) in a dose dependent manner after 6 h treatment. This suggests that chrysolysin might act to promote the host's inflammatory response after P. chrysogenum exposures.