Rhabdoid variant of urothelial carcinoma of the urinary bladder: a case report with emphasis on immunohistochemical analysis regarding the formation of rhabdoid morphology.
Various histological variants of urothelial carcinoma (UC) have been described. They are associated with different clinical outcomes and/or therapeutic approaches; in addition, recognition of these histological variants is also important in preventing diagnostic misinterpretations. Histological variants based on cytoplasmic features, such as plasmacytoid, rhabdoid, clear-cell, and lipoid-rich variants, have been described in invasive UC. Herein, we report an exceedingly rare case of a rhabdoid variant of UC arising in the urinary bladder of a 61-year-old man. Including UC, the presence of rhabdoid cells has been described in various types of malignant tumors. These tumors are regarded as more aggressive neoplasms than those without rhabdoid cells. It has been previously found that non-degraded aggregation of intermediate filaments and membrane proteins conjugated with ubiquitin and p62 is a noticeable finding in the formation of rhabdoid morphology. We have validated the existence of this mechanism in a rhabdoid variant of UC by extensive immunohistochemical analysis.


Clinical implications of the basic defects in Cockayne syndrome and xeroderma pigmentosum and the DNA lesions responsible for cancer, neurodegeneration and aging.
Cancer, aging, and neurodegeneration are all associated with DNA damage and repair in complex fashions. Aging appears to be a cell and tissue-wide process linked to the insulin-dependent pathway in several DNA repair deficient disorders, especially in mice. Cancer and neurodegeneration appear to have complementary relationships to DNA damage and repair. Cancer arises from surviving cells, or even stem cells, that have down-regulated many pathways, including apoptosis, that regulate genomic stability in a multi-step process. Neurodegeneration however occurs in nondividing neurons in which the persistence of apoptosis in response to reactive oxygen species is, itself, pathological. Questions that remain open concern: sources and chemical nature of naturally occurring DNA damaging agents, especially whether mitochondria are the true source; the target tissues for DNA damage and repair; do the human DNA repair deficient diseases delineate specific pathways of DNA damage relevant to clinical outcomes; if naturally occurring reactive oxygen species are pathological in human repair deficient disease, would anti-oxidants or anti-apoptotic agents be feasible therapeutic agent?


Burnout and the Sexual Assault Nurse Examiner: Who Is Experiencing Burnout and Why?
A sexual assault nurse examiner role exemplifies the high-stress and highly emotional patient interactions that are often associated with burnout. The purpose of this study was to examine the frequency of burnout among sexual assault nurse examiners in North Carolina. This cross-sectional study was an anonymous survey of practicing sexual assault nurse examiners within North Carolina using the Maslach Burnout Inventory and additional demographics. Results were analyzed with odds ratios, confidence intervals, Fisher exact, chi-square, and Kruskal Wallis tests as appropriate. Among 95 respondents, burnout was more frequent in sexual assault nurse examiners who stopped both emergency and nurse examiner work (55.6%, odds ratio 4.41, 95% confidence interval 1.07-18.06) and in dual function nurses (both emergency and nurse examiner work, 35.7%, odds ratio 2.71, 95% confidence interval 1.04-7.06). Sexual assault nurse examiners who had a high percentage of pediatric cases (above the median of 40%) were more likely to meet burnout thresholds for emotional exhaustion scores > 26 (48.78% vs 25.93%, χ2 = 5.30, P = .02) and more likely to meet burnout thresholds for depersonalization scores > 9 (48.78% vs 24.07%, χ2 = 6.28, P = .01). Higher frequency of burnout threshold criteria was found in those people who worked concurrently as a sexual assault nurse examiner and an emergency nurse and in those who had retired from both specialties. We also found that sexual assault nurse examiners with a higher case mix of pediatric cases had higher emotional exhaustion scores and higher depersonalization scores.


Analysis of mebendazole polymorphs by Fourier transform IR spectrometry using chemometric methods.
A diffuse reflectance IR Fourier transform IR spectrometry (DRIFTS) method was developed for the rapid, direct measurement of mebendazole in drugs. Conventional KBr spectra and DRIFTS spectra were compared for the best determination of the active substance in the drug formulations. Two chemometric approaches were used in the data processing: multicomponent partial least squares (PLS2) and principal component regression. The best results were obtained with the PLS2 method.


Thin film complementary metal oxide semiconductor (CMOS) device using a single-step deposition of the channel layer.
We report, for the first time, the use of a single step deposition of semiconductor channel layer to simultaneously achieve both n- and p-type transport in transparent oxide thin film transistors (TFTs). This effect is achieved by controlling the concentration of hydroxyl groups (OH-groups) in the underlying gate dielectrics. The semiconducting tin oxide layer was deposited at room temperature, and the maximum device fabrication temperature was 350 °C. Both n and p-type TFTs showed fairly comparable performance. A functional CMOS inverter was fabricated using this novel scheme, indicating the potential use of our approach for various practical applications.


Combinative ligand-receptor interactions: epinephrine depresses RAW264 macrophage antibody-dependent phagocytosis in the absence and presence of met-enkephalin.
Macrophages have been shown to possess cell surface receptors for opiates and catecholamines. The abilities of these ligands to affect RAW264 macrophage antibody-dependent effector activity directed against sheep red blood cells were tested. Phagocytosis was measured by the uptake of 51Cr labeled erythrocytes and optical microscopy. Cytolysis was measured by 51Cr-release assays. Met-enkephalin increased specific antibody-dependent phagocytosis in a dose-dependent fashion; the optimal dose was found to be 10(-8) M. Epinephrine diminished phagocytosis in a dose-dependent manner exhibiting a maximal inhibition at 10(-4)-10(-5) M. This inhibition can be blocked by propranolol. The combined effects of simultaneous treatment with met-enkephalin and epinephrine were measured. At the several doses tested, the combined effects of these two ligands on the amount of phagocytosis were equivalent to or more inhibitory than epinephrine alone. Thioglycolate-elicited murine peritoneal macrophages demonstrated similar responses to epinephrine, met-enkephalin, and their combination. Therefore, in vitro models more closely approximating in vivo neuroregulation of macrophage function demonstrate phagocytic inhibition.


Effects of a 3-month weight-bearing and resistance exercise training on circulating osteogenic cells and bone formation markers in postmenopausal women with low bone mass.
Osteoporosis is a health issue in postmenopausal women. Physical activity is recommended in these subjects, since it has positive effects on bone mass. Cellular mechanisms underlying this effect are still unclear. Osteogenic cells, released after physical exertion, could be a key factor in exercise-induced bone formation. The aim of our research was to explore if a weight-bearing and resistance exercise program could positively affect circulating osteogenic cells (OCs), markers of bone formation and quality of life (QoL) in osteopenic postmenopausal women. We recruited 33 postmenopausal women with a T-score at lumbar spine or femoral neck between - 1 and - 2.5 SD. Anthropometric and fitness parameters, bone-remodeling markers, OCs, and QoL were evaluated at the time of enrolment, after 1-month run-in period, and after 3 months of weight-bearing and resistance exercise. After 3 months of training, the pro-collagen type 1 N-terminal peptide (P1NP) and the number of OCs were significantly increased, with no significant increase of the type 1 collagen cross-linked C-telopeptide (sCTX). We also observed a significant increase in body height, one-repetition maximum (1RM) on the pull-down lat machine and leg press, and mean VO2max. The increase of immature circulating OCs was significantly correlated with the improvement of 1RM both of the upper and lower limbs. Moreover, QoL was significantly improved with regard to pain, physical function, mental function, and general QoL. The improvement in QoL, namely in the overall score and in the pain score, was significantly correlated with the increase in height. The exercise program we trialed is able to increase the markers of bone formation and the commitment of immature OCs with no significant increase in the markers of bone resorption. Our results confirm that combined weight-bearing and resistance physical activity is an effective tool to improve QoL of postmenopausal women with low bone mass. NCT03195517.


Vaginal template implant for cervical carcinoma with vaginal stenosis or inadvertent diagnosis after hysterectomy.
For cervical carcinoma patients with poor geometry for conventional intracavitary radiotherapy, a simple vaginal template for interstitial implantation as a substitute was used. This template has also been used to treat patients who had hysterectomy done without knowledge of an early tumor in the cervix, and for patients with recurrent disease. This is a report of the treatment results. A total of 21 patients were treated over from July 1987 to June 1991 with this vaginal template implant forming part of the treatment, 12 of these were performed for vaginal stenosis. The applicator consists of a front piece and an end piece. Holes were drilled in the front piece to guide the implantation of the cervix or vaginal vault. The diameter of applicators varied from 2 cm to 3.5 cm. Depending on the diameter of the applicators, six to eight needles on the periphery, or eight peripheral plus one central needle were used. The activity of the needles were around 8 mCi with a total length of 5.5 cm. The end piece was locked onto the front piece by a bayonet-type locking device. The purpose of the end piece was two-fold: to make up the length of the whole applicator to fit the vagina and to keep the implanted needles in place without being extruded. The implantation was performed under general anesthesia. One of the twelve patients treated with the vaginal template implant for vaginal stenosis had relapsed centrally but subsequently died of intercurrent disease. Two other patients died of intercurrent disease at 26.2 and 41.9 months, respectively, without evidence of relapse. Nine other patients had been followed with no evidence of local relapse for 23.7 to 54.6 months. This vaginal template implantation is a satisfactory means of treating patients with vaginal stenosis and those who had hysterectomy done without knowledge of an early tumor in cervix.


Feline calicivirus capsid protein expression and capsid assembly in cultured feline cells.
The capsid protein of feline calicivirus (FCV) was expressed by using plasmids containing cytomegalovirus, simian virus 40, or T7 promoters. The strongest expression was achieved with the T7 promoter and coinfection with vaccinia virus expressing the T7 RNA polymerase (MVA/T7pol). The FCV precursor capsid protein was processed to the mature-size protein, and these proteins were assembled in to virus-like particles.


Extracellular Actin in Health and Disease.
This review considers the functions of extracellular actin - cell surface bound, associated with extracellular matrix, or freely circulating. The role of this protein in different pathological processes is analyzed: its toxic effects and involvement in autoimmune diseases as an autoantigen. The extracellular actin clearance system and its role in protection against the negative effects of actin are characterized. Levels of free-circulating actin, anti-actin immunoglobulins, and components of the actin clearance system as prognostic biomarkers for different diseases are reviewed. Experimental approaches to protection against excessive amounts of free-circulating F-actin are discussed.


Karyopherin beta3: a new cellular target for the HPV-16 E5 oncoprotein.
Epidemiological and experimental studies have shown that high-risk human papillomaviruses (HPVs) are the causative agents of cervical cancer worldwide, and that HPV-16 is associated with more than half of these cases. In addition to the well-characterized E6 and E7 oncoproteins of HPV-16, recent evidence increasingly has implicated the HPV-16 E5 protein (16E5) as an important mediator of oncogenic transformation. Since 16E5 has no known intrinsic enzymatic activity, its effects on infected cells are most likely mediated by interactions with various cellular proteins and/or its documented association with lipid rafts. In the present study, we describe a new cellular target that binds to 16E5 in COS cells and in stable human ectocervical cell lines. This target is karyopherin beta3, a member of the nuclear import receptor family with critical roles in the nuclear import of ribosomal proteins and in the secretory pathway.


Proteomic identification of gender molecular markers in Bothrops jararaca venom.
Variation in the snake venom proteome is a well-documented phenomenon; however, sex-based variation in the venom proteome/peptidome is poorly understood. Bothrops jararaca shows significant sexual size dimorphism and here we report a comparative proteomic/peptidomic analysis of venoms from male and female specimens and correlate it with the evaluation of important venom features. We demonstrate that adult male and female venoms have distinct profiles of proteolytic activity upon fibrinogen and gelatin. These differences were clearly reflected in their different profiles of SDS-PAGE, two-dimensional electrophoresis and glycosylated proteins. Identification of differential protein bands and spots between male or female venoms revealed gender-specific molecular markers. However, the proteome comparison by in-solution trypsin digestion and label-free quantification analysis showed that the overall profiles of male and female venoms are similar at the polypeptide chain level but show striking variation regarding their attached carbohydrate moieties. The analysis of the peptidomes of male and female venoms revealed different contents of peptides, while the bradykinin potentiating peptides (BPPs) showed rather similar profiles. Furthermore we confirmed the ubiquitous presence of four BPPs that lack the C-terminal Q-I-P-P sequence only in the female venom as gender molecular markers. As a result of these studies we demonstrate that the sexual size dimorphism is associated with differences in the venom proteome/peptidome in B. jararaca species. Moreover, gender-based variations contributed by different glycosylation levels in toxins impact venom complexity. Bothrops jararaca is primarily a nocturnal and generalist snake species, however, it exhibits a notable ontogenetic shift in diet and in venom proteome upon neonate to adult transition. As is common in the Bothrops genus, B. jararaca shows significant sexual dimorphism in snout-vent length and weight, with females being larger than males. This sexual size dimorphism suggests the tendency for female specimens to feed on larger prey, and for male specimens to go on a diet similar to that of juveniles. Variation in the snake venom proteome is a ubiquitous phenomenon occurring at all taxonomic levels. At the intraspecific variation level, the individual contribution to the venom proteome is important but effects contributed by age and feeding habits may also affect the proteome phenotype. Whether sex-based factors play a role in venom variation of a species that shows sexual size dimorphism is poorly known. The use of proteomic strategies supported by transcriptomic data allows a more comprehensive assessment of venom proteomes uncovering components that are gender-specific.


Human bocavirus infections in hospitalized Greek children.
The epidemiology of human bocavirus (HBoV) infections has not been described in Greece, a south-eastern European country. To define the epidemiological profile and the clinical characteristics associated with HBoV infection in a population of children hospitalized with respiratory tract infection. During a one-year period throat swab samples were collected from 370 previously healthy children, aged 14 days to 13 years, admitted to two different paediatric wards because of respiratory tract infection. Samples were tested for HBoV by PCR amplifying a part of the NS1 gene. Human bocavirus was detected in 12 children (3.2%). Four of the 12 cases were co-infections, 3 of them with influenza A and 1 with coronavirus OC43. Cases were observed only during the cold months. The mean age of children was 1.8 years (range 2 months to 4 years). The most common symptoms were fever, cough and various degrees of respiratory distress. All children were clinically diagnosed as having lower respiratory tract infections, mainly pneumonia and acute laryngotracheobronchitis, and recovered uneventfully. HBoV infections occur in Greece mostly among very young children. They accounted for 3.2% of children hospitalized with acute respiratory disease. Cases were observed only in late autumn to early spring.


HLA-antigens and tuberculosis in the Egyptian population.
Peripheral blood lymphocytes from 42 unrelated Egyptian patients with tuberculosis and 156 healthy persons were HLA phenotyped for the A,B and DR loci using the NIH lymphocytotoxicity tests. Statistical analysis of the results showed that A2 and B5 had significantly increased frequencies among patients with tuberculosis compared with the controls. Patients with A2 had more severe disease than did patients without A2 and the number with B5 antigen was less than those without the antigen. This observation suggests that A2 and B5 may influence susceptibility to tuberculosis, but not the course of the disease. Furthermore, a linkage disequilibrium was found between A2 and B5 antigen among tuberculous patients, but their association bore no relation to the severity of the disease.


Predictive Contribution of the Superficial Neck Muscles to Short-Latency Rate of Force Development of the Head and Neck.
To evaluate the contribution of splenius capitis, sternocleidomastoid, and upper fibers of trapezius activation to the gains in rate of force development (RFD) of the head and neck during maximum voluntary ballistic contractions. RFD gain was facilitated by a single-session intervention for maximum voluntary ballistic contractions in the anterior direction, oriented at 45° to the midsagittal plane, which require active restraint of axial rotation. Muscle activation for the agonist (sternocleidomastoid) and 2 antagonists (splenius capitis and upper fibers of trapezius) was evaluated. The study sample included 12 physically active men (mean age, 22.6 y). RFD (N·m·s-1; 0-100 ms) and integrated muscle activity (50 ms before and 100 ms after force onset) were measured at 10 minutes, 20 minutes, and 2 days postintervention, relative to baseline. Muscle activation predictive of RFD gains was evaluated by linear regression analysis. RFD reproducibility was evaluated using the coefficient of variation of the typical error. The intervention yielded a 1.95- to 2.39-fold RFD gain (P ≤ .05), with greater RFD gain for participants with a lower peak moment of force (<10.9 N·m) than those with a higher peak moment (≥10.9 N·m) at baseline (P ≤ .002). For the low group, 65% to 74% of the RFD gain was predicted by ipsilateral sternocleidomastoid activation, with ipsilateral splenius capitis activation predicting 77% to 92% of RFD gain for the high group. Absolute peak and impulse of static force were greater for the high than for the low group (P ≤ .04). RFD reproducibility was high (coefficient of variation of the typical error ≤ 14.4%). The agonist- and antagonist-focused synergies might reflect different functional priorities, higher RFD gain compared with higher head-neck force.


Automated microextraction by packed sorbent of endocrine disruptors in wastewater using a high-throughput robotic platform followed by liquid chromatography-tandem mass spectrometry.
An automated microextraction by packed sorbent followed by liquid chromatography-tandem mass spectrometry (MEPS-LC-MS/MS) method was developed for the determination of four endocrine disruptors-parabens, benzophenones, and synthetic phenolic antioxidants-in wastewater samples. The method utilizes a lab-made repackable MEPS device and a multi-syringe robotic platform that provides flexibility to test small quantities (2 mg) of multiple extraction phases and enables high-throughput capabilities for efficient method development. The overall performance of the MEPS procedure, including the investigation of influencing variables and the optimization of operational parameters for the robotic platform, was comprehensively studied through univariate and multivariate experiments. Under optimized conditions, the target analytes were effectively extracted from a small sample volume of 1.5 mL, with competitive detectability and analytical confidence. The limits of detection ranged from 0.15 to 0.30 ng L-1, and the intra-day and inter-day relative standard deviations were between 3 and 21%. The method's applicability was successfully demonstrated by determining methylparaben, propylparaben, butylated hydroxyanisole, and oxybenzone in wastewater samples collected from the São Carlos (SP, Brazil) river. Overall, the developed method proved to be a fast, sensitive, reliable, and environmentally friendly analytical tool for water quality monitoring.


Alternative RNA splicing of the murine alpha A-crystallin gene: protein-coding information within an intron.
The eye lens contains a structural protein (alpha-crystallin), composed of two homologous primary gene products, alpha A2 and alpha B2. In certain rodents, there is another minor alpha-crystallin polypeptide, alpha Ains, which is identical to alpha A2 except for a 22 amino acid insert between residues 63 and 64 of the alpha A2 chain. Here we show that the mouse contains a single alpha A-crystallin gene, which has a 1376 bp intron separating codons 63 and 64 of the alpha A2-crystallin mRNA. A sequence encoding a 23 amino acid insert peptide was found 266 bp into the intron. The nucleotide borders of this sequence deviate from the AGGT consensus sequence. The DNA sequence encoding the insert peptide hybridizes to a cytoplasmic 14S RNA, demonstrating that it is transcribed in the lens. We propose that the murine alpha A2-crystallin gene generates both the alpha A2 and the alpha Ains mRNAs by alternative splicing.


An effective and simplified pH-stat control strategy for the industrial fermentation of vitamin B(12) by Pseudomonas denitrificans.
In order to improve the productivity of vitamin B(12) by Pseudomonas denitrificans carried out in a 120-m(3) fermenter, the effect of pH on vitamin B(12) biosynthesis was investigated. Results obtained from shake flask experiments showed that the feeding of carbon source (beet molasses or glucose) and methyl-group donor (betaine or choline chloride) significantly influenced the pH and the biosynthesis of vitamin B(12). In contrast to beet molasses or choline chloride, using glucose as a feed medium and betaine as a methyl-group donor, pH could be maintained at a stable range. As a result, higher vitamin B(12) production was achieved. Accordingly, an effective and simplified pH-stat control strategy was established for the fermentation of vitamin B(12) in a 120-m(3) industrial fermenter. When the new pH control strategy was applied, pH was stably kept in the range of 7.15-7.30 during fermentation. Thus, 214.3 microg/mL of vitamin B(12) was achieved.


Novel hydroxycarotenoids with improved antioxidative properties produced by gene combination in Escherichia coli.
We have used combinatorial biosynthesis to synthesize novel lipophilic carotenoids that are powerful cellular antioxidants. By co-expressing three different carotenoid desaturases in combination with a carotenoid hydratase, a cyclase, and a hydroxylase on compatible plasmids in Escherichia coli, we synthesized four novel carotenoids not previously detected in biological material or chemically synthesized. Their identification was based on their relative retention times on HPLC, spectroscopic properties, molecular weights, number of hydroxy groups, and 1H-NMR spectra. The carotenoids were designated as 1-HO-3', 4'-didehydrolycopene, 3, 1'-(HO)2-gamma-carotene, 1,1'-(HO)2-3, 4, 3', 4'-tetradehydrolycopene, and 1, 1'-(HO)2-3, 4-didehydrolycopene. These novel acyclic derivatives differ from structurally related compounds by extension of the conjugated polyene chain as well as additional hydroxy groups at position C-1'. We determined their antioxidative activity in a liposome-membrane model system, which showed that their ability to protect against photooxidation and radical-mediated peroxidation reactions was linked to the length of the conjugated double-bond system and the presence of a single hydroxy group. The protection of membrane degradation was superior to the related 1-HO and 1, 1'-(HO)2 lycopene derivatives, making them interesting pharmaceutical candidates.


Sequence of a chicken cDNA encoding a GRB2 protein.
The nucleotide (nt) sequence of a chicken cDNA encoding a protein homologous to the human GRB2 (growth factor receptor-binding protein) was determined. Remarkably high identities were found on the nt (88%) and deduced amino acid sequence (96%) levels.


Rater Wealth Predicts Perceptions of Outgroup Competence.
National income has a pervasive influence on the perception of ingroup stereotypes, with high status and wealthy targets perceived as more competent. In two studies we investigated the degree to which economic wealth of raters related to perceptions of outgroup competence. Raters' economic wealth predicted trait ratings when 1) raters in 48 other cultures rated Americans' competence and 2) Mexican Americans rated Anglo Americans' competence. Rater wealth also predicted ratings of interpersonal warmth on the culture level. In conclusion, raters' economic wealth, either nationally or individually, is significantly associated with perception of outgroup members, supporting the notion that ingroup conditions or stereotypes function as frames of reference in evaluating outgroup traits.


Improved Photoacoustic-Based Oxygen Saturation Estimation With SNR-Regularized Local Fluence Correction.
As photoacoustic (PA) imaging makes its way into the clinic, the accuracy of PA-based metrics becomes increasingly important. To address this need, a method combining finite-element-based local fluence correction (LFC) with signal-to-noise-ratio (SNR) regularization was developed and validated to accurately estimate oxygen saturation (SO2) in tissue. With data from a Vevo LAZR system, performance of our LFC approach was assessed in ex vivo blood targets (37.6%-99.6% SO2) and in vivo rat arteries. Estimation error of absolute SO2 and change in SO2 reduced from 10.1% and 6.4%, respectively, without LFC to 2.8% and 2.0%, respectively, with LFC, while the accuracy of the LFC method was correlated with the number of wavelengths acquired. This paper demonstrates the need for an SNR-regularized LFC to accurately quantify SO2 with PA imaging.


Funnel-like organization in sequence space determines the distributions of protein stability and folding rate preferred by evolution.
To understand the physical and evolutionary determinants of protein folding, we map out the complete organization of thermodynamic and kinetic properties for protein sequences that share the same fold. The exhaustive nature of our study necessitates using simplified models of protein folding. We obtain a stability map and a folding rate map in sequence space. Comparison of the two maps reveals a common organizational principle: optimality decreases more or less uniformly with distance from the optimal sequence in the sequence space. This gives a funnel-shaped optimality surface. Evolutionary dynamics of a sequence population on these two maps reveal how the simple organization of sequence space affects the distributions of stability and folding rate preferred by evolution.


Imaging of Patients with Complex Hemodialysis Arterio-Venous Fistulas using Time-Resolved Dynamic CT Angiography: Comparison with Duplex Ultrasound.
To evaluate the feasibility and potential on therapy management of time-resolved dynamic computed tomography angiography (dCTA) in patients with forearm arterio-venous fistula (AVF)/arterio-venous grafts (AVG). Thirty-five patients with complex failing forearm AVF/AVGs were examined with ultrasound and a dCTA protocol. Diagnosis and therapy management was evaluated versus duplex ultrasound (DUS) in three different readouts: 1. all dCTA datasets; 2. one arterial phase of the dCTA dataset; 3. one arterial and one venous dataset out of the dCTA dataset. All reads were performed >30 days apart from each other. Using all data of the dCTA examination, 20 patients were classified as having a stenosis >50%, 12 high-shunt flow, 11 partial thrombosis, 5 venous aneurysms and 5 complete thrombosis of their AVF/AVG grafts. This lead to 13 additional pathologic findings not visible on DUS and reclassification as normal in one patient with suspected AVF stenosis and complete thrombus on DUS. These additional findings lead to a direct change of therapeutic management in 8 patients. Compared to readout 1 (53 pathologies), readout number 2 and 3 revealed only 33 and 41 pathologies, respectively. dCTA provides additional information, improving diagnostic confidence and leading to changes in therapy management when compared to DUS alone.


Gabaergic stimulation inhibits central actions of angiotensin II: pressor responses, drinking and release of vasopressin.
In conscious rats, intracerebroventricular (i.c.v.) treatment with the GABA agonist muscimol (1-100 ng) suppressed the pressor responses to ANG II (100 ng i.c.v.) in a dose-dependent and reversible fashion. Treatment i.c.v. with GABA (1-500 micrograms) produced a similar but shorter inhibition. Inhibition of endogenous GABA degradation with amino-oxyacetic acid (AOAA, 30 mg/kg i.p.) markedly reduced the pressor responses to ANG II (10-1000 ng i.c.v.) with a recovery period of 24 h. This inhibition was reversed by the GABA antagonist bicuculline (1 microgram i.c.v.). Muscimol (100 ng i.c.v.) did not significantly attenuate the pressor responses to i.c.v. histamine (10 micrograms). Pretreatment with muscimol (100 ng i.c.v.) drastically reduced the drinking responses to i.c.v. ANG II (500 ng) and increased the latency to drink. Muscimol also suppressed drinking induced by carbachol (50 ng i.c.v.). Muscimol (10-1000 ng i.c.v.) inhibited the ANG II (100 ng i.c.v.)-induced release of AVP from the pituitary gland with complete suppression of the response at the highest dose. Our results demonstrate that the GABAergic system exerts an inhibitory control on pathways mediating the various central actions of ANG II, which appears to be most specific for the ANG II-induced pressor responses.


Pineal gland hydroxyindole-O-methyl transferase (HIOMT) from rainbow trout (Salmo gairdneri) subject to temperature-dependent substrate inhibition by N-acetylserotonin.
The effect of different incubation temperatures on the activity of trout hydroxyindole-O-methyltransferase (HIOMT) was determined in order to study the potential effects of changes in environmental temperature on production of melatonin. In the study, N-acetylserotonin (NAS) was found to inhibit trout pineal HIOMT, inhibitory concentration decreasing with a decrease in incubation temperature. An increase in incubation temperature was accompanied by an increase in maximal velocity (Vm) and a decrease in affinity of the substrate NAS for the enzyme, while the affinity of the inhibitor NAS remained more or less constant with increase in temperature up to 30 degrees C. It was interesting to note that at subinhibitory concentrations of NAS, possibly concentrations within the normal physiological range, there was a balance between increased velocity and decreased affinity for substrate, with increase in temperature resulting in a constant rate of melatonin production. On the basis of these results it is suggested that substrate inhibition might be involved in modulation of melatonin production and that trout pineal HIOMT can maintain a constant rate of melatonin production over a wide range of temperatures by a combination of compensatory changes in enzyme kinetic parameters.


Cyclic-AMP content in Escherichia coli B/b as affected by N1-(delta 2-isopentyl)adenine.
N6-(delta 2-isopentenyl)adenine, like other cytokinins, does not detectably modify Escherichia coli growth, but strongly affects cellular levels of cAMP. A substantial delay of the highest level of intracellular cAMP, a reduction to about one half of such maximum level, and a slight increase of cAMP secreted into the medium are reported.


Discourse analysis and the doctor-patient relationship.
This article describes a system of discourse analysis, called a "taxonomy of verbal response modes," which can be applied to medical interviews. The taxonomy identifies eight basic categories: disclosure, question, edification, acknowledgement, advisement, interpretation, confirmation, and reflection, which are defined by three principles of classification. The categories are mutually exclusive and exhaustive. Each mode conveys a particular interpersonal intent and also has a characteristic grammatical form. With eight forms and eight intents, the taxonomy includes sixty-four possible verbal response modes, eight "pure modes," in which form and intent coincide, and fifty-six "mixed modes," in which form and intent differ. The taxonomy has yielded fine-grained descriptions of patient-physician interaction and has identified particular types of utterances and verbal exchanges that are associated with patients' satisfaction with their medical interviews. The system provides a detailed descriptive vocabulary that may be useful for teaching interviewing skills.


Serum metabolomics study of patients with allergic rhinitis.
Allergic rhinitis (AR) negatively affects the healthy lives of many individuals. Most previous studies on AR focused on the expression of cytokines, with only a few analyzing cytokine expression from a metabolomics viewpoint. Therefore, it is worthwhile to study AR at the metabolic level. Consequently, we aimed to identify differential serum biomarkers by metabolomics. In this study, the orthogonal partial least squares discriminant analysis (OPLS-DA) model was applied to characterize the differences in serum samples collected from patients with AR and healthy volunteers. Ten metabolites (except hexadecanoic acid) were found to be altered significantly (p < .05) in the former group, according to results of principal component analysis and OPLS-DA, indicating that these metabolites could be potential biomarkers. MetaboAnalyst 4.0 and pathway enrichment analysis showed that these changes in metabolites mainly involved three pathways, namely, porphyrin and chlorophyll metabolism, arachidonic acid metabolism, and purine metabolism. Our findings may contribute to a better understanding of the potential pathogenesis mechanisms and provide a metabolic evidence for in-depth studies of AR.


Coupling of a viral K+-channel with a glutamate-binding-domain highlights the modular design of ionotropic glutamate-receptors.
Ionotropic glutamate receptors (iGluRs) mediate excitatory neuronal signaling in the mammalian CNS. These receptors are critically involved in diverse physiological processes; including learning and memory formation, as well as neuronal damage associated with neurological diseases. Based on partial sequence and structural similarities, these complex cation-permeable iGluRs are thought to descend from simple bacterial proteins emerging from a fusion of a substrate binding protein (SBP) and an inverted potassium (K+)-channel. Here, we fuse the pore module of the viral K+-channel KcvATCV-1 to the isolated glutamate-binding domain of the mammalian iGluR subunit GluA1 which is structural homolog to SBPs. The resulting chimera (GluATCV*) is functional and displays the ligand recognition characteristics of GluA1 and the K+-selectivity of KcvATCV-1. These results are consistent with a conserved activation mechanism between a glutamate-binding domain and the pore-module of a K+-channel and support the expected phylogenetic link between the two protein families.


R483 and F plasmid genes promoting RNA degradation: comparative restriction mapping.
The gene promoting nucleic-acid degradation (pnd) on IncIa plasmid R483 was cloned into pBR322. It is located on a 0.85 kilobase (kb) EcoRI-SalI fragment and is close to Tn7. The pnd gene has similar properties to the srnB gene on the F plasmid. A cleavage map of the 0.85 kb pnd fragment was constructed and compared with that of the 1.18 kb EcoRI-BamHI fragment containing the srnB gene. These two regions showed marked heterogeneity as evidenced by their distinctly different restriction maps. This result suggests separate paths of evolution of the two genes for stable RNA degradation.


Aldehyde dehydrogenase and aldehyde reductase in isolated bovine brain microvessels.
The activities of aldehyde dehydrogenase and aldehyde reductase were measured in microvessel fractions isolated from bovine brain. Aldehyde dehydrogenase specific activity in microvessels was enriched 1.5-fold over that measured in grey matter. The specific activity of aldehyde reductase was significantly lower in parenchymal vessles and microvessels than in grey matter. The presence of aldehyde dehydrogenase and aldehyde reductase in cerebral microvasculature is consistent with previous reports of enrichment of other enzymes involved in synthesis and degradation of monoamines. The enrichment of aldehyde dehydrogenase in cerebral microvasculature provides additional evidence for an enzymatic blood-brain barrier for biogenic aldehydes or ethanol-derived acetaldehyde.


Intracranial hypertension in Behçet's disease: demonstration of sinus occlusion with use of digital subtraction angiography.
We describe two patients with Behçet's disease who had symptomatic intracranial hypertension due to cerebral venous sinus thrombosis. The sinus thrombosis was demonstrated by digital subtraction angiography. In those patients with Behçet's disease who have unexplained headaches, papilledema, and elevated cerebrospinal fluid pressure, venous digital subtraction angiography is an expedient, accurate, and safe procedure for demonstrating intracranial venous thrombosis.


Cortical superficial siderosis is associated with reactive astrogliosis in cerebral amyloid angiopathy.
Cortical superficial siderosis (cSS) has recently emerged as one of the most important predictors of symptomatic intracerebral hemorrhage and is a risk factor for post-stroke dementia in cerebral amyloid angiopathy (CAA). However, it remains unknown whether cSS is just a marker of severe CAA pathology or may itself contribute to intracerebral hemorrhage risk and cognitive decline. cSS is a chronic manifestation of convexal subarachnoid hemorrhage and is neuropathologically characterized by iron deposits in the superficial cortical layers. We hypothesized that these iron deposits lead to local neuroinflammation, a potentially contributory pathway towards secondary tissue injury. Accordingly, we assessed the distribution of inflammatory markers in relation to cortical iron deposits in post-mortem tissue from CAA cases. Serial sections from the frontal, parietal, temporal, and occipital lobes of nineteen autopsy cases with CAA were stained with Perls' Prussian blue (iron) and underwent immunohistochemistry against glial fibrillary acidic protein (GFAP, reactive astrocytes) and cluster of differentiation 68 (CD68, activated microglia/macrophages). Digitized sections were uploaded to the cloud-based Aiforia® platform, where deep-learning algorithms were utilized to detect tissue, iron deposits, and GFAP-positive and CD68-positive cells. We observed a strong local relationship between cortical iron deposits and reactive astrocytes. Like cSS-related iron, reactive astrocytes were mainly found in the most superficial layers of the cortex. Although we observed iron within both astrocytes and activated microglia/macrophages on co-stains, there was no clear local relationship between the density of microglia/macrophages and the density of iron deposits. Iron deposition resulting from cSS is associated with local reactive astrogliosis.


Lower esophageal sphincter dysfunction in esophageal atresia: nocturnal regurgitation and aspiration pneumonia.
The association between lower esophageal sphincter (LES) incompetence, gastroesophageal reflux, and recurrent pneumonia in patients who have undergone successful repair of esophageal atresia (EA) and tracheoesophageal fistula is demonstrated in this study. The efficacy of esophageal manometric examination in the evaluation of the LES in young children after EA repair is documented. This study also provides evidence that infants and children with LES incompetence associated with EA may have LES responsiveness to bethanechol. Once established by manometry, this responsiveness may be used to manage the patient until surgical repair would be advantageous.


Inflammatory status predicts contact lens discomfort under adverse environmental conditions.
To characterize and predict the clinical and tear molecular response of contact lens (CL) wearers exposed to a controlled adverse desiccating environment (CADE). Objective and subjective variables and tear cytokine levels were evaluated of monthly silicone hydrogel CL wearers pre- and post-90 min of CADE exposure. Unsupervised hierarchical agglomerative clustering based on relative change from baseline values was used to identify response profiles (clusters). A multiple logistic regression model was used to identify cluster membership predictors. Forty-seven CL wearers were divided into 3 clusters having similar age (mean: 27.7 ± 7.7 years) and sex distribution. All of them showed a significant (p ≤ 0.05) increase in limbal hyperemia and staining after CADE exposure. Additionally, Cluster-1 (n = 22, 46.8%) membership was characterized by a significant (p ≤ 0.05) higher worsening of corneal and limbal staining, increased CL wear symptoms, and reduced epidermal-growth-factor and increased interleukin (IL)-4 and IL-6 tear levels. Cluster-2 (n = 22, 46.8%) showed no changes (p > 0.05) in symptoms after CADE; however, their IL-12p70, monocyte-chemoattractant-protein-1 and regulated-on-activation, normal-T-cell-expressed-and-secreted (RANTES) post-exposure tear levels significantly (p ≤ 0.05) increased. Finally, Cluster-3 (n = 3, 6.4%) mainly showed significant higher blink rate (78.1 ± 21.7) during CADE. Corneal staining and tear IL-12p70 levels were identified as Cluster-1 membership predictors. Most of silicone hydrogel CL wearers exposed to CADE showed a worsening of the ocular surface integrity and an upregulated tear inflammatory status. However, only half of them reported worsening of CL wear symptoms. These CL wearers were detected based on corneal integrity and tear inflammatory status. These findings can help reduce CL wear discontinuation and drop out.


Dithranol-mediated, dose-dependent priming and activation of luminol-enhanced chemiluminescence responses of human neutrophils in vitro.
At concentrations of 10 micrograms/ml and greater, dithranol (anthralin) caused an intense, dose-related activation of luminol-enhanced chemiluminescence (LECL) of human neutrophils and also increased oxygen consumption by these cells. Activation of LECL, which was maximal with 40 micrograms/ml dithranol, occurred promptly, peaked at 3-5 min, then declined. Dithranol-mediated stimulation of LECL was inhibited by catalase and by the protein kinase C inhibitor, H-7. Neutrophils from individuals with chronic granulomatous disease were unresponsive to the pro-oxidative effects of dithranol. At concentrations of 2.5 micrograms/ml and less, dithranol did not directly activate the LECL responses of neutrophils. However pre-treatment of neutrophils with dithranol at concentrations of 0.5-2.5 micrograms/ml increased the LECL-responses of cells subsequently stimulated with calcium ionophore and opsonized zymosan. These observations demonstrate two distinct dose-related, pro-oxidative interactions of dithranol with human neutrophils: low-dose priming and high-dose activation of oxidant generation. Since phagocyte-derived reactive oxidants are immunosuppressive, these pro-oxidative interactions of dithranol with human neutrophils may contribute to the pharmacotherapeutic mechanisms of this agent.


Qualitative Study to Design an Online Community for Patients with Psoriasis.
The aim of this research was to understand the needs of patients with psoriasis and the concerns of specialists in this pathology. Interviews with adults with psoriasis and dermatologists were conducted. We found 4 main dimensions: Frequently asked questions; Social stigma; Education to patients and Patient empowerment. The findings represent a first approach to understand the needs and perceptions of patients and dermatologists, for the design of a virtual community.


Effects of the CRF1 antagonist SSR125543A on aggressive behaviors in hamsters.
Corticotropin-releasing factor (CRF) and its receptor subtypes have been implicated in endocrine and behavioral responsivity to stress and emotion, including fear, anxiety, and aggression. SSR125543A is a new nonpeptide selective antagonist at the CRF1 receptor that has been shown to produce an anxiolytic-like effect in a number of animal models of anxiety. The present study investigated effects of an oral dose of 10, or 30 mg/kg of SSR125543A on aggressive behaviors of resident male Syrian hamsters toward male intruders. The high dose (30 mg/kg) of the CRF1 receptor antagonist produced a higher latency to bite and lower lateral attack frequencies and chase durations, indicating a reduction in aggression toward intruders in resident hamsters. The same dose of SSR125543A also enhanced frequency and duration of olfactory investigation, indicating that neither avoidance of the opponent nor deficiency in social activity is responsible for the reduction in aggression seen in these animals.


Mitochondrial genome of Opsaridium microlepis (cypriniformes: cyprinidae).
The complete mitochondrial genome of Opsaridium microlepis was determined by using llumina sequencing method. The genome is 168 25 bp in length, comprising 13 protein-coding genes, 20 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes and one control region (D-loop region). Most genes are encoded on the heavy strand (H-strand), except for nad6 and eight tRNA genes. Base composition of the H-strand are A (27.97%), C (26.34%), G (18.37%) and T (27.32%) with AT bias of 55.29%.


Behavior-associated changes in blood pressure during heroin self-administration.
Changes in arterial blood pressure (ABP) were studied in freely moving rats during the development and performance of operant IV heroin self-administration (SA) behavior (100 micrograms/kg/injection). Heroin SA was accompanied by bidirectional, phasic ABP fluctuations in the absence of significant alterations in long-term, basal ABP levels. In trained rats, ABP gradually increased starting 5 min before each lever-press for the drug, reached a peak at the moment of lever-press and abruptly decreased after heroin infusion. This biphasic pattern corresponded to a preresponding behavioral activation followed by a postheroin sedation. These ABP fluctuations were absent during the initial heroin self-injections in drug-naive rats and during the first self-injections of a session in trained rats. A slight hypo- and hypertension were seen in these cases, respectively. The postdrug ABP decrease became more pronounced after rats received a double dose of heroin. With training, nonreinforced lever-presses and sound stimulation previously associated with heroin self-injections also significantly decreased ABP. Thus, a gradual ABP increase appears to be an essential correlate of drug-seeking and -taking behavior, while a subsequent ABP decrease may be related to the alleviation of these behaviors by heroin and possibly correlate to its rewarding (euphorigenic) action.


Comparison of chemometric methods for the screening of comprehensive two-dimensional liquid chromatographic analysis of wine.
Two chemometric methods are compared for the rapid screening of comprehensive two-dimensional liquid chromatographic (LC×LC) analysis of wine. The similarity index and Fisher ratio methods were both found to be able to distinguish geographical variability and to determine potentially significant peaks for further quantitative and qualitative study. An experimental data set consisting of five different wine samples and multiple simulated data sets were analyzed in the investigation of the screening methods. Several statistical analyses are employed in the understanding and verification of the results from the similarity index and Fisher ratio methods. The sum rank difference (SRD) method was used to compare the rankings of the two different methods as applied to the different data sets and to determine the amount of variability associated with the ranking of the peak differences. The major advantage the similarity index method offers is that it is an unsupervised method; no a priori knowledge of the samples (i.e., class identification) is required, while the Fisher ratio method is supervised. Both methods are rapid and require little user intervention other than the determination of a threshold for inclusion/exclusion of compounds from further analysis.


The association of upward economic mobility with textbook outcomes among patients undergoing general and cardiovascular surgery.
While disadvantaged neighborhoods may be associated with worse outcomes and earlier death, the relationship between economic opportunity and surgical outcomes remains unexplored. Medicare beneficiaries who underwent AAA, CABG, colectomy or cholecystectomy were identified and stratified into quintiles based on upward economic mobility. Risk-adjusted probability of adverse postoperative outcomes were examined relative to economic mobility. Among 1,081,745 Medicare beneficiaries (age: 75.5 years, female: 43.0%, White: 91.3%), risk-adjusted 30-day postoperative mortality decreased in a stepwise fashion from 6.0%(5.9-6.1) in the lowest quintile of upward economic mobility to 5.3%(5.2-5.4) in highest upward economic mobility (lowest vs. highest economic mobilityobility OR:1.14 (95%CI:1.11-1.17)). Similar associations were noted for postoperative complications (OR:1.04, 95%CI:1.02-1.06), extended length-of-stay (OR:1.07, 95%CI:1.06-1.09), and 30-day readmission (OR:1.04, 95%CI:1.02-1.05). Black beneficiaries had a higher risk of post-operative mortality across upward economic mobility quintiles except within the highest upward mobility group (referent, White patients, OR:0.93, 95%CI:0.79-1.09, p=0.355). Economic upward mobility was associated with post-operative outcomes. Race-based differences were mitigated at the highest levels of upward economic mobility, highlighting the importance of socioeconomics as a health equity lever.


Acute Myocarditis Related to COVID-19: Comparison to SARS and MERS.
Myocardial involvement has been described during previous SARS and MERS outbreaks. Infection by SARS-CoV-2 (COVID-19) can range from asymptomatic to life-threatening multi-system disease. Heart involvement most commonly occurs during severe COVID-19 infection. Myocardial injury, based on elevated levels of myocardial enzymes, has been noted in up to 30% of patients with COVID-19 infection and could be a marker for worse prognosis. A few cases of possible myocarditis due to SARS-CoV-2 have been described, providing variable degree of evidence of direct myocardial involvement. We reviewed in detail those cases in comparison to relevant literature on SARS and MERS and attempted to draw initial conclusions in regard to clinical presentation, treatment and prognosis.


Enzymatic synthesis of isoamyl butyrate catalyzed by immobilized lipase on poly-methacrylate particles: optimization, reusability and mass transfer studies.
Isoamyl butyrate (banana flavor) was synthesized by esterification reaction of isoamyl alcohol and butyric acid in heptane medium. Immobilized Thermomyces lanuginosus lipase (TLL) prepared via physical adsorption on mesoporous poly-methacrylate particles (PMA) was used as biocatalyst. The factors that affect the esterification reaction were optimized by response surface methodology (RSM). Under optimal experimental conditions, maximum ester conversion percentage of 96.1 and 73.6% was reached after 50 and 90 min, respectively, for esterification reaction performed at equimolar ratio alcohol:acid at 500 and 2000 mM of each substrate. Under these experimental conditions, the esterification reaction was not controlled by external and intra-particle mass transfer effects. The product (isoamyl butyrate) was confirmed by proton nuclear magnetic resonance ((1)H NMR) spectroscopy. Reusability tests showed that the biocatalyst retained around 96 and 31% of its initial activity after eight successive esterification cycles performed at 500 and 2000 mM, respectively. The application of the biocatalyst prepared showed to be a promising strategy to catalyze flavor ester synthesis in a non-aqueous medium.


Selective serotonin reuptake inhibitors for unipolar depression: a systematic review of classic long-term randomized controlled trials.
Selective serotonin reuptake inhibitors are increasingly used in the long-term treatment of depression. Much of the supporting evidence about the effects of these drugs comes from discontinuation trials, a variant of randomized controlled trials whose design is problematic to interpret. We conducted a systematic review to examine the efficacy and acceptability of long-term therapy with selective serotonin reuptake inhibitors relative to placebo in the treatment of unipolar depression. We identified placebo-controlled randomized trials with a treatment duration of at least 6 months by searching MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials to update a recently published systematic review. Efficacy was defined in terms of response to treatment (50% improvement in depression score relative to baseline) and remission (score of 7 or below on the Hamilton rating scale for depression). Key secondary outcomes included quality of life, return to work, need for additional treatment and self-harm. Overall acceptability was defined in terms of dropouts for any reason over a course of treatment. Of the 2693 records identified initially, we included 6 randomized controlled trials that met our eligibility criteria. These studies had a moderate risk of bias, had assigned a total of 1299 participants with depression to either treatment or placebo and had followed both groups for 6-8 months. We observed statistically significant improvements in response to treatment (odds ratio [OR] 1.66, 95% confidence interval [CI] 1.12-2.48), but not in remission (OR 1.46, 95% CI 0.92-2.32) or acceptability (OR 0.87, 95% CI 0.67-1.14). The effects appeared greater among patients without comorbidities. There is a lack of classic randomized controlled trials of serotonin reuptake inhibitors lasting more than 1 year for the treatment of depression. The results of our systematic review support current recommendations for 6-8 months of antidepressant treatment following initial recovery but provide no guidance for longer treatment.


Toward zero effective phacoemulsification time using femtosecond laser pretreatment.
To compare effective phacoemulsification time after femtosecond laser pretreatment with conventional phacoemulsification and the associated effect on visual outcomes and endothelial cell loss. Prospective, consecutive, single-surgeon case-control study. Controls underwent phacoemulsification cataract extraction plus insertion of an intraocular lens (IOL). Cases underwent pretreatment with the femtosecond laser followed by phacoemulsification cataract extraction and IOL insertion. Two hundred one eyes underwent cataract surgery between April 2012 and July 2012. Data collected included patient demographics, preoperative characteristics, femtosecond lens fragmentation method, effective phacoemulsification time (EPT), intraoperative complications, and postoperative outcomes. Effective phacoemulsification time, intraoperative complications, corneal endothelial cell loss, as well as postoperative best-corrected visual acuity, intraocular pressure, and refractive outcomes. Patient demographics were similar between groups. There was no difference between baseline cataract grades (2.59 ± 0.71 vs. 2.52 ± 0.72, not significant). One hundred percent of cases pretreated with the femtosecond laser had complete capsulotomy. Mean EPT was reduced by 83.6% in the femtosecond pretreatment group (P<0.0001) when compared with controls, with 30% having 0 EPT (P<0.0001). Effective phacoemulsification time was reduced 28.6% within the femtosecond group using improved lens fragmentation algorithms, and a further 72.8% reduction was achieved with a 20-gauge phacoemulsification tip. Overall, there was a 96.2% reduction in EPT between controls and the optimized femtosecond pretreatment group. This was associated with a 36.1% reduction in endothelial cell loss in the femtosecond group. Visual and refractive outcomes were similar to those of conventional cataract surgery. Femtosecond laser pretreatment results in a significant reduction in effective phacoemulsification time, including the possibility of 0 EPT. Further reductions may be achieved using optimization of lens fragmentation patterns and surgical technique. The author(s) have no proprietary or commercial interest in any materials discussed in this article.


Transforming growth factor beta increases the expression of HIV-1 gene in transfected human mesangial cells.
Human immunodeficiency virus type 1 (HIV) infection is often complicated by focal glomerulosclerosis (FGS) and other renal lesions collectively termed HIV associated nephropathy (HIVAN). FGS is characterized by glomerular mesangial expansion and increased synthesis of matrix components. The molecular pathogenic mechanisms associated with the development of HIV associated nephropathy are unknown. Experimental animal models suggest a role for cytokines and growth factors, particularly transforming growth factor beta (TGF-beta), in the pathogenesis of glomerulosclerosis. Patients with AIDS have elevated plasma and tissue levels of TGF-beta. We carried out experiments to determine whether primary human mesangial cells (HMC) in culture can be transfected with HIV-1 genes. HMC were transfected with a chloramphenicol acetyl transferase (CAT) reporter construct containing HIV-1 acetyl transferase (CAT) reporter construct containing HIV-1 LTR sequences. Our results show successful transfection of HMC with HIV-1 LTR gene. HMC transfected with LTR gene are responsive to the HIV-1 regulatory gene product Tat. To study whether TGF-beta can modulate the expression of HIV-1 LTR gene in HMC, HMC transfected with an HIV-1 LTR CAT plasmid were treated with TGF-beta and other growth factors two hours before harvest. TGF-beta specifically increased the expression of the HIV-1 gene in HMC in a dose dependent manner. We further studied whether up-regulation of HIV-1 LTR expression in HMC was mediated by the effect of TGF-beta on the interaction of transcription factors to their binding sites.(ABSTRACT TRUNCATED AT 250 WORDS)


Development and validation of RP-HPLC method for sildenafil citrate in rat plasma-application to pharmacokinetic studies.
Sildenafil citrate (SIL) is used in the treatment of erectile dysfunction and other chronic disorders. For the pharmacokinetic investigation of SIL we developed a simple and sensitive method for the estimation of SIL in rat plasma by reverse phase high-performance liquid chromatography (RP-HPLC). The drug samples were extracted by liquid-liquid extraction with 300 μl of acetonitrile and 5 ml of diethyl ether. Chromatographic separation was achieved on C18 column using methanol:water (85:15 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 230 nm. The retention time of SIL was found to be 4.0 min having a separation time less than 5 min. The developed method was validated for accuracy, precision, linearity and recovery. Linearity studies were found to be acceptable over the range of 0.1-6 μg/ml. The method was successfully applied for the analysis of rat plasma sample for the application in pharmacokinetic study, drug interaction, bioavailability and bioequivalence.


Secondary (intermediate) alveolar bone grafting.
Secondary bone grafting of the maxilla and the residual alveolar clefts at the stage of transitional dentition, and in conjunction with orthodontic treatment, has become a well-accepted treatment modality. This article discusses the goals of the procedure, the timing of the bone graft, and the design of the flap for soft-tissue coverage of the bone graft.


Identification of TIAR as a protein binding to the translational regulatory AU-rich element of tumor necrosis factor alpha mRNA.
In monocyte/macrophages, the translation of tumor necrosis factor alpha (TNF-alpha) mRNA is tightly regulated. In unstimulated cells, translation of TNF-alpha mRNA is blocked. Upon stimulation with lipopolysaccharides, this repression is overcome, and the mRNA becomes efficiently translated. The key element in this regulation is the AU-rich element (ARE). We have previously reported the binding of two cytosolic protein complexes to the TNF-alpha mRNA ARE. One of these complexes (complex 1) forms with extracts of both unstimulated and lipopolysaccharide-stimulated macrophages and requires a large fragment of the ARE containing clustered AUUUA pentamers. The other complex (complex 2) is only detected after cell activation, binds to a minimal UUAUUUAUU nonamer, and is composed of a 55-kDa protein. Here, we report the identification of the RNA-binding protein TIAR as a protein involved in complex 1. The RNA sequence bound by TIAR and the cytoplasmic localization of this protein in macrophages argue for an involvement of TIAR in TNF mRNA posttranscriptional regulation.


Refined Disease Risk Index for Hematological Malignancies, Including Rare Disorders, After Allogeneic Stem Cell Transplantation.
The refined disease risk index (R-DRI) is a well-designed prognostic parameter that is based on only the disease type and status and is used for stratifying patients undergoing allogeneic hematopoietic stem cell transplantation (allo HSCT) into 4 risk groups. However, the application of the R-DRI for rare diseases has remained unclear. We evaluated 135 patients who underwent allo HSCT for hematological malignancies including rare diseases, such as acute leukemia of ambiguous lineage, acute T-cell leukemia/lymphoma, extranodal natural killer T-cell lymphoma, and lymphoblastic lymphoma, at our institute. According to the R-DRI, overall survival (OS) and progression-free survival at 2 years for patients with the low, intermediate, high, and very high groups were 66.7% and 66.7%, 60.8% and 56.0%, 27.1% and 23.7%, and 5.9% and 5.1%, respectively (P < .0001 and P < .0001, respectively). OS showed no significant difference between B-cell non-Hodgkin lymphoma (B-NHL) and T-cell non-Hodgkin lymphoma (T-NHL) (P = .71). Moreover, OS at 1 year was 80%, 14.3%, 60%, and 0% for the intermediate risk group, the very high-risk group of B-NHL, the intermediate risk group, and the high-risk group of T-NHL, respectively (P = .035). We showed the applicability of the R-DRI for hematological malignancies, including rare disorders. However, we suggest that T-NHL patients may be better to be assigned between the nodal group and the extranodal group in the R-DRI.


Myelin basic protein deposition in the optic and sciatic nerves of dysmyelinating mutants quaking, jimpy, Trembler, mld, and shiverer during development.
An ontogenetic survey of the basic protein of myelin, common to both central and peripheral nervous systems, was carried out on normal C57Bl and five dysmyelinating mutant mice. Myelin basic protein (MBP) was quantified by radioimmunoassay in the optic and sciatic nerves of mice from birth to adult stages, giving special attention to the premyelinating and early myelination periods. In the optic nerves of normal mice, MBP was already detectable at birth but the active period of myelin deposition was shown to occur after day 10 postnatal. The timing and rate of accumulation of MBP were normal in Trembler. In contrast, they were abnormal in the other mutants. In the quaking mouse, the active period of MBP deposition was delayed, and its final concentration represented no more than 12% of normal in the adult. No active period of MBP deposition was observed in the other mutants. In the jimpy mouse, a slow accumulation of MBP resulted in a final concentration reaching 2% of the normal value at 25 days. In mild and shiverer mice, the MBP was hardly detectable. In the sciatic nerves of normal mice, the active period of MBP deposition occurred between days 3 and 12 postnatal. No substantial changes occurred in the period of 2 months--2 years.(ABSTRACT TRUNCATED AT 250 WORDS)


Repair of DNA single-strand breaks in X-irradiated yeast. I. Use of the DNA-unwinding method to measure DNA strand breaks.
The DNA-unwinding method developed by Ahnström and his coworkers to measure DNA strand breaks in mammalian cells was used to measure single-strand breaks (SSB) in the DNA of intact yeast cells. DNA unwinding, which took place inside the rigid cell wall of yeast, was investigated as a function of time, radiation dose, and of pH and salt concentration of the alkaline solution. After DNA unwinding had taken place, the cell wall was destroyed by partial enzymatic digestion and sonication in the presence of detergents. Fragments of single- and double-stranded DNA were separated using hydroxylapatite chromatography. In this way the most suitable conditions for DNA unwinding within the cell wall were established. The results show that SSB and double-strand breaks (DSB) give rise to different kinetics of DNA unwinding.


Use of dementia care mapping in the care for older people with intellectual disabilities: A mixed-method study.
The ageing of people with intellectual disabilities, with associated morbidity like dementia, calls for new types of care. Person-centred methods may support care staff in providing this, an example being Dementia Care Mapping (DCM). DCM has been shown to be feasible in ID-care. We examined the experiences of ID-professionals in using DCM. We performed a mixed-methods study, using quantitative data from care staff (N = 136) and qualitative data (focus-groups, individual interviews) from care staff, group home managers and DCM-in-intellectual disabilities mappers (N = 53). DCM provided new insights into the behaviours of clients, enabled professional reflection and gave new knowledge and skills regarding dementia and person-centred care. Appreciation of DCM further increased after the second cycle of application. DCM is perceived as valuable in ID-care. Further assessment is needed of its effectiveness in ID-care with respect to quality of care, staff-client interactions and job performance.


Correlates of prenatal HIV testing in women with undocumented status at delivery.
To determine factors associated with prenatal HIV testing in women who accepted rapid testing at delivery. The mother-infant rapid intervention at delivery (MIRIAD) protocol offered counseling and voluntary HIV testing in six US cities including New York City (NYC). These hospitals are required to document the HIV status of pregnant women or their infants. From January 2002 to January 2005, 653 HIV-negative women were interviewed post-partum. 63% of women reported prior HIV testing during the index pregnancy, although their results were not available at delivery. Multivariate logistic modeling identified receipt of prenatal care and delivery in NYC as being associated with having been offered prenatal HIV testing. In a model restricted to women receiving medical care, emergency department (ED) use and delivery outside of NYC were associated with not having been offered testing. In a model restricted to women who were offered prenatal testing, acceptance was associated with delivery outside of NYC. Improved documentation of prenatal test results, expanded prenatal testing in non-traditional settings like EDs, and routine voluntary "opt-out" testing during pregnancy may further reduce perinatal HIV transmission.


[Composite lymphoma: Case report of a coexisting follicular and mantle cell lymphoma in situ in a cervical node].
Composite lymphoma represents 1-4% of lymphomas. Only 8 case reports concerned coexisting follicular lymphoma and mantle cell lymphoma. Here, we report the case of an 81 years old man who has been diagnosed with a composite follicular and in situ mantle cell lymphoma. The use of a large panel of immunohistochemical stains associated with the flow cytometry results have allowed us to make this particular diagnosis. We highlight here a common clonal origin of the composite lymphoma's two entities, as described in previous publications.


Activation of glycogen phosphorylase by serotonin and 3,4-methylenedioxymethamphetamine in astroglial-rich primary cultures: involvement of the 5-HT2A receptor.
Neurotransmitters, neuropeptides, and ions regulate glycogen levels in the brain by modulating the activity of glycogen synthase (GSase) and glycogen phosphorylase (GPase). GPase is co-localized with glial fibrillary acidic protein (GFAP), an astroglia-specific marker, suggesting that glycogen is localized in astroglial cells. Additionally, functional serotonin (5-HT) receptors are found in both neurons and glia, and 5-HT is known to stimulate glycogenolysis. It is reported that 3,4-methylenedioxymethamphetamine (MDMA), a drug of abuse, stimulates the release and inhibits the reuptake of 5-HT, and selectively inhibits the activity of MAO-A. These biochemical consequences of MDMA lead to increased extra-cellular 5-HT levels. This study investigates the effects of MDMA(+) and serotonin (5-HT) on glycogen metabolism in the rat brain. A histochemical method was designed to visualize active glycogen phosphorylase (GPase) in an astroglial-rich primary culture. Serotonin activated GPase in a concentration-dependent manner (100 nM-100 microM). Maximal activation by 5-HT was achieved by 50 microM and resulted in a 167% increase in the number of reactive sites (P < 0.001). MDMA(+) (500 nM-50 microM) directly stimulated GPase activity with maximal activation induced by 5 microM, which caused a 70% increase in the number of reactive sites (P < 0.001). The 5-HT2 receptor agonist, 1-(2,5-dimethoxy-4-bromophenyl)-2-aminopropane (DOB), also displayed a concentration-dependent increase in the number of GPase reactive sites. Maximal stimulation by DOB occurred at 100 nM which increased the number of reactive sites by 166% (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


The cytoskeletal framework of sea urchin eggs and embryos: developmental changes in the association of messenger RNA.
Extraction of sea urchin eggs and embryos with Triton X-100 generated a cytoskeletal framework (CSK) composed of a cortical filamentous network and an internal system of filaments associated with ribosomes. The CSK contained only 10-20% of the cellular protein, RNA, and lipid. A specific subset of proteins was enriched in the CSK. Several lines of evidence suggest that mRNA is a component of the CSK of both eggs and embryos. First, the CSK contained poly(A) sequences which hybridized with [3H]poly(U). Second, the CSK contained polyribosomes. Finally, RNA extracted from the CSK showed translational activity in an in vitro system. The nonhistone messages present in the CSK were qualitatively similar to those solubilized by detergent, as determined by separation on polyacrylamide gels of the products of in vitro translation. In the unfertilized egg, most mRNA was present as nonpolyribosomal messenger ribonucleoprotein complexes which, along with monoribosomes, were efficiently extracted by Triton X-100. The converse was found in blastulae, as most of the mRNA was present as polyribosomes associated with the CSK, although monoribosomes were still efficiently extracted by detergent. These results indicate a correlation between the activation of protein synthesis in eggs and the association of polyribosomes with the CSK.


Association of specific cell-surface glycoproteins with a triton X-100-resistant complex of plasma membrane proteins isolated from T-lymphoma cells (P1798).
A non-ionic detergent-resistant complex of membrane-associated proteins and cell-surface glycoproteins has been isolated by gel filtration and isopyknic centrifugation of purified plasma membranes from the murine T-lymphoma P 1798. This complex elutes as a high molecular weight peak (greater than 15 X 10(6) D) and contains two specific sets of (1) cell-surface glycoproteins; (2) membrane-associated proteins. The cell-surface glycoproteins consist of two vectorially labelled major components present in a fixed molar ratio: The Thy-1 glycoprotein and a non-H-2 glycoprotein of 55 kD. Minor but significant amounts of the class I histocompatibility antigen Qa-2 are also contained in the detergent-resistant complex. The membrane-associated proteins are not vectorially labelled, and form a complex group of proteins in the 30-70 kD range. Since actin is not detectable among these polypeptides, they probably constitute a plasma membrane-associated structure that is distinct from actin-containing, submembranous cytoskeletal elements.


Difference in the diastolic left ventricular wall motion velocities between aortic and mitral regurgitation by pulsed tissue Doppler imaging.
We evaluated the difference in the diastolic left ventricular (LV) wall motion velocity between chronic isolated aortic and mitral regurgitation (AR and MR, respectively) by recording subendocardial motion velocity patterns at the middle site of the LV posterior wall in the parasternal (along the short axis) and apical (along the long axis) long-axis views of the left ventricle with pulsed tissue Doppler imaging. We studied 33 patients with AR and 35 with MR, showing moderate to severe regurgitation, and 34 healthy controls (C). The end-diastolic LV dimension along the short axis was greater in the AR and MR groups than in the C group, and that along the long axis was greater in the AR group than in the MR and C groups. There were no significant differences in percent LV fractional shortening along the short axis among the 3 groups, whereas that along the long axis was significantly smaller in the AR group than in the MR and C groups. The peak early diastolic wall motion velocity (Ew) and the time to Ew from the aortic component of the second heart sound (S2 -Ew) along the long axis were significantly lower and longer, respectively, in patients with AR than in the 2 other groups. The Ew and S2 -Ew along both the short and long axes were significantly higher and shorter, respectively, in patients with MR than in the 2 other groups. The peak early diastolic velocity of the transmitral flow correlated positively with Ew along the short axis in all patients with AR and correlated positively with Ews along the long and short axes in all patients with MR. In conclusion, early diastolic LV filling was associated with expansion of the LV wall along the short axis but with decreased excursion along the long axis in patients with AR, whereas that in patients with MR was associated with expansion of the LV wall along both the long and short axes. Pulsed tissue Doppler imaging was useful for evaluation of diastolic LV function along the long and short axes in patients with diastolic LV volume overload.


Mesoporous silicas in materials engineering: Nanodevices for bionanotechnologies.
In this review, the most valuable opportunities offered by mesoporous silica nanoparticles in the field of development of nanodevices for bionanotechnology applications are reviewed. The state of the art is critically discussed with particular emphasis on cancer-related application, paying attention to all the aspects of the design and development of the process that engineers the selective administration of an anticancer agent to cancer tissues. The analyses of the critical factors that limit this process are taken into account and the technical solutions proposed to face these factors are discussed. Furthermore, targeting to difficult tissues and forefront applications such as cancer immunotherapy, diagnostic, theranostic, and gene therapy are considered. Lastly, the authors provide their opinion on the reasons according to which the translation of this generation of nanodevices from laboratory research into practical clinical and eventually into the market is possible.


Assessment of maize and beans storage insect pest in major grain markets, Morogoro-Tanzania.
Post-harvest losses for maize and beans are common storage challenges affecting Tanzania's smallholder and large-scale farmers' storage facilities. Consequently, this leads to a reduction of over 50% of harvested grain qualitatively and quantitively per year. Therefore, this experimental study was conducted at the entomological laboratory of the Sokoine University of Agriculture Tanzania, where the research was aimed to assess different common storage insect pests of beans and maize at Morogoro major markets (i.e., Kihonda, Mazimbu and Morogoro Central Market) with similar storage conditions in Morogoro municipality. The complete block design (CBD) with three experimental replications was used. Where locations (Kihonda, Mazimbu and Central market) markets were considered as experimental treatments. The results obtained show that there were no significant differences in number of insect pests (such as maize weevils and bean bruchids) identified between the Kihonda and Central markets (P > 0.05). However, there is a significant difference (P< 0.05) between Mazimbu and other markets regarding the number of insect pest multiplication resulting from the storage condition of those grains in a particular market. Nevertheless, continuous winnowing of stored grains was observed to significantly intensify the infestation of the storage pest in maize and common beans. Therefore, alternatively, to reduce insect pest infestation to maize and common beans in storage facilities, it is important to avoid constant winnowing of stored grains.


Glycosaminoglycan chain analysis and characterization (glycosylation/epimerization).
Glycosaminoglycans (GAGs) including chondroitin sulfate (CS), dermatan sulfate (DS), and heparan sulfate/heparin (HS/Hep) are linear polysaccharides and involved in the regulation of various biological events through interaction with functional proteins. GAGs are modified by sulfation at various positions of each saccharide residue and the epimerization of uronic acid residues during the chain's biosynthesis, resulting in enormous structural diversity. This structural diversity is the basis for the wide range of biological activities of GAGs. Thus, the structural analysis of GAGs is key to understanding their biological functions. This chapter describes detailed instructions for the extraction and structural analysis of GAGs from cultured cells and tissues using a combination of GAG-degrading enzymes and high-performance liquid chromatography.


Bilateral clavicle fracture external fixation.
Fractures of the middle third of the clavicle are frequent and their conservative treatment ends in bone union in nearly 95% of cases. Surgical treatment is unanimously indicated with open fractures or in cases of cutaneous damage, neurovascular complications, and impaction of the shoulder stump syndromes. We report herein a case of bilateral fractures of the clavicle that required double stabilization with an external fixator following major cutaneous damage appearing after the initial conservative management. The intraoperative discovery of Propionibacterium acnes infection and bone union obtained within the classical time frame, with a satisfactory functional result, all retrospectively proved the soundness of this indication.


Management of ovarian cysts in infants.
To discuss the experience of diagnosis and treatment of ovarian cyst in infants. A retrospective review was conducted on 20 infants who suffered from ovarian cyst. There were no dysplasia ovarian was found in children which were preoperatively diagnosed simplex cyst. Within thirteen children preoperatively detected mixed cystic-solid lesion, six cases ovarian cysts disappeared and two cases underwent poor blood supply in the following time. Adverse effects for ovarian cyst in infants can be prevented by agressive surgical intervention. Harmful effects of ovarian cyst can be prevented by positive surgical intervention despite the diagnostic difficulties in children with clinical symptoms of this condition.


Modelling the inhibitory effect of copper sulfate on the growth of Penicillium expansum and Botrytis cinerea.
This study aimed to investigate the effect of copper sulfate (from 0 to 8 mmol kg(-1)) on radial growth rate and lag time of two moulds responsible for vine grapes spoilage: Penicillium expansum strain 25·03 and Botrytis cinerea, strains BC1 and BC2. A new model was developed to describe tailing and shoulders in the inhibition curves. Because of tailing, the minimum inhibitory concentration (MIC), was not defined as the concentration at which no growth was observed, but as the concentration at which the lag time was infinite. The concentrations at which μ = μ(opt)/2, (Cu50), were in the range of 2·2-2·6 mmol kg(-1). Radial growth rate of P. expansum and the reciprocal of the lag time were linearly correlated (r = 0·84). In contrast, in the range 0-4 mmol kg(-1), an inhibition of growth of B. cinerea was observed whereas germination remained unaffected (i.e. the lag time was constant). In the range 4-8 mmol kg(-1) , the radial growth rate of B. cinerea was almost constant (c. 1 mm day(-1)), but germination was inhibited (i.e. the lag time was increased). The MIC values were 4·7 mmol kg(-1) for P. expansum, 8·2 and 7·3 mmol kg(-1) for B. cinerea strain BC1 and BC2, respectively, demonstrating that some isolates of these moulds are resistant to copper. Copper concentrations at 4 mmol kg(-1) would be sufficient to control the development of these isolates, but the toxicity of copper should be extended to other isolates and evaluated in vineyards.


The end product of transglutaminase crosslinking: simultaneous quantitation of [Nepsilon-(gamma-glutamyl) lysine] and lysine by HPLC-MS3.
Transglutaminases catalyze the formation of Nepsilon-(gamma-glutamyl) isodipeptide crosslinks between proteins. These enzymes are thought to participate in a number of diseases, including neurological disease and cancer. A method associating liquid chromatography and multiple stage mass spectrometry has been developed for the simultaneous quantitation of [Nepsilon-(gamma-glutamyl) lysine] isodipeptide and lysine on an ion trap mass spectrometer. Highly specific detection has been achieved in MS3 mode. The method includes a derivatization step consisting of butylation of carboxylic groups and acetylation of amide groups, a liquid-liquid extraction, and a 19-min separation on a 100x2.1-mm Beta-basic C18 column with an acetonitrile gradient elution. 13C6-(15)N2 isotopes of the isodipeptide and the lysine serve as internal standards. The assay was linear in the range of 50 pmol/ml to 75 nmol/ml for the isodipeptide and the range of 10 nmol/ml to 3.5 micromol/ml for the lysine, with correlation coefficients greater than 0.99 for both ions. Intra- and inter-day coefficients of variation ranged from 3.5 to 15.9%. The method was successfully applied to human biological samples known to be crosslinked by transglutaminase such as cornified envelopes of epidermis, fibrin, and normal and Huntington disease brain.


Madelung's disease associated with polyneuropathy and symptomatic hypokalemia.
Madelung's disease (multiple symmetric lipomatosis) is a rare disease characterized by abnormal diffuse lipomatosis in proximal upper limbs and neck. Previous reports have shown that this disease is associated with alcoholism, polyneuropathy, mitochondrial disease, and glucose intolerance. Here, we describe a 46-year-old man having Madelung's disease associated with polyneuropathy and symptomatic hypokalemia. He presented with insidious-onset weakness and numbness in lower limbs for 7 years and recent deterioration of symptoms. Proximal weakness improved with potassium supplement. Our observation may extend the phenotype of Madelung's disease to hypokalemic periodic paralysis.


Association of combined engagement in cognitive and physical activity with domain-specific cognitive function: The Shanghai Aging Study.
This cross-sectional study aimed to examine the association of the combined engagement in cognitive activity (CA) and physical activity (PA) with domain-specific cognition in community-dwelling older adults. We analysed data from 3192 dementia-free participants aged ≥50 years in the Shanghai Aging Study. CA was assessed using Shanghai Cognitive Activities Scale. PA was determined based on questionnaires and further transformed into metabolic equivalent values. We used multivariate linear and logistic regression models to estimate the β and odds ratio of CA, PA, or combined CA and PA and each neuropsychological test. A high level of CA was associated with a better performance in most of the tests, except for the conflicting instructions task (CIT) and stick test (ST). In contrast, PA displayed no significant associations with any test. Engagement in high CA and high PA was associated with the best performance in Mini-Mental State Examination, recall in ST, categorisation in Modified Common Objects Sorting Test (MCOST), immediate recall, delayed recall, and recognition in Auditory Verbal Learning Test. Participants with "high CA and low PA" had the lowest risk of impairment in Go/No-Go correct tapping in CIT, rotate in ST, item naming, and category naming in MCOST, Trail Making Test (TMT)-A, and TMT-B. Our study suggests that engagement in both high CA and high PA may be the most efficacious way to maintain various domains of cognition. A higher level of CA may help to preserve cognition among older individuals who have difficulties in performing PA.


Comparision of anti-HLA antibodies of kidney transplant candidates with chronic renal failure by two different methods: Flow-PRA and Luminex PRA.
The aim of the study was to compare anti-HLA antibodies examined as panel-reactive antibody (PRA) in kidney transplant candidates with chronic renal failure (CRF) with the use of 2 methods: Flow-PRA and Luminex-PRA. CRF patients displaying class I PRA (n = 34) and/or class II PRA (n = 41) were tested by the 2 different methods from April 2012 to September 2012, using antigen-coated beads. Eleven (32.3%) 34 patients tested for class I PRA were female and 23 (67.7%) male; 17 (41.5%) 41 patients tested for class II PRA were female and 24 (58.5%) male. Only 2 patients were preemptive, the others had been subjected to dialysis. The concordance ratio of class I PRA test results between Flow-PRA and Luminex PRA was 67.6%. Whereas 13 samples (38.2%) were positive by Flow-PRA, 22 (64.7%) were positive by Luminex-PRA. Two of the 3 patients not previously immunized were found to be positive only by Luminex PRA; 1 was noted to be positive only by Flow-PRA. Regarding class II PRA screening, the concordance between Flow-PRA and Luminex PRA was 70.7%. Whereas 14 (34.1%) samples were positive for Flow-PRA; 24 (58.5%) were positive for Luminex-PRA. The 2 patients not previously immunized were positive only in Luminex PRA. We speculated that the reason for the low concordance ratios was due to the use of sera that had been previously found to be indeterminate in PRA tests. We also speculated that the low concordance ratios were due to the coating procedure for the beads, which may cause changes in antigenic epitopes and decrease concordance between Flow-PRA and Luminex-PRA.


Sentinel node mapping affects intraoperative pulse oximetric recordings during breast cancer surgery.
In invasive breast cancer lymphatic mapping with patent blue vital dye (PBV) is used intraoperatively to identify the sentinel lymph nodes: the first axillary node draining the mammary lymphatic basin and first involved by the metastatic growth in breast cancer. Patent blue vital dye spreads to tissues giving a bluish tinge to patients. We have noted the possibility that intraoperative peripheral pulse oximetric (SpO2) values are artificially low when intradermal PBV is used. Twenty patients with normal pulmonary function undergoing breast cancer surgery in standardized anesthesia either did or did not receive intradermal PBV sentinel node marking. The radial artery was cannulated for blood-gas-analysis; arterial oxygen tension (PaO2); and arterial oxygen saturation (SaO2). Peripheral oxygen saturation was measured using the light absorption technique. Red and infrared light (660 and 900 nm), used by pulseoxymetry, is partially absorbed when passing through the tissue. The amount of light absorbed is sensed and saturation calculated. The color of the skin was evaluated. Peripheral oxygen saturation decreased only immediately after the injection of PBV, and remained at a significantly lower level (P<0.001) throughout the operation and up to 90 min postoperatively. Arterial oxygen tension and SaO2 values did not decrease after intradermal PBV. Patent blue vital dye made patients' skin more bluish (P<0.001). No changes in SpO2, PaO2 and SaO2 were found in control patients. The spectrum of PBV has a peak absorption at 640 nm, thus making the SpO2 values incorrect. Peripheral oxygen saturation values are falsely low and true arterial oxygenation is not impaired when PBV is used during sentinel node mapping.


Different-batch metabolome analysis of Saccharomyces cerevisiae based on gas chromatography/mass spectrometry.
Each experimental step in metabolomics based on mass spectrometry for microorganisms, such as cultivation, sampling, extraction of metabolites, analysis, and data processing includes different systematic errors. Even if the same protocol is used, it is difficult to compare the data from different cultivation days or different analysis days. To obtain reliable quantitative data, it is necessary to develop an analytical workflow that can reduce errors from different batch of cultivation and analysis days. We compared metabolomics methods for Saccharomyces cerevisiae in terms of reproducibility to optimize the analytical workflow, particularly quenching and data processing. Our data also showed that reproducible data could be obtained with high signal to noise ratio. Therefore, we optimized a time segmented selective ion monitoring (SIM) method for high sensitive analysis with low-risk of false positives. The optimized workflow was applied to metabolome analysis of single transcription factor deletion mutants. As a result, we obtained clusters that were independent of cultivation day and analysis day but were strain-dependent. This study can help to implement large-scale or long-term studies, in which samples are divided among several laboratories because of the high number of samples.


High-speed preparative reversed-phase high-performance liquid chromatography of synthetic oligonucleotides.
A rapid method is described for the purification and analysis of synthetic oligonucleotides, based on reversed-phase high-performance liquid chromatography. Volatile buffers and a short column (40 mm X 4.6 mm) packed with Nucleosil 300-5 C4 were employed. Monitoring the column effluent with an UV detector provides an excellent means of controlling product quality. The method is suitable for the purification of crude synthesis products, as well as for desalting and removing gel contaminants from oligonucleotides eluted from polyacrylamide gels. The total time required per sample is less than 25 min.


Chlorido{N-[(E)-2-(diphenyl-phosphan-yl)benzyl-idene]-2-(thio-phen-2-yl)ethan-amine-κP}gold(I).
The title compound, [AuCl(C(25)H(22)NPS)], crystallizes with two independent mol-ecules in the asymmetric unit in which the thio-phene fragments are disordered over two sets of sites with 0.537 (10):0.463 (10) and 0.701 (9):0.299 (9) occupancy ratios. In both cases, the thio-phene ring is rotated by approximately 180° for the second component. Important geometrical parameters include Au-P = 2.235 (2) and 2.237 (2) Å, Au-Cl = 2.286 (2) and 2.292 (2) Å, and P-Au-Cl = 177.39 (8) and 172.63 (7)°. Weak inter-molecular C-H⋯Cl inter-actions are observed in the crystal structure.


Hypnosis within a psychospiritual approach in the case of a woman diagnosed with dystonia.
This case report describes a psychospiritual intervention utilizing hypnosis with a client seeking help for problems with involuntary movement of the right arm and hand, accompanied by a great deal of pain. Doctors had diagnosed her condition as dystonia with the presumption that it was primarily physical in nature. She came to Carolinas Integrative Health seeking an integrative approach to her problem, one that brings together conventional and complementary medicine. The client was referred to the author by the Medical Director for a mind/body/spirit approach, including hypnosis. Initial evaluation suggested there might be a psychogenic component to the problem. The client also revealed a spiritual worldview, theistic in nature. Utilizing hypnosis, together with a process the author calls "Centering in Self," the client's spiritual beliefs and resources were accessed and utilized in bringing about resolution of pain and almost complete cessation of involuntary movements.


Cognitive deficits associated with restless legs syndrome (RLS).
Restless legs syndrome produces significant chronic sleep loss, which despite not causing expected profound sleepiness, might nonetheless produce cognitive deficits similar to those seen with acute sleep deprivation, i.e. involving mostly pre-frontal cortical (PFC) functioning. Sixteen patients off RLS treatment for at least 2 weeks and 15 age- and gender-matched control subjects had polysomnograms (PSGs) on two consecutive nights. Cognitive tests were given in the morning after the second night. Six cognitive tests were used: two Verbal Fluency tests and the Trail Making tests were selected to be particularly sensitive to PFC function and sleep loss. Porteus Mazes and the Stroop Test were selected to reflect more general frontal and executive function. The Colored Progressive Matrices were used to assess general cognitive skills. RLS patients compared to controls showed significant (P<0.05) and sizeable (20-40%) deficits on two of the three PFC tests and marginally non-significant deficit (P<0.1) on the third. The other three tests showed no significant differences. The results indicate that RLS patients show cognitive deficits similar to that reported for one night of sleep loss.


Mapping the genomic diaspora of gastric cancer.
Gastric cancer (GC) is a leading contributor to global cancer incidence and mortality. Pioneering genomic studies, focusing largely on primary GCs, revealed driver alterations in genes such as ERBB2, FGFR2, TP53 and ARID1A as well as multiple molecular subtypes. However, clinical efforts targeting these alterations have produced variable results, hampered by complex co-alteration patterns in molecular profiles and intra-patient genomic heterogeneity. In this Review, we highlight foundational and translational advances in dissecting the genomic cartography of GC, including non-coding variants, epigenomic aberrations and transcriptomic alterations, and describe how these alterations interplay with environmental influences, germline factors and the tumour microenvironment. Mapping of these alterations over the GC life cycle in normal gastric tissues, metaplasia, primary carcinoma and distant metastasis will improve our understanding of biological mechanisms driving GC development and promoting cancer hallmarks. On the translational front, integrative genomic approaches are identifying diverse mechanisms of GC therapy resistance and emerging preclinical targets, enabled by technologies such as single-cell sequencing and liquid biopsies. Validating these insights will require specifically designed GC cohorts, converging multi-modal genomic data with longitudinal data on therapeutic challenges and patient outcomes. Genomic findings from these studies will facilitate 'next-generation' clinical initiatives in GC precision oncology and prevention.


Photoreceptive net in the mammalian retina. This mesh of cells may explain how some blind mice can still tell day from night.
We have discovered an expansive photoreceptive 'net' in the mouse inner retina, visualized by using an antiserum against melanopsin, a likely photopigment. This immunoreactivity is evident in a subset of retinal ganglion cells that morphologically resemble those that project to the suprachiasmatic nucleus (SCN), the site of the primary circadian pacemaker. Our results indicate that this bilayered photoreceptive net is anatomically distinct from the rod and cone photoreceptors of the outer retina, and suggest that it may mediate non-visual photoreceptive tasks such as the regulation of circadian rhythms.