Quantitative improvements in peptide recovery at elevated chromatographic temperatures from microcapillary liquid chromatography-mass spectrometry analyses of brain using selected reaction monitoring.
Elevated chromatographic temperatures are well recognized to provide beneficial analytical effects. Previously, we demonstrated that elevated chromatographic temperature enhances the identification of hydrophobic peptides from enriched membrane samples. Here, we quantitatively assess and compare the recovery of peptide analytes from both simple and complex tryptic peptide matrices using selected reaction monitoring (SRM) mass spectrometry. Our study demonstrates that elevated chromatographic temperature results in significant improvements in the magnitude of peptide recovery for both hydrophilic and hydrophobic peptides from both simple and complex peptide matrices. Importantly, the analytical benefits for quantitative measurements in mouse whole brain matrix are highlighted, suggesting broad utility in the proteomic analyses of complex mammalian tissues. Any improvement in peptide recovery from chromatographic separations translates directly to the apparent sensitivity of downstream mass analysis in microcapillary liquid chromatography-mass spectrometry (muLC-MS) based proteomic applications. Therefore, the incorporation of elevated chromatographic temperatures should result in significant improvements in peptide quantification as well as detection and identification.


Evaluating on-line continuing education for nurses.
The rapid increase in web sites for nursing continuing education demands critical appraisal by the user. The level of anonymity of the web site authors and sponsors may lead to a laxity of accountability found in no other media. This article discusses criteria for web site evaluation and provides a rating instrument to assist learners in determining the quality and usability of an on-line nursing continuing education offering. Using the rating instrument, two web sites are reviewed. A list of selected nursing continuing education web sites is included.


Path integral Monte Carlo simulation of the absorption spectra of an Al atom embedded in helium.
We use a multilevel path integral Monte-Carlo (PIMC) method to simulate the arrangement of He atoms around a single Al atom doped in a He cluster. High-level ab initio Al-He pair potentials and a Balling and Wright pairwise Hamiltonian model are used to describe the full potential and the electronic asymmetry arising from the open-shell character of the Al atom in its ground and excited electronic states. Our calculations show that the doping of the Al 3p electron strongly influences the He packing. The results of the PIMC simulation are used to predict the electronic excitation spectrum of an Al atom embedded in He clusters. With inclusion of tail corrections for the ground and excited states potentials, the calculated 3d<--3p spectrum agrees reasonably well with the experimental spectrum. The blueshift of the calculated spectrum associated with the 4s<--3p transition of solvated Al is about 25 nm (2000 cm-1) larger than seen in experiments on Al embedded in bulk liquid He. We predict that the spectrum associated with the 4p<--3p transition will be blueshifted by approximately 7000 cm-1 (nearly 1 eV).


p53 protein subcellular localization and apoptosis in rodent corneal epithelium cell culture following ultraviolet irradiation.
The tumor-suppressor gene p53 encodes a phosphoprotein involved in the control of cell growth. p53 expression and function have been documented in malignancy, apoptosis and the aging processes. Recently, p53 has been mapped and characterized in the normal cornea across different species. In the present study, high levels of cytoplasmic p53 protein were noted in normal primary corneal epithelium cultures by immunohistochemistry and western blot analysis. Following ultraviolet (UV) irradiation, the level of cytoplasmic p53 protein expression was increased beginning from 30 min and lasting until 6 h post-irradiation and then returned close to control levels by 24 h. Cytoplasmic p53 phosphorylation was detected from 30 min following UV treatment until 6 h post-irradiation. p53 protein became apparent in the nucleus in a fraction of these cultured cells beginning 30 min following UV irradiation and was still present 24 h later. We also found that p53 colocalized with mitochondria 2 h following UV irradiation in some of the cells and remained there up to 24 h. As the expression levels of p53 transcription following UV irradiation were not significantly altered, the increase in cytoplasmic p53 protein expression may be conditional only upon post-translational stabilization. We also observed that the apoptotic index increased following UV irradiation in the same time frame as the p53 nuclear transfer and was partially suppressed by pifithrin-α, which is a reversible inhibitor of p53-mediated apoptosis and p53-dependent gene transcription. The present study offers new evidence suggesting that cytoplasmic p53 in rodent corneal epithelium is functionally active.


Observations on the ability of repeated, light exposures to Schistosoma mansoni cercariae to induce resistance to reinfection in Kenyan baboons (Papio anubis).
Using groups of 4 baboons (Papio anubis), 21.0% of a trickle exposure of 480 Schistosoma mansoni cercariae/baboon (10/week for 48 weeks) were recovered as adult worms (Group A) compared with 72.6% from a single massive exposure of 2500 cercariae/baboon (Group C). Group B, exposed first to the trickle and then to the massive exposure simultaneously with Groups A and C, yielded a worm recovery of 54% which was slightly but significantly less than Group C. The trickle infection apparently induced a substantial resistance to light re-exposure but only a partial, although significant, resistance to a massive re-exposure. In Group B, however, there was a marked reduction in the degree of gross pathology, despite high tissue egg production by the mature challenge worms. The parasitological results, in terms of resistance to reinfection by and development of pathology to a trickle infection, are not materially different from those obtained in studies using conventional heavy primary and challenge exposures with S. mansoni. The significance of these findings is discussed in relation to the epidemiology of human schistosomiasis mansoni in which heavy exposures are believed to be the exception rather than the rule.


Measurement of the rates of basal pinocytosis of horseradish peroxidase and internalization of heat-aggregated IgG by macrophages from normal and streptozotocin-induced diabetic rats.
The rates of basal pinocytosis and internalization of Fc receptor-bound model immune complexes by macrophages from control (Group 1, n = 9), insulin-treated non-diabetic (2, n = 9), insulin-deficient diabetic (3, n = 8) and insulin-treated diabetic (4, n = 8) rats were measured. Pinocytic rates, as determined by uptake of horseradish peroxidase (HRP), were comparable for all experimental groups (1, 19.6 +/- 5.3; 2, 18.6 +/- 6.0; 3, 18.7 +/- 5.5; 4, 24.5 +/- 9.1; mean +/- 1 SD, pg per min per 10(6) cells, analysis of variance P greater than 0.05). The rates of internalization of Fc receptor-bound model immune complexes were decreased in insulin-treated non-diabetic rats (2, 41.7 +/- 10) and both groups of diabetic rats (3, 39 +/- 5.6; 4, 44.6 +/- 6.9) compared with control animals (1, 54.4 +/- 7.2; mean +/- 1 SD, percentage internalized per 10 min per 10(6) cells, analysis of variance P less than 0.01). Under the conditions of study, comparable amounts of model immune complexes were bound by macrophages from each of the groups; thus, the amount of internalized material was decreased in all three experimental groups (2, 3 and 4). These data suggest that insulin treatment, as well as the diabetic environment, can contribute to a decreased rate of internalization of Fc receptor-bound immune complexes, and may thereby contribute to impaired phagocytosis that has been demonstrated to occur in diabetes. These changes appear to be specific to Fc receptor-mediated internalization, as no differences in the rates of basal pinocytosis were observed.


If You Don't Have Valence, Ask Your Neighbor: Evaluation of Neutral Words as a Function of Affective Semantic Associates.
How do humans perform difficult forced-choice evaluations, e.g., of words that have been previously rated as being neutral? Here we tested the hypothesis that in this case, the valence of semantic associates is of significant influence. From corpus based co-occurrence statistics as a measure of association strength we computed individual neighborhoods for single neutral words comprised of the 10 words with the largest association strength. We then selected neutral words according to the valence of the associated words included in the neighborhoods, which were either mostly positive, mostly negative, mostly neutral or mixed positive and negative, and tested them using a valence decision task (VDT). The data showed that the valence of semantic neighbors can predict valence judgments to neutral words. However, all but the positive neighborhood items revealed a high tendency to elicit negative responses. For the positive and negative neighborhood categories responses congruent with the neighborhood's valence were faster than incongruent responses. We interpret this effect as a semantic network process that supports the evaluation of neutral words by assessing the valence of the associative semantic neighborhood. In this perspective, valence is considered a semantic super-feature, at least partially represented in associative activation patterns of semantic networks.


alpha-fetoprotein binding specificity for arachidonate, bilirubin, docosahexaenoate, and palmitate. A spin label study.
A dianionic spin label, 1-L-glutamate-5-N-(1-oxyl-2,2,6,6-tetramethyl-4-aminopiperidinyl)-i,4-dinitrobenzene, has been used to probe the relative binding specificity of a single anionic ligand site on bovine alpha-fetoprotein (AFP) to arachidonate, bilirubin, docosahexaenoate, and plamitate. The binding isotherm of the spin label with AFP, as shown by a Scatchard plot, indicates the presence of a single high affinity binding site. The site-site relationship of the four endogenous ligands, arachidonate, bilirubin, docosahexaenoate, and palmitate, was determined by studying their effectiveness in competing for this anionic ligand binding site on AFP. Scatchard plots of the spin label in the presence of 1 to 3 molar equivalents of arachidonate, bilirubin, and docosahexaenoate and up to 6 molar equivalents of palmitate have been determined. The effectiveness of the four endogenous ligands in displacing the spin label from its primary binding site is bilirubin greater than or equal arachidonate approximately equal to docosahexaenoate greater than palmitate. These results indicate that polyunsaturated essential fatty acids and bilirubin share a high affinity binding site on AFP. We propose that the function of this anionic ligand binding site on AFP is for the transport of bilirubin and polyunsaturated fatty acids in fetal serum, as well as for the cross-placental transfer of this metabolite and of essential fatty acids.


[Municipal wastewater treatment using the up-biological aerated filter].
Up Biological Aerated Filter(UBAF) was used for the advanced wastewater treatment. The average removal rate of CODCr, BOD5, SS, NH4(+)-N and turbidity was 22.2%, 55%, 84.4%, 49%, 75.6% respectively. The hydraulic and COD load of influent influenced directly the quality of effluent.


Budesonide in the management of patients with Crohn's disease.
Modern medical therapy is increasingly based on evidence. The evidence presented here is that budesonide (Entocort, Astra Pharma) 9 mg/day is superior to placebo and equivalent to systemically active glucocorticosteroids in achieving disease remission in patients with active Crohn's disease, and in prolonging the recurrence time of symptomatic disease. Budesonide causes less disturbance to adrenal function than prednisone or prednisolone and may cause fewer steroid-associated symptoms. Thus, budesonide is the safer, more effective steroid of choice to treat patients with Crohn's disease.


Validation of the acetylene rebreathing method for measurement of cardiac output at rest and during high-intensity exercise.
The use of the acetylene rebreathing method to estimate cardiac output (CO) during high-intensity exercise, which may be influenced by recirculation of acetylene, has not been validated. This study was designed to validate the acetylene rebreathing method to measure CO during high-intensity exercise using the direct Fick method. CO was measured at rest and during exercise at 25%, 50%, 75% and 90% of the nine subjects' maximum oxygen uptake (VO2max) by the direct Fick and acetylene rebreathing method. CO measured by the acetylene rebreathing method correlated with work rate (r = 0.90, P < 0.01) and with oxygen uptake (r = 0.94, P < 0.01). The correlation coefficient of CO between both methods was r = 0.91 (P < 0.01). There was no significant difference in CO measured by each method at rest as well as at each work rate. The difference in CO between each method was greater at lower CO than at higher CO. At 90% of VO2max, the CO measured by acetylene rebreathing was nearly identical to that measured by the Fick method. It can be concluded that acetylene rebreathing for measurement of CO is valid not only at rest but also during exercise, especially during high-intensity exercise.


Effect of levels of starch, fiber, and lactose on digestion and growth performance of early-weaned rabbits.
The effect of source of carbohydrate on gut histology, digestion efficiency, and growth performance in early-weaned (25 d) rabbits at the starter period (25 to 39 d) was investigated. Six diets were factorially arranged to study the effect of partial substitution of starch (0, 25, or 50%) by lactose at two levels of fiber (30 or 36% NDF). Diets were formulated to meet or exceed essential nutrient requirements of growing rabbits. A feeding trial was conducted to measure the effect of treatments on growth performance in 252 rabbits that were fed the experimental diets in the starter period and thereafter received a common feed until 60 d of age. Fecal apparent digestibility was determined at 35 d of age in nine animals per diet. The four diets with extreme lactose content were used to determine ileal apparent digestibility of starch and lactose (nine replicates per diet), weights of stomach and cecum, stomach pH, cecal fermentation traits, amylase and disaccharidase activities (10 animals per diet), and jejunal morphology (six animals per diet). Weaning increased (P < 0.001) amylase activity by 59% but decreased (P < 0.001) maltase, sucrase, and lactase activities by 30, 48, and 72%, in parallel with a reduction of villus height by 19%. Dietary NDF level did not affect either jejunal morphology or sucrase and lactase activities but increased amylase (P = 0.05) and maltase (P < 0.001) activities by 22 and 92%, respectively. Substitution of starch by lactose had no effect on jejunal morphology or enzymatic activity. Ileal lactose and starch digestibility were not affected by dietary NDF or lactose level and averaged 73.8 and 90.8%, respectively. Substitution of starch by fiber and lactose affected ileal flux of starch plus lactose (by -0.5 and +1.7 g/d) and cecal pH (by +2.1 and -2.8%, respectively). Fecal NDF digestibility was relatively low (23.1% on average) and was not affected by treatments, whereas that of lactose and starch was almost complete. An increase of dietary NDF level led to an impairment of ADG and feed efficiency in the starter (P < 0.002) and in the overall (P < 0.03) fattening period. Substitution of starch by lactose linearly decreased (P < 0.001) feed efficiency in the starter period and linearly increased (P < 0.001) diarrhea incidence in the fattening period. The results indicate that digestive capability of early-weaned rabbits is limited and should be taken into account to establish optimal levels and sources of carbohydrates in the starter diet.


Raman Spectroscopic Fingerprints of Atomically Precise Ligand Protected Noble Metal Clusters: Au38 (PET)24 and Au38-x Agx (PET)24.
Distinct Raman spectroscopic signatures of the metal core of atomically precise, ligand-protected noble metal nanoclusters are reported using Au38 (PET)24 and Au38-x Agx (PET)24 (PET = 2-phenylethanethiolate, -SC2 H4 C6 H5 ) as model systems. The fingerprint Raman features (occurring <200 cm-1 ) of these clusters arise due to the vibrations involving metal atoms of their Au23 or Au23-x Agx cores. A distinct core breathing vibrational mode of the Au23 core has been observed at 90 cm-1 . Whereas the breathing mode shifts to higher frequencies with increasing Ag content of the cluster, the vibrational signatures due to the outer metal-ligand staple motifs (between 200 and 500 cm-1 ) do not shift significantly. DFT calculations furthermore reveal weak Raman bands at higher frequencies compared to the breathing mode, which are associated mostly with the rattling of two central gold atoms of the bi-icosahedral Au23 core. These vibrations are also observed in the experimental spectrum. The study indicates that low-frequency Raman spectra are a characteristic fingerprint of atomically precise clusters, just as electronic absorption spectroscopy, in contrast to the spectrum associated with the ligand shell, which is observed at higher frequencies.


Intraoperative ultrasound margin evaluation as a tool to reduce positive superficial margins in nipple and skin sparing mastectomy in breast cancer patients.
Intraoperative ultrasound (IOUS) guided conservative surgery has been shown to reduce rates of positive margins in breast cancer. The aim of the study is to evaluate the feasibility of using IOUS to assess superficial/anterior margins in nipple and skin sparing mastectomy (NSM/SSM) and its impact on reducing rates of positive margins. This prospective study includes all breast cancer patients who had an indication for NSM/SSM at our Institution. Superficial margin width was measured by IOUS before surgery and the area marked on the skin. Same measurement was performed afterwards in the mastectomy specimen. Any superficial margin < 5 mm was re-excised intraoperatively following the mark on the skin. Fifty-nine patients were included, 47 patients (79.7%) underwent NSM, and 12 patients (20.3%) a SSM. Of the 59 patients, 23 (38.98%) had margins ≥5 mm and 36 patients (61.02%) had margins of ≤5 mm. Of the 36 patients with superficial margins ≤5 mm, 20 had margins <2 mm, and 6 of them had intraoperative involved superficial margins in final pathology. However, after IOUS-guided re-excision, final pathology showed no involved margins. A 2 mm margin was set as the cut-off point for performing an intraoperative re-excision. IOUS guided re-excisions for intraoperative margins ≤2 mm significantly reduced the risk of close/positive margins in final pathology, p < 0.0001. The results showed that IOUS margin evaluation significantly reduced the rate of superficial positive margins in NSM/SSM. It is feasible and effective and may avoid challenging reoperations and/or additional radiation therapy for positive margins.


[Immunohistochemical study on the keratin expression in generating rat hair tissues using anti-hair keratin monoclonal antibodies].
To investigate keratin expressions of generating rat hair tissues, monoclonal antibodies, HKN-2, 4, 5 and 6, were immunohistochemically applied on the skin specimens from 19-day-old rat fetus, newborn, 3-day-old and 7-day-old rats. In anagen hair tissues of 7-day-old rats, HKN-2 reacted with developing cells of the medulla, cortex, hair cuticle and inner and outer root sheaths. In the epidermis, HKN-2 reacted with spinous through horny cells. HKN-4 displayed a broad reactivity with all of the skin epithelial cells including hair tissue. These reactions were first seen in hair peg stage. HKN-2 and 4 first showed strong reactions with the innermost cell layer of the outer root sheath in anagen hair tissues of newborn rats. Reactivities by HKN-5 and 6 were localized in hair tissues in rats; however, HKN-5 showed no reaction with the innermost cell layer of the outer root sheath. HKN-5 and 6 showed no reaction with hair tissue before anagen. It is concluded that hair germ is different from epidermal basal cells in keratin expressions, that hair-specific keratins seem to be produced after the morphological differentiation of each layer, and that the innermost cell layer may differ from other outer root sheath since hair generation.


Inhibition of human immunodeficiency virus-1 reverse transcriptase activity by rubromycins: competitive interaction at the template.primer site.
Rubromycins, a class of quinone antibacterials, were discovered to selectively inhibit human immunodeficiency virus-1 (HIV-1) RNA-directed DNA polymerase (reverse transcriptase) (RT) activity more potently than cellular DNA polymerase alpha. beta- and gamma-rubromycin each inhibited equipotently HIV-1 RT and avian myeloblastosis virus RT, in a concentration-dependent manner, and were significantly weaker as inhibitors of calf thymus DNA polymerase alpha. These agents inhibited HIV-1 RT reversibly, were competitive with respect to template.primer, and were noncompetitive with respect to TTP. Dixon analyses yielded HIV RT Ki values of 0.27 +/- 0.014 and 0.13 +/- 0.012 microM for beta- and gamma-rubromycin, respectively. Similarly, using DNA polymerase alpha, the Ki values were 25.1 +/- 4.3 and 3.9 +/- 0.6 microM for beta- and gamma-rubromycin, respectively. Because these agents were toxic to noninfected human T lymphoid cells using concentrations at or above 6 microM, HIV-1 infectivity studies were carried out at 0.8-6 microM. At these concentrations, which are below the range expected to provide protection, no significant antiviral activity was observed. Although beta- and gamma-rubromycins did not possess sufficient HIV RT inhibitory potency or selectivity versus mammalian DNA polymerase to demonstrate antiviral activities, these studies support the hypothesis that specific molecules containing quinone functional groups can selectively inhibit viral polymerase activities over cellular polymerase activities. In addition, these studies suggest that rubromycins may be lead structures for the development of more potent and selective agents.


Sebaceous gland carcinoma of the eyelid presenting as a conjunctival papilloma.
An unusual presentation of sebaceous carcinoma of the eyelid is described in a 96-year-old man who presented with a large papillomatous palpebral conjunctival lesion in the left upper eyelid. The patient underwent a shave excision of the lesion, followed by a full thickness excision with paraffin section margin control. Histopathology revealed a sebaceous gland carcinoma with no evidence of pagetoid spread. Although rare, sebaceous gland carcinoma should be considered in the differential diagnosis of a conjunctival papilloma.


Takayasu's arteritis treated by percutaneous transluminal angioplasty with stenting in the descending aorta.
A 17-yr-old young woman was referred to our hospital with a 2-yr history of claudication of the lower extremities and severe arterial hypertension. Physical examination revealed significantly different blood pressures between both arms (160/92 and 180/95 mmHg) and legs (92/61 and 82/57 mmHg). The hematological and biochemical values were within their normal ranges, except for the increased erythrocyte sedimentation rate (83 mm/hr) and C-reactive protein (6.19 mg/L). On 3-dimensional computed tomographic angiography, the ascending aorta, the aortic arch and its branches, and the thoracic and, descending aorta, but not the renal artery, were shown to be stenotic. The diagnosis of type IIb Takayasu's arteritis was made according to the new angiographic classification of Takayasu's arteritis, Takayasu conference 1994. Percutaneous transluminal angioplasty with stenting was performed on the thoracic and abdominal aorta. After the interventional procedures, the upper extremity blood pressure improved from 162/101 mmHg to 132/85 mmHg, respectively. She has been free of claudication and there have been no cardiac events during 2-yr of clinical follow-up.


The pH mediated effects of initial glucose concentration on the transitory occurrence of extracellular metabolites, gas exchange and growth yields of aerobic batch cultures of Klebsiella pneumoniae.
The changes in growth kinetics in aerobic batch cultures of Klebsiella pneumoniae were followed by measurements of extracellular metabolites, rates of gas exchange, dissolved oxygen tension, pH, and carbon balance at all stages of growth. When the initial growth-limiting glucose concentration in media without pH control was increased from 1.0 g carbon L(-1) to 2.2 g carbon L(-1), the number of different, mainly acidic, extracellular metabolites of glucose at the end of exponential growth increased, while the proportion of acetate decreased. During the postexponential growth phase, the extracellular metabolites were oxidized, resulting in an increasing complexity of changes in pH, gas exchange, and dissolved oxygen tension with increasing initial substrate concentration. All these parameters showed concomitant stepwise changes. This pattern was independent of the dissolved oxygen tension in the range 30-200 muM. When pH was kept constant, the number, slope, and relative magnitude of the steps in gas exchange and dissolved oxygen tension were pH-dependent, being most complex at low pH. Detailed carbon balances showed that 20% of the initial glucose was converted into extracellular metabolites at the end of exponential growth at neutral pH. In the postexponential phase, pyruvate (2%) was reoxidized first followed by acetate (13%). The observed molar growth yield coefficient (Y(ATP)) was 8.4 if the transitory occurrence of pyruvate and acetate was accounted for, and 6.4 if it was neglected. The corrected observed molar growth yield coefficient (Y'(ATP)) was 9.4 and compared well with the true molar growth yield coefficient (Y(Max) (ATP)), which was found to be 11.0. Specific in situ respiration rates of the exponential growth phase of cultures grown at different controlled pH values compared well with in situ values for energy-limited chemostat grown cells at the same growth rates, suggesting that growth in the batch culture was energy-limited throughout the exponential growth phase. This view was supported by low levels of intracellular glycogen and exopolysaccharides of all cultures, by the value of Y'(ATP) of 9.4, and by a constant specific production rate of the extracellular metabolites throughout exponential growth. It was concluded that even under strictly aerobic conditions, control of pH is as important as control of dissolved oxygen tension during growth of enterobacteriaceae in batch cultures.


Extended delivery of the antimitotic agent colchicine from thermoresponsive N-isopropylacrylamide-based copolymer films to human vascular smooth muscle cells.
The aim of this study was to establish the capacity of thermoresponsive poly(N-isopropylacrylamide) copolymer films to deliver bioactive concentrations of an antimitotic agent to human vascular smooth muscle cells (HASMC) over an extended period of time. Copolymer films were prepared using a 50:50 (w/w) ratio of N-isopropylacrylamide (NiPAAm) monomer to the more hydrophobic N-tert-butylacrylamide (NtBAAm) and loaded with the antimitotic agent colchicine (0.1 micromol per film) at room temperature. Colchicine release from films was sustained over a 14-day period. At 24 h postloading, the concentration of colchicine in the medium overlying films was 2.12 +/- 0.16 microM; this fell to 0.20 +/- 0.01 microM at 7 days and decreased further to 0.12 +/- 0.01 microM after 14 days. Colchicine released from copolymer films inhibited proliferation when subsequently placed on HASMC: at 0.1 microM, released colchicine reduced proliferation to 18.5 +/- 0.8% of control cells (p < 0.001, n = 9). The antiproliferative effect of released colchicine was comparable to that of native colchicine, as observed in separate experiments. Furthermore, colchicine released from 50:50 polymer films inhibited the proliferation of cells grown in the same environment as the copolymer. Inhibition of cell proliferation was not due to the release of cytotoxic particles from the copolymer because medium incubated with copolymer for 5 days and then applied to HASMC did not alter cell viability. In conclusion, this study demonstrates that 50:50 NiPAAm:NtBAAm copolymers can deliver bioactive concentrations of the antimitotic agent colchicine to human vascular cells over an extended period of time.


Effects of ambient conditions on solar assisted heat pump systems: a review.
Solar assisted heat pump (SAHP) systems aim to increase the performance of heat pumps by supporting with solar energy using various heating modes, whose performances depend on ambient conditions. In the literature, SAHP systems are classified by structure types, and the heating mode options are not considered as the main priority of the design but a result of the structure of the system. This paper aimed to investigate the effect of ambient conditions on heating modes, and identify the preferable ambient condition ranges for each SAHP system depends on their heating modes, by using a narrative review of 47 recent studies, that shed light on the problem. For this purpose, direct solar heating (DSH), air source heat pump (ASHP), solar source heat pump (SSHP) and solar-air source heat pump (S/ASHP) modes were stated as the basic heating modes of SAHP systems. In accordance with the literature, SAHP systems were classified as direct expansion solar assisted heat pump (DX-SAHP) and series, parallel and dual source indirect expansion solar assisted heat pumps (IDX-SAHP). Solar irradiation, ambient temperature, relative humidity and wind speed has been taken as major ambient conditions to investigate. Reviewed studies indicated that, parallel and series IDX-SAHP are preferable in high solar irradiations about 800 W/m2 with their DSH mode options. Frosting on evaporator is prevented in DX-SAHP with its S/ASHP mode, and even in the presence of frosting, unlike IDX-SAHPs ASHP mode, freezing is a factor that improves performance on flat evaporator in cold and humid conditions. This study indicates that there are obscure areas for future studies to focus on for a better comparison between SAHP types. Moreover, proposed novel designs of this paper, such as solar preheating of air in IDX-SAHP systems to add S/ASHP mode as an option, might enhance the performance and applicability of SAHP systems.


PlanTEnrichment: A tool for enrichment analysis of transposable elements in plants.
Transposable elements (TEs) are mobile DNA sequences that play roles in gene regulation, and have a potential to influence the expression of nearby genes by functioning as cis-regulatory sequences. However, bioinformatics tools facilitating analysis of the associations between TEs and nearby genes in plants are still lacking. We therefore reanalyzed the comprehensive annotation data of gene models and TEs of 11 plant species available in Ensembl Plants database, and built an up-to-date, unique tool called PlanTEnrichment, enabling enrichment analysis of TEs located within the upstream regions of a given gene list. PlanTEnrichment takes, for example, a group of differentially expressed genes under a particular biological condition as input and returns the list of TEs associated with those genes, along with their calculated enrichment scores and statistical significances. PlanTEnrichment is freely available at http://tools.ibg.deu.edu.tr/plantenrichment/ and is likely to substantially enhance our understanding of the role of TEs in diverse biological processes.


The Stress-Sensing TORC2 Complex Activates Yeast AGC-Family Protein Kinase Ypk1 at Multiple Novel Sites.
Yeast (Saccharomyces cerevisiae) target of rapamycin (TOR) complex 2 (TORC2) is a multi-subunit plasma membrane-associated protein kinase and vital growth regulator. Its essential functions are exerted via phosphorylation and stimulation of downstream protein kinase Ypk1 (and its paralog Ypk2). Ypk1 phosphorylates multiple substrates to regulate plasma membrane lipid and protein composition. Ypk1 function requires phosphorylation of Thr504 in its activation loop by eisosome-associated Pkh1 (and its paralog Pkh2). For cell survival under certain stresses, however, Ypk1 activity requires further stimulation by TORC2-mediated phosphorylation at C-terminal sites, dubbed the "turn" (Ser644) and "hydrophobic" (Thr662) motifs. Here we show that four additional C-terminal sites are phosphorylated in a TORC2-dependent manner, collectively defining a minimal consensus. We found that the newly identified sites are as important for Ypk1 activity, stability, and biological function as Ser644 and Thr662. Ala substitutions at the four new sites abrogated the ability of Ypk1 to rescue the phenotypes of Ypk1 deficiency, whereas Glu substitutions had no ill effect. Combining the Ala substitutions with an N-terminal mutation (D242A), which has been demonstrated to bypass the need for TORC2-mediated phosphorylation, restored the ability to complement a Ypk1-deficient cell. These findings provide new insights about the molecular basis for TORC2-dependent activation of Ypk1.


The stability and steady-state kinetics of vanadium chloroperoxidase from the fungus Curvularia inaequalis.
In this article we report on the steady-state kinetics of the chlorination and the stability of the vanadium chloroperoxidase from the fungus Curvularia inaequalis. The data show that the kinetics of this enzyme resemble that of the vanadium bromoperoxidase from the seaweed Ascophyllum nodosum. At low pH, chloride inhibited the enzyme, but the inhibition was of a dual nature. At pH 4.1 a mixed type of inhibition by chloride with respect to hydrogen peroxide was observed whereas at pH 3.1 the nature of the inhibition became competitive. The log Km for hydrogen peroxide decreased linearly with pH with a slope of -1 in the pH range 3-5. A reaction mechanism is presented to explain the observed data. We also showed that this class of enzymes is inhibited by nitrate. At pH 5.5, nitrate inhibits the chlorination reaction competitively with respect to chloride (Ki = 2 mM) and uncompetitively with respect to hydrogen peroxide. Furthermore, we showed that the enzyme produces HOCl as a reaction product. The enzyme exhibited a high thermostability (tm of 90 degrees C) and displayed high stability in organic solvents (solutions of 40% methanol, ethanol or 2-propanol) and moderate stability in the presence of the chaotropic agent guanidine/HCl (G1/2, the concentration of guanidine/HCl at which the enzyme activity was half the original activity was 3.7 M).


miR-30a regulates the proliferation and invasion of breast cancer cells by targeting Snail.
The present study aims to investigate the effect of miR-30a on the proliferative and invasive abilities of breast cancer cells, and to observe the role of miR-30a in the pathogenesis of breast cancer. With the increase of pathological grade and malignant degree of breast cancer cells, the miR-30a expression level gradually decreased (P<0.01). Transfection with miR-30a mimic significantly inhibited the proliferative and invasive ability of SK-BR-3 cells (P<0.01), while transfection with anti-miR-30a significantly improved the proliferative and invasive ability of these cells (P<0.01). It was revealed using bioinformatic methods that Snail was the functional target gene of miR-30a, and this was verified by the results of a luciferase reporter gene assay. The results of analysis of Snail expression in breast cancer tissues and breast cancer cells revealed that with the increase in pathological grade and malignant degree of breast cancer cells, Snail expression levels gradually increased (P<0.01). Western blotting revealed that miR-30a significantly inhibited Snail expression in SK-BR-3 cells, upregulated the expression of EMT-associated E-cadherin, and downregulated the expression of EMT-associated N-cadherin and Vimentin. MiR-30a was able to affect the proliferation and invasion of breast cancer cells by regulating Snail expression, and therefore has a regulatory effect on the occurrence and development of breast cancer.


Complete Oxidation of Toluene Over Supported Palladium Catalysts: Effect of Support.
The catalytic oxidation of toluene was carried out to investigate the effect of the support on the properties and performance of a supported palladium catalyst (1 wt% loading). γ-Al2O3), SiO2 and TiO2 were used as supports. The properties of the prepared catalysts were characterized by XRD, BET, PT, TPD, TPR, XPS and FE-TEM analyses. The experimental results showed that the order of catalytic performance was 1 wt% Pd/TiO2 > 1 wt% Pd/SiO2 > 1 wt% Pd/γ-Al2O3. A strong interaction between the palladium and support was found, which affected the palladium surface concentration and particle size. The higher catalytic activity of the Pd/TiO2 catalyst was attributed to the higher palladium surface concentration and larger particle size.


Does stem length matter?
There are still some controversial issues regarding total hip arthroplasty including cemented vs. cementless, short stem vs. long stem. A literature review on the currently published survival results of short stems is encouraging and appears to be comparable with that of traditional uncemented stems. However, only few mid-term and long-term follow-up studies are available; studies with more patients and longer follow-up periods are needed.


Implicit nonpolar solvent models.
We have systematically analyzed a new nonpolar solvent model that separates nonpolar solvation free energy into repulsive and attractive components. Our analysis shows that either molecular surfaces or volumes can be used to correlate with repulsive free energies of tested molecules in explicit solvent with correlation coefficients higher than 0.99. In addition, the attractive free energies in explicit solvent can also be reproduced with the new model with a correlation coefficient higher than 0.999. Given each component optimized, the new nonpolar solvent model is found to reproduce monomer nonpolar solvation free energies in explicit solvent very well. However, the overall accuracy of the nonpolar solvation free energies is lower than that of each component. In the more challenging dimer test cases, the agreement of the new model with explicit solvent is less impressive. Nevertheless, it is found that the new model works reasonably well for reproducing the relative nonpolar free energy landscapes near the global minimum of the dimer complexes.


Multiple foreign bodies in a neonate.
We report a rare instance of nine foreign bodies in a neonate that included a coin, safety pin, screw, cotton piece, polythene piece, and four glass pieces. Of these, six foreign bodies were removed by esophagoscopy and endoscopy, two glass pieces were passed in feces and one could not be removed. The child died 5 days after admission.


Transient carcinoembryonic antigen elevations during adjuvant chemotherapy for colorectal cancer reflect the burden of residual micrometastatic disease.
Carcinoembryonic antigen (CEA) testing is routinely used to monitor the progress of patients with advanced cancer on treatment, or else to detect relapse during follow-up, particularly in colorectal cancer (CRC). Although CEA levels have been reported to rise during adjuvant drug therapies, the mechanism of such 'surges' is not clear. This study was conducted to clarify the clinical significance of this phenomenon. We conducted a retrospective analysis of CEA levels in 88 consecutive patients receiving adjuvant chemotherapy in our center: 39 patients with primary CRC and a comparison cohort of 49 patients with breast cancer treated with adjuvant chemotherapy. In the event of 2 serial CEA increases, endoscopic and/or imaging investigations were performed to exclude recurrence. Subset analyses were based on nodal status and primary tumor type. Primary resection was associated with significant CEA decline in patients with CRC but not in those with breast cancer. Forty-three patients (48.9%) experienced CEA fluctuations exceeding 0.5 ng/mL during adjuvant chemotherapy; CEA increases indicated true recurrence in 2 patients (4.7%). Adjuvant CEA surges occurred both more often and more extensively in disease associated with >or= 4 positive nodes in patients with CRC but not in patients with breast cancer (P < .05). Both the frequency and extent of CEA surges during adjuvant chemotherapy parallel the severity of preoperative nodal involvement in CRC but not in breast cancer, suggesting that such surges reflect tumorilytic effects on occult disease in patients with CRC only. However, whether these CEA surges predict survival that is inferior (ie, because of greater burden of residual disease) or superior (ie, because of greater tumorilytic efficacy) to that of stagematched 'nonsurge' patients, remains to be determined by larger, prospective CRC studies.


Conversion of Similar Xenochemicals to Dissimilar Products: Exploiting Competing Reactions in Whole-Cell Catalysis.
Many enzymes have latent activities that can be used in the conversion of non-natural reactants for novel organic conversions. A classic example is the conversion of benzaldehyde to a phenylacetyl carbinol, a precursor for ephedrine manufacture. It is often tacitly assumed that purified enzymes are more promising catalysts than whole cells, despite the lower cost and easier maintenance of the latter. Competing substrates inside the cell have been known to elicit currently hard-to-predict selectivities that are not easily measured inside the living cell. We employ NMR spectroscopic assays to rationally combine isomers for selective reactions in commercial S. cerevisiae. This approach uses internal competition between alternative pathways of aldehyde clearance in yeast, leading to altered selectivities compared to catalysis with the purified enzyme. In this manner, 4-fluorobenzyl alcohol and 2-fluorophenylacetyl carbinol can be formed with selectivities in the order of 90%. Modification of the cellular redox state can be used to tune product composition further. Hyperpolarized NMR shows that the cellular reaction and pathway usage are affected by the xenochemical. Overall, we find that the rational construction of ternary or more complex substrate mixtures can be used for in-cell NMR spectroscopy to optimize the upgrading of similar xenochemicals to dissimilar products with cheap whole-cell catalysts.


Primary malignant lymphomas of the central nervous system in man.
Sixty-eight primary malignant lymphomas of the CNS exclusively confined to the brain and its leptomeninges from a series of about 8000 intracranial neoplasms (incidence 0.85%) were examined and classified according to current histopathologic criteria. Average age at onset of symptoms was 55 years, mean duration of illness to time of diagnosis was 3 months. Survival averaged 1,8 months with supportive care, but 17,2 months with surgery, radiation and/or chemotherapy. CSF cytology was useful and reliable tool for clinical diagnosis. The cerebral hemispheres were affected in about 50%, the basal ganglia in 18%, posterior fossa in 10%, while multifocal lesions amounted to 22%. All CNS tumors were of the diffuse type of non-Hodgkin's lymphomas; no follicular (germinal center) lymphomas were observed. Three histological patterns comparable to extraneural lymphomas were distinguished: Immunoblastoma (reticulosarcoma) occurred most frequently (58.8%), lympho-plasmacytoid immunocytoma constituted 28 percent, while lymphoblastic lymphoma occurred least frequently (13.2%). There were no significant differences with regard to onset, location, growth pattern or clinical course except for a much poorer prognosis of lymphoblastic lymphoma. Although there are no definite cytological differences between malignant lymphomas arising in extraneural sites or as primary lesions in the CNS, the latter showed a much greater proportion of phagocyting histiocytes (and microglia) and a frequent occurrence of plasmacytes and their precursors which apparently exceded pure host reaction. The prognostic value of modern classification schemes for CNS lymphomas needs further critical evaluation.


Growing skull fracture after cranial vault reshaping in infancy.
Growing skull fractures are reported in three infants undergoing cranial vault surgery for craniosynostosis. Although primary dural injury was overt in only one patient, an extensive dural defect with spatially coincident bony absence was identified in all patients at late exploration. The coincidence of dural injury, deficient overlying bone, and a pulsatile growing brain are necessary conditions for the development of growing skull fractures. With the advent of increasingly radical cranial vault procedures during infancy, the potential for this complication to arise may increase. Definitive correction requires autogenous tissue reconstruction of the dural and bony defect.


Fifteen years follow-up of hip arthroplasties with a cemented monoblock femoral stem and a flanged acetabular component.
Our study evaluated the survival and complication rate of 147 total hip arthroplasties at least fifteen years after index surgery. All patients treated in our institution in 1986 with a cemented monoblock prosthesis and a flanged acetabular component were identified and a phone interview could be conducted with 144 (147 hips). No revisions for aseptic loosening were required and only 3 revisions of the index arthroplasty took place: one for infection (after 2 years), one for recurrent dislocation (after 13 years) and one for trochanteric bursitis where the wire was removed after 6 years and revision because of septic loosening was done after 14 years. Heterotopic ossification (HO) graded by means of the last follow-up radiograph showed that more than 50 % of the hips had grade I HO. These results indicate that the technique used for total hip replacement in this study is durable.


A study on the structure, mechanism, and biochemistry of kanamycin B dioxygenase (KanJ)-an enzyme with a broad range of substrates.
Kanamycin A is an aminoglycoside antibiotic isolated from Streptomyces kanamyceticus and used against a wide spectrum of bacteria, including Mycobacterium tuberculosis. Biosynthesis of kanamycin involves an oxidative deamination step catalyzed by kanamycin B dioxygenase (KanJ), thereby the C2' position of kanamycin B is transformed into a keto group upon release of ammonia. Here, we present for the first time, structural models of KanJ with several ligands, which along with the results of ITC binding assays and HPLC activity tests explain substrate specificity of the enzyme. The large size of the binding pocket suggests that KanJ can accept a broad range of substrates, which was confirmed by activity tests. Specificity of the enzyme with respect to its substrate is determined by the hydrogen bond interactions between the methylamino group of the antibiotic and highly conserved Asp134 and Cys150 as well as between hydroxyl groups of the substrate and Asn120 and Gln80. Upon antibiotic binding, the C terminus loop is significantly rearranged and Gln80 and Asn120, which are directly involved in substrate recognition, change their conformations. Based on reaction energy profiles obtained by density functional theory (DFT) simulations, we propose a mechanism of ketone formation involving the reactive FeIV = O and proceeding either via OH rebound, which yields a hemiaminal intermediate or by abstraction of two hydrogen atoms, which leads to an imine species. At acidic pH, the latter involves a lower barrier than the OH rebound, whereas at basic pH, the barrier leading to an imine vanishes completely. DATABASES: Structural data are available in PDB database under the accession numbers: 6S0R, 6S0T, 6S0U, 6S0W, 6S0V, 6S0S. Diffraction images are available at the Integrated Resource for Reproducibility in Macromolecular Crystallography at http://proteindiffraction.org under DOIs: 10.18430/m36s0t, 10.18430/m36s0u, 10.18430/m36s0r, 10.18430/m36s0s, 10.18430/m36s0v, 10.18430/m36s0w. A data set collection of computational results is available in the Mendeley Data database under DOI: 10.17632/sbyzssjmp3.1 and in the ioChem-BD database under DOI: 10.19061/iochem-bd-4-18.


Effect of low-thermal dissection device versus conventional electrocautery in mastectomy for female-to-male transgender patients.
Subcutaneous mastectomy, the first step in sexual reassignment surgery of female-to-male transsexuals, is associated with high rates of complication and revision surgery. Also, conventional electrosurgery and the associated thermal tissue damage may compromise outcome. This retrospective randomised clinical study evaluated the effect of low-thermal plasma dissection device (PEAK PlasmaBlade, Medtronic, Minneapolis, Minnesota) in comparison with conventional electrosurgery. A total of 17 female-to-male transsexuals undergoing mastectomy were randomised to PEAK PlasmaBlade on one breast side and to monopolar electrosurgery on the other side of the same patient. Wounds of 17 patients were examined histologically for acute thermal injury. Significantly less total volume of drain output (58.8 ± 37.4 mL vs 98.5 ± 76.4 mL; P = .012) was found on the PEAK PlasmaBlade side compared with the electrosurgery side. Duration of drain was significantly shorter on the PEAK PlasmaBlade side (2.5 ± 0.7 days vs 3.2 ± 0.6 days; P = .010). Furthermore, the PEAK PlasmaBlade side showed fewer thermal damages (41.2% vs 82.4%; P = .039) and thermal injury depth from PEAK PlasmaBlade side was less (3170 vs 4060 μm). PEAK PlasmaBlade appears to be superior to monopolar electrosurgery for mastectomy in female-to-male transsexuals, because it demonstrated less thermal tissue damage, less total volume of drain output, and shorter duration of drain, resulting in faster wound healing.


[Etiology of purulent infections in the Tiumen' region based on hospital data].
A study of laboratory findings for 474 examined patients, who were discharged from the Surgical Department of the Tyumen Regional Clinical Hospital, showed that S. aureus caused the infectious processes in 31.7% of the examinees. Oxacyllinresistant strain S aureus made up 9.7%.e examinees, and 22.6% of pathologies were caused in them by E. coli. E. coli was frequently found to be combined with polyresistant strains E. faecalis, E. faecium, K. pneumoniae and P. aeruginosa, which could be explained by the circulation of hospital strains within a hospital and by the selection of microorganisms with a high resistance level to antibacterial drugs.


Phenotypic Characteristics and Genetic Diversity of Salmonella enterica Serotype Derby Isolated from Human Patients and Foods of Animal Origin.
Salmonella enterica serotype Derby is among the three most common serotypes of nontyphoidal Salmonella isolated from patients with diarrhea in China. In this study, 133 Salmonella Derby isolates from human patients (n = 74) and foods of animal origin (n = 59) in Shanghai, China, between September 2013 and December 2014, were selected to study its phenotypic characteristics and genetic diversity. The isolates were subjected to antimicrobial susceptibility testing, plasmid replicon typing, virulence profile determination, and molecular subtyping by pulsed-field gel electrophoresis (PFGE). Isolates were frequently resistant to tetracycline (87.22%), sulfisoxazole (74.44%), and streptomycin (62.41%), and a low frequency of resistance was found toward ofloxacin (3.01%), ceftazidime (2.26%), and cefepime (1.50%); in addition, 93 (69.92%) isolates were multidrug resistant. The most common plasmid incompatibility replicon types were the IncF family (FIA, 51.31%; FIC, 27.82%; and FIB, 21.80%) and IncP types (35.34%): these plasmid types may be associated with the spread of antibiotic resistance and virulence genes. All isolates were positive for the Salmonella pathogenicity island (SPI) gene avrA and the fimbrial gene bcfC from among the 10 virulence genes detected, and most of them carried ssaQ (99.25%), mgtC (97.74%), siiD (98.50%), sopB (97.74%), and sopE (96.99%). PFGE showed 68 patterns in nine main clusters at an 85% similarity threshold. Most of the isolates from different sources possessed the same fingerprints or molecular profiles in each cluster, which strongly suggests the possibility that foods of animal origin, especially pork, serve as an important source for human infection. Moreover, this diversity may suggest strains originating from multiple clones. Therefore, surveillance on this serotype should be strengthened to prevent transmission of Salmonella Derby from foods of animal origin, especially pork, to humans.


Monte Carlo Dosimetry of the 60Co BEBIG High Dose Rate for Brachytherapy.
The use of high-dose-rate brachytherapy is currently a widespread practice worldwide. The most common isotope source is 192Ir, but 60Co is also becoming available for HDR. One of main advantages of 60Co compared to 192Ir is the economic and practical benefit because of its longer half-live, which is 5.27 years. Recently, Eckert & Ziegler BEBIG, Germany, introduced a new afterloading brachytherapy machine (MultiSource®); it has the option to use either the 60Co or 192Ir HDR source. The source for the Monte Carlo calculations is the new 60Co source (model Co0.A86), which is referred to as the new BEBIG 60Co HDR source and is a modified version of the 60Co source (model GK60M21), which is also from BEBIG. The purpose of this work is to obtain the dosimetry parameters in accordance with the AAPM TG-43U1 formalism with Monte Carlo calculations regarding the BEBIG 60Co high-dose-rate brachytherapy to investigate the required treatment-planning parameters. The geometric design and material details of the source was provided by the manufacturer and was used to define the Monte Carlo geometry. To validate the source geometry, a few dosimetry parameters had to be calculated according to the AAPM TG-43U1 formalism. The dosimetry studies included the calculation of the air kerma strength Sk, collision kerma in water along the transverse axis with an unbounded phantom, dose rate constant and radial dose function. The Monte Carlo code system that was used was EGSnrc with a new cavity code, which is a part of EGS++ that allows calculating the radial dose function around the source. The spectrum to simulate 60Co was composed of two photon energies, 1.17 and 1.33 MeV. Only the gamma part of the spectrum was used; the contribution of the electrons to the dose is negligible because of the full absorption by the stainless-steel wall around the metallic 60Co. The XCOM photon cross-section library was used in subsequent simulations, and the photoelectric effect, pair production, Rayleigh scattering and bound Compton scattering were included in the simulation. Variance reduction techniques were used to speed up the calculation and to considerably reduce the computer time. The cut-off energy was 10 keV for electrons and photons. To obtain the dose rate distributions of the source in an unbounded liquid water phantom, the source was immersed at the center of a cube phantom of 100 cm3. The liquid water density was 0.998 g/cm3, and photon histories of up to 1010 were used to obtain the results with a standard deviation of less than 0.5% (k = 1). The obtained dose rate constant for the BEBIG 60Co source was 1.108±0.001 cGyh-1U-1, which is consistent with the values in the literature. The radial dose functions were compared with the values of the consensus data set in the literature, and they are consistent with the published data for this energy range.


Branched-chain amino acid aminotransferase of Escherichia coli: overproduction and properties.
ilvE gene of Escherichia coli was inserted into the region downstream of the tac promotor. As a result, the branched-chain amino acid aminotransferase was overproduced by about a hundred-fold in E. coli W3110. The overproduced aminotransferase was purified from cell extracts about 40-fold to homogeneity. Chemical and physicochemical analyses confirmed that it was a product of the ilvE gene. The enzyme existed in a hexamer with a subunit molecular weight of 34,000; the double trimer model of the enzyme presumed by the previous chemical cross-linking experiments (Lee-Peng, F.-C. et al. (1979) J. bacteriol. 139, 339-345) was supported by electron micrographs. The circular dichroic (CD) spectrum of branch-chain amino acid aminotransferase had double negative maxima at 210 and 220 nm. The alpha-helical content was estimated to be about 40% from the CD spectrum in the region of 200 to 250 nm. The absorption spectrum of the enzyme showed two peaks at 330 and 410 nm. There was no pH-dependent spectral shift. The CD spectrum of the coenzyme, pyridoxal 5'-phosphate, had negative peaks at 330 and 410 nm. These spectral properties of branched-chain amino acid aminotransferase were quite different from those of E. coli aspartate aminotransferase. Each subunit bound approximately 1 mol of pyridoxal 5'-phosphate. A lysyl residue, which forms a Schiff base with the aldehyde group of the pyridoxal 5'-phosphate, was identified in the primary structure of the enzyme.


Skin calcium-binding protein in squamous metaplasia of human uterine cervix.
The distribution of skin calcium-binding protein in squamous cell metaplasia of human endocervix, in normal human skin, and in ovarian cancer was determined by the immunofluorescence technique. A rabbit antiserum specific to rat SCaBP was characterized by Ouchterlony immunodiffusion and by immunoprecipitation of 125I-labeled SCaBP. The specificity of antibody labeling was demonstrated by using preimmune rabbit serum and SCaBP antiserum competitively absorbed with purified SCaBP. In normal human skin SCaBP was found exclusively in the basal layer cell cytoplasm. This protein was not detected in normal columnar epithelium of endocervix. Epithelial tissues in the zone of transition between the cylindrical epithelium of the endocervical mucosa and the stratified squamous epithelium of the exocervix were obtained from 14 patients with a wide variety of squamous cell metaplasia. In the early stage of metaplasia SCaBP was detected exclusively in the cytoplasm of reserve undifferentiated cells. In the terminal stage of metaplasia the SCaBP was present only in the basal cell layer. SCaBP was found in several layers of dysplastic tissue, and this distribution appeared to be related to the loss of normal maturation of the epithelium. SCaBP was also present in squamous cell carcinoma of endocervix especially in the least differentiated regions of the tumor. No SCaBP was detected in any ovarian cancer cells. These findings are potentially useful as a means of early detection of squamous metaplasia and of distinguishing premalignant anaplastic lesions from those that are benign and nonproliferative. In addition, the presence of SCaBP in tumors derived from metaplastic epithelia and its absence in the ovarian cancer indicate that immunohistochemical search for this protein might be of value in tumors in which an epidermoid origin is a possibility.


Analysis on clinical effects of dilation and curettage guided by ultrasonography versus hysteroscopy after uterine artery embolization in the treatment of cesarean scar pregnancy.
The purpose of this study was to investigate the efficiency, complications, and subsequent fertility outcomes of two methods in treating patients with cesarean scar pregnancy (CSP) after receiving uterine artery embolization (UAE) treatment. A total of 62 CSP patients who visited our hospital and underwent UAE from January 2013 to January 2018 were retrospectively analyzed. Patients were either treated by dilation and curettage (D&C) guided by ultrasonography or hysteroscopy. The differences of related clinical indicators, clinical efficacy, complications, and subsequent fertility outcomes between the two groups were analyzed. The rates of therapeutic success of the ultrasonography group and hysteroscopy group were 84.6% and 95.7%, respectively, the difference was not statistically significant (P=0.243). However, the intraoperative blood loss, duration of hospitalization, and overall complications were significantly lower in hysteroscopy group compared with D&C guided by ultrasonography group (P<0.05 for all). Meanwhile, hysteroscopy had the advantage of discovering potential diverticulum in the lower segment of anterior wall of uterus (P<0.001). D&C guided by ultrasonography or hysteroscopy for the treatment of CSP after UAE resulted in similarly good clinical outcomes. Compared with treatment of D&C guided by ultrasonography, hysteroscopy had less complications and had the advantages of discovering diverticulum. It can be used as an effective way for the treatment of CSP.


Modelling the influence of host community composition in a sylvatic Trypanosoma cruzi system.
Species composition of wild reservoir hosts can influence the transmission and maintenance of multi-host vector borne pathogens. The 'pace of life' hypothesis proposes that the life history strategy of reservoir hosts can influence pathogen transmission of vector borne generalist pathogens. We use empirical data to parameterize a mathematical model that investigates the impacts of host life history traits on vector transmission dynamics of the vector-borne multi-host parasite Trypanosoma cruzi in habitats characterized by different degrees of deforestation and varying host community structure. The model considers susceptible and infected vector and host populations. When comparing the proportion of vectors infected with T. cruzi predicted by the model with empirical data, we found a trend of increasing vector infection as anthropogenic landscape disturbance increases for both data and model output. The model's vector infection rates were significantly lower than empirical results, but when incorporating host congenital transmission in the model, vector infection approaches field data. We conclude that intervened habitats associated with r-selected host species communities predict higher proportions of infected vectors.


Measuring differential treatment benefit across marker specific subgroups: The choice of outcome scale.
Clinical and epidemiological studies of anticancer therapies increasingly seek to identify predictive biomarkers to obtain insights into variation in treatment benefit. For time to event endpoints, a predictive biomarker is typically assessed using the interaction between the biomarker and treatment in a proportional hazards model. Interactions are contrasts of summaries of outcomes and depend upon the choice of the outcome scale. In this paper, we investigate interaction contrasts under three scales - the natural logarithm of hazard ratio, the natural logarithm of survival probability, and survival probability at a pre-specified time. We illustrate that we can have a non-zero interaction on survival or logarithm of survival probability scales even when there is no interaction on the logarithm of hazard ratio scale. Since survival probabilities have clinically useful interpretation and are easier to convey to patients than hazard ratios, we recommend evaluating a predictive biomarker using survival probabilities. We provide empirical illustration of the three scales of interaction for evaluating a predictive biomarker using reconstructed data from a published melanoma study.


Expression of the B8.7 antigen on hairy cells and relation with the LMW-BCGF response.
Hairy cells are classified as B cell tumors at a preplasma cell stage of differentiation and are believed to represent cells undergoing a switch process. These cells are stimulated in vitro to DNA synthesis and multiplication in the presence of the lymphokine LMW-BCGF. We have tested the level of expression on these cells of the newly described B8.7 activation marker which has been reported to be associated with the capacity of various B cells to respond to LMW-BCGF. The presence of this marker has been readily detected on the hairy cells of 10 of the 12 patients tested in this study; interestingly, for one of the negative cases, the tumor cells were unable to proliferate in response to LMW-BCGF. As on normal B cells, a marked inhibition of the LMW-BCGF dependent response could be achieved in the presence of a monoclonal anti-B8.7 antibody, sustaining the proposal that the B8.7 molecule is involved in the signaling pathway of this growth factor. IFN-alpha is highly efficient in the therapy of hairy cell leukemia (HCL), and we confirm in the present study that IFN-alpha also inhibits the LMW-BCGF dependent proliferation of hairy cells in vitro. In addition, we show that this inhibition is independent of a significant modulation of the B8.7 antigen, a molecule putatively associated with the LMW-BCGF receptor.


Structure and evolution of the Smith-Magenis syndrome repeat gene clusters, SMS-REPs.
An approximately 4-Mb genomic segment on chromosome 17p11.2, commonly deleted in patients with the Smith-Magenis syndrome (SMS) and duplicated in patients with dup(17)(p11.2p11.2) syndrome, is flanked by large, complex low-copy repeats (LCRs), termed proximal and distal SMS-REP. A third copy, the middle SMS-REP, is located between them. SMS-REPs are believed to mediate nonallelic homologous recombination, resulting in both SMS deletions and reciprocal duplications. To delineate the genomic structure and evolutionary origin of SMS-REPs, we constructed a bacterial artificial chromosome/P1 artificial chromosome contig spanning the entire SMS region, including the SMS-REPs, determined its genomic sequence, and used fluorescence in situ hybridization to study the evolution of SMS-REP in several primate species. Our analysis shows that both the proximal SMS-REP (approximately 256 kb) and the distal copy (approximately 176 kb) are located in the same orientation and derived from a progenitor copy, whereas the middle SMS-REP (approximately 241 kb) is inverted and appears to have been derived from the proximal copy. The SMS-REP LCRs are highly homologous (>98%) and contain at least 14 genes/pseudogenes each. SMS-REPs are not present in mice and were duplicated after the divergence of New World monkeys from pre-monkeys approximately 40-65 million years ago. Our findings potentially explain why the vast majority of SMS deletions and dup(17)(p11.2p11.2) occur at proximal and distal SMS-REPs and further support previous observations that higher-order genomic architecture involving LCRs arose recently during primate speciation and may predispose the human genome to both meiotic and mitotic rearrangements.


[Classification and management of sternal wound complications after cardiac surgery].
To define the classification of sternal wound complications after cardiac surgery and to explore the appropriate surgical treatment. Between July 2008 and January 2014, 260 patients with sternal wound complications after cardiac surgery were treated. There were 124 males and 136 females, aged 11-75 years (mean, 49.5 years). The disease duration was 13-365 days (mean, 26.6 days) with a wound length of 1-25 cm (mean, 13.4 cm). The wounds were divided into type I (n = 70), type II (n = 64), type III (n = 42), type IV (n = 78), and type V (n = 6) according to self-generated classification for sternal wound complications after cardiac surgery. After debridement, wounds of type I and type II were repaired with local flap transplantation; wounds of type III were repaired with local flap transplantation combined with butterfly sternal fixation (n = 28), with bilateral pectoralis muscle flap combined with butterfly sternal fixation (n = 11), and with bilateral pectoralis muscle flap (n = 3); wounds of type IV were repaired with bilateral pectoralis muscle flap (n = 65), rectus abdominis muscle flap (n = 5), and pedicled omental flap (n = 8); and wounds of type V were repaired with pedicled omental flap. All the operations were successfully performed. Three patients died after pedicled omental flap repair, including 1 case of type IV and 2 cases of type V. The hospitalization time were 4-86 days (mean, 18.3 days). Primary wound healing was obtained in 248 cases (96.5%); poor healing occurred in 9 patients, which were cured after second surgery in 8 cases and after the third surgery in 1 case. The surgical treatment based on self-generated classification is appropriate to sternal wound complications after cardiac surgery. It can provide clinical evidence for the choice of subsequent operation.


Whole proteome copy number dataset in primary mouse cortical neurons.
The functional diversity of neurons is specified through their proteome resulting in elaborate and tightly regulated protein interaction networks and signalling that regulates neuronal processes. Dysregulation of these dynamic networks in development or in adulthood lead to neurodevelopmental or neurological disorders respectively. Over the past few decades, mass spectrometry has become a powerful tool for quantifying and resolving any proteome, including complex tissues such as the brain proteome, with technological advances leading to higher levels of resolution and throughput than traditional biochemical techniques. In this article, we provide a proteomic reference dataset that has been generated to identify proteins and quantify their level of expression in primary mouse cortical neurons. It represents a summary analysis of previously published data in (Antico et al., 2021). Mouse cortical neurons were isolated from E16.5 C57Bl/6J mice and cultured for 21 days in vitro (DIV). We employed the mitochondrial uncouplers AntimycinA/Oligomycin (AO) to induce mitochondrial depolarisation that is a well-established paradigm to assess mitophagic signalling. Total lysates from mouse primary cortical neurons were subjected to label-free quantitative proteomic analysis using both data dependent acquisition (DDA) and data independent acquisition (DIA) modes. DDA proteomic analysis identified a total dataset of 9367 proteins in mouse cortical neurons and absolute abundance of proteins was calculated as copy numbers per cell. DDA dataset was also processed to generate a reference spectral library to fit in and quantify MS spectra generated in DIA mode. Quantitative DIA analysis identified more than 6000 protein groups and statistical comparison of the two analysed groups (untreated and AO-treated) revealed that the neuronal proteome was largely unchanged post mitochondrial depolarisation for 5 hours. To our knowledge, these files represent the most comprehensive DDA and DIA reference datasets of fully functional maturated mouse primary cortical neurons and serve as a valuable resource for further investigating the role of specific proteins involved in neurobiology and neurological disorders such as Alzheimer's disease (AD), Parkinson's disease (PD) and Autism Spectrum Disorders (ASD).


Reference values of within-district intraclass correlations of academic achievement by district characteristics: results from a meta-analysis of district-specific values.
Randomized experiments are often considered the strongest designs to study the impact of educational interventions. Perhaps the most prevalent class of designs used in large-scale education experiments is the cluster randomized design in which entire schools are assigned to treatments. In cluster randomized trials that assign schools to treatments within a set of school districts, the statistical power of the test for treatment effects depends on the within-district school-level intraclass correlation (ICC). Hedges and Hedberg (2014) recently computed within-district ICC values in 11 states using three-level models (students in schools in districts) that pooled results across all the districts within each state. Although values from these analyses are useful when working with a representative sample of districts, they may be misleading for other samples of districts because the magnitude of ICCs appears to be related to district size. To plan studies with small or nonrepresentative samples of districts, better information are needed about the relation of within-district school-level ICCs to district size. Our objective is to explore the relation between district size and within-district ICCs to provide reference values for math and reading achievement for Grades 3-8 by district size, poverty level, and urbanicity level. These values are not derived from pooling across all districts within a state as in previous work but are based on the direct calculation of within-district school-level ICCs for each school district. We use mixed models to estimate over 7,000 district-specific ICCs for math and reading achievement in 11 states and for Grades 3-8. We then perform a random effects meta-analysis on the estimated within-district ICCs. Our analysis is performed by grade and subject for different strata designated by district size (number of schools), urbanicity, and poverty rates.


Molecular basis for the substrate specificity of NIMA-related kinase-6 (NEK6). Evidence that NEK6 does not phosphorylate the hydrophobic motif of ribosomal S6 protein kinase and serum- and glucocorticoid-induced protein kinase in vivo.
The AGC family of protein kinases, which includes isoforms of protein kinase B (also known as Akt), ribosomal S6 protein kinase (S6K), and serum- and glucocorticoid-induced protein kinase (SGK) are activated in response to many extracellular signals and play key roles in regulating diverse cellular processes. They are activated by the phosphorylation of the T loop of their kinase domain by the 3-phosphoinositide-dependent protein kinase-1 and by phosphorylation of a residue located C-terminal to the kinase domain in a region termed the hydrophobic motif. Recent work has implicated the NIMA (never in mitosis, gene A)-related kinase-6 (NEK6) as the enzyme that phosphorylates the hydrophobic motif of S6K1 in vivo. Here we demonstrate that in addition to phosphorylating S6K1 and SGK1 at their hydrophobic motif, NEK6 also phosphorylates S6K1 at two other sites and phosphorylates SGK1 at one other site in vitro. Employing the Jerini pepSTAR method in combination with kinetic analysis of phosphorylation of variant peptides, we establish the key substrate specificity determinants for NEK6. Our analysis indicates that NEK6 has a strong preference for Leu 3 residues N-terminal to the site of phosphorylation. Its mutation to either Ile or Val severely reduced the efficacy with which NEK6-phosphorylated peptide substrates, and moreover, mutation of the equivalent Leu residue in S6K1 or SGK1 prevented phosphorylation of their hydrophobic motifs by NEK6 in vitro. However, these mutants of S6K1 or SGK1 still became phosphorylated at their hydrophobic motif following insulin-like growth factor-1 stimulation of transfected 293 cells. This study provides the first description of the basis for the substrate specificity of NEK6 and indicates that NEK6 is unlikely to be responsible for the IGF1-induced phosphorylation of the hydrophobic motif of S6K, SGK, and protein kinase B isoforms in vivo.


Measuring identification with the mothering role.
Three summated rating scales entitled Feelings of Motherliness (FOM), Conception of the Fetus as a Person (CFP), and Appropriateness of Fantasies about the Baby-to be (AFB) were constructed. During instrument development, data were collected from a sample of 169 women selected using the criteria of gravidity, ethnicity, and experience. Thirty-four of these women were used to test the final versions of the scales. Coefficient alphas for FOM, CFP, and AFB were.87, .89, and .67, respectively. Coefficient alphas for the subscales ranged from .88 to 1.0. Content validity was built into the scales. The data supported the concept validity of FOM and CFP, but not AFB. Three subscales of the Parental Attitude Research Instrument were used as criterion measures; criterion validity was not established. Construct validity was assessed using convergent and discriminant measures. Data provided support for convergent validity, but not for discriminant validity. Further modification and testing of the scales are needed.


Interactive effects of ammonia and light intensity on hematochemical variables in broiler chickens.
This study examined the influence of early atmospheric ammonia exposure, light intensity throughout rearing, and their interaction on blood gases, electrolytes, and acid-base balance in broiler chickens under environmentally controlled conditions. The experiment consisted of a 3 x 3 factorial arranged in a randomized complete block design, with trials being replicated over time. The 9 treatments consisted of 3 levels (0, 25, and 50 ppm) of ammonia concentrations for 14 d and levels (0.2, 2.0, and 20 lx) of light intensities from 8 to 36 d of age. Venous blood samples were collected on d 6, 11, 14, and 35. On d 6, partial pressure of CO2 and Na+ increased significantly (P <or= 0.05), whereas partial pressure of O(2), pH, and K+ decreased with increasing ammonia concentration. As light intensity increased, pO2 and K+ were significantly (P <or= 0.05) reduced. Ammonia x light intensity interactions were observed for hemoglobin, hematocrit, K+, and BW. The interaction of ammonia and light intensity for 7 d further exacerbated physiological variables. The main effect of ammonia was more pronounced than that of light intensity. These conditions worsened as the duration of ammonia concentration exposure and light intensity increased from d 7 to 14 of exposure. However, all affected variables returned to near normal at later time points in the exposed chickens so that the apparent effects were lost. Plasma corticosterone and glucose concentrations were not significantly altered by exposure to differing levels of ammonia or light intensity, suggesting an absence of stress related to ammonia, light intensity, or their interaction. It was concluded that exposure of broiler chickens to aerial ammonia concentrations of 0 to 50 ppm from d 1 to 14 posthatch in the presence of light intensities ranging from 0.2 to 20 lx had no direct effect on some physiological blood variables and did not induce stress in broilers.


[The Role of Transcriptional Factor Gene TBX5 in Development of Disorders of Cadiac Conduction].
In order to study relationship between development of idiopathic atrioventricular (AV) and intraventricular disorders of cardiac conduction (DCC) with single nucleotide polymorphism (SNP) of TBX5 gene we examined 260 persons with primary DCC (71 patients with abnormal AV conduction, 84 and 105 patients with disordered conduction along right and left brunches of His bundle, respectively) as well as 257 individuals without cardiovascular diseases (control group). Patients were divided into subgroups depending on nosology, age, and sex. Diagnosis was verified by standard cardiological methods and retrospective analysis of available results of previous examinations. Molecular-genetic study of DNA was used for identification of genotype of TBX5 gene SNP. The results indicated significant preponderance of rare GG genotype (CNP-marker rs3825214) of TBX5 gene in the group of patients with left bundle branch block and in the subgroup of women with this pathology. These data suggest that presence of GG genotype (rs3825214) of TBX5 gene increases probability of development of idiopathic DCC along left bundle branch mainly in women.


Ligand size dependence of U-N and U-O bond character in a series of uranyl hexaphyrin complexes: quantum chemical simulation and density based analysis.
A series of uranyl complexes with hexaphyrin ligands are investigated at the density functional level of theory and analysed using a variety of density-based techniques. A relationship is identified between the size of the ligand and the stability of the complex, controlled by the presence of meso-carbon centres in the porphyrin ring. The complex with the smallest ligand, cyclo[6]pyrrole, is found to have enhanced covalent character in equatorial U-N bonds as defined by the quantum theory of atoms in molecules (QTAIM), as well as enhanced stability, compared to the larger complexes. QTAIM data are supported by electron density difference distributions, integrated electronic properties and analysis of the reduced density gradient (RDG), which all show unambiguous evidence of electron sharing in all U-N bonds. In all complexes, a weakening of the covalent axial U-Oyl interaction in comparison to free uranyl is found, with evidence for a separation of electronic charge resulting in a more ionic interaction. A relationship between covalent character in the U-N bonds and the magnitude of uranyl charge redistribution is identified, where the greater the covalent character of the U-N interaction, the more ionic the U-Oyl interaction appears. The complex with the largest ligand, hexaphyrin(18.104.22.168.1.1), is found to have additional interactions with the uranyl oxygen centres, perturbing the U-Oyl interaction.


Endocrine actions of myostatin: systemic regulation of the IGF and IGF binding protein axis.
Myostatin's inhibitory actions on striated muscle growth are believed to be directly mediated by locally produced myostatin and possibly by IGF binding proteins (IGFBPs). We therefore measured skeletal muscle, heart, and liver expression, in neonates and adults, and circulating levels of various IGF axis components (IGF-I, IGFBP-1 to IGFBP-3, and acid labile subunit) in wild-type and mstn-/- mice. Compared with wild type, differences in muscle expression were tissue specific, although IGF-I receptor expression was higher in all mstn-/- neonatal tissues and in adult gastrocnemius. Liver expression of several components also differed between genotype as IGF-I receptor, IGFBP-3 and IGFBP-5 expression was higher in mstn-/- neonates and IGF-I and IGFBP-3 in adults. Circulating IGF-I levels were also higher in mstn-/- adults, whereas IGFBP-1 and IGFBP-2 levels were lower. Comparing IGF-I:IGFBP molar ratios suggested that the relative IGF-binding capacity was potentially lower in mstn-/- mice, and thus, total and "free" IGF-I levels may be elevated. This in turn may increase negative feedback control on GH, because mstn-/- liver weights were lower. Bone growth was similar in both genotypes, suggesting that changes in circulating IGF-I may be more important to muscle, whose mass is enhanced in mstn-/- mice, than to bone. Myostatin receptors, but not myostatin itself, are expressed in the liver. Changes in hepatic production of circulating IGF axis components could therefore result from the loss of endocrine myostatin. Thus, myostatin may inhibit striated muscle growth directly at the cellular level and indirectly through systemic effects on the IGF axis.


[Anglo-Saxon guidelines for the treatment of community-acquired pneumonia applicable in The Netherlands as well].
There are three Anglo-Saxon guidelines for the management of patients with a community-acquired pneumonia: an American, a British and a Canadian one. The guidelines correspond fairly well. There is a subdivision into categories according to whether the patients are treated at home (formerly healthy patients younger than 60 years versus patients with pre-existent disease or aged 60 years and more) or in the hospital (patients not needing intensive care versus those who do need it). For each category the most common causative micro-organisms are listed together with recommended antibiotic treatment. The Canadian guidelines have nursing home patients as a separate category because of slightly different causative organisms due to frequent microaspiration. The guidelines are applicable to the situation in the Netherlands, with a few exceptions: antibiotic resistance is not a major problem in the Netherlands (as yet), and contrary to what the guidelines state an agent with activity against Pseudomonas aeruginosa is not necessary; the same agents as in category III can be prescribed in these patients. A macrolide or azalide antibiotic is advisable for intensive care patients in view of the possibility of infection with Legionella pneumophila or Mycoplasma pneumoniae.


Chirped Optical Solitons in Single-mode Birefringent Fibers.
The trapping behavior of two chirped solitons forming a bound state in a single-mode birefringent fiber is investigated on the basis of a model of coupled nonlinear Schroedinger equations. The positive initial chirp plays an important role in controlling the threshold amplitude for soliton trapping without causing excessive pulse broadening.


Mitomycin C in large infrequent doses in breast cancer.
Seventy-one women with far-advanced breast cancer resistant to standard chemotherapeutic agents were administered mitomycin C using an intermittent high dose schedule. One group consisted of 54 patients with measurable metastatic tumor; a second group consisted of 18 patients with nonmeasurable osseous metastases. Objective response rate in group 1 was 26% for an average duration of 2 1/2 months. Subjective response rate in group 2 was 44% for an average duration of 3 months. Response and toxicity data were similar to those of studies employing the less convenient protracted low-dose schedule. Prior treatment with other alkylating agents did not adversely affect response. Further investigation into the role of mitomycin in combination chemotherapy programs is recommended.


Developmental Changes in Pretend Play from 22- to 34-Months in Younger Siblings of Children with Autism Spectrum Disorder.
Developmental trajectories of children's pretend play and social engagement, as well as parent sensitivity and stimulation, were examined in toddlers with an older sibling with autism spectrum disorder (ASD, high risk; HR) and toddlers with typically-developing older siblings (low risk; LR). Children (N = 168, 97 boys, 71 girls) were observed at 22, 28, and 34 months during free play with a parent and elicited pretend play with an examiner. At 28 and 34 months, children were asked to imagine the consequences of actions pantomimed by the examiner on a pretend transformation task. At 36 months children were assessed for ASD, yielding 3 groups for comparison: HR children with ASD, HR children without ASD (HR-noASD), and LR children. Children in all 3 groups showed developmental changes, engaging in more bouts of pretend play and obtaining higher scores on the elicited pretend and transformation tasks with age, but children with ASD lagged behind the other 2 groups on most measures. Children with ASD were also less engaged with their parents or the examiner during play interactions than either LR or HR-noASD children, with minimal developmental change evident. Parents, regardless of group, were highly engaged with their children, but parents of HR-noASD children received somewhat higher ratings on stimulation than parents of LR children. Most group differences were not accounted for by cognitive functioning. Instead, lower social engagement appears to be an important correlate of less advanced pretend skills, with implications for understanding the early development of children with ASD and for early intervention.


Antitumor activity of cytotoxic T lymphocytes engineered to target vascular endothelial growth factor receptors.
The demonstration that angiogenesis is required for the growth of solid tumors has fueled an intense interest in the development of new therapeutic strategies that target the tumor vasculature. Here we report the development of an immune-based antiangiogenic strategy that is based on the generation of T lymphocytes that possess a killing specificity for cells expressing vascular endothelial growth factor receptors (VEGFRs). To target VEGFR-expressing cells, recombinant retroviral vectors were generated that encoded a chimeric T cell receptor comprised of VEGF sequences linked to intracellular signaling sequences derived from the zeta chain of the T cell receptor. After transduction of primary murine CD8 lymphocytes by such vectors, the transduced cells were shown to possess an efficient killing specificity for cells expressing the VEGF receptor, Flk-1, as measured by in vitro cytotoxicity assays. After adoptive transfer into tumor-bearing mice, the genetically modified cytotoxic T lymphocytes strongly inhibited the growth of a variety of syngeneic murine tumors and human tumor xenografts. An increased effect on in vivo tumor growth inhibition was seen when this therapy was combined with the systemic administration of TNP-470, a conventional angiogenesis inhibitor. The utilization of the immune system to target angiogenic markers expressed on tumor vasculature may prove to be a powerful means for controlling tumor growth.


Toxicity profiles in rats treated with tumorigenic and nontumorigenic triazole conazole fungicides: Propiconazole, triadimefon, and myclobutanil.
Conazoles are a class of azole based fungicides used in agriculture and as pharmaceutical products. They have a common mode of antifungal action through inhibition of ergosterol biosynthesis. Some members of this class have been shown to be hepatotoxic and will induce mouse hepatocellular tumors and/or rat thyroid follicular cell tumors. The particular mode of toxic and tumorigenic action for these compounds is not known, however it has been proposed that triadimefon-induced rat thyroid tumors arise through the specific mechanism of increased TSH. The present study was designed to identify commonalities of effects across the different conazoles and to determine unique features of the tissue responses that suggest a toxicity pathway and a mode of action for the observed thyroid response for triadimefon. Male Wistar/Han rats were treated with triadimefon (100, 500, 1800 ppm), propiconazole (100, 500, 2500 ppm), or myclobutanil (100, 500, 2000 ppm) in feed for 4, 30, or 90 days. The rats were evaluated for clinical signs, body and liver weight, histopathology of thyroid and liver, hepatic metabolizing enzyme activity, and serum T3, T4, TSH, and cholesterol levels. There was a dose-dependent increase in liver weight but not body weight for all treatments. The indication of cytochrome induction, pentoxyresorufin O-dealkylation (PROD) activity, had a dose-related increase at all time points for all conazoles. Uridine diphopho-glucuronosyl transferase (UDPGT), the T4 metabolizing enzyme measured as glucuronidation of 1-naphthol, was induced to the same extent after 30 and 90 days for all three conazoles. Livers from all high dose treated rats had centrilobular hepatocyte hypertrophy after 4 days, while only triadimefon and propiconazole treated rats had hepatocyte hypertrophy after 30 days, and only triadimefon treated rats had hepatocyte hypertrophy after 90 days. Thyroid follicular cell hypertrophy, increased follicular cell proliferation, and colloid depletion were present only after 30 days in rats treated with the high dose of triadimefon. A dose-dependent decrease in T4 was present after 4 days with all 3 compounds but only the high doses of propiconazole and triadimefon produced decreased T4 after 30 days. T3 was decreased after high-dose triadimefon after 4 days and in a dose-dependent manner for all compounds after 30 days. Thyroid hormone levels did not differ from control values after 90 days and TSH was not increased in any exposure group. A unique pattern of toxic responses was not identified for each conazole and the hypothesized mode of action for triadimefon-induced thyroid gland tumors was not supported by the data.


Recurrent gynandroblastoma of ovary-A case report: a molecular and immunohistochemical analysis.
Gynandroblastoma is a rare ovarian tumor that is composed of both Sertoli cells and granulosa cells. Only 23 cases have been reported in the literature, and recurrence has never been described. We report the first case of a recurrent gynandroblastoma along with its molecular analysis and immunohistochemical studies. A 49-year-old Gravida 0 woman with a 10-year prior diagnosis of ovarian-mixed stromal tissue tumor (well-differentiated Sertoli cell and granulosa cell tumor) and staging laparotomy, presented now with a retroperitoneal mass and an elevated inhibin level. CT scan was suspicious for recurrence. The patient had no prior adjuvant therapy. The histomorphological features of the recurrent tumor had both Sertoli cell and granulosa cell tumor. The molecular analysis of both primary and recurrent tumor showed minor genetic instability in the 17q12.2 gene locus with no dedifferentiation or progression, which is consistent with a low-grade tumor. The immunohistochemical staining profile showed positivity for CD99, inhibin, calretinin, and vimentin; focal positivity for cytokeratin AE1/AE3 and negative for EMA and melan-A. All the previously mentioned immunostainings support the diagnosis. We report the first case of a recurrent gynandroblastoma 10 years after initial presentation along with its molecular analysis and immunohistochemical studies.


Adapting to the 30-degree visual perspective by emulating the angled laparoscope: a simple and low-cost solution for basic surgical training.
The ability to handle and adapt to the visual perspectives generated by angled laparoscopes is crucial for skilled laparoscopic surgery. However, the control of the visual work space depends on the ability of the operator of the camera, who is often not the most experienced member of the surgical team. Here, we present a simple, low-cost option for surgical training that challenges the learner with static and dynamic visual perspectives at 30 degrees using a system that emulates the angled laparoscope. A system was developed using a low-cost camera and readily available materials to emulate the angled laparoscope. Nine participants undertook 3 tasks to test spatial adaptation to the static and dynamic visual perspectives at 30 degrees. Completing each task to a predefined satisfactory level ensured precision of execution of the tasks. Associated metrics (time and error rate) were recorded, and the performance of participants were determined. A total of 450 repetitions were performed by 9 residents at various stages of training. All the tasks were performed with a visual perspective of 30 degrees using the system. Junior residents were more proficient than senior residents. This system is a viable and low-cost alternative for developing the basic psychomotor skills necessary for the handling and adaptation to visual perspectives of 30 degrees, without depending on a laparoscopic tower, in junior residents. More advanced skills may then be acquired by other means, such as in the operating theater or through clinical experience.


Accuracy and safety of percutaneous US-guided needle biopsies in liver metastasis and hemangiomas.
The aim of this study was to compare small size aspiration and large size cutting needles on the diagnostic yield in the biopsies of liver and especially to investigate the reliability on percutaneous biopsy of liver hemangiomas. Nine hundred fifty patients with percutaneous ultrasonography-guided liver biopsies were evaluated retrospectively. This study enrolled 841 patients biopsied with either large size cutting needle (Tru-cut 14G-18G) or small size aspiration needle (WestCott 20G). Further evaluation was performed in 312 patients with metastasis and 48 patients with hemangiomas. Diagnostic yield was higher in the large size cutting needle group (96.8%, 150/155) than in the small size aspiration needle group (84.1%, 132/157) in liver metastasis (P<0.001). There was no significant difference among diagnostic accuracies of Tru-cut 14G, 16G, and 18 G needles in metastasis (P=0.255). Accuracy rate was 77.9% (208/267) in benign and 89.5% (514/574) in malignant diseases. Sensitivity, specificity, and accuracy were 81.2% (514/633), 100% (208/208), and 85.8% (722/841), respectively. Only two major complications were found (0.16%) with small needles. The authors suggest the use of large size cutting needles, because they provide more accurate diagnosis, and should be used in liver metastasis instead of small aspiration needles, if there is no on-site pathologist at aspiration biopsies or a more specific diagnosis is required. Among them, 18G cutting needle should be chosen. All the needles, including the large cutting type, were found safe in the biopsies of liver hemangiomas.


Clinical, cytogenetic and molecular findings in nine Moroccan patients with Fanconi anemia.
Fanconi anemia (FA) is a rare inherited hematological disease due to a defect in the DNA repair pathway resulting in congenital abnormalities and high susceptibility to develop cancers. The cytogenetic analysis using alkylating agents is still a reference test to establish the diagnosis. Despite the genetic heterogeneity, the identification of the causal mutation is actually performed especially after the development of next generation sequencing (NGS). we report here nine Moroccan patients referred to the department of Medical Genetics for suspicion of FA. We realized a genetic consultation to establish a clinical record with biological data before carrying out the genetic analysis. Karyotyping with mitomycin was performed for all the probands before elaborating molecular study. We used massively parallel sequencing to analyse the three most frequent mutated genes FANCA, FANCC, and FANCG, representing 84% of all genes involved in FA. all the patients showed hematological signs associated with at least one extra-hematological congenital anomaly. The chromosomal breaks were significantly higher for the nine patients, compared to the controls. The molecular diagnosis was confirmed in 8 of the 9 families tested (88.8%) with 4 novel mutations. The next generation based sequencing identified 9 variations: 6 in the FANCA gene (66.6%), 3 in the FANCG gene (33.3%) and no FANCC variation was found. Of those, 7 were homozygous and 2 were compounds heterozygous. to the best of our knowledge, this is the first molecular report of Moroccan patients with FA suggesting the predominance of two genes without any recurrent mutation. The molecular analysis of FANCA and FANCG genes should be offered first for all patients in Morocco.