An efficient method to detect calcitonin mRNA in normal and neoplastic rat C-cells (medullary thyroid carcinoma) by in situ hybridization using a digoxigenin-labeled synthetic oligodeoxyribonucleotide probe.
We report here an efficient and rapid method for the specific detection of calcitonin in tumor C-cells of medullary thyroid carcinoma (MTC). This occasionally aggressive tumor arises from the endocrine thyroid C-cells. Its principal marker is calcitonin, the predominant C-cell secretion, which is detected in patients and in our animal model by radioimmunoassay of the plasma, as well as by immunohistochemistry of thyroid tissues. Although calcitonin is easily detectable in normal C-cells, its content is greatly reduced in tumor cells owing to the disappearance of the secretory granules that store the mature peptide. This finding suggests cell dedifferentiation correlated with an increasing aggressivity of the tumor. We therefore developed a rapid detection of calcitonin mRNA by in situ hybridization on routine paraffin sections, using a synthetic oligodeoxyribonucleotide probe labeled with digoxigenin-dUTP. The reaction was detected with an anti-digoxigenin antibody conjugated with alkaline phosphatase, and the enzyme catalyzed the appearance of a dark blue color. The signal was exclusively restricted to the normal, hyperplastic, and tumor C-cells. It was specific, as increasing concentrations of the unlabeled oligonucleotide led to progressive disappearance of the reaction. Its sensitivity was slightly diminished as compared with corresponding frozen sections, but the intensity of the signal was quite acceptable. High levels of calcitonin mRNA were found in all normal and hyperplastic C-cells. They were increased in most of the tumor MTC cells, which did not correlate with the amount of intracellular peptide stores but explained the abnormally high basal levels of circulating calcitonin of the tumor-bearing rats. ISH is therefore of greater value than ICC for an early anatomopathological detection of this tumor. Our data show that the tumor cells are not "dedifferentiated." They only lack the granular compartment storing the mature peptide before exocytosis, but CT biosynthesis and the rest of the secretory process seem to be complete. Our results suggest that factors expressed in malignant C-cells affect basic cell mechanisms involved in the storage of the mature calcitonin, rather than the expression of the CALC gene.


Seventh international symposium on neural regeneration
Copyright


Activating Graphyne Nanosheet via sp-Hybridized Boron Modulation for Electrochemical Nitrogen Fixation.
Exploring new metal-free catalysts with high activity for nitrogen reduction reaction (NRR) is highly desirable but remains a big challenge. Graphyne (GY) is a typical two-dimensional carbon material with many excellent properties. However, the NRR has rarely been envisaged on a GY-based metal-free catalyst up to now. Density functional theory calculations reveal that although pristine GY is inactive for N2 reduction, boron modulation can endow it with efficient activity toward NRR. Natural bond orbitals analysis, spin/charge density distributions, and free energy change diagrams are performed and discussed. Three boron doping formats including sp2-substituted, sp-substituted, and adsorbed configuration are considered. The obtained data show sp-substitution will induce local moderate spin and charge densities at the boron site on the GY surface, which is convenient for N2 adsorption and activation, and conductive to N-related intermediates formation and transformation. Moreover, the incorporated sp-hybridized boron can provide one empty p orbital and one occupied p orbital around itself, which plays a key role as an electron reservoir to accept electrons from and donate electrons to the adsorbed N-related species, and thus facilitate N2 reduction and ammonia synthesis. Henceforth, it provides more opportunities for preparing GY and other carbon materials as efficient catalysts toward renewable energy conversion and storage.


Status of management effort in 153 marine protected areas across the English Channel.
A conceptual framework was developed for assessing the sub-level of protection in 185 multiple-use marine protected areas (MPAs) in the English Channel through a survey on management effort. Data were retrieved from 153 MPAs: 4.56% were assigned low management effort, 83.70% were assigned medium management effort, and 11.76% were assigned high management effort. Overall, French MPAs performed better in terms of management effort than English MPAs and lack of consistency in ratings by different management bodies in England was found. Lack of correlation between management effort and conservation status within an available subset of 13 MPAs suggests that management may not be as influential a factor for the effective conservation of MPAs, especially in marine environments under heavy human pressure such as the English Channel. It is suggested that MPAs in such areas may therefore require an upgrade of their legal level of protection to be effective.


Prevalence of anemia in children 1 to 12 years of age. Results from a nationwide probabilistic survey in Mexico.
To describe the epidemiology and analyze factors associated with iron deficiency anemia in a probabilistic sample of the Encuesta Nacional de Nutrición 1999 (ENN-99) [National Nutritional Survey 1999 (NNS-99)]. The sample included 8,111 children aged 1 to 12 years, and was nationally representative by rural and urban strata and by four geographical regions. Capillary hemoglobin was measured using a portable photometer (HemoCue). The analysis of the determining factors of anemia was performed by odds ratios derived from a logistic regression model and multiple regression models. The prevalence of anemia was 50% in infants < 2 years of age, with no significant differences between urban and rural strata or among regions. It varied between 14 and 22% in 6-11 year-old children and was higher in the South region and among the indigenous children. Dietary intake of iron was 50% of the recommended daily allowance in children < 2 years of age, but not in older children. Phytate ( approximately 500-800 mg/d) and tannin (approximately 19 mg/d) intakes were very high in children over 7 years of age. Hemoglobin was positively associated with nutritional status of children (p = 0.01), socioeconomic status (p range 0.05-0.001), duration of lactation in children under 2 years of age (p = 0.1), and iron and calcium intake (p = 0.02), but not with folic acid or vitamin B12 intake. Hemoglobin was negatively associated with maternal education (p = 0.01) in older children, but not in those under 2 years of age. We present evidence of an alarming national epidemic of anemia, particularly marked in children 12 to 24 months of age. The control of anemia should be considered as an urgent national concern given its grave consequences on the physical and mental development of these children and on their long-term health. The English version of this paper is available too at: http://www.insp.mx/salud/index.html.


A Cre-reporter transgenic mouse expressing the far-red fluorescent protein Katushka.
Cre/loxP-dependent expression of fluorescent proteins represents a powerful biological tool for cell lineage, fate-mapping, and genetic analysis. Live tissue imaging has significantly improved with the development of far-red fluorescent proteins, with optimized spectral characteristics for in vivo applications. Here, we report the generation of the first transgenic mouse line expressing the far-red fluorescent protein Katushka, driven by the hybrid CAG promoter upon Cre-mediated recombination. After germ line or tissue-specific Cre-driven reporter activation, Katushka expression is strong and ubiquitous, without toxic effects, allowing fluorescence detection in fresh and fixed samples from all tissues examined. Moreover, fluorescence can be detected by in vivo noninvasive whole-body imaging when Katuhska is expressed exclusively in a specific cell population deep within the animal body such as pancreatic beta cells. Thus, this reporter model enables early, widespread, and sensitive in vivo detection of Cre activity and should provide a versatile tool for a wide spectrum of fluorescence and live-imaging applications.


Late and unusual intestinal features in cystic fibrosis-pseudotumoral intestinal wall thickening.
12 C.F. patients, 3 males and 9 females, aged 7 y to 36 y (mean 15 y) have had radiological studies of the G. I. tract, either by barium meal or barium enema, resulting in the following findings: - thickened mucosal folds of the duodenum in 2 cases, - a defect of the medial part of the caecum with or without propensity to intussusception, due to a large appendix with hypertrophic mucus glands in 7 cases; - tiny defects of the walls of the caecum and of the colon due to enlarged mucus glands in 5 cases, - a pseudotumoral thickening of the walls of the caecum and of the right colon, confirmed by CT and endoscopy in a woman aged 36 y. Several of these cases have been histologically confirmed either after surgery or by endoscopy. The lesions of the appendix with propensity to intussusception account for the intermittent colicky pains of these patients; radiological evaluation may result in eventual surgery.


5-Hydroxytryptamine and rat passive peritoneal anaphylaxis.
Antigen challenge of rats, sensitized by intraperitoneal injection with rat anti-serum, did not result in a detectable increase in the 5-hydroxytryptamine (5-HT) levels in their peritoneal fluids over the background level induced by sensitisation alone. The maximum amount of extravasation produced by intraperitoneal injection of 5-HT into passively sensitised rats was less than that produced by antigen or histamine, and the doses of 5-HT producing these levels of extravasation produced an produced an increase of 5-HT concentrations in the peritoneal fluids. Therefore, 5-HT is unlikely to make much direct contribution to the extravasation produced during rat passive peritoneal anaphylaxis. However, when given intraperitoneally to rats, 5-HT potentiates the extravasation produced by histamine.


Material properties estimation of layered soft tissue based on MR observation and iterative FE simulation.
In order to calculate deformation of soft tissue under arbitrary loading conditions, we have to take both non-linear material characteristics and subcutaneous structures into considerations. The estimation method of material properties presented in this paper accounts for these issues. It employs a compression test inside MRI in order to visualize deformation of hypodermic layered structure of living tissue, and an FE model of the compressed tissue in which non-linear material model is assigned. The FE analysis is iterated with updated material constant until the difference between the displacement field observed from MR images and calculated by FEM is minimized. The presented method has been applied to a 3-layered silicon rubber phantom. The results show the excellent performance of our method. The accuracy of the estimation is better than 15%, and the reproducibility of the deformation is better than 0.4 mm even for an FE analysis with different boundary condition.


[Bowel imaging: a reassessment. Part 2: CT and MRI].
This is the second part of a review of bowel imaging. While the first part addressed conventional X-ray techniques and ultrasonography, the second part discusses the diagnostic features of computed tomography (CT) and magnetic resonance imaging (MRI) including virtual colonography and PET-CT in the diagnosis of bowel disorders. Indications, performance and the diagnostic impact of the different methods are presented and discussed in the context of competitive methods such as (capsule-)endoscopy.


Single-cell chromatin immunocleavage sequencing (scChIC-seq) to profile histone modification.
Our method for analyzing histone modifications, scChIC-seq (single-cell chromatin immunocleavage sequencing), involves targeting of the micrococcal nuclease (MNase) to a histone mark of choice by tethering to a specific antibody. Cleaved target sites are then selectively PCR amplified. We show that scChIC-seq reliably detects H3K4me3 and H3K27me3 target sites in single human white blood cells. The resulting data are used for clustering of blood cell types.


Intrauterine device-associated actinomycosis simulating pelvic malignancy.
We describe pelvic and rectal actinomycosis in a patient with an intrauterine contraceptive device. The presentation was that of an indolent, noninfectious process which mimicked a pelvic malignancy. As in many cases of pelvic actinomycosis, the diagnosis was not suspected preoperatively. Moreover, suppurative disease progressed despite removal of the intrauterine device. In patients who have intrauterine contraceptive devices or who have had them removed recently, abdominal pain, recurrent vaginal bleeding or adnexal masses should prompt a diligent search for potentially pathogenic actinomyces in the genital tract.


The application of stop-flow microwave technology to scaling-out SNAr reactions using a soluble organic base.
A model S(N)Ar reaction which gives a range of substituted diaryl ethers has been re-developed to function with the soluble organic base DBU in the place of insoluble potassium carbonate. Manufacture of these diaryl ethers has then been achieved by scaling-out in an automated stop-flow microwave reactor to give productivities of >0.5 kg per day. Analogous reaction partners have also been scaled up in this reactor to extend the scope of the study. Brief comparison in a scale-up batch microwave reactor is also made. Lastly, continuous 24 h processing is reported in this small microwave stop-flow reactor, which requires no manual intervention once started.


What Should We Expect? A Comparison of the Community Benefit and Projected Government Support of Maryland Hospitals.
Designation as a tax-exempt, not-for-profit entity carries with it specific tax benefits. In exchange for tax exemptions, not-for-profit entities are expected to provide benefits to their communities. To evaluate whether hospitals provide community benefits (CBs) equivalent to the financial subsidies and advantages extended to them, tax liabilities and financial support were projected for all Maryland acute care hospitals between 2010 and 2012 and in the aggregate over the 3 years of this study. A comparison was then made between the provision of CBs and the financial support that governments provide to the hospitals. The results indicate that hospitals provide significantly and substantially more CBs than the material financial support they receive. Even after modeling changes in CB activities and the associated tax liabilities that may result from transitioning to taxable status, the benefits that hospitals provide to the communities they serve continue to exceed the potential government tax revenues.


Anti-SGPG antibody in CIDP: nosological position of IgM anti-MAG/SGPG antibody-associated neuropathy.
Polyneuropathy with monoclonal gammopathy usually is considered a nosological entity different from chronic inflammatory demyelinating polyneuropathy (CIDP). Criteria proposed by the American Academy of Neurology AIDS Task Force (1991), however, show monoclonal gammopathy to be a condition concurrent with CIDP. The purpose of this study was to clarify the nosological relationship between CIDP and IgM anti-myelin-associated glycoprotein (MAG)/sulfated glucuronyl paragloboside (SGPG)-associated polyneuropathy. We investigated IgM anti-MAG/SGPG antibody in 85 CIDP patients by various methods, then examined the relation of M-protein to the presence of IgM anti-MAG/SGPG antibody. In our large study, 17 (20%) of 85 CIDP patients had high IgM anti-SGPG antibody titers in the enzyme-linked immunosorbent assay. This was confirmed by thin-layer chromatography-immunostaining for IgM anti-SGPG antibody and immunoblotting for IgM anti-MAG antibody. Immunoelectrophoresis and immunofixation, respectively, detected IgM M-protein in 6 (35%) and 13 (76%) of the 17 CIDP patients. We conclude that some patients with IgM anti-MAG/SGPG antibody with or without monoclonal gammopathy may be diagnosed as having CIDP, when patients are diagnosed according to the current CIDP criteria.


G-scan--mobile multiview 3-D measuring system for the analysis of the face.
The development of optical 3-D measuring techniques and their use in industrial quality assurance, in design, and for rapid prototyping has experienced strong growth. A large number of optical 3-D measuring methods and systems are on the market in dentistry. CAD/CAM production has become firmly established in dental medicine, not least due to the systematic introduction of the Cerec technique and the digiDent method. The scanners on which these technologies are based are designed for a relatively small measuring area. To be able to measure and three-dimensionally assess the face--and the numerous changes in the face/forehead/neck region--it was necessary to design and develop a self-calibrating measuring system with gray code for clinical use: the G-Scan measuring system. Objects up to a size of 500 x 500 x 400 mm can be acquired three-dimensionally with it, with a measuring inaccuracy of 10 to 70 microm in a typical measuring time of 15 s. The present article describes the measuring principle, the system parameters, and the features of the new measuring system, and illustrates the measuring results on 3-D displays of the face in static occlusion and in functional occlusion positions.


Behavioral consequences of repeated nicotine during adolescence in alcohol-preferring AA and alcohol-avoiding ANA rats.
Epidemiological studies suggest that exposure to nicotine at adolescent age is associated with increased potential to use alcohol and that genetic predisposition may further increase the risk. The present study addressed adolescent vulnerability to repeated nicotine exposure and its influence on subsequent ethanol self-administration by investigating interactions between nicotine-induced behavioral sensitization and voluntary ethanol consumption in alcohol preferring AA (Alko Alcohol) and alcohol nonpreferring ANA (Alko Non-Alcohol) rat lines selected for differential ethanol intake. Adolescent and adult rats received 10 injections of nicotine (0.5 mg/kg s.c.), given every second day from postnatal day (Pnd) 27 and 75, respectively. Nicotine-induced (0.5 mg/kg) locomotor activity was measured acutely after the first injection, and after the repeated treatment with nicotine on Pnds 52 and 86 in the adolescent groups and on Pnd 99 in the adult groups. After this, acquisition of voluntary ethanol (10% v/v) consumption as well as nicotine-induced (0.5 mg/kg) ethanol intake was measured in the AA rats. Adolescent AA rats were more sensitive than adolescent ANA rats to the locomotor effects of nicotine. They were also stimulated more than adult AA rats, but such a difference was not found among ANA rats. Adolescent and adult rats did not differ in their susceptibility to nicotine-induced behavioral sensitization. Genetic predisposition to ethanol self-administration did not interact with development of behavioral sensitization in either adolescents or adults. Acquisition of ethanol intake was enhanced in the adolescent groups relative to the adult groups in a manner that was independent of the nicotine treatment. An increase in ethanol intake was found after challenging animals with nicotine, and this effect was enhanced in the nicotine-treated adolescent group. These findings provide no or little support for the views that adolescent animals are more sensitive to the neurobehavioral effects of repeated exposure to nicotine and that exposure to nicotine in adolescence may contribute to enhanced vulnerability to ethanol abuse. Furthermore, genetic predisposition to high or low ethanol self-administration does not seem to be a factor that influences individual vulnerability to the neurobehavioral effects of repeated administration of nicotine.


Directed microbial biosynthesis of deuterated biosurfactants and potential future application to other bioactive molecules.
Deuterated rhamnolipids were produced using strain AD7 of Pseudomonas aeruginosa, which was progressively adapted to increasing levels of deuterium in D(2)O and carbon substrates. Fourteen different deuterated rhamnolipid structures, including structural isomers, were produced which is similar to normal protonated structures. There were two main products monorhamnolipid Rha-C(10)-C(10) and dirhamnolipid Rha(2)-C(10)-C(10). The levels of deuteration varied from 16% with 25% D(2)O + h-glycerol to 90% with 100% D(2)O + d-glycerol. When d-tetradecane was used with H(2)O, virtually all the deuterium appeared in the lipid chains while using h-tetradecane + D(2)O led to the majority of deuterium in the sugars. The adaptation to growth in deuterium appeared to be metabolic since no genetic changes could be found in the key rhamnolipid biosynthetic genes, the rhamnosyl transferases RhlB and RhlC. Deuterated sophorolipids were similarly produced using Candida bombicola and Candida apicola although in this case, no adaptation process was necessary. Up to 40 different sophorolipids were produced by these yeasts. However, unlike the rhamnolipids, use of D(2)O did not lead to any deuteration of the lipid chains, but direct incorporation into the lipid was achieved using d-isostearic acid. The results from these experiments show the feasibility of producing deuterated bioactive compounds from microorganisms coupled with the possibility of manipulating the pattern of labelling through judicious use of different deuterated substrates.


Effect of daily high doses of paracetamol given orally during spermatogenesis in the rat testes.
Daily treatment of male rats with high doses of Paracetamol (500 mg/kg) during spermatogenesis provokes a significant decrease of testes weight (p is less than 0.001), a significant increase of testicular cytosol glutathione transferase activity ( p is less than 0.02) and of lipid peroxides in the whole homogenate (p is less than 0.01) but not in the microsomes. These alterations could be related to a stimulation of testicular metabolism but cannot produce a biochemical lesion such as glutathione depletion since its testicular content is not modified in these experimental conditions.


"Brain-muscle loop" in the fragility of older persons: from pathophysiology to new organizing models.
The imperative action of the geriatric medicine is to prevent disability in older persons. Many epidemiological studies have been conducted in the last decades for improving knowledge of the aging process and their interactions with age-related diseases, especially for the identification of the relationship between sarcopenia and loss of mobility. Factors influencing muscle integrity can be classified into six main physiologic subsystems, but the central nervous system certainly plays a crucial role for maintaining muscle integrity in older persons. Recent data show that the reduced muscle strength and not muscle mass could be considered the core of the fragility in predicting changes of gait velocity and mobility and conferring a higher risk of mortality in older persons. Sarcopenia and cognitive decline could, therefore, produce slow gait velocity in older persons, with devastating effect and consequences. Perhaps the most notorious corollary is falling, which is often caused by an underlying gait problem. Injuries caused by accidental falls range from relatively innocent bruises to major fractures or head trauma. Another important consequence is reduced mobility, which leads to loss of independence. This immobility is often compounded by a fear of falling, which further immobilises patients and affects their quality of life and physical performance. When we search the association between brain pathology and muscle function in older persons, we amazingly find that established composite measure of physical frailty is associated with brain pathology. Sarcopenia, which produces muscle dysfunction, slow gait velocity and cognitive decline, could share a strong bidirectional relationship, and this suggests the coexistence of both cognitive and motor dysfunctions in older persons to characterize a new syndrome characterized by slow gait and cognitive complaints, the motoric-cognitive risk syndrome (MRC). In this review, we want to emphasize the relationship between memory complaints with muscle function integrating cognitive and physical evaluation, even with amyloid PET study, to identify older patients at high risk of cognitive and physical decline.


Control of influenza in the long-term-care facility: a review of established approaches and newer options.
Influenza infections pose a serious threat to residents of nursing homes and other long-term-care facilities. Annual vaccination of residents and staff with the currently licensed inactivated influenza vaccine continues to be the mainstay of prevention. Live attenuated influenza vaccine, which is expected to be licensed in the United States in the near future, may offer added protection for elderly persons when administered in conjunction with inactivated vaccine. Antiviral agents also can be useful as an adjunct to vaccination, especially for control of institutional outbreaks. Two new antiviral agents that appear to be less toxic than amantadine and rimantadine have recently been approved.


[An experimental study of the effect of perfusion of duodenal juices in the vagally innervated antral pouch upon gastric endocrine and exocrine function].
Vagally innervated antral pouches were created in adult mongrel dogs, the interiors of these antral pouches were perfused with glycocholate, taurocholate, canine bile and human pancreatic juice, and antral function was investigated from the viewpoint of gastric endocrine and exocrine response. Perfusion with glycocholate and taurocholate of 20 mM induced a significant increase in gastric acid secretion as well as serum gastrin secretory response. Perfusion with canine bile and human pancreatic juice induced a pronounced increase in serum gastrin levels as well as gastric acid secretion starting immediately after the beginning of perfusion, and serum gastrin secretory response also displayed a marked increase. These results demonstrated that the regurgitation into the stomach of duodenal fluid containing bile and pancreatic juice, constitutes one factor causing increased serum gastrin levels and gastric acid secretion.


[Toxic epidermal necrolysis].
Toxic epidermal necrolysis is the prototype of a proapoptotic disease characterized by system CD95 dysrregulation. Drugs constitute the main antigenic triggers. Hystopatologically it is characterized by epidermis detachment and necrosis with apoptotic keratinocytes. Clinical presentation includes erithematous-ampullous lesions in the skin and mucous membranes. It is associated with serious complications such as severe sepsis and septic shock. The management in the intensive care unit includes support treatment and specific treatment with immunoglobulins that alter disease course. Recombinant activated Factor VII is effective to control the associated microvascular haemorraghe.


[Microbial contamination of air by activated sludge units (author's transl)].
Measurements of the germ emissions in various activated sludge units with different ventilations systems have shown that the lowest density occurs with small bubble ventilation; with medium bubble and brush ventilation rather high values were measured; the spray devices used for foam elimination had a particularly unfavorable influence. In great germ densities above the pools (50-100 000 germs/m3) - depending on the weather - between 500 and 1500 germs/m3 were measured at a distance of 50 and 100 meters. At distances of 200 and 400 m the number of germs was about the same as normally measured in the outside air (100-500 germs/m3). In a closed pool altogether considerably higher values were measured: In the immediate neighbourhood of the activated sludge unit they amounted to abt. 10 000-25 000 germs/m3; in the hall 3000-4000 germs/m3 were measured, whereby the part of coliform bacteria amounted to 1-2% and that of enterococci to 2-4%. It has to be taken into account that besides the measured intestinal flora further pathogenous agents are present in the aerosols. In closed units, therefore, a greater risk of aerogenous infections exists, whereas in open-air pools this danger can be judged as very small.


Calcium homeostasis in Trypanosoma brucei. Identification of a pH-sensitive non-mitochondrial calcium pool.
The objective of this study was to characterize mechanisms which maintain intracellular calcium homeostasis in bloodstream forms of Trypanosoma brucei. The identification of homeostatic pathways is required to understand signal transduction in these organisms. The fluorescent probes Fura-2, 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, and bisoxonal were used to measure intracellular calcium ([Ca2+]i), intracellular pH (pHi), and membrane potential, respectively. Homeostatic pathways maintained [Ca2+]i at 98 +/- 12 nM in the presence of 1.8 mM extracellular calcium despite a steady leak of calcium into the cell. The addition of 2.7 microM nigericin acidified the cytosol, depolarized the plasma membrane, and induced an approximate 3-fold increase in [Ca2+]i. The rise in [Ca2+]i could not be induced with valinomycin or gramicidin D under conditions where membrane depolarization occurred. By contrast, the proton ionophore, carbonyl cyanide p-trifluoromethoxyphenylhydrazone, elevated [Ca2+]i in a manner that was not additive with nigericin. Changes in pHi appeared to regulate [Ca2+]i since: 1) stepwise addition of K+ to nigericin-treated cells generated and incremental increase in pHi and concomitant decrease in [Ca]i; 2) addition of serum to nigericin-treated cells allowed simultaneous recovery of pHi and [Ca2+]i; and 3) addition of NH4Cl to untreated cells resulted in a biphasic change in pHi with corresponding biphasic change in [Ca2+]i. The rise in [Ca2+]i was derived from an intracellular pool which was not dependent on functional cytochrome oxidase, mitochondrial alternative oxidase, or the F0F1-ATPase. These data demonstrate that large quantities of calcium are reversibly stored in a non-mitochondrial pH-sensitive intracellular pool in T. brucei. We conclude that changes in pHi can serve to trigger calcium signals in these organisms.


Nutritional status and depression, sleep disorder, and quality of life in hemodialysis patients.
The malnutrition-inflammation score (MIS) is a scoring system that measures malnutrition and inflammation. We sought to explore its associations with depression, sleep disturbance, and quality of life. This was a cross-sectional study. This study took place at the Baskent University Outpatient Hemodialysis Unit (Ankara, Turkey). We enrolled 67 hemodialysis patients (male/female, 34/33; age, 47.7 +/- 11.4 years [mean +/- SD]; hemodialysis duration, 103.7 +/- 59.1 months [mean +/- SD]). We retrospectively recorded patients' monthly clinical and laboratory findings from the previous 6 months. The same physician calculated MIS scores. We interviewed all patients, and each completed a Beck Depression Inventory (BDI) assessment. We used the Pittsburgh Sleep Quality Index (PSQI) to assess quality of sleep, and the Medical Outcomes Study 36-item short form (SF-36) questionnaire to evaluate health-related quality of life. The main outcome measures involved the univariate and multivariate relationships of the MIS with BDI, PSQI, and SF-36. Patients with PSQI scores of < or = 5 ("good sleepers") had lower MIS scores than did poor sleepers (6.8 +/- 2.5 vs. 8.8 +/- 3.2, P < .05). Patients with moderate-to-severe depression (BDI score > or = 19) had higher MIS scores (9.0 +/- 3.2 vs. 6.5 +/- 2.5, P = .005) and higher PSQI scores (7.6 +/- 2.1 vs. 4.7 +/- 1.8, P = .001), compared with patients with BDI scores < 19. Increased MIS scores were correlated with increased comorbidity (P = .01) and poor SF-36 scores (P = .009). Increased MIS is significantly associated with the presence of depression, sleep disorders, and poor quality of life. This close relationship may help establish the MIS as an important determinant of the increased morbidity and mortality of hemodialysis patients.


Magnetic resonance imaging of the pancreas.
In this article, the author discusses his experience using a superconducting magnet for evaluation of the pancreas. The discussion includes evaluation of neoplastic and inflammatory disease of the pancreas and comparison of MRI with CT scanning.


Lack of effects of a lyposterolic extract of Serenoa repens on plasma levels of testosterone, follicle-stimulating hormone, and luteinizing hormone.
Twenty men, aged 50 to 75 years (mean, 67 years), suffering from benign prostatic hypertrophy received 160 mg of a lyposterolic extract of Serenoa repens, twice daily for 30 days. Before and at the end of treatment, plasma levels of testosterone, follicle-stimulating hormone, and luteinizing hormone were determined. No changes in plasma hormone levels occurred as a result of treatment. It is concluded that Serenoa extract, which is useful in the treatment of benign prostatic hypertrophy, does not act via systemic changes of hormone levels.


Quadrupolar nutation NMR to discriminate central and satellite transitions: Spectral assignments for a Ziegler-Natta catalyst.
In this contribution we used solid state 35Cl (I=3/2) quadrupolar NMR to study a MgCl2/2,2-dimethyl-1,3-dimethoxypropane (DMDOMe) adduct that serves as a model system for Ziegler-Natta catalysis. Employing large Radio-Frequency (RF) field strengths we observe three spectral features with strongly varying line widths. The assignment of the spectra is complicated because of the large difference in quadrupolar interactions experienced by the different sites in the system. The satellite transitions (ST) of relatively well-defined bulk Cl sites are partially excited and may overlap with the central transition (CT) resonances of more distorted surface sites. We show that nutation NMR of the ST of I=3/2 spins yields a unique pattern that makes a clear distinction between an extensively broadened central transition and the satellite transitions of a component with a smaller quadrupolar interaction. This allows us to unambiguously unravel the spectra of the MgCl2 adduct showing that we observe CT and ST of the bulk phase of MgCl2-nanoparticles with a CQ of 4.6MHz together with the CT of surface sites displaying an average CQ of ∼10MHz.


Toxic levels of some heavy metals in drinking groundwater in Dakahlyia Governorate, Egypt in the year 2010.
Fifty-four drinking groundwater samples were collected in April 2010 from some districts of the Dakahlyia governorate, Egypt. The water samples were analyzed by atomic absorption spectrophotometer for iron, manganese, lead, nickel, chromium, zinc, copper, cobalt and cadmium concentrations. All samples but two were found suitable for drinking; from the water sample from Aga district showed slightly higher levels of cadmium and nickel (Ikhtab and Feshbena) than other areas; the concentrations were higher than the permissible limits of Egyptian Ministry of Health and World Health Organization.


Impact of newer unidirectional and bidirectional barbed suture on vesicourethral anastomosis during robot-assisted radical prostatectomy and its comparison with polyglecaprone-25 suture: an initial experience.
To evaluate impact of unidirectional barbed suture on vesicourethral anastomosis (VUA) during robot-assisted radical prostatectomy (RARP) and its comparison with Polyglecaprone-25 suture. The study was initiated as pilot study; the participants were grouped into three based on the suture material used for VUA, i.e., monofilament Polyglecaprone-25 (group 1), unidirectional barbed Glycolic acid-trimethylene carbonate (group 2), and bidirectional barbed Polyglycolic acid-polycaprolactone (group 3), respectively. Group 1 was included retrospectively and the latter two prospectively. All cases were operated upon by the same surgeon, proficient in RARP. Patient-demographics, intraoperative and peri-operative data were collected. Our technique of mucosa-to-mucosa VUA is carried out in a choreographed manner using unibarbed suture. Fifty-five patients were included; 25 each in group 1 and 2. The group 3 was prematurely closed after 5 cases due to perceived inappropriateness of needle characteristics of the suture material. Therefore, the statistical analysis was performed between group 1 and 2 only. Preoperative characteristics including age, PSA, clinical stage, and biopsy grade were similar between the groups. The anastomosis time was significantly less in group 2 (8.4 ± 1.7 min vs. 14.3 ± 4.8 min; P = 0.0001; t test). Postoperative hospital stay was less in group 2 (2.7 ± 1.1 days vs. 1.9 ± 0.8 days; P = 0.023; Mann-Whitney U). None of the patient had presented with urinary leaks, urinary retention, or anastomotic strictures at follow-up of 6 months. VUA with unidirectional barbed suture is safe and takes less time compared to monofilament suture as repeated cinching; help of assistance and knot tying are not required by virtue of its self-retaining characteristics.


A sensitive chromatographic method for the detection of pyruvyl groups in microbial polymers from sediments.
A method was developed for the quantitation of pyruvyl groups in microbial polymers using mild acid hydrolysis, o-phenylenediamine labeling, reversed-phase high-performance liquid chromatography (RP-HPLC), and fluorescence detection. The method was used to determine the pyruvate content of various microbial exopolysaccharides and to estimate the abundance of polymeric pyruvate in freshwater sediments. The results of this method were compared with those of several other pyruvate assays. The detection limit of the method was 1.6 nmol pyruvate. As little as 3.7μg of the bacterial polysaccharide xanthan gum, or from 5 to 22 mg of sediment (depending on polymeric pyruvate content), were needed for detection and quantitation of polymeric pyruvate. The results should be useful in determining the contribution of polymeric pyruvate to total metal-binding ligands in sediments.


Effect of time on the viability of ischemic skin flaps treated with vascular endothelial growth factor (VEGF) cDNA.
This study examined the efficacy of gene therapy on wound healing. The authors investigated whether delivery of the gene encoding a particular cytokine, known to be important in angiogenesis, could affect ischemic skin flaps. Anterior abdominal skin flaps, based solely on the epigastric artery and vein, were created in the Sprague-Dawley rat model. At the time of elevation, the arterial pedicle supplying each flap was infused either with the gene for vascular endothelial growth factor (VEGF) or physiologic saline alone. The flaps were resutured into place and observed for a period of either 4 or 3 days, at which time the pedicle was ligated. Twenty minutes following ligation, blood flow in the flaps was measured by dye fluorescence. Tissue viability of the flaps was subsequently measured by planimetry after a period of 7 days. Flaps that received the VEGF gene and were ligated at 4 days had an average dye fluorescence index (DFI) of 31.1 following ligation, and 93.9 percent viable tissue after 7 days. Flaps that received saline alone, and were ligated following a similar interval, had an average DFI of 14.0 and 31.9 percent viable tissue. Among the subjects that were ligated at 3 days, only a single, gene-infused flap had any noticeable viable tissue after 7 days. The DFI of these groups was 11.0 for the gene-infused group and 22.1 for the saline-infused group. The results suggest that delivery of the gene for VEGF can improve the survival of ischemic skin flaps, but that the effect of gene therapy is not limitless.


The role of interleukin-33 in chronic rhinosinusitis.
Interleukin (IL)-33, a new member of the IL-1 family, is constitutively expressed in epithelial tissues and lymphoid organs and plays an important role in the pathogenesis of allergic disease. However, the role of IL-33 in chronic rhinosinusitis with nasal polyps (CRSwNP) remains unclear. To investigate the role of IL-33 in the pathophysiology of CRSwNP. We investigated IL-33 expression and its cellular origins in the nasal polyps (NPs) of human subjects by immunohistochemistry (IHC), quantitative reverse transcription PCR (qRT-PCR), and multiplex cytokine assays. Correlations between IL-33 expression and other inflammatory markers were also explored. To investigate the role of IL-33 in CRSwNP, anti-IL-33 antibody was used in a murine model of CRS. Uncinate process tissues from control (19), CRSsNP (61), CRSwNP (69) and NP tissues (71) were used in this study. Increased expression of IL-33 mRNA and protein in patients with CRSwNP compared with controls was observed. The concentration of IL-33 protein in CRSwNP was positively correlated with the number of neutrophils and the expression of several Th1 and Th17 inflammatory markers, including interferon (IFN)-γ, IL-1β, tumour necrosis factor (TNF)-α, IL-17A, IL-22, and various markers for neutrophil recruitment. However, protein levels of IL-5 and quantity of eosinophils were inversely correlated with levels of IL-33. The expression of tissue inhibitor of metalloproteinase (TIMP)-1 was negatively correlated with IL-33 protein levels, while the expression of matrix metalloproteinase (MMP)-2 and MMP-9 was positively correlated with IL-33 protein levels. In animal studies, IL-33 expression was upregulated in the CRSwNP group compared with controls. Anti-IL-33 treatment reduced the thickness of oedematous mucosa, subepithelial collagen deposition, and infiltration of neutrophils, but infiltration of eosinophils was not reduced. This treatment also inhibited the expression of neutrophilic inflammatory cytokines, but not IL-4. In addition, the expression of intracellular adhesion molecule 1, vascular adhesion molecule 1 and CXCL-2 in the nasal mucosa was suppressed in mice treated with anti-IL-33 antibody. Our data suggest a role for IL-33 in the pathogenesis of CRSwNP via neutrophil recruitment. Therefore, anti-IL-33 may provide a new treatment strategy to target infiltrating neutrophils in CRSwNP.


Chronic diabetic complications in patients with MODY3 diabetes.
MODY3 diabetes, which is caused by a mutation in the hepatocyte nuclear factor-1alpha gene (HNF-1alpha) on chromosome 12, represents a relatively common monogenic form of diabetes in Finland. Age at onset of the disease can vary from 10 to 60 years, but little is known about the natural course of the disease, particularly the development of diabetes-related chronic complications. The availability of genetic markers now allows description of the clinical course of the disease. In order to examine the prevalence of chronic diabetic complications in MODY3, we examined 57 carriers with HNF-1alpha mutations for the presence of micro- and macrovascular complications. Thirty-four percent of the MODY patients had mild and 13% had severe non-proliferative or proliferative retinopathy; this figure did not differ from the figures in insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM) patients matched for duration and glycaemic control but not for age. Neither did the prevalence of microalbuminuria differ between MODY3 and IDDM or NIDDM patients (19 vs 24 and 23%). Neuropathy was observed with the same frequency as previously reported in IDDM. Hypertension was less frequent in MODY3 and IDDM than in NIDDM (24.5 and 19 vs 53.7%; p < 0.001). Coronary heart disease was more common in MODY3 than in IDDM (16 vs 4.5%; p < 0.02) but less common than in the older NIDDM patients (33.3%; p < 0.02). In a multiple logistic regression analysis, poor glycaemic control was an independent risk factor for retinopathy (p = 0.03), microalbuminuria (p < 0.04) and neuropathy (p = 0.03). In conclusion, microangiopathic complications are observed with the same frequency in patients with MODY3 diabetes as in IDDM and NIDDM and are strongly related to poor glycaemic control.


Rheumatoid meningitis: a localized immune process.
Rheumatoid pachymeningitis is a rare complication of rheumatoid arthritis. This disease was confined to the dura and pia-arachnoid of the lumbar cord in our patient. Her neurologic deficits responded to surgical decompression and corticosteroid therapy. Radiologic evidence and the differences in cell count, protein, and glucose content between lumbar and cisternal cerebrospinal fluid indicate that rheumatoid pachymeningitis can be localized to a discrete region of the central nervous system. Elevated immunoglobulins, IgM and IgG rheumatoid factors, low molecular weight IgM, and immune complexes were found in the cerebrospinal fluid and implicate an immune reaction in the pathogenesis of this disease, which is probably similar to inflammatory processes involving other organs in rheumatoid arthritis.


[Cervical cystic metastasis: differential diagnosis with branchiogenic carcinoma].
This paper try to call you attention about the cystic metastatic lesion and the differential diagnostic with the branchiogenic carcinoma. We presented a case of cystic metastases in the neck, and this case has prompted us to discuss the clinic features of these metastases.


Evaluation of a diagnostic algorithm for sputum smear-negative pulmonary tuberculosis in HIV-infected adults.
The Revised National TB Control Program bases diagnosis of tuberculosis (TB) on sputum smear examination and response to a course of antibiotics, whereas World Health Organization recommends early chest radiography [chest x-ray (CXR)] for HIV-infected symptomatic patients. We evaluated the utility of initial CXR in the diagnostic algorithm for symptomatic HIV-infected patients with negative sputum smears. HIV-infected ambulatory patients with cough or fever of ≥2 weeks and 3 sputum smears negative for acid-fast bacilli were enrolled in Chennai and Pune, India, between 2007 and 2009. After a CXR and 2 sputum cultures, a course of broad-spectrum antibiotics was given and patients were reviewed after 14 days. Sensitivity, specificity, positive and negative predictive values of symptoms, CXR, and various combinations for diagnosing pulmonary tuberculosis (PTB) were determined, using sputum culture as gold standard. Five hundred four patients (330 males; mean age: 35 years; median CD4: 175 cells per cubic millimeter) were enrolled. CXR had a sensitivity and specificity of 72% and 57%, respectively, with positive predictive value (PPV) of 21% and negative predictive value (NPV) of 93% to diagnose PTB. TB culture was positive in 49 of 235 patients (21%) with an abnormal initial CXR and 19 of 269 patients (7%) with a normal CXR (P < 0.001). Sensitivity and specificity of cough ≥2 weeks for predicting PTB was 97% and 6%, with PPV and NPV of 14% and 94%, respectively. Although moderately sensitive, basing a diagnosis of TB on initial CXR leads to overdiagnosis. An absence of weight loss had a high NPV, whereas none of the combinations had a good PPV. A rapid and accurate diagnostic test is required for HIV-infected chest symptomatic.


Results of kidney retransplantation.
Sixty-two patients had a second renal transplant at the University of Florida, Gainesville , between Nov 30, 1966 and June 26, 1982. Two-year graft function for all 62 patients was 53.2%, and patient survival was 83.9%. The 51 recipients of cadaveric grafts had a two-year graft survival of 47%. Eleven patients who received second grafts from related donors had a two-year graft survival of 81.8%. Source of first grafts and length of survival of first grafts did not seem to be important in determining the outcome of second grafts. Recipients of cadaveric second kidneys had a significantly better two-year graft survival if they waited longer than six months between loss of the first kidney and retransplantation (52.9%) than if they waited less than six months (35.3%). We believe the results of kidney retransplantation justify second transplants in patients who lost their first grafts.


Methylation status of EXT1 and EXT2 promoters and two mutations of EXT2 in chondrosarcoma.
Germline mutation and functional loss of EXT1 or EXT2 are commonly found in multiple osteochondromas and predispose to the development of chondrosarcoma. Mutations of EXT1 and EXT2 have rarely been detected in sporadic secondary chondrosarcomas from osteochondroma; these frequently display loss of heterozygosity at the EXT1 and EXT2 loci, but primary chondrosarcomas typically do not. To evaluate promoter methylation (which is an epigenetic gene silencing mechanism) of EXT1 and EXT2, we performed methylation-specific polymerase chain reaction (PCR) for 20 chondrosarcoma cases (12 primary, 3 secondary to osteochondroma, 2 secondary to enchondromatosis, 2 extraskeletal ordinary, and 1 clear cell) and in five cell lines. In addition, mutation analysis of the EXT1 and EXT2 coding regions was performed using PCR-single-strand conformation polymorphism and sequencing analysis for 12 of the 20 chondrosarcoma cases (8 primary, 1 secondary to enchondromatosis, 1 secondary to osteochondroma, and 2 extraskeletal ordinary) and five cell lines. Promoter methylation of EXT1 and EXT2 was not detected in any of the cases, and both EXT1 and EXT2 were expressed in all cell lines. Two missense mutations in EXT2 (D227E and R299H) were detected among the chondrosarcoma cases. When considering tumor development in primary chondrosarcoma, we should include mutations in EXT2, along with the status of other members of the EXT gene family.


An isotype-specific spot-ELISA for the enumeration of antibody-secreting hybridomas and the determination of isotype switch variants.
A solid-phase spot enzyme-linked immunosorbent assay (spot-ELISA) using rat monoclonal antibodies (MAbs) and an image-processing system is described. This isotype-specific spot-ELISA permits the enumeration of antibody-secreting cells irrespective of the specificity of the secreted antibodies. When used in combination with an ELISA, the antibody production per cell can also be evaluated. In addition, isotype switch variants, which arise spontaneously in antibody-producing cell lines, can be determined. This study compared four assays: three antigen-specific spot-ELISAs, using enzyme-conjugated polyclonal antibodies as well as rat MAbs; and an isotype-specific spot-ELISA using rat MAbs. There were no significant differences between these four spot-ELISA systems. For one tested cell line (alpha huIgA1/gamma 1), the number of antibody-secreting cells fluctuated between 60% and 95% during several passages. For the other tested cell line (alpha huIgA1/gamma 2b), the number of antibody-secreting cells decreased from 90% to 70% after several passages. The results of the spot-ELISA were in agreement with flow cytometric (FC) analysis of cytoplasmic IgG. This indicates that for these two cell lines, the synthesized IgG was also secreted into the culture fluid. Using the isotype-specific spot-ELISA, the switch frequency of five murine hybridomas (alpha huIgA1/gamma 1, alpha huIgA1/gamma 2b, alpha HRP, RIV6, MN12) was determined. The switch frequencies varied from 1/82,000 for the alpha HRP cell line to 1/660,000 for the alpha huIgA1/gamma 2b cell line.


Psychiatric, neuropediatric, and neuropsychological symptoms in a case of hypomelanosis of Ito.
This case report presents a thirteen year-old boy who was diagnosed as having Hypomelanosis of Ito. The developmental history includes severe failure to thrive, and moderate atypical autism as well as diverse clinical and neuropsychological symptoms are present. The pattern of neuropsychological functioning, which can be partially related to the neurophysiological findings, is discussed within the context of existing neuropsychological theories about autistic disorders.


Toxicity and antitumor activity of the vitamin D analogs PRI-1906 and PRI-1907 in combined treatment with cyclophosphamide in a mouse mammary cancer model.
Active and less toxic vitamin D analogs could be useful for clinical applications. In the present study, we evaluated the toxicity and antitumor effect of two new synthetic analogs of vitamin D, namely PRI-1906 [(24E)-24a-Homo-(1S)-1,25-dihydroxyergocalciferol] and its side-chain unsaturated homo analog PRI-1907. The toxicity and calcemic activity, as well as antitumor effect of calcitriol analogs was investigated in vivo. The studies were performed in a mouse mammary 16/C cancer model. Since calcitriol and its analogs inhibited 16/C tumor growth only slightly, we applied them in the combined therapy with cyclophosphamide (CY). Moreover, cell cycle analysis and VDR and p27 expression were investigated. The LD50 values after five daily subcutaneous (s.c.) injections were 7.8, 10.0 and 2.4 microg/kg per day for calcitriol, PRI-1906 and PRI-1907, respectively. The serum calcium level increased to 40, 23 and 63% over the control for these compounds. We also compare the antitumor activity of the PRI-1906 with the calcitriol and previously studied PRI-2191 (1,24-dihydroxyvitamin D3, tacalcitol). Statistically significant inhibition of tumor growth by calcitriol up to the eighth day was observed in all schedules applied. PRI-1906 inhibited the tumor growth at doses 1 and 5 microg/kg per day, and PRI-2191 only at the dose 5 microg/kg per day. Addition of vitamin D analogs increased the antitumor effect of CY. PRI-1906 exhibited toxicity higher than PRI-2191 but lower than calcitriol and antitumor activity similar to both PRI-2191 and calcitriol. This new analog seems to be a good candidate for the combined treatment of mammary cancer.


Ultrastructure of sperm, spermiogenesis, and sperm-egg interactions in selected invertebrates and lower vertebrates which use external fertilization.
This review discusses the ultrastructure of sperm with reference to their development, the surface morphology of the egg, and the processes of sperm binding and penetration during fertilization. These topics are treated for selected invertebrates and lower vertebrates which live in aquatic environments and fertilize their eggs externally. Specifically, sperm eggs from cnidarians, echinoderms, decapod crustaceans, ascidians, lampreys, bony fishes, and amphibians are discussed. Sperm from the majority of these groups exhibit the classical head-midregion-tail configuration characteristic of primitive sperm. Specific variations within this general morphology have been described. The notable exceptions to the primitive-sperm paradigm are the sperm of decapod crustaceans and amphibians. Eggs from all of the animals considered are covered by complex vitelline envelopes except those of cnidarians. In general, the ultrastructural analysis of these egg envelopes shows that they are composed of fibrous subunits. Sperm bind to the vitelline envelope and then penetrate through it to fertilize the egg in all groups reviewed except fishes. In sperm ultrastructure which occur during penetration of the egg envelopes in both flagellated and non-flagellated sperm. These changes, which involve membrane fusion and reorganization as well as movement of membranous organelles, aid the sperm in reaching the actual site of gamete fusion.


New hemodynamic approach to angiogenesis: color and pulsed Doppler ultrasonography.
To determine the capacity of color and spectral Doppler ultrasonography (US) to quantify angiogenesis in vivo and to characterize low-resistance intratumor blood flow. Thirty-two tumors, xenografted into mice, were studied with Doppler US. The number of intratumor vessels visualized with color Doppler US was compared with the density of microvessels and the number of vessels >100 microm determined by histologic examination. The resistance index and the peak systolic velocities were evaluated. The number of intratumor vessels visualized by color Doppler US was correlated with the number of vessels >100 microm (P<0.001) determined histologically. When vessel density was >30, intratumor vessels were always detected by color Dopper US. The resistance index and peak systolic velocities were significantly lower in intratumor than in peritumor vessels. Color Doppler US evaluated tumor angiogenesis accurately. Spectral analysis confirmed the low resistance of intratumor blood flow.


The glia-derived protease nexin 1 persists for over 1 year in rat brain areas selectively lesioned by transient global ischaemia.
The re-expression of the developmentally regulated serine protease inhibitor glia-derived nexin (GDN) was investigated 1 year after transient global ischaemia induced by the four-vessel occlusion technique in rats. The CA1 sector of the hippocampus was severely shrunken due to the absence of pyramidal cells, but still clearly discernible due to the continued presence of the parvalbumin-containing GABAergic neurons. In this partially neuron-depleted hippocampus, GDN immunoreactivity was found in reactive astrocytes containing glial fibrillary acidic protein. GDN-positive astrocytes were also found in other lesioned areas, the reticular thalamic nucleus and the cerebellar cortex. Thus, the re-expression of GDN in the adult excitotoxically lesioned brain described previously in the gerbil model of ischaemia persists. The continued presence of the protease inhibitor might disturb the proteolytic balance and lead to the deposition of pathological breakdown products of proteins, e.g. beta-amyloid.


Golimumab in the treatment of psoriatic arthritis.
Psoriatic arthritis (PsA) is an inflammatory arthritis that can be aggressive and destructive, resulting in significant morbidity. While many new agents have been approved for the treatment of PsA over the past decade, TNF inhibitors (TNFi)remain an anchor treatment, in part based on extensive clinical experience. Recently, the TNFi golimumab was approved for intravenous use in PsA. Areas covered: This expert review presents an overview of the currently available treatment options for PsA with a focus on the evidence from clinical trials supporting the use of golimumab in PsA. This information is placed in context with recent advances in the understanding of PsA pathogenesis and treatment. Expert commentary: The rapid growth of treatment options available for PsA has brought the prospect of personalized treatment selection closer to reality but improved understanding of the domains of PsA and new biomarkers may be needed before such changes reach the clinic. Until patients who are most likely to benefit from a given agent can be identified and then treated accordingly, TNFi will likely remain a central option and their further development, such as the introduction of an intravenous form of golimumab, remains an important part of treatment improvement.


Calcium phosphate cements as drug delivery materials.
Calcium phosphate cements are used as synthetic bone grafts, with several advantages, such as their osteoconductivity and injectability. Moreover, their low-temperature setting reaction and intrinsic porosity allow for the incorporation of drugs and active principles in the material. It is the aim of the present work to: a) provide an overview of the different approaches taken in the application of calcium phosphate cements for drug delivery in the skeletal system, and b) identify the most significant achievements. The drugs or active principles associated to calcium phosphate cements are classified in three groups, i) low molecular weight drugs; ii) high molecular weight biomolecules; and iii) ions.


Fluorescence lifetimes and spectral properties of protochlorophyllide in organic solvents in relation to the respective parameters in vivo.
In this work, absorption and fluorescence spectra of protochlorophyllide (Pchlide), as well as its fluorescence lifetime, were investigated in organic solvents having different physical properties. The obtained Pchlide spectral features are discussed in relation to the parameters describing solvent properties (refractive index and dielectric constant) and taking into account the specific solvent-Pchlide interaction. The correlation of Pchlide Qy and Soret absorption bands with solvent polarizability function ((n2 - 1)/(n2 + 2)) has been found; however, the dispersion of the observed points was rather high. A small Stokes shift of a magnitude between 50 and 300 cm(-1) was found, which indicates low sensitivity of Pchlide to nonspecific solvation. The fluorescence decay of Pchlide was single exponential in all the investigated solvents, with the lifetime value ranging from 5.2 ns for dioxane to 3.5 ns for methanol. Dependence of the obtained fluorescence lifetimes on the solvent orientation polarizability, a parameter being the function of both refractive index and dielectric constant, was discussed. In water-methanol mixtures, a further decrease of the fluorescence lifetime was observed, giving values of 2.9 ns for 25% methanol. Double-exponential decay of Pchlide fluorescence was found for Pchlide in a solution of 15% methanol with the lifetimes of 4.5 +/- 0.5 ns and 1.2 +/- 0.3 ns and in pure water with the lifetimes of 2.5 +/- 0.5 ns and 0.4 +/- 0.1 ns. The obtained results are discussed in relation to spectroscopic properties of Pchlide in vivo.


A structural-energetic basis for B-cell epitope prediction.
Structural-energetic analysis of peptide and protein antigens in the context of binding to antibody reveals fundamental differences between the cross-reactions of antipeptide antibody with protein and antiprotein antibody with peptide, providing a physicochemical basis for B-cell epitope prediction as applied to the development of peptide-based vaccines and immunodiagnostics.


Adsorption thermodynamic characteristics of Chlorella vulgaris with organic polymer adsorbent cationic starch: Effect of temperature on adsorption capacity and rate.
Aiming at optimizing the adsorption process of Chlorella vulgaris and cationic starch, the adsorption thermodynamic characteristics were evaluated. Different from inorganic calcium salt adsorbent, the adsorption nature of organic polymer cationic starch is exothermic (ΔH° < 0) and spontaneous (ΔG° < 0). Besides, the adsorption capacity and rate can be well described by Langmiur isotherm and pseudo second kinetic models. As results of exothermic nature and great driving force of lower temperature, the adsorption capacity and rate declined with the rising temperature. The maximal values of them were obtained at 278.15 K, which were 9148.14 mg microalgae (g cationic starch)-1 and 8.74 × 10-6 mg g-1 min-1. Additionally, with insufficient adsorbent, the highest adsorption efficiency (96.37%) was achieved at 278.15 K for stirring 150 min. For 288.15, 298.15, 308.15 and 318.15 K, the adsorption efficiency decreased to 93.77%, 86.75%, 83.32% and 81.57% and the time consumed were at least 40 min longer.


Extra-articular reconstructive surgery for combined anterolateral-anteromedial rotatory instability.
The knees of 14 patients (12 male and 2 female) were treated surgically by extra-articular reconstruction for combined anterolateral-anteromedial rotatory instability. Healing and rehabilitation required approximately 8 months after the operations. Only after adequate rehabilitation, including return of full confidence, were the patients released to full activity (although contact sports are not recommended). Of the group, 12 patients returned for follow-up evaluations and one returned a questionnaire. One patient was lost to follow-up. One patient did not return to full activity because in the 3rd month postoperatively, he stepped in a hole while jogging, reinjured his knee, and refused further surgery. The functional improvement in this series by our extra-articular procedure (a combination of an anteromedial capsulorrhaphy and pes transfer medially with Ellison's modification of an iliotibial band rerouting and partial biceps advancement laterally) compares favorably with the reported anterior cruciate ligament reconstructions. Further comparisons with longer periods of time after surgical procedures are needed to determine whether extra-articular repair with interarticular reconstruction of the anterior cruciate ligament is significantly better than this technically easier extra-articular repair alone.


Changing a state mental health system through litigation: the Arizona experiment.
Litigation may be a viable means to change mental health systems for the chronically mentally ill. Court orders achieved in the 1970s have been ignored or proven inefficient over the years. However, in Arizona, a strategy was designed by a team of lawyers and psychiatrists to deal with failures seen elsewhere in court actions seeking adequate services for chronically mentally ill patients. This novel approach produced a favorable court outcome and triggered a chain reaction at the executive and legislative levels to reform radically the system of care for the chronically mentally ill in Arizona.


Muscle-specific RING finger-2 (MURF-2) is important for microtubule, intermediate filament and sarcomeric M-line maintenance in striated muscle development.
The efficient functioning of striated muscle is dependent upon the structure of several cytoskeletal networks including myofibrils, microtubules, and intermediate filaments. However, little is known about how these networks function together during muscle differentiation and maintenance. In vitro studies suggest that members of the muscle-specific RING finger protein family (MURF-1, 2, and 3) act as cytoskeletal adaptors and signaling molecules by associating with myofibril components (including the giant protein, titin), microtubules and/or nuclear factors. We investigated the role of MURF-2, the least-characterized family member, in primary cultures of embryonic chick skeletal and cardiac myocytes. MURF-2 is detected as two species (approximately 55 kDa and approximately 60 kDa) in embryonic muscle, which are down-regulated in adult muscle. Although predominantly located diffusely in the cytoplasm, MURF-2 also colocalizes with a sub-group of microtubules and the M-line region of titin. Reducing MURF-2 levels in cardiac myocytes using antisense oligonucleotides perturbed the structure of stable microtubule populations, the intermediate filament proteins desmin and vimentin, and the sarcomeric M-line region. In contrast, other sarcomeric regions and dynamic microtubules remained unaffected. MURF-2 knock-down studies in skeletal myoblasts also delayed myoblast fusion and myofibrillogenesis. Furthermore, contractile activity was also affected. We speculate that some of the roles of MURF-2 are modulated via titin-based mechanisms.


Radiation therapy salvage of Hodgkin's disease following chemotherapy failure.
Between 1972 and 1984, 17 patients with advanced Hodgkin's disease failing intensive combination chemotherapy in previously unirradiated nodal and/or pulmonary sites were treated with salvage radiotherapy. Treatment consisted of comprehensive wide field radiotherapy to all known areas of disease. Doses administered to these fields ranged from 1,700 to 5,000 rad, with only three patients (18%) receiving less than 3,000 rad to any field. With a median follow-up of over 4 years, 88% achieved a complete response, with median actuarial disease-free survival (DFS) of 19 months (range, 4 to 61+). Actuarial median survival was 64 months, with a range of 4 to 134+ months. Nine patients (53%) are currently alive with three (18%) in continuous complete remission (CR) for 24, 30, and 61 months. In addition, four patients relapsing after salvage radiotherapy are now in CR following additional therapy. Patients younger than 35 years of age had a significantly increased overall survival when compared with older patients (P less than .005). An initial complete response to chemotherapy lasting 12 or more months appeared to be a favorable prognostic factor, although small patient numbers preclude statistical significance. Comprehensive salvage radiotherapy is of significant benefit in patients with advanced Hodgkin's disease relapsing after combination chemotherapy in nodal and/or pulmonary sites.


Prorocentrol, a polyoxy linear carbon chain compound isolated from the toxic dinoflagellate Prorocentrum hoffmannianum.
A polyoxy linear carbon chain compound, prorocentrol (1), was isolated from cultured cells of the dinoflagellate Prorocentrum hoffmannianum, which produces a polyether carboxylic acid, okadaic acid. The structure of 1 was elucidated by detailed analyses of 2D NMR spectra. Compound 1 possesses 30 hydroxy groups, 1 ketone, and 8 double bonds on the C65-linear carbon chain. Its partial relative configuration was deduced by the proton-proton and long-range carbon-proton coupling constants, and compound 1 showed moderate cytotoxicity and antidiatom activity.


Generation of lipid neutrophil chemoattractant by irradiated bovine aortic endothelial cells.
Radiation injury to blood vessels is associated with an acute inflammatory process. We investigated the capacity of cultured bovine aortic endothelial cells (BAEC) to produce chemotactic factors after radiation injury. BAEC in serum-free media were irradiated with a cobalt-60 Gammacell 220 and the cell supernatants were assayed for chemotactic activity for human neutrophils in a Boyden chamber. There was a rapid release of chemotactic activity into the BAEC supernatants which was dependent both on the dose of radiation (5 to 40 Gy) and the time between irradiation and sample collection. In contrast, isolation of BAEC lysates by freeze-thawing was not associated with the presence of similar chemotactic activity. The chemotactic activity released from the irradiated BAEC was not destroyed by boiling nor by treatment with trypsin. The release of the chemotactic activity was, however, inhibited by the addition of a lipoxygenase inhibitor but not by the addition of a cyclooxygenase inhibitor before the irradiation. The chemotactic activity was recovered from the cell supernatants in the lipid phase after extraction with chloroform/methanol. Furthermore, the chloroform/methanol extracts co-eluted with authentic leukotriene B4 when the BAEC were prelabeled with [14C] arachidonic acid. However, we were unable to detect endogenous leukotriene B4 with RIA. Instead, the only detectable endogenous lipid present in the supernatants was 13-hydroxyoctadecadienoic acid which is derived from linoleic acid via the lipoxygenase pathway. 13-Hydroxyoctadecadienoic acid, however, had no chemotactic activity. These findings suggest that endothelial cells rapidly release a chemotactic agent after irradiation, the release of which is associated with a lipoxygenase pathway. The release of this chemotactic activity may account in part for the acute inflammatory response that is observed after ionizing irradiation.


A retrospective comparison of allogeneic peripheral blood stem cell and bone marrow transplantation results from a single center: a focus on the incidence of graft-vs.-host disease and relapse.
To detect the effect of the stem cell source, allogeneic peripheral blood stem cell transplantations (alloPBSCTs) performed between 1995 and 1997 from human leukocyte antigen (HLA)-identical siblings in 40 patients with acute and chronic hematological disorders were compared with a historical group of 40 patients with similar variables who had received allogeneic bone marrow transplants (alloBMTs) between 1993 and 1995. Patients in both groups were identical except that both the recipient and the donor ages were, on average, higher in the alloPBSCT group (26 vs. 36 [p = 0.005] and 27 vs. 32 [p = 0.024], respectively). Patients received similar therapy excluding posttransplant granulocyte colony-stimulating factor administration (97% in alloBMT vs. 12.5% in alloPBSCT). The median time to reach neutrophil counts >0.5 x 10(9)/L and platelet counts >20 x 10(9)/L was 13 and 14 days, respectively, in patients receiving alloPBSCTs compared with 19 and 27 days in patients receiving alloBMTs (p = 0.0014 and p = 0.0002). The alloPBSCT group required similar transfusions of red blood cells or platelets. The incidence of grade II-IV acute graft-vs.-host disease (aGVHD) was similar in both groups. However, chronic GVHD (cGVHD) of all grades developed in 78.1% of patients in the alloPBSCT group after a median follow-up period of 12.5 (range 0.5-34) months. In alloBMT recipients, cGVHD of all grades developed in 21.4% after a median follow-up period of 38 (range 0.5-62) months (p = 0.00001). Day 100 transplant-related mortality was also similar: 20% (8 of 40) in the alloBMT patients and 17.5% (7 of 40) in the alloPBSCT group. Although not statistically significant, a relatively higher relapse rate occurred in the alloBMT group (21.4 vs. 10.7%). The estimated disease-free survival in month 24 was 51.3% for alloBMT and 54.6% for alloPBSCT, and the estimated overall survival in month 24 was 56.1% for alloBMT and 64.6% for alloPBSCT. In conclusion, this retrospective comparison suggests that alloPBSCT from HLA-identical donors is associated with faster engraftment, fewer transfusions, and no greater incidence of aGVHD, but a high incidence of cGVHD.


Biotransformation of Ni-substituted hydrous ferric oxide by an Fe(III)-reducing bacterium.
The reductive biotransformation of a Ni(2+)-substituted (5 mol %) hydrous ferric oxide (NiHFO) by Shewanella putrefaciens, strain CN32, was investigated under anoxic conditions at circumneutral pH. Our objectives were to define the influence of Ni2+ substitution on the bioreducibility of the HFO and the biomineralization products formed and to identify biogeochemical factors controlling the phase distribution of Ni2+ during bioreduction. Incubations with CN32 and NiHFO were sampled after 14 and 32 d, and both aqueous chemistry and solid phases were characterized. By comparison of these results with a previous study (Fredrickson, J. K.; Zachara, J. M.; Kennedy, D. W.; Dong, H.; Onstott, T. C.; Hinman, N. W.; Li, S. W. Geochim. Cosmochim. Acta 1998, 62, 3239-3257), it was concluded that coprecipitated/sorbed Ni2+ inhibited the bioreduction of HFO through an undefined chemical mechanism. Mössbauer spectroscopy allowed analysis of the residual HFO phase and the identity and approximate mass percent of biogenic mineral phases. The presence of AQDS, a soluble electron shuttle that obviates need for cell--oxide contact, was found to counteract the inhibiting effect of Ni2+. Nickel was generally mobilized during bioreduction in a trend that correlated with final pH, except in cases where PO4(3-) was present and vivianite precipitation occurred. CN32 promoted the formation of Ni(2+)-substituted magnetite (Fe2IIIFe(1-x)IINixIIO4) in media with AQDS but without PO4(3-). The formation of this biogenic coprecipitate, however, had little discernible impact on final aqueous Ni2+ concentrations. These results demonstrate that coprecipitated Ni can inhibit dissimilatory microbial reduction of amorphous iron oxide, but the presence of humic acids may facilitate the immobilization of Ni within the crystal structure of biogenic magnetite.


Effect of 4 weeks of Pilates on the body composition of young girls.
There is a need to find ways to increase the physical activity levels and improve the body composition and blood pressure of girls. Thirty 11-year-old girls were recruited from two after school programs in Houston Texas in Spring 2005. Participants from one program (16) were randomly assigned to intervention, the other (14) served as controls. BMI, BMI percentile, waist circumference and blood pressure were assessed before and after the intervention. Pilates classes were provided free of charge for an hour per day at the intervention site, 5 days a week, for 4 weeks. Four participants wore heart rate monitors during every session and completed enjoyment and perceived exertion questionnaires. Repeated measures analysis of variance with time (within) and group (between) as factors was performed. Mean attendance was 75%, mean heart rate 104 bpm, mean perceived exertion 5.9 (1-10 scale) and enjoyment 4.4 (1-5 scale). There was a significant (P = 0.039) time by group interaction for BMI percentile. Graphs indicated that this difference was influenced by large reductions in the BMI percentile of healthy girls. Girls enjoyed Pilates, and participation for 4 weeks lowered BMI percentile. Pilates holds promise as a means of reducing obesity.


Changes in nutritional status in nursing home residents and associated factors in nutritional status decline: a secondary data analysis.
The aim of this study was to describe changes in the nutritional status of nursing home residents over a period of 1 year and identify factors associated with a decline in nutritional status. The maintenance of good nutritional status is important for nursing home residents. Therefore, it is essential to identify risk factors that indicate a decline in nutritional status to take early prevention steps. Secondary data analysis of repeated cross-sectional studies. Data collection was performed between 2009-2013 using the International Prevalence Measurement of Care Problems questionnaire. Data from three purposively selected nursing homes that included 157 residents were analysed. The comparison between baseline data and data collected 1 year later showed that the nutritional status declined in 22·8% of the residents and improved in 6·5% of the cases. The body mass index and changes in the body mass index were significantly different between residents with stable/improved and declined nutritional status. The multivariate logistic regression analysis showed that care dependency, length of stay, changes in body mass index and malnutrition risk at baseline were significantly associated with a decline in nutritional status. This study showed that the baseline risk of malnutrition is the most important risk factor indicating a decline in nutritional status. Therefore, healthcare professionals should identify the malnutrition risk and take action as early as possible.


In vivo BDNF modulation of adult functional and morphological synaptic plasticity at hippocampal mossy fibers.
Brain-derived neurotrophic factor (BDNF) has been proposed as a key regulator and mediator of long-term synaptic modifications related to learning and memory maintenance. Our previous studies show that application of high-frequency stimulation (HFS) sufficient to elicit LTP at the dentate gyrus (DG)-CA3 pathway produces mossy fiber structural modifications 7 days after tetanic stimulation. In the present study, we show that acute intrahippocampal microinfusion of BDNF induces a lasting potentiation of synaptic efficacy in the DG-CA3 projection of anesthetized adult rats. Furthermore, we show that BDNF functional modifications in synaptic efficacy are accompanied by a presynaptic structural long-lasting reorganization at the hippocampal mossy fiber pathway. These findings support the idea that BDNF plays an important role as synaptic messenger of activity-dependent synaptic plasticity in the adult mammalian brain, in vivo.


Absorption and disposition of ethambutol in rabbits.
The absorption and disposition of ethambutol was examined in six rabbits in a three-way crossover study. Each rabbit received 45-mg/kg doses of ethambutol in three treatments: one intravenous injections and two oral solutions, ethambutol alone and ethambutol in the presence of aluminum hydroxide (40 mg/kg). Half-lives of ethambutol ranged from 2.26 to 5.20 h when administered alone and 2.18 to 4.00 h when coadministered with the antacid; the difference was not significant (p greater than 0.3). Mean clearance after the oral administrations (189.2 mL/min/kg) was significantly greater than the mean intravenous clearance (43.7 mL/min/kg) (p less than 0.01), suggesting a first-pass metabolism of ethambutol when administered nonparenterally to rabbits. The volume of distribution ranged from 5.5 to 17.8 L/kg, suggesting an extensive distribution of ethambutol outside the central compartment and, possibly, a localized deposit within the body tissues. Mean bioavailability of ethambutol was approximately 28% and was not affected by the presence of aluminum hydroxide. The rate of ethambutol absorption, however, was slightly delayed by the antacid.


The exon-3-encoded domain of IL-15ralpha contributes to IL-15 high-affinity binding and is crucial for the IL-15 antagonistic effect of soluble IL-15Ralpha.
We previously showed that a natural soluble form of interleukin-15 (IL-15) Ralpha corresponding to the full-length ectodomain of IL-15Ralpha behaved as a potent antagonist of IL-15 action through IL-15Ralpha/beta/gamma, whereas a recombinant soluble IL-15Ralpha sushi domain did not, but instead acted as an agonist of IL-15 action through IL-15Rbeta/gamma. In order to determine precisely the molecular basis governing these antagonistic versus agonistic actions, we compared the binding properties and biological effects of recombinant soluble IL-15Ralpha (sIL-15Ralpha) species containing the sushi domain and different remaining parts of the ectodomain. We first demonstrate that the exon-3-encoded domain and, more particularly, its N-terminal 13-amino-acid (aa) peptide are important, in addition to the adjacent exon-2-encoded sushi domain, for the stabilization of the high-affinity IL-15.IL-15Ralpha complex by slowing down its dissociation rate and by contributing to about 10-20% of the free energy of interaction. We next show that all sushi-containing sIL-15Ralpha are agonists on IL-15Rbeta/gamma, coordinately increasing IL-15 binding and IL-15-induced proliferation. Their agonistic potencies are proportional to their respective affinities for IL-15. We then show that the antagonistic effect of sIL-15Ralpha in the context of IL-15Ralpha/beta/gamma is due to the 13-aa peptide that creates a sterical constraint impeding the binding of the sIL-15Ralpha.IL-15 complex to the membrane-anchored IL-15Ralpha/beta/gamma. In the frame of the soluble IL-15Ralpha sushi domain-IL-15 fusion protein that contains the 13-aa peptide, this constraint is alleviated as a result of a conformational effect due to the covalent linking of the 13-aa peptide to the N-terminus of IL-15. The soluble IL-15Ralpha sushi domain-IL-15 fusion protein is therefore able to bind and activate both the IL-15Rbeta/gamma and the IL-15Ralpha/beta/gamma receptors.


A test of the auricular surface aging technique.
A recently developed technique used in the estimation of age at death is based upon the metamorphosis of the auricular surface of the ilium. This technique was designed to be used in seriation to estimate the age distribution of a skeletal sample. However, the technique has also been used in forensic science cases, which must be analyzed on a case-by-case basis. The present study examines the use of the auricular surface technique as the single aging factor. Two hypotheses are tested using a sample of 189 individuals drawn from the Terry Collection, housed at the Smithsonian Institution. The hypotheses are (1) is the method equally applicable across race and sex, and (2) how well does the method perform as the single aging factor? The results indicate that the amount of degenerative change in the auricular surface is not dependent upon race or sex in any given age category. However, the rate of degenerative change is too variable to be used as a single criterion for the estimation of age; the range of estimation error is simply too large for forensic science purposes.


Minimally invasive mitral valve surgery is associated with equivalent cost and shorter hospital stay when compared with traditional sternotomy.
Mitral valve surgery is increasingly performed through minimally invasive approaches. There are limited data regarding the cost of minimally invasive mitral valve surgery. Moreover, there are no data on the specific costs associated with mitral valve surgery. We undertook this study to compare the costs (total and subcomponent) of minimally invasive mitral valve surgery relative to traditional sternotomy. All isolated mitral valve repairs performed in our health system from March 2012 through September 2013 were analyzed. To ensure like sets of patients, only those patients who underwent isolated mitral valve repairs with preoperative Society of Thoracic Surgeons scores of less than 4 were included in this study. A total of 159 patients were identified (sternotomy, 68; mini, 91). Total incurred direct cost was obtained from hospital financial records. Analysis demonstrated no difference in total cost (operative and postoperative) of mitral valve repair between mini and sternotomy ($25,515 ± $7598 vs $26,049 ± $11,737; P = .74). Operative costs were higher for the mini cohort, whereas postoperative costs were significantly lower. Postoperative intensive care unit and total hospital stays were both significantly shorter for the mini cohort. There were no differences in postoperative complications or survival between groups. Minimally invasive mitral valve surgery can be performed with overall equivalent cost and shorter hospital stay relative to traditional sternotomy. There is greater operative cost associated with minimally invasive mitral valve surgery that is offset by shorter intensive care unit and hospital stays.


Auditory-somatosensory multisensory processing in auditory association cortex: an fMRI study.
Using high-field (3 Tesla) functional magnetic resonance imaging (fMRI), we demonstrate that auditory and somatosensory inputs converge in a subregion of human auditory cortex along the superior temporal gyrus. Further, simultaneous stimulation in both sensory modalities resulted in activity exceeding that predicted by summing the responses to the unisensory inputs, thereby showing multisensory integration in this convergence region. Recently, intracranial recordings in macaque monkeys have shown similar auditory-somatosensory convergence in a subregion of auditory cortex directly caudomedial to primary auditory cortex (area CM). The multisensory region identified in the present investigation may be the human homologue of CM. Our finding of auditory-somatosensory convergence in early auditory cortices contributes to mounting evidence for multisensory integration early in the cortical processing hierarchy, in brain regions that were previously assumed to be unisensory.


Evolution of the androgen receptor pathway during progression of prostate cancer.
The present work focused on the potential involvement of selective adaptations of the androgen receptor pathway in the initiation and progression of prostate cancer. We defined the androgen receptor pathway by selecting 200 genes that were androgen responsive in prostate cancer cell lines and/or xenografts. This androgen receptor pathway gene signature was then used for profiling prostate cancer xenografts and patient-derived samples. Approximately half of the androgen receptor pathway genes were up-regulated in well-differentiated prostate cancer compared with normal prostate. Functionally distinct parts of the androgen receptor pathway were specifically down-regulated in high-grade cancers. Unexpectedly, metastases have down-regulated the vast majority of androgen receptor pathway genes. The significance of this progressive down-regulation of androgen receptor pathway genes was shown for a few androgen receptor-regulated genes. Lower mRNA expression of HERPUD1, STK39, DHCR24, and SOCS2 in primary prostate tumors was correlated with a higher incidence of metastases after radical prostatectomy. HERPUD1 mRNA expression predicted the occurrence of metastases almost perfectly. In vitro experiments showed that overexpression of the stress response gene HERPUD1 rapidly induces apoptosis. Based on the functions of the genes within the distinct subsets, we propose the following model. Enhanced androgen receptor activity is involved in the early stages of prostate cancer. In well-differentiated prostate cancer, the androgen receptor activates growth-promoting as well as growth-inhibiting and cell differentiation genes resulting in a low growth rate. The progression from low-grade to high-grade prostate carcinoma and metastases is mediated by a selective down-regulation of the androgen receptor target genes that inhibit proliferation, induce differentiation, or mediate apoptosis.


Evaluation of early enteral feeding in children less than 3 years old with smaller burns (8-25 per cent TBSA).
Early enteral feeding and high protein nutrition have been advocated for burned patients. The safety and efficacy of early high protein nasogastric feeding (NG) have not been evaluated in very young children. The present study evaluated such feeding in children less than 3 years old with smaller burns (8-25 per cent of total body surface area). Children (n = 10) were able to tolerate high protein intake without detrimental effects. The incidences of gastrointestinal complications related to NG feeding were low. The children needed approximately 2 weeks of supplemental NG feeding which provided two-thirds of total energy intake and three-fourths of protein intake. In spite of smaller burns, the mean measured resting metabolic expenditure (REE) was 1.3 x predicted REE. The mean energy intake of 92 per cent of recommended daily allowances (RDA) for energy or 1.7 x predicted REE was able to maintain body weight. The mean protein intake was 4.3 g/kg/day with a non-protein calorie ratio of 114:1. During the first week postburn, plasma concentrations of prealbumin, albumin and transferrin were low. The high protein intake was able to raise these visceral proteins to normal ranges. These results indicate that early NG feeding is safe and efficacious for achieving increased energy intake and improved protein status in very young children.


Serially subcultivated cells as substrates for poliovirus production for vaccine.
Four human diploid cell cultures, WI-38, MRC-5, HEL 299, and IMR-90, and the aneuploid simian cell culture LLC-MK2 were considered in this study as alternatives to primary monkey kidney cells in the production of Inactivated Poliomyelitis Vaccine (IPV). The composition of the basal media used for cell growth and viral replication was found to be critical. Medium CMRL-1969 required a 50% increase in the concentrations of arginine, cystine, glutamine, isoleucine, leucine, methionine, serine, and threonine to produce optimum cell and viral antigen yields, particularly in the Multi-Surface-Cell-Propagator (MSCP) and in the microcarrier suspension culture systems. Also, Medium 199, with Earle's Basic Salt Solution (BSS) for the human diploid cultures, and Hanks' BSS for the simian aneuploid culture, increased the D-antigen yields when compared with other virus growth media deficient in nucleic acid precursors. Approximately 3--4 cell population doublings were obtained in microcarrier culture systems. The average D-antigen yields for poliovirus Types 1,2 and 3 were about 110, 25 and 35, respectively. The physiological requirements for the four human diploid cell cultures were similar indicating easy interchange. The major advantage of the aneuploid LLC-MK2 cells is the reduced serum requirement. The microcarrier culture system is suitable for these cell substrates in the production of IPV provided the microcarriers can be kept in "single-bead" suspension at low rotational speeds of, initially, 10 rpm, with slight increases as the cell density increases.


Insulinoma: Diagnosis and surgical treatment. Retrospective analysis of 31 cases.
Insulinomas are rare tumors that are usually benign, single and curable by simple surgical excision. They can present problems in diagnosis and localization. Retrospective analysis of patients with insulinoma managed during a 13-year period (1992-2005) at a tertiary-level institution. 31 patients (mean age 38.4 [SD 13.3] years; 16 men) presented with hypoglycemic symptoms for 4.6 (5.5) years. In 22 (71%) patients, the lesion was successfully localized pre-operatively. Of various pre-operative localization techniques, CT angiography (5/6; 83%), intra-arterial digital subtraction angiography (11/17; 65%), dual-phase CT (8/14; 57%) and conventional MRI (4/13; 31%) had high rates of successful tumor localization. Intra-operative palpation and ultrasonography also had localization success rates (22/30 [76%] and 11/12 [92%], respectively); each identified one lesion that the other procedure did not localize. Of the 30 patients who underwent surgery, 28 had solitary tumor. Pre-operative investigations to localize insulinoma are helpful despite the availability of intra-operative ultrasound. Dual-phase CT should be the non-invasive investigation of first choice.


Silver-Mediated Trifluoromethylthiolation-Iodination of Arynes.
A one-pot trifluoromethylthiolation-iodination of arynes with trifluoromethylthiosilver (AgSCF3) and 1-iodophenylacetylene is described. This protocol allows rapid construction of o-trifluoromethylthiolated arene building blocks in moderate yields. These products were found to be excellent precursors for Yagupolskii-Umemoto-type electrophilic trifluoromethylation reagents.


[Comparison of management of ambulatory emergency patients in the family practice and the hospital emergency center].
The increase in health care cost is broadly discussed in the public, either in the hospital or ambulatory setting. With the present study we compared the invoice amount generated by a first doctor's advice in the general practitioner to the one in the emergency room. In addition, we compared the diagnostic procedures used in the general practice to the one in the hospital, and whether the choice of diagnostic procedures would substantially influence the invoice amount. The invoice amount in the hospital setting was on average CHF 197.-, i. e. 134%, higher compared to the general practice. The difference in the invoice amount was largely explained by more diagnostic procedures in the hospital. Moreover, part of the higher invoice amount was generated by the extensive physical examination by doctors in the hospital.


Identification of bloodmeals in haematophagous Diptera by cytochrome B heteroduplex analysis.
We developed a DNA assay for bloodmeal identification in haematophagous insects. Specific host cytochrome B gene sequences were amplified by PCR and classified on the basis of their mobility in a heteroduplex assay. In the blackfly Simulium damnosum s.l. (Diptera: Simuliidae), human cytochrome B DNA sequences were identifiable up to 3 days following ingestion of the bloodmeal. In the tsetse Glossina palpalis (Diptera: Glossinidae) collected from tsetse traps in Ivory Coast, bloodmeals were identified as taken from domestic pigs on the basis of their heteroduplex pattern and DNA sequence. Evidently the cytochrome B sequence shows sufficient interspecific variation to distinguish between mammalian host samples, while exhibiting minimal intraspecific variation. The stability of DNA in bloodmeals, for several days post-ingestion by haematophagous insects, allows PCR-HDA assays to be used reliably for host identification.


Management information system for leprosy eradication programme--an alternative information system.
For efficient monitoring of multidrug therapy programmes for leprosy both at microlevel (individual patient monitoring) as well as microlevel (programme monitoring), DANIDA decided to develop an alternative, simple and quick information system using a computer. A patient data base system was designed using dBase III Plus package. The field workers of the National Leprosy Eradication Programme were trained in transcribing data on to coded data sheets. The data of 1750 patients of six leprosy control units from the 4 MDT districts were processed and feedback reports were sent to paramedical workers and programme managers. The initial experience in the field over the past year has shown that a computerized management information system is feasible and well accepted by the field staff for the purpose of improving monitoring.


Monitoring of low molecular weight heparin anticoagulation during haemodialysis with a Sonoclot Analyzer.
Sonoclot was used in this study to monitor low molecular weight heparin (LMWH) during haemodialysis. Two different intravenous doses (standard / half-dose) of dalteparin were studied in eight patients. Blood was sampled for coagulation analyses with Sonoclot, thrombin-antithrombin (TAT) and anti-Xa. A visual fibrin deposition score (VFS) in the venous drip chamber was also evaluated. All patients completed their dialysis. There was a progressive increase in TAT levels, which correlated to the dalteparin dose. Significant differences (p<0.05) were found for TAT, VFS and Sonoclot celite-activated clotting time (SonACT) between the different LMWH dosages. TAT and Sonoclot correlated to each other, but not to the VFS. SonACT was significantly increased at two hours, with the high dalteparin dose compared to the lower dose. Both Sonoclot and TAT failed to predict the VFS. No patient had any clinical clotting events and all dialyses were completed successfully.


Radiographic assessment of coxarthrosis following slipped capital femoral epiphysis. A 32-year follow-up study of 151 hips.
In all, 108 patients (151 hips) previously treated for slipped capital femoral epiphysis (SCFE), were evaluated radiographically for assessment of coxarthrosis at an average age of 47 years. Five projections were obtained of each hip (supine a.p., supine modified frog lateral, standing a.p., standing anterolateral oblique, and standing posterolateral oblique) and the superior, anterior, and posterior joint space heights were measured (abnormal: superior < or = 3.0 mm and/or anterior and/or posterior < or = 2.5 mm). Sixty-three hips (42%) had joint space narrowing in at least one projection. The standing a.p. and 2 lateral oblique views identified 54 hips as abnormal, and the supine a.p. and modified frog lateral views uncovered an additional 9. The supine a.p. view was the least helpful; 12 abnormal hips on the standing a.p. view were normal on the supine a.p. view. Isolated narrowing in the anterior and/or posterior joint space was found in 9 hips (6%). These changes could only be assessed by the 2 lateral oblique views. Our results illustrate that a combination of standing radiographs, visualising the circumference of the articular space of the hip joint, and modified frog lateral projections is necessary to fully identify coxarthrosis in hips previously treated for SCFE.