Top-down therapy: is the evidence strong enough?
Crohn's disease (CD) has usually been managed in an escalation manner, introducing more powerful (and toxic) drugs only once those with a better safety profile had failed. However, the natural history of CD under conventional therapeutic strategies results in high intestinal resection requirements and high rates of clinical relapse and steroid dependence. Indirect data seem to point at an improved efficacy of drugs when they are introduced early after disease diagnosis. The spreading use of immunomodulators and the appearance of biological agents prompted the idea of their early introduction in order to change the natural history of the disease. By now, only thiopurines have been shown to reduce steroid requirements, relapse rates, and even surgical requirements, at least in pediatric CD. However, many other 'top-down' treatment strategies have not yet been evaluated. In addition, there is a risk of overtreating those 10-30% of patients that will have a benign course of the disease; that's the reason why the implementation of top-down strategies remains as a matter of debate.


[Risk factors of gastropathy development in workers exposed to dust].
Systemic leucocyte-induced oxidative stress in prolonged occupational exposure to dust modifies oxidative metabolism of neutrophils and RBC, causes inflammatory and destructive, autoimmune processes in gastric lining and possible subsequent occupational gastropathies. To verify gastropathies caused by technogenic dust aerosols, search of preclinical biologic markers of gastric lining damage is essential. The article presents main steps in occupational gastropathies formation, trigger mechanisms, leading pathogenetic factors of gastric lining damage and of atrophic fundus gastritis. Using the mentioned complex of biomarkers in gastric lining state monitoring among workers at periodic and thorough medical examination is main item in forecasting individual risk of occupational gastropathies and in possible approach to their prevention and treatment.


Co-culturing monocytes with smooth muscle cells improves cell distribution within a degradable polyurethane scaffold and reduces inflammatory cytokines.
Activated monocytes can promote inflammation or wound repair, depending on the nature of the implant environment. Recent work showed that a degradable, polar-hydrophobic-ionic polyurethane (D-PHI) induced an anti-inflammatory monocyte phenotype. In the current study it is hypothesized that wound-healing phenotype monocytes (activated by D-PHI material chemistry) will promote human vascular smooth muscle cells (hVSMC) to attach and migrate into porous D-PHI scaffolds. hVSMC migration is necessary for hVSMC population of the scaffold and tissue formation to occur, and then, once tissue formation is complete, the monocyte should promote contractile phenotype markers in the hVSMC. hVSMC were cultured for up to 28 days with or without monocytes and analyzed for cell viability, attachment (DNA) and migration. Lysates were analyzed for the hVSMC contractile phenotype markers calponin and α-smooth muscle actin (α-SMA) as well as urokinase plasminogen activator (uPA; pro-migration marker) using immunoblotting analysis. Histological staining showed that hVSMC alone remained around the perimeter of the scaffold, whereas co-culture samples had co-localization of monocytes with hVSMC in the pores, a more even cell distribution throughout the scaffold and increased total cell attachment (P<0.05). Co-culture samples had higher cell numbers and more DNA than the addition of both single cell cultures. The water-soluble tetrazolium-1 data suggested that cells were not dying over the 28 day culture period. Calponin, also linked to cell motility, was maintained up to 28 days in the co-culture and hVSMC alone, whereas α-SMA disappeared after 7 days. Co-cultures on D-PHI showed that monocytes were activated to a wound-healing phenotype (low TNF-α, elevated IL-10), while promoting uPA expression. In summary, this study showed that, by co-culturing monocytes with hVSMC, the latter showed increased total cell attachment and infiltration into the D-PHI scaffold compared with hVSMC alone, suggesting that monocytes may promote hVSMC migration, a condition necessary for ultimately achieving uniform tissue formation in porous scaffolds.


Approach to acute headache in adults.
Approximately one-half of the adult population worldwide is affected by a headache disorder. The International Headache Society classification and diagnostic criteria can help physicians differentiate primary headaches (e.g., tension, migraine, cluster) from secondary headaches (e.g., those caused by infection or vascular disease). A thorough history and physical examination, and an understanding of the typical features of primary headaches, can reduce the need for neuroimaging, lumbar puncture, or other studies. Some red flag signs and symptoms identified in the history or during a physical examination can indicate serious underlying pathology and will require neuroimaging or other testing to evaluate the cause of headache. Red flag signs and symptoms include focal neurologic signs, papilledema, neck stiffness, an immunocompromised state, sudden onset of the worst headache in the patient's life, personality changes, headache after trauma, and headache that is worse with exercise. If an intracranial hemorrhage is suspected, head computed tomography without contrast media is recommended. For most other dangerous causes of headache, magnetic resonance imaging or computed tomography is acceptable.


[Common health disorders: self-care and self-medication].
Community level. "Santa María de Benquerencia" Health District, Toledo. People over 15, chosen by random sampling from the municipal census, who stated that they had suffered some health disorder over the previous fortnight. 212 surveys were accepted as valid (average age 42.75, 51.89% male and 48.11% female). 73% had suffered one of the disorders included in the survey during the previous fortnight. The commonest response was self-medication (39.84%), especially when dealing with pain and Pyrosis. Pyrazolones and Salicylates were the most commonly used drugs. The instructions were only read in 48.64% of cases. No measure was adopted in 34.56% of the disorders. The doctor was only consulted in 6.86% of cases. Both these responses were more common in men and young people. Non-pharmacological self-care was adopted in 30.07% of cases. This was generally of a dietetic or physical nature and was commoner in women and elderly people. Self-care, whether pharmacological or not, is the most common response to the perception of some symptom. These practices (especially self-medication because of its possible attendant problems) should be directed and used as one tool more in health care.


Experience applying the UK meticillin-resistant Staphylococcus aureus (MRSA) guidelines in a tertiary referral hospital in the United Arab Emirates 1999-2002: same guidelines, different cultures.
Meticillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial pathogen worldwide. There are few reports concerning MRSA in the United Arab Emirates (UAE). We report our experience with MRSA in a 400-bed tertiary referral hospital in the UAE, which followed the UK MRSA guidelines. MRSA data were reviewed to include demographic and clinical data on all new MRSA cases; a review of the ward environment including number of single rooms, sinks, toilets and bathrooms, frequency of cleaning; cultural observations; and number of infection control personnel per beds. MRSA was an uncommon pathogen. There were 90 new MRSA cases from 1999 to 2002 including two clusters of MRSA. While the procedures followed were the same as those in the UK, there were differences in the hospital environment compared with the UK and in cultural aspects of the patients' behaviour. At least 70% of inpatients were in single rooms with ensuite bathrooms. It was rare for more than two patients to share a toilet or bathroom. There were fewer than recommended infection control personnel and no antibiotic restriction policy in the hospital. Cleaners were on the wards for >100 h per week and were available 24 h per day for rapid response. We conclude that there are many factors that influence the management and control of MRSA, including cultural and social behaviour.


Modulation Transfer Function of AZ111 Photoresist.
For the utilization of photoresist materials it is desirable to describe quantitatively the photoresist image and thereby introduce control over photoresist processing. The photoresist AZ111 has been shown to be a linear image recording medium and a modulation transfer function (MTF) defined and measured. Measured and calculated image profiles in this resist are quite comparable.


[An unfolding model for confusion matrices].
An unfolding model is proposed for analyzing the confusion matrices obtained from stimulus identification experiments. In the model, row and column stimuli in the matrices are represented as two sets of points in a multidimensional space, and confusion probabilities are expressed as a function of the distances between the row and the column points. The coordinates of the points are estimated by the maximum likelihood method under a constraint. The proposed model enables us to explore interesting aspects of the perceptual representations of the stimuli, as illustrated by two examples. Some advantages of the proposed model are discussed in regard to its psychological implication.


Author Correction: Progress in multiple sclerosis - from diagnosis to therapy.
In the originally published article, Maria Pia Amato's affiliation was incomplete - it should have included IRCCS Fondazione Don Carlo Gnocchi, Florence, Italy. This correction notice has been appended to the PDF version of the manuscript.


Isolated extraocular muscle involvement as the ophthalmic manifestation of leukaemia.
Clinical and imaging features of patients with orbital leukaemia primarily involving extraocular muscles were evaluated. This retrospective case series includes patients with leukaemia whose only ophthalmic manifestation was extraocular muscle enlargement. Demographic data, clinical information on the systemic disease, prominent ocular signs and symptoms, computed tomography and magnetic resonance imaging characteristics, treatments applied and the outcomes were collected. Five patients were diagnosed as leukaemic infiltration of extraocular muscle between 1995 and 2008. The age at presentation ranged between 3 and 61 years. Acute myeloid leukaemia was the diagnosis in two patients, and chronic lymphocytic leukaemia, chronic myeloid leukaemia and biphenotypic acute leukaemia were found in one patient each, respectively. One patient had bilateral involvement. The lateral rectus muscle was affected in four patients and the superior rectus muscle in one case. Restricted ocular motility was the most common finding. In one patient who had no prior history of leukaemia, an incisional biopsy established the diagnosis. All patients received multi-agent chemotherapy. Four patients expired after a rapid decline of the systemic status within a mean period of 7 months. Leukaemic infiltration of extraocular muscles is a rare and late manifestation of the advanced disease associated with relapse and there seems to be a predilection for the lateral rectus muscle. Systemic prognosis remains dismal despite intensive chemotherapy.


Continuously-variable series-elastic actuator.
Actuator efficiency is an important factor in the design of powered leg prostheses, orthoses, exoskeletons, and legged robots. A continuously-variable series-elastic actuator (CV-SEA) is presented as an efficient actuator for legged locomotion. The CV-SEA implements a continuously-variable transmission (CVT) between a motor and series elastic element. The CVT reduces the torque seen at the motor and allows the motor to operate in speed regimes of higher efficiency, while the series-elastic element efficiently stores and releases mechanical energy, reducing motor work requirements for actuator applications where an elastic response is sought. An energy efficient control strategy for the CV-SEA was developed using a Monte-Carlo minimization method that randomly generates transmission profiles and converges on those that minimize the electrical energy consumption of the motor. The CV-SEA is compared to a standard SEA and an infinitely variable series elastic actuator (IV-SEA). Simulations suggest that a CV-SEA will require less energy that an SEA or IV-SEA when used in a knee prosthesis during level-ground walking.


Excited-state proton transfer in gas-expanded liquids: the roles of pressure and composition in supercritical CO2/methanol mixtures.
Excited-state proton transfer from 5,8-dicyano-2-naphthol to methanol takes place in CO2/methanol mixtures, in the pressure and temperature ranges of supercritical CO2. The efficiency of the proton-transfer step decreases with the pressure. This is assigned to the perturbation of the methanol clusters solvating the naphthol.


Public health practice in schools of public health: is there a fit?
Public health practice has been a part of schools of public health for a long time and yet it is still an emerging field of scholarship. Practice faculty often are isolated in their schools and not part of the mainstream. This article explores the elements of culture that are necessary to support a practice focus in a school of public health.


Arthroplasty for old tuberculosis of the knee.
We have reviewed six patients with old tuberculosis of the knee treated by total replacement an average of 35 years after the primary infection. Three patients had no antituberculous prophylaxis and three had drugs for two to three weeks before and three weeks after the operation. One patient with a missed primary diagnosis had a relapse of the tuberculous arthritis 18 months after his arthroplasty and was successfully treated with antituberculous drugs for one year. At an average follow-up of 6.3 years all the patients were markedly improved. Old tuberculosis of the knee can be treated successfully with arthroplasty but there is a risk of reactivation of disease and prophylactic drugs are recommended.


A review of the use of methadone for the treatment of chronic noncancer pain.
Methadone, although having been available for approximately half a century, is now receiving increasing attention in the management of chronic pain. This is due to recent research showing that methadone exhibits at least three different mechanisms of action including potent opioid agonism, N-methyl-D-aspartate antagonism and monoaminergic effects. This, along with methadone's excellent oral and rectal absorption, high bioavailability, long duration of action and low cost, make it a very attractive option for the treatment of chronic pain. The disadvantages of significant interindividual variation in pharmacokinetics, graduated dose equivalency ratios based on prerotation opioid dose when switching from another opioid, and the requirement for special exemption for prescribing methadone make it more complicated to use. The present review is intended to educate physicians interested in adding methadone to their armamentarium for assisting patients with moderate to severe pain.


Toll-like receptor 3 gene expression in Egyptian patients with glomerulonephritis and hepatitis C virus infection.
Hepatitis C virus (HCV) infection is associated with both chronic liver disorders and several extrahepatic manifestations including renal disease. Toll-like receptor 3 (TLR3), a component of the innate immune system, is a pathogen recognition receptor that recognizes viral double-stranded RNA. This work investigated TLR3 expression in peripheral blood mononuclear cells from HCV-positive patients with glomerulonephritis. One hundred and thirty patients with glomerulonephritis were initially enrolled in the study. After exclusion of 52 patients with secondary glomerulonephritis, 78 patients were screened for HCV infection. TLR3 expression in peripheral blood mononuclear cells was studied in 46 patients with HCV-positive glomerulonephritis and 32 patients with HCV-negative glomerulonephritis using a real-time PCR comparative quantitation approach and results were compared to a control group of 20 healthy subjects. TLR3 expression was significantly higher in patients with HCV-positive glomerulonephritis than in HCV negative patients and controls (p < 0.0001). TLR3 expression correlated positively with HCV viral load, interleukin-1β, serum creatinine and inversely with creatinine clearance in patients with HCV-positive glomerulonephritis. This work shows that TLR3 expression is upregulated in HCV-positive patients with glomerulonephritis. Overexpression is associated with reduced renal function and increased interleukin 1β level.


Daily symptom burden in end-stage chronic organ failure: a systematic review.
Chronic diseases are nowadays the major cause of morbidity and mortality worldwide. Patients with end-stage chronic organ failure may suffer daily from distressful physical and psychological symptoms. The objective of the present study is to systematically review studies that examined daily symptom prevalence in patients with end-stage chronic organ failure, with attention to those that included patients with either congestive heart failure (CHF), chronic obstructive pulmonary disease (COPD) or chronic renal failure (CRF). Thirty-nine articles (8 CHF, 7 COPD, 2 CHF and COPD, 22 CRF) have been included. The included studies used various study designs. There was a wide range of daily symptom prevalence that may be due to the heterogeneity in methodology used. Nevertheless, findings suggest significant symptom burden in these patients. This review highlights the need for further prospective and longitudinal research on symptom prevalence in patients with end-stage CHF, COPD and CRF to facilitate the development of patient-centred palliative care programs.


High-resolution recombinational map of mouse chromosome 16.
Five intersubspecific backcrosses and an intercross were used to establish a sex-averaged recombinational map spanning 56 cM across most of mouse Chromosome 16 (Chr 16). A total of 123 markers were ordered using an interval mapping approach to identify 425 recombination sites in a collection of 1154 meioses from 1155 progeny generated in the six crosses. The markers include the 10 "classic" Chr 16 reference markers, 26 additional genes or transcripts including two phenotypic markers (Pit1dw and Kcnj6wv), and 87 simple sequence length polymorphisms (SSLPs). One set of monozygotic twins was detected among the 304 meioses mapped to highest resolution. The reference markers and SSLPs allow the map to be well integrated with existing maps of Chr 16. The average distance between crossover sites is less than 500 kb for most chromosomes, making this collection of recombinant chromosomes useful as a binning and ordering resource for YAC-based physical map assembly on Chr 16.


Proceedings of a consensus conference: Risk-Based Decision Making for Blood Safety.
Blood safety decision making has become increasingly complex, and a framework for risk-based decision making is, thus, needed. The purpose of this consensus conference was to bring together international experts in an effort to develop the foundations for such a framework. These proceedings are described with a view to making available to the transfusion medicine community the considerable amount of information and insight that was presented and that emerged through debate by the experts, panel members, and delegates.


Epinephrine dosage effects on cerebral and myocardial blood flow in an infant swine model of cardiopulmonary resuscitation.
Although epinephrine increases cerebral blood flow (CBF) and left ventricular blood flow (LVBF) during cardiopulmonary resuscitation (CPR), the effects of high dosages on LVBF and CBF and cerebral O2 uptake have not been examined during prolonged CPR. We determined whether log increment dosages of epinephrine would enhance LVBF and CBF and cerebral O2 uptake in an infant swine CPR model. We compared these responses with epinephrine to those with the alpha-adrenergic agonist, phenylephrine. CPR was performed in five groups (n = 6) of pentobarbital-anesthetized piglets (3.5-5.6 kg) receiving a continuous epinephrine infusion (0, 1, 10, and 100 micrograms.kg-1.min-1) or phenylephrine infusion (40 micrograms.kg-1.min-1). Plasma epinephrine concentrations increased 10-100-fold in the control group during CPR and in a stepwise manner such that concentrations were increased by more than 10(4) in the 100 micrograms.kg-1.min-1 epinephrine group. In the control group with no epinephrine infusion, LVBF decreased to less than 10 ml.min-1.100 g-1 by 5 min of CPR. With epinephrine in dosages of 10 and 100 micrograms.kg-1.min-1, LVBF at 5 min was 75 +/- 19 and 44 +/- 15 ml.min-1.100 g-1, respectively, which was significantly greater than values in the control group. With more prolonged CPR, LVBF remained significantly greater than that in the control group but only at 10 micrograms.kg-1.min-1 of epinephrine. Phenylephrine also increased LVBF for 10 min of CPR when compared with the control group. All dosages of epinephrine and phenylephrine maintained CBF close to prearrest values for 20 min of CPR. With prolonged CPR, 10 and 100 micrograms.kg-1.min-1 epinephrine resulted in significantly greater CBF than that in the control group. Incremental dosages of epinephrine did not statistically increase cerebral O2 uptake or lower the cerebral fractional O2 extraction when compared with the control group, despite the higher CBF that was generated. In this immature animal CPR model, 10 micrograms.kg-1.min-1 epinephrine is an optimal dosage for maximizing both CBF and LVBF, a dosage that substantially exceeds the current recommended epinephrine dosage for human infant CPR. In addition, for short periods of CPR, 40 micrograms.kg-1.min-1 phenylephrine increases CBF and LVBF to levels similar to those generated by high dosages of epinephrine.


Salicylates inhibit NF-kappaB activation and enhance TNF-alpha-induced apoptosis in human pancreatic cancer cells.
Tumor necrosis factor (TNF-alpha)-induced apoptosis is limited by its coactivation of nuclear factor kappa B (NF-kappaB)-dependent anti-apoptotic genes. Sodium salicylate (NaSal) inhibits NF-kappaB activation by limiting phosphorylation and degradation of its bound inhibitor protein, IkappaB-alpha. We examined whether NaSal enhances TNF-alpha-induced apoptosis in cultured human pancreatic cancer cell lines. Two cultured human pancreatic cancer cell lines were studied. PANC-1 and BxPC-3 cells were serum-starved for 12 h, pretreated or not for 1 h with NaSal (5-20 mM), and then stimulated with recombinant human TNF-alpha (400 units/ml). Western blots of cytoplasmic lysates were performed to demonstrate IkappaB-alpha phosphorylation and degradation. Western blots of nuclear extracts were performed to assess nuclear translocation of NF-kappaB. In separate cultures, apoptosis was measured 4.5 h after TNF-alpha stimulation by both ELISA detection of interhistone DNA fragments and flow cytometry with propidium iodide staining. TNF-alpha induced IkappaB-alpha phosphorylation and degradation, which was inhibited by NaSal in both cell lines. TNF-alpha-induced apoptosis (DNA fragmentation) increased significantly when BxPC-3 cells were pretreated with NaSal. Flow cytometry confirmed this, demonstrating increases in apoptotic cell fractions: 8.5% (untreated), 9.3% (TNF-alpha alone), 14.9% (15 mM NaSal), and 22.9% (NaSal and TNF-alpha). In contrast, no increases in apoptosis were measured in the PANC-1 cell line among the various treatment groups. NaSal enhances TNF-alpha-induced apoptosis while inhibiting IkappaB-alpha phosphorylation and degradation in BxPC-3 human pancreatic cancer cells.


EC4 European Syllabus for Post-Graduate Training in Clinical Chemistry and Laboratory Medicine: version 3 - 2005.
The EC4 Syllabus for Postgraduate Training is the basis for the European Register of Specialists in Clinical Chemistry and Laboratory Medicine. The syllabus: Indicates the level of requirements in postgraduate training to harmonise the postgraduate education in the European Union (EU); Indicates the level of content of national training programmes to obtain adequate knowledge and experience; Is approved by all EU societies for clinical chemistry and laboratory medicine. The syllabus is not primarily meant to be a training guide, but on the basis of the overview given (common minimal programme), national societies should formulate programmes that indicate where knowledge and experience is needed. The main points of this programme are: Indicates the level of requirements in postgraduate training to harmonise the postgraduate education in the European Union (EU); Indicates the level of content of national training programmes to obtain adequate knowledge and experience; Is approved by all EU societies for clinical chemistry and laboratory medicine. Knowledge in biochemistry, haematology, immunology, etc.; Pre-analytical conditions; Evaluation of results; Interpretations (post-analytical phase); Laboratory management; and Quality insurance management. The aim of this version of the syllabus is to be in accordance with the Directive of Professional Qualifications published on 30 September 2005. To prepare the common platforms planned in this directive, the disciplines are divided into four categories: Indicates the level of requirements in postgraduate training to harmonise the postgraduate education in the European Union (EU); Indicates the level of content of national training programmes to obtain adequate knowledge and experience; Is approved by all EU societies for clinical chemistry and laboratory medicine. Knowledge in biochemistry, haematology, immunology, etc.; Pre-analytical conditions; Evaluation of results; Interpretations (post-analytical phase); Laboratory management; and Quality insurance management. General chemistry, encompassing biochemistry, endocrinology, chemical (humoral), immunology, toxicology, and therapeutic drug monitoring; Haematology, covering cells, transfusion serology, coagulation, and cellular immunology; Microbiology, involving bacteriology, virology, parasitology, and mycology; Genetics and IVF.


Detection of Plasmodium falciparum using a cloned DNA probe: a simple procedure suitable for field application.
A simple procedure was developed for spotting blood samples directly onto nylon membrane filter, without the necessity to treat samples with pronase or proteinase K, followed by hybridizing with 32P-labelled DNA probe, pUNK1-45. This probe detected specifically P. falciparum DNA and did not cross react with DNA from man, P. knowlesi, P. chabaudi or P. cynomolgi. The probe was sensitive to detect a parasitemia of 0.001% in 20 microliters of blood.


Bleeding from the small intestine and aortic regurgitation in Noonan syndrome.
A 62-year-old man was admitted to our hospital because of melena. On admission physical examination revealed that he had typical features of Noonan syndrome (NS). Investigation via upper endoscopy with the single balloon demonstrated oozing from the small intestine. Bleeding sometimes occurs in patients with NS. We speculated that coagulation defects or vascular malformations might have been present at the first visit in this case. However, coagulation function was normal. By upper endoscopy with the single balloon we clearly revealed the angioectasia in the small intestine. This case documents the first association among NS, aortic regurgitation and angioectasia in the small intestine.


The levels of 12 cytokines and growth factors in tears: hyperthyreosis vs euthyreosis.
Simultaneous analyses of the contents and ratios of 12 cytokines and growth factors in single samples of human tears were performed, and the results were compared between a group of healthy subjects and a group of patients with Graves' hyperthyreosis (GH) without thyroid-associated orbitopathy (TAO). Determinations and concentration measurements of interleukins (IL-2, IL4, IL-6, IL-8, IL-10, IL-1α, and IL-1β) interferon (IFN-γ), tumor necrosis factor (TNF-α), monocyte chemoattractant protein (MCP-1), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) were performed with single tear samples from 21 patients with hyperthyreosis and 22 healthy subjects. The analyses were performed using a Randox microchip with an Evidence Biochip Array Analyzer. We found significant differences between the healthy donor group and the hyperthyreosis group in the levels of IL-6, IL-10, VEGF, IL-1α, and MCP-1. The concentration of IL-6 was considerably higher in the hyperthyreosis group, IL-10 was higher in the healthy donor group, and VEGF and MPC-1 were higher in the hyperthyreosis group. The IL-8 and IFN-γ levels were higher in the hyperthyreosis group. The ratios of all of the cytokines to anti-inflammatory IL-10 were significantly elevated in the hyperthyreosis group. There are clear differences in the levels of cytokines and growth factors in the tears of healthy subjects and patients with GH without TAO. Tear cytokine changes and related dysfunctional tear syndrome (DTS) could be an early sign of occult TAO in Graves' hyperthyreosis patients.


Cryptococcal meningitis in AIDS patients - a report of two cases.
A fifty-five year old gentleman with HIV infection was investigated for meningitis.Cryptococcus neoformans was isolated.Second case was a lady of 42 years, with HIV infection, was also investigated for meningitis. Cryptococcus neoformans was isolated. Antigen was detected in CSF as well as serum in both the cases.


In vitro comparison of human fibroblasts from intact and ruptured ACL for use in tissue engineering.
The present study compares fibroblasts extracted from intact and ruptured human anterior cruciate ligaments (ACL) for creation of a tissue engineered ACL-construct, made of porcine small intestinal submucosal extracellular matrix (SIS-ECM) seeded with these ACL cells. The comparison is based on histological, immunohistochemical and RT-PCR analyses. Differences were observed between cells in a ruptured ACL (rACL) and cells in an intact ACL (iACL), particularly with regard to the expression of integrin subunits and smooth muscle actin (SMA). Despite these differences in the cell source, both cell populations behaved similarly when seeded on an SIS-ECM scaffold, with similar cell morphology, connective tissue organization and composition, SMA and integrin expression. This study shows the usefulness of naturally occurring scaffolds such as SIS-ECM for the study of cell behaviour in vitro, and illustrates the possibility to use autologous cells extracted from ruptured ACL biopsies as a source for tissue engineered ACL constructs.


Cathepsin B-like cysteine proteases and Caenorhabditis elegans homologues dominate gene products expressed in adult Haemonchus contortus intestine.
Proteins expressed by nematode intestinal cells are potential targets for parasite control by immune or chemical based strategies. To expand our knowledge on nematode intestinal proteins, expressed sequence tags were generated for 131 cDNA clones from the intestine of adult female Haemonchus contortus. An estimated 55 distinct protein genes or gene families were identified. Predicted proteins represented diverse functions. Several predicted polypeptides were related to H. contortus proteins implicated in inducing protective immunity against challenge infections of this parasite. The dominant intestinal transcripts were represented by cathepsin B-like cysteine protease genes (cbl) (17% of protein coding expressed sequence tags (ESTs) analyzed). An estimated 11 previously undescribed cbl genes were identified, doubling the recognized members of this gene family. Multiple C-type lectin sequences were identified. Other notable sequences included a predicted Y-box binding protein, serine/threonine kinases and a cyclin E-like sequence. Predicted protein homologues were found in Caenorhabditis elegans for all but one H. contortus sequence (99%), while fewer homologues from other parasitic nematodes were found. Many of the proteases, lipase and C-type lectin homologues in C. elegans had apparent signal peptides, suggesting that they are secreted. Several gene products had no obvious similarity outside the phylum Nematoda. The ESTs identified intestinal genes with potential application to immune control, understanding of basic intestinal regulatory processes and refinement of nematode genomic resources.


Influence of adhesive application methods and rebonding agent application on sealing effectiveness of all-in-one self-etching adhesives.
The choice of adhesive application methods could affect the microleakage of self-etch adhesives. To evaluate the effect of acid-etching, doubling adhesive application time, doubling adhesive coating, and rebonding agent application on microleakage of self-etch adhesives in Class V cavities. Seventy human third molars with Class V cavities assigned to five groups according to different adhesive application protocols for the three dentin adhesives (Clearfil S3 Bond, Kuraray Medical, Okayama, Japan; Optibond All-in-One, Kerr Corporation Orange, CA, USA; G-Aenial Bond, GC Corporation, Tokyo, Japan): group 1, manufacturer's recommendations; group 2, prior acid-etching of cavities; group 3, double application time; group 4, two consecutive coats of the adhesives; group 5, rebonding application on restoration margins. After bonding, the cavities were filled with a resin composite (Filtek Supreme XT, 3M ESPE Dental Products, St. Paul, MN, USA). The teeth were thermocycled, and the specimens were examined for microleakage using methylene blue as a marker. For Clearfil S3 Bond and Optibond All-in-One, microleakage in groups 2 and 5 were significantly lower than other groups' enamel margins. In groups 1, 2, 4, and 5, Clearfil S3 Bond exhibited significantly more leakage than the other dentin bonding agents in dentin margins. Microleakage was significantly higher on dentinal margins compared with the enamel margins for Clearfil S3 Bond in all of the groups. Optibond All-in-One showed significantly lower microleakage in dentin margins in all groups except groups 2 and 5. Acid-etching usually promoted the reduction of microleakage in enamel margins. On the other hand, rebonding application usually contributed to the reduction of microleakage more than other methods in enamel and dentin margins. Acid-etching or rebonding application may contribute to reduction of microleakage of all-in-one self-etching adhesives.


New Iminodiacetate-Thiosemicarbazone Hybrids and Their Copper(II) Complexes Are Potential Ribonucleotide Reductase R2 Inhibitors with High Antiproliferative Activity.
As ribonucleotide reductase (RNR) plays a crucial role in nucleic acid metabolism, it is an important target for anticancer therapy. The thiosemicarbazone Triapine is an efficient R2 inhibitor, which has entered ∼20 clinical trials. Thiosemicarbazones are supposed to exert their biological effects through effectively binding transition-metal ions. In this study, six iminodiacetate-thiosemicarbazones able to form transition-metal complexes, as well as six dicopper(II) complexes, were synthesized and fully characterized by analytical, spectroscopic techniques (IR, UV-vis; 1H and 13C NMR), electrospray ionization mass spectrometry, and X-ray diffraction. The antiproliferative effects were examined in several human cancer and one noncancerous cell lines. Several of the compounds showed high cytotoxicity and marked selectivity for cancer cells. On the basis of this, and on molecular docking calculations one lead dicopper(II) complex and one thiosemicarbazone were chosen for in vitro analysis as potential R2 inhibitors. Their interaction with R2 and effect on the Fe(III)2-Y· cofactor were characterized by microscale thermophoresis, and two spectroscopic techniques, namely, electron paramagnetic resonance and UV-vis spectroscopy. Our findings suggest that several of the synthesized proligands and copper(II) complexes are effective antiproliferative agents in several cancer cell lines, targeting RNR, which deserve further investigation as potential anticancer drugs.


Mechanical response of single myoblasts to various stretching patterns visualized by scanning probe microscopy.
The mechanical memory effect of single cells was reported in our recent study. In order to clarify this effect, various sequential stimuli of uniaxial deformation were applied to cells by deformable culture dishes and a deformation device, and the local stiffness distribution of single C2C12 myoblasts was visualized by scanning probe microscopy. Following a single step stretching, cellular stiffness first increased steeply and then gradually decreased for two hours. By a single step stretching 30 min after a long pulse-like deformation with a pulse duration of 30 min, the cells responded in the same way. On the other hand, they did not respond to a single step stretching 30 min after a short pulse-like deformation with a pulse duration of 0.5 min. These results indicated that cellular mechanical response to external deformation is affected strongly by a preceding deformation and that the duration time of the preceding deformation is an important factor in the change in mechanical response. We consider that the change in mechanical response contributes to a regulatory mechanism of cellular contractile force.


Magnolol Nanoparticles Exhibit Improved Water Solubility and Suppress TNF-α-Induced VCAM-1 Expression in Endothelial Cells.
The expression of the adhesion molecule vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells enables the attachment of leukocytes to the endothelium, which may lead to inflammation and the development of atherosclerosis. Magnolol is a major bioactive compound derived from the plant species Magnolia officinalis. In this study, we synthesized a novel nanoparticle formulation of magnolol to improve its water solubility and physicochemical properties, evaluated its effects on TNF-α-induced VCAM-1 expression in endothelial cells, and determined the signal transduction pathways involved. Our findings demonstrated that the magnolol nanoparticle system showed great improvements in physicochemical properties and water solubility owing to a reduction in particle size, transformation from a crystalline to amorphous structure, and the formation of hydrogen bonds with the nanoparticle carriers. In terms of its biological actions, magnolol nanoparticles attenuated TNF-α-induced VCAM-1 protein expression, promoter activity, and mRNA expression in endothelial cells in vitro. This was found to be mediated by the ERK, AKT, and NF-κB signaling pathways. In addition, magnolol nanoparticles inhibited TNF-α-induced leukocyte adhesion to endothelial cells, and suppressed TNF-α-induced VCAM-1 expression in the aortic endothelium of mice. In summary, since magnolol nanoparticles inhibit endothelial VCAM-1 expression and leukocyte adhesion to endothelial cells, this novel drug formulation may be a potentially useful therapeutic formulation to prevent the development of atherosclerosis and inflammatory diseases.


Rapid preparation of mycobacterium N-glycolyl Lipid I and Lipid II derivatives: a biocatalytic approach.
Breaking down barriers: A rapid, inexpensive preparation of the structurally complex mycobacterial N-glycolyl Lipid I, Lipid II, and their analogues from a range of different synthetic N-glycolyl and N-glycinyl Park's nucleotides is described (see scheme). The biotransformations were catalyzed by a readily available biocatalyst obtained from a bacterial cell-free membrane fraction. The unnatural N-glycinyl Lipid II was found to be a substrate of Mycobacterium tuberculosis (Mtb) transglycosylase, PonA, and N-glycolyl Lipid I was a weak inhibitor against PonA.


Breaking the code.
When a nurse is found guilty of professional misconduct it stays on her or his career record for ten years and will almost certainly hinder that nurse's attempts to find a new job.


[Spleen-preserving distal pancreatectomy in treatment of solid-pseudopapillary neoplasm].
To investigate the feasibility and safety of spleen-preserving distal pancreatectomy with conservation of the splenic arteries and veins in treatment of benign neoplasms of distal pancreas. Four patients with solid-pseudopapillary neoplasm, 1 males and 3 females, aged 30 (17 - 37), underwent laparotomy. The distal pancreas with tumor was cut. The perforating branches between the splenic artery and vein and the distal pancreas were isolated, ligated, and cut. The splenic artery and vein and the short and left gastric vessels were all preserved. The spleen remained intact. The mean operation time was 208 minutes +/- 52 minutes, and the mean blood loss was 475 ml +/- 96 ml. Blood transfusion was not necessary. The mean post-operative hospital stay was 18 days +/- 13 days. No post-operative complications were found except for pancreatic leakage occurring in one patient that was cured by conservative treatment. No metastasis and recurrence was found. Safe and feasible, spleen-preserving distal pancreatectomy with conservation of the splenic arteries and veins is one of the best choices for treatment of benign and borderline tumors of pancreas.


Strategies to modulate cellular energetic metabolism during sepsis.
Growing evidence suggests that mitochondrial inhibition plays a major role in the development of multiple organ failure during sepsis. Early correction of tissue hypoxia, strict control of glycaemia and modulation of oxidative and nitrosative stress may protect mitochondria during the acute inflammatory response. Once mitochondrial dysfunction has developed, the regulated induction of a hypometabolic state, analogous to hibernation, may protect the cells from severe bioenergetic failure and a critical fall in ATP. Though this is clinically manifest as organ dysfunction, it may actually represent an adaptive response to a prolonged, severe inflammatory stress. Repair of damaged organelles, stimulation of mitochondrial biogenesis and re-activation of cellular metabolism may accelerate the recovery phase and thus improve clinical outcomes. The aim of this review is to discuss putative interventions aimed at preventing or reversing mitochondrial dysfunction that may have possible clinical relevance, and to stress the importance of the correct timing of intervention.


The complete mitochondrial genome of the sandbar shark Carcharhinus plumbeus.
The sandbar shark, Carcharhinus plumbeus, a major representative species in shark fisheries worldwide is now considered vulnerable to overfishing. A pool of 774,234 Roche 454 shotgun sequences from one individual were assembled into a 16,706 bp mitogenome with 33× average coverage depth. It comprised 13 protein coding genes, 22 transfer RNA's, 2 ribosomal genes and 2 non-coding regions, typical of a vertebrate mitogenome. As expected for sharks, an A-T nucleotide bias was evident. This adds to rapidly growing number of mitogenome assemblies for the economically important Carcharhinidae family. The C. plumbeus mitogenome will assist researchers, fisheries and conservation managers interested in shark molecular systematics, phylogeography, conservation genetics, population and stock structure.


The entry of antibiotics into human monocytes.
Effective therapy of infections due to facultative intracellular micro-organisms, which persist after ingestion by mononuclear phagocytic cells, requires the use of antibiotics with the ability to inactivate these intra-phagocytic bacteria. Since entry of antibiotics into mononuclear phagocytes is a pre-requisite for activity against such intracellular organisms, we have determined the uptake of 11 radiolabelled antibiotics by human peripheral blood monocytes. beta-Lactams (penicillin G, cefamandole and cefotaxime), gentamicin, and metronidazole had a limited ability to enter monocytes, achieving cellular concentrations which were equal to or less than extracellular levels (C/E less than or equal to 1). Imipenem, a novel beta-lactam antibiotic, rapidly bound to monocytes, but cell-associated drug progressively declined during further incubation. Chloramphenicol, a lipid-soluble drug, and trimethoprim were concentrated three-fold by human monocytes. In comparison with the other antibiotics, roxithromycin (C/E = 14), clindamycin (C/E = 6 to 7) and erythromycin propionate (C/E = 4 to 5) were markedly concentrated by human monocytes. In contrast to our findings in other phagocytes, there was no evidence that active membrane transport was involved in monocyte uptake of clindamycin, erythromycin and roxithromycin. We have demonstrated that several antibiotics are highly concentrated within human monocytes, and it will be important to evaluate the effects of this antibiotic uptake on various monocyte functions, including intra-phagocytic antibacterial activity.


Gp130 and ras mediated signaling in human plasma cell line INA-6: a cytokine-regulated tumor model for plasmacytoma.
Cytokines of the gp130-family, particularly interleukin(IL)-6, play a crucial role in the propagation of malignant plasma cells. The role of IL-6 and other gp130-cytokines was studied in the human plasma cell line INA-6 in vitro and in INA-6 xenografts. The proliferative response to gp130-cytokines was evaluated and activated components of gp130-signaling pathways were identified by Western blotting and DNA binding studies. Specifically, expression of IL-6 and receptors for IL-6 and leukemia inhibitory factor were analysed by RT-PCR and ELISA. The plasma cell line INA-6 was cultured for several years remaining strictly dependent on exogenous IL-6. Other gp130-cytokines had no significant effect on INA-6 cell proliferation in vitro. Due to an activating mutation in the N-ras gene, mitogen-activated protein kinases (MAPK) were constitutively phosphorylated. In contrast, signal transducer and activator of transcription(STAT)-3 activation was dependent on stimulation with IL-6. Blocking of either one of these pathways resulted in a significant decrease of INA-6 cell proliferation. Remarkably, INA-6 xenografts did not require exogeneous IL-6 for proliferation in vivo. Instead, an autocrine IL-6 loop and, in certain tumor sublines, responsiveness to additional gp130-cytokines was induced during in vivo growth. Activation of the gp130 signal transducer is mandatory for INA-6 cell growth in vitro and in vivo. Both the MAPK and the Jak/STAT pathway are operative in malignant plasma cells and either one is essential for plasma cell growth. The INA-6 cell line provides a preclinical model to study growth regulation of human plasmacytoma cells and to evaluate novel therapeutic strategies.


Driving change in rural workforce planning: the medical schools outcomes database.
The Medical Schools Outcomes Database (MSOD) is an ongoing longitudinal tracking project ofmedical students from all medical schools in Australia and New Zealand. It was established in 2005 to track the career trajectories of medical students and will directly help develop models of workforce flow, particularly with respect to rural and remote shortages. This paper briefly outlines the MSOD project and reports on key methodological factors in tracking medical students. Finally, the potential impact of the MSOD on understanding changes in rural practice intentions is illustrated using data from the 2005 pilot cohort (n = 112). Rural placements were associated with a shift towards rural practice intentions, while those who intended to practice rurally at both the start and end of medical school tended to be older and interested in a generalist career. Continuing work will track these and future students as they progress through the workforce, as well as exploring issues such as the career trajectories of international fee-paying students, workforce succession planning, and the evaluation of medical education initiatives.


Chemical Vapor Deposited Few-Layer Graphene as an Electron Field Emitter.
Chemical vapor deposition (CVD) growth of graphene on polycrystalline copper (Cu) foil in a low pressure conditions has been presented, aiming to achieve the highest quality with large-scale fabrications, which requires comprehensive understanding and effective controlling of the growth process. Herein, few-layer graphene (FLG) films with large-domain sizes were grown on Cu metal catalyst substrates using a vertical mass-flow hot-filament CVD reactor, with the intention of large scale production, by optimizing the CVD system and three of the process parameters: (i) gas flow compositions, (ii) substrate annealing time and (iii) graphene deposition time. The detailed scanning electron microscope and Raman spectroscopy analysis indicate that all the above mentioned process parameters affect growth of FLG film on Cu substrate. The presence of two intense peaks, G and 2D-band at 1583.6 and 2702.6 cm⁻¹ for synthesized sample at optimized conditions (H₂/CH₄ ratio of 50:1 at graphene deposition time of 10 minutes and substrate annealed time for 20 minutes) revealed the formation of FLG films with large domain size. These graphene films on Cu have shown the room temperature field electron emission characteristics, hence appears to be prospective candidate for vacuum nanoelectronics.


Noble metal nanoparticle-induced oxidative stress modulates tumor associated macrophages (TAMs) from an M2 to M1 phenotype: An in vitro approach.
Diagnosis of cancer and photothermal therapy using optoelectronic properties of noble metal nanoparticles (NPs) has established a new therapeutic approach for treating cancer. Here we address the intrinsic properties of noble metal NPs (gold and silver) as well as the mechanism of their potential antitumor activity. For this, the study addresses the functional characterization of tumor associated macrophages (TAMs) isolated from murine fibrosarcoma induced by a chemical carcinogen, 3-methylcholanthrene (MCA). We have previously shown antitumor activity of both gold nanoparticles (AuNPs) and silver nanoparticle (AgNPs) in vivo in a murine fibrosarcoma model. In the present study, it has been seen that AuNPs and AgNPs modulate the reactive oxygen species (ROS) and reactive nitrogen species (RNS) production, suppressing the antioxidant system of cells (TAMs). Moreover, the antioxidant-mimetic action of these NPs maintain the ROS and RNS levels in TAMs which act as second messengers to activate the proinflammatory signaling cascades. Thus, while there is a downregulation of tumor necrosis factor-α (TNF-α) and Interleukin-10 (IL-10) in the TAMs, the proinflammatory cytokine Interleukin-12 (IL-12) is upregulated resulting in a polarization of TAMs from M2 (anti-inflammatory) to M1 (pro-inflammatory) nature.


Evaluation of a dipstick for candida.
Differentiation of the infecting organism is important in determining effective therapy for vaginitis. The utility of a simple, dehydrated culture strip in detecting Candida infections was assessed by culturing specimens from 203 patients, 83 with symptoms of vaginitis. Parallel culture (Sabouraud's medium) and identification were performed in every fourth case by an independent clinical laboratory; KOH preparations for Candida were examined in every case. In 48 of 50 cases, the strip agreed with Sabouraud's culture; one culture was lost due to laboratory error, and one strip did not become positive until after 48 hours' incubation. Only 70% of KOH preparations revealed Candida when the strip culture was positive. Diagnostically superior to the KOH preparation and nearly equal to a Sabouraud's culture, the strip was simple to use, inexpensive, and reliable; as such, it is a useful addition in the ob-gyn office or clinic for routine screening for vaginal candidiasis.


A survey of women urology residents regarding career choice and practice challenges.
To determine the reasons why urology has become an increasingly popular career choice for women medical students and to uncover challenges they face during their training period. A mailed survey was sent to all 176 female urology residents. Fifty-five surveys were received, for a return rate of 31%. General surgery was the most common alternative specialty choice considered by female urology trainees, although there was wide representation from other disciplines. Diversity of procedures was the most frequently cited reason for choosing urology. The majority of respondents interested in fellowships expect to seek training in pediatric or female/reconstructive urology. More than half of these residents have been confronted with negative behavior by male patients and by male colleagues in relation to their gender. Understanding the perspectives, concerns, and predilections of women urology applicants and trainees is essential to maximize the contributions women will make to the specialty and its patients.


Levels of intracellular adhesion molecule-1 and vascular cell adhesion molecule-1 in children with Henoch-Schönlein purpura.
Henoch-Schönlein purpura (HSP) is a small vessel vasculitis. Soluble adhesion molecules play a very important role in the immuno-inflammatory reaction of damaged vascular tissues. This study investigated the prognostic and diagnostic potential of soluble intracellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in HSP. Serum levels of sICAM-1 and sVCAM-1 were studied in 26 children with HSP. Paired blood samples (during acute and convalescent stages) were collected from 17 of the children and assayed by enzyme-linked immunosorbent assay. Correlations with clinical manifestations were examined. Seventeen healthy children served as controls. Both sICAM-1 and sVCAM-1 were significantly elevated at the acute stage compared with the remission stage of HSP patients versus controls (p=0.006 and p=0.0173, respectively). Although the levels of sICAM-1 and sVCAM-1 were not correlated with the severity of clinical manifestations in HSP, these soluble adhesion molecules may serve as diagnostic markers.


Rapid response of circulating myeloid dendritic cells to inhaled allergen in asthmatic subjects.
Dendritic cells (DC) are thought to play a key role in the initiation and maintenance of T cell immunity to inhaled antigens. While the density of DC within the bronchial mucosa is increased in stable asthma, there is little information currently available concerning the effects of allergen inhalation on DC in subjects with asthma. To enumerate changes in the numbers of circulating CD33(+) myeloid DC in asthmatics, before and after allergen challenge. Blood DC numbers were enumerated by flow cytometry before and at 3, 6 and 24 h after inhaled allergen and diluent in 10 mild, allergic asthmatic subjects. Blood DC numbers rapidly fell from 3.42 +/- 0.30 x 10(7)/L at baseline, to 2.10 +/- 0.17 x 10(7)/L by 3 h post-challenge (P < 0.01), and remained significantly below baseline values at both 6 and 24 h following allergen challenge. No such changes in DC numbers were noted after diluent challenge. A similar, early fall in circulating lymphocytes was also noted post-allergen challenge, whereas changes in circulating eosinophil and neutrophil numbers occurred more slowly. A significant proportion of myeloid DC rapidly 'disappear' from the circulation following allergen inhalation, suggesting that margination of circulating myeloid DC, and their recruitment into the airway mucosa, is an important feature of the immune response to inhaled allergen.


Experimental tests of the adaptive significance of sexual recombination.
Numerous theories have been proposed to explain the advantages of sexual recombination the exchange of hereditary material between different genomes or homologous chromosomes. Many of these candidate benefits have been evaluated in controlled laboratory experiments, which, collectively, strongly indicate that sexual recombination provides important long-term advantages.


Phenotypic and genetic relationships of common health disorders with milk and fat yield persistencies from producer-recorded health data and test-day yields.
The objective of this study was to investigate phenotypic and genetic relationships of common health disorders in dairy cows with milk (PMY) and fat (PFY) yield persistencies. Health and production data from 398 commercial dairy herds were used. Disease traits were defined in binary form for individual lactations considering mastitis only during the first 100 d in milk (MAST1), only after 100 d in milk (MAST2), and at any stage of lactation (MAST), and reproductive disorders (REPRO), metabolic disorders (METAB), and lameness (LAME). The persistencies were defined to be uncorrelated with 305-d yields. Impact of the diseases on PMY and PFY were investigated separately in first (FL) and later (LL) lactations. Phenotypic associations of PMY and PFY with likelihood of diseases in current and subsequent lactations were examined using odds ratios from a logistic regression model. Linear-threshold sire-maternal grandsire models were used to estimate genetic correlations of displaced abomasums (DA), ketosis (KET), metritis (MET), MAST, MAST1, and MAST2 with PMY and PFY across parities. Metabolic diseases and REPRO had significantly positive relationships with PMY and PFY in both FL and LL. Significantly greater PMY and PFY were associated with MAST1 in LL. Significantly lower PMY and PFY were related to MAST2 in both FL and LL, whereas cows affected by MAST had significantly less persistent lactations. Incidence of MAST and MAST2 decreased with increasing PMY and PFY in the present and previous lactation. Heritability of disease incidences were 0.03 (DA), 0.01 (KET), 0.10 (MAST), 0.02 to 0.05 (MAST1), 0.02 (MAST2), and 0.04 to 0.10 (MET). Displaced abomasum, KET, MAST, MAST1, and MET had unfavorable genetic correlations of 0.35, 0.46, 0.17, 0.02, and 0.27 with PMY, and 0.16, 0.21, 0.07, 0.06, and 0.12 with PFY, respectively. Favorable genetic correlations were found for MAST2 with PMY (-0.24) and PFY (-0.04). Results suggest that diseases in early lactation increase persistency of milk and fat yield. Selection for greater lactation persistency must consider these antagonistic relationships.


Social exclusion in clients with comorbid mental health and substance misuse problems.
The concept of comorbid mental health problems and substance misuse has gained prominence in the last two decades, due in part to the closure of large psychiatric hospitals and to the increasing prevalence of drug use in the community. This client group has a dual requirement for both medical and social care needs and is at risk for social exclusion. A retrospective matched case-control study to examine aspects of social exclusion between service users who have comorbid diagnoses and those with a single diagnosis. Samples were drawn from the service users of a mental health Trust in the South-East of England, from both Adult Mental Health (n = 400) and Drug and Alcohol services (n = 190). Data were collected from Care Programme Approach assessment forms and medical records. McNemar's chi(2) and odds ratios via a conditional logit regression model are used to test for differences in the social exclusion indicators. There were significant differences in social exclusion between the comorbid and singly diagnosed clients of the Adult Mental Health service, but differences were less pronounced between the comorbid and singly diagnosed clients of the specialist Drug and Alcohol service. Recent Government policy advocates treating comorbid clients within mainstream mental health services. Health care workers need to recognise the likelihood of high levels of social exclusion among clients with comorbid problems.


Tooth surface loss among people exposed to cement and stone dust in the work environment in Tanzania.
The effect of cement and stone dust on teeth was explored in a cross-sectional study, using blind dental examinations. The sample consisted of 36 workers who had been exposed to to the dust and 62 control workers. Tooth surface loss was observed in 72.2 per cent of the exposed workers and in 48.4 per cent of the controls (P less than 0.03). In both the maxillae (P less than 0.001) and the mandible (P less than 0.02) the amount of tooth surface loss was greater in the exposed workers than in the controls. Both anterior and posterior teeth were affected. These findings indicate that tooth surface loss caused by work-related dust should be considered an occupational hazard.


A Behavioural Case Study of Early Social Isolation of a Subadult White-Handed Gibbon (Hylobates lar).
Our aim in this study was to analyse the effects of early social isolation on the behaviour of a white-handed gibbon (Hylobates lar) and at the same time to improve his level of welfare. The subject was a 6-year-old male, isolated from conspecific as well as other non-human primates since he was 3 months old. We presented the gibbon with a series of species-specific vocalisations, and we then introduced a 23-year-old conspecific female into his cage. Our subject did not respond to playbacks, whereas he immediately interacted positively with the conspecific female. After 2 days of presentation, the pair started to spend time in proximity to each other and initiated grooming through the wire-mesh dividing the cages. Four days later we recorded vocal duets. No obvious ste-reotypic behaviours were observed, and the prolonged isolation did not seem to compromise the ability of the young gibbon to socialise with the female conspecific. It appears that prolonged isolation does not always compromise the possibility of recovering socially in a satisfactory manner.


Long-term clinical outcomes of supracricoid partial laryngectomy with cricohyoidoepiglottopexy for glottic carcinoma.
Background: Laryngeal carcinoma should be treated with the intent of organ-sparing, and supracricoid partial laryngectomy with cricohyoidoepiglottopexy (CHEP) might be an important option. Aims/objectives: The purpose of this study was to evaluate the clinical outcomes of glottic carcinoma patients treated with CHEP. Materials and methods: A series of 164 cases with glottic carcinoma undergoing CHEP from 2006 to 2010 was retrospectively analyzed. Results: The 10-year overall survival (OS) rate, disease-specific survival (DSS) rate, and disease-free survival (DFS) rate were 77.6%, 78.8%, 74.1%, respectively. The OS, DSS, and DFS of patients with stage T1 were higher than patients with stages T2 and T3. Patients with locoregional recurrence and distant metastases had lower OS and DFS than patients with neither recurrence nor metastasis. The DFS of patients with advanced laryngeal carcinoma was worse than that of patients with early-stage carcinoma. T2 and T3 stages, locoregional recurrence, and distant metastases had predictive value regarding patient survival. Additionally, the decannulation rate of postoperative patients was 95.1%, and the nasogastric feeding tube removal rate was 100%. Conclusions and Significance: CHEP provided reliable oncologic and functional outcomes, and it should be considered as a standard function-sparing option for glottic T1b, T2, and selected T3 carcinoma patients.


Association of pulmonary atresia with intact ventricular septum and aortic valve stenosis. Prenatal diagnosis.
A rare association of pulmonary atresia with an intact septum was diagnosed through echocardiography in a fetus 32 weeks of gestational age. The diagnosis was later confirmed by echocardiography of the newborn infant and further on autopsy. The aortic valve was bicuspid with a pressure gradient of 81 mmHg, and the right ventricle was hypoplastic, as were the pulmonary trunk and arteries, and the blood flow was totally dependent on the ductus arteriosus.


Alanine and aspartate aminotransferases in normal and denervated skeletal muscle.
Activities of alanine and aspartate aminotransferases are maintained during the first 5 weeks of growth, but decrease subsequently in normal chick gastrocnemii. In sciatectomized muscles, a 5-fold elevation in these enzymes reveals increased utilization of amino acids as a compensatory metabolic support during denervation atrophy.


[Immunohistochemical study on vitrectomy specimens of proliferative retinal diseases].
To evaluate the characteristics of cellular proliferation in different retinal diseases. Three specific antibodies [to cytokeratin (CK), glial fibrillary acidic protein (GFAP) and actin, respectively] were used. Vitrectomy specimens from 28 cases with different proliferative retinal diseases were studied immunohistochemically. In the vitreous with proliferative vetreoretinopathy (PVR), most of the proliferative cells were derived from glial and retinal pigment epithelial (RPE) cells. In the vitreous of proliferative diabetic retinopathy (PDR), RPE cells might enhance the contraction of proliferative membranes. Increase in actin in firovascular membranes with PDR may play a positive role on the pericytes dropping out from microvessels. The cell proliferation in the vitreous associated with the increase of levels of growth factors in pathological vitreous, possibly, is one of the mechanisms for the deterioration of proliferative retinal diseases.


Structure of complex cell wall polysaccharides isolated from Trichoderma and Hypocrea species.
The structure of fungal polysaccharides isolated from the cell wall of Trichoderma reesei, T. koningii, and Hypocrea psychrophila, have been investigated by means of chemical analyses and 1D and 2D NMR spectroscopy. The polysaccharides have an irregular structure, idealized as follows: [formula: see text] The proportions of the different side chains vary from a species to another, being n above some three times larger in H. psychrophila than in T. reesei or T. koningii.


Experiences in the measurement of RBC-bound IgG as markers of cell age.
An immunologically mediated pathway has been largely accepted to be one of the mechanisms involved in the clearance of senescent or prematurely damaged RBC. According to this pathway, RBC removal is mediated by binding of naturally occurring IgG to clustered integral membrane proteins, followed by complement deposition. The validation of an immunoenzymatic method for the detection of RBC-bound autologous IgG is presented. The use of RBC-bound IgG as an index related to red cell age was evaluated by measuring IgG binding in RBC treated with the clustering agent ZnCl2, in density fractionated RBC and in a selected group of patients expected to have an altered RBC life span. The immunoenzymatic method for IgG detection resulted to be reproducible (CV = 3.4%). IgG binding to in vitro clustered RBC was found to be enhanced to a very great extent, about 20 times higher with respect to untreated RBC. A slight but significant increase (about 1.8-fold) in membrane-bound IgG was observed in the highest density fraction of normal RBC, which constituted 1% of the total cells. A significantly greater number of RBC-bound IgG was measured in splenectomized beta-thalassemia intermedia patients and in subjects with secondary decreases in the C3 complement fraction concentration.


Putative down-stream signaling molecule of GTPase in Porphyromonas gingivalis.
Porphyromonas gingivalis is a strict anaerobic bacterium mainly responsible for periodontal disease in oral cavity. Putative GTPase gene (pgp) of this bacterium was cloned and its recombinant protein (rPGP) was produced in Escherichia coli. Based on the amino acid sequence of SGP that is a GTP-binding protein of Streptococcus mutans, putative GTPase amino acid sequence was deduced in the data base of genome sequences of Porphyromonas gingivalis. A 900-bp PCR fragment was amplified with P. gingivalis genomic DNA as a template and cloned into E. coli JM 109. Then pgp was transferred into pQE-30 expression vector to make pQE-PGP for production of rPGR. This protein was produced and purified by Ni-NTA affinity column chromatography. Anti-PGP antibody was also produced in Sprague Dawley rats. Using Westernblot analysis with this antibody, it was confirmed that the rPGP produced in E. coli was identical to that of donor strain. Furthermore, by Southernblot analysis it was revealed that the pgp was originated from P. gingivalis. By immunoprecipitation with anti-PGP antibody and N-terminal amino acid sequence analysis it was found that PGP was able to bind to acetate kinase, which was reported to be a secondary signaling-molecule in anaerobic microorganisms. Therefore, these results imply that P. gingivalis produces putative GTPase and this protein might play a potential role in signaling pathway in oral biofilm formation.


Damping by bulk and shear viscosity of confined acoustic phonons for nanostructures in aqueous solution.
A nanoparticle in aqueous solution is modeled as a homogeneous elastic isotropic continuum sphere in contact with an infinite viscous compressible Newtonian fluid. The frequencies and damping of the confined vibrational modes of the sphere are calculated for the material parameters of a CdSe nanoparticle in water and a poly(methyl methacrylate) nanosphere in water. Although the effects of viscosity are found to be negligible for macroscopic objects, for nanoscale objects, both the frequency and damping of the vibrational modes are significantly affected by the viscosity of the liquid. Furthermore, both shear viscosity and bulk viscosity play an important role. A model of the spherical satellite tobacco mosaic virus consisting of outer solid layers with a water core is also investigated, and the viscosity of the water core is found to significantly damp the free vibrational modes. The same approach can be applied for nonspherical geometries and also to viscoelastic nanoparticles.


Beta-amyloid peptides inhibit acetylcholine release from cholinergic presynaptic nerve endings isolated from an electric ray.
We investigated the effects of beta-amyloid (Abeta) peptides on cholinergic synaptosomes isolated from the electric organ of the Japanese marine ray Narke japonica. Fresh and pre-incubated solutions of Abeta(1-42) inhibited acetylcholine (ACh) release from the synaptosomes evoked by high [K+] depolarization when incubated with synaptosomes for 10 min before the depolarizing stimulus. A freshly prepared solution of Abeta(1-40) did not inhibit the evoked ACh release, but prolonged pre-incubation of Abeta(1-40) solution caused the inhibition. Abeta(1-15) neither in fresh nor pre-incubated solution inhibited. These results have demonstrated that Abeta peptides can acutely inhibit the depolarization-evoked release of ACh by acting directly on cholinergic presynaptic nerve endings. The electrophoresis analysis showed a strong correlation between Abeta aggregation and its inhibition for ACh release.


Heat-shock gene expression in alcoholic liver disease in the rat is related to the severity of liver injury and lipid peroxidation.
To evaluate the relationship between heat-shock gene expression and the severity of pathologic liver injury and lipid peroxidation in experimental alcoholic liver disease, we used the intragastric-feeding rat model. Six groups of male Wistar rats weighing between 225 and 250 g were fed liquid diets containing different dietary fats (saturated fat, corn oil, and fish oil) and ethanol or dextrose for 1 month. Pathologic injury, mRNA and protein levels of HSP70, microsomal conjugated dienes, and hydrogen peroxide were evaluated at sacrifice. Fish oil-ethanol fed rats developed the most severe injury, while the saturated fat-ethanol group showed no liver injury. The highest levels of HSP70 mRNA were seen in the fish oil-ethanol group. There was no difference in HSP70 protein levels between the groups. The levels of HSP70 mRNA correlated significantly with microsomal conjugated dienes (r = 0.87, P < 0.01) and hydrogen peroxide (r = 0.90, P < 0.01). Increased centrilobular HSP70 expression was seen in rats showing liver injury. The close relationship seen between oxidant stress and HSP70 mRNA but not protein suggest that the binding of HSP70 protein to other damaged proteins reduces free HSP70 protein leading to increased HSP70 expression. HSP70 expression may also be a cellular adaptive response to ethanol-induced oxidative stress.


Gastrointestinal bleeding under dabigatran.
Dabigatran-absorption is dependent on the intestinal P-glycoprotein (P-gp)-system, and P-gp activity is modulated by several drugs. We report an 83-old female with atrial fibrillation who developed gastrointestinal bleeding. She was under a therapy with non-steroidal anti-inflammatory drugs (NSAID) and P-gp-modulating drugs and renal function was impaired. We conclude that NSAID and P-gp-modulating drugs should be avoided in dabigatran-treated patients. If renal function deteriorates the dabigatran-dosage should be reduced or the therapy should be stopped. There is an urgent need to increase knowledge about drug interactions with dabigatran.


DNA methylation profiling can classify HIV-associated lymphomas.
HIV-positive patients have a 60-fold to 200-fold increased incidence of non-Hodgkin lymphomas, including Burkitt lymphoma, diffuse large B-cell lymphoma, and primary central nervous system lymphoma. HIV-associated lymphomas frequently have features such as extranodal involvement, decreased responses to standard chemotherapy, and high relapse rates, which indicate a poor prognosis. General pathological features do not clearly differentiate HIV-associated lymphomas from non-HIV lymphomas. To investigate the features of HIV-associated lymphomas, we performed genome-wide DNA methylation profiling of HIV and non-HIV lymphomas using Illumina GoldenGate Methylation Cancer Panel I and Illumina Infinium HumanMethylation450 BeadChip microarrays. DNA methylation profiles in HIV-associated and non-HIV lymphomas were characterized using unsupervised hierarchical clustering analyses. The analyses of promoter regions revealed unique DNA methylation profiles in HIV-associated lymphomas, suggesting profile differences compared with non-HIV lymphomas, which implies specific gene regulation in HIV-associated lymphoma involving DNA methylation. Based on HumanMethylation450 BeadChip data, 2541 target sites were selected as differing significantly in comparisons between HIV-associated and non-HIV-associated lymphomas using Wilcoxon's rank-sum test (P <0.05) and Δβ values more than 0.30. Recurrent cases of HIV-associated lymphoma had different profiles compared with nonrecurrent HIV lymphomas. DNA methylation profiling indicated that 2541 target sites differed significantly in HIV-associated lymphoma, which may partly explain the poor prognosis. Our data indicate that the methylation profiles of target genes have potential in elucidating HIV-associated lymphomagenesis and can serve as new prognostic markers.


Expression of completely gamma-carboxylated recombinant human prothrombin.
Human prothrombin cDNA has been expressed in mammalian cells to yield biologically active, fully gamma-carboxylated prothrombin. A 2.0-kilobase cDNA encoding full-length prothrombin was isolated from a human fetal liver library using a cDNA fragment recovered from a lambda gt11 human hepatoma expression library. Prothrombin cDNA was cloned into a mammalian expression vector and transfected into Chinese hamster ovary cells. Selection for expression of dihydrofolate reductase yielded cell lines secreting up to 0.55 microgram/ml of prothrombin. Recombinant prothrombin synthesized in the presence of vitamin K was quantitatively recovered from tissue culture medium by affinity chromatography using conformation-specific antibodies directed against the metal-stabilized, gamma-carboxylated conformer. The purified material migrated as a single band on denaturing polyacrylamide gels with an electrophoretic mobility equivalent to that of plasma-derived human prothrombin. Automated Edman degradation of recombinant prothrombin revealed a single amino-terminal sequence identical to that of plasma-derived prothrombin. Recombinant and plasma-derived prothrombin interacted similarly with antibodies specific for total prothrombin, abnormal des-gamma-carboxyprothrombin, and two metal-stabilized conformers of prothrombin. Recombinant prothrombin exhibited a specific coagulant activity equivalent to that of plasma-derived prothrombin. The gamma-carboxyglutamic acid analysis of recombinant prothrombin demonstrated 9.9 +/- 0.4 mol of gamma-carboxyglutamic acid/mol of prothrombin. These results represent the first description of the expression of a recombinant vitamin K-dependent protein in which all of the expressed protein is gamma-carboxylated.


Does omental pedicle flap reduce anastomotic leak and septic complications after rectal cancer surgery?
Whether creation of omental pedicle flap (OPF) to reinforce bowel anastomosis can reduce septic outcomes remains controversial. The aim of this study was to investigate the role of this technique on anastomotic leak and septic complications after rectal cancer surgery. Patients who underwent rectal cancer surgery from 01/2008 to 12/2013 were identified and categorized into two groups based on OPF creation versus no-OPF creation. Clinical, operative characteristics and postoperative anastomotic leak and surgical site infections within 30 days after surgery were compared between the groups. There were 65 (14%) and 403 (86%) patients in OPF and no-OPF group, respectively. In multivariate analysis, OPF was not found to be associated with anastomotic leak (p = 0.35), organ/space infections (p = 0.99) and overall surgical site infections (p = 0.65). Three hundred and sixty eight (78.6%) patients underwent diversion. OPF did not reduce septic complications irrespective of the stoma status (p > 0.05). There were no differences between the two groups in terms of operative (p = 0.46) and non-operative management (p = 0.14). OPF neither reduced the incidence of anastomotic leak and surgical site infections nor had any impact on the management of anastomotic leak.


Unilateral palatal and abducens palsy in childhood hepatitis A virus infection.
Isolated cranial nerve paresis in childhood hepatitis A virus infection is rare. The authors report an instance of concomitant right-hand side palatal and abducens palsy, developing in the course of an otherwise uncomplicated hepatitis A virus infection in a 5-year-old girl. The neurological complications were transient, and she recovered completely with supportive therapy.


[Combination chemotherapy including cisplatin in lung cancer by bronchial artery infusion].
Chemotherapy with bronchial artery infusion (BAI) was given to 34 patients with primary lung cancer. Treatment regimens usually employed cis-diammine-dichloroplatinum (CDDP) plus peplomycin for squamous cell carcinoma, and CDDP plus vindesine for adenocarcinoma. The provisional therapeutic effects were evaluated roentgenographically with reference to histological type, T factor and degree of vascularization. Out of 10 cases of squamous cell carcinoma, 7 cases (70%) showed tumor regression greater than 50%, in contrast to 4 of 17 cases (23.5%) of adenocarcinoma. The effects in cases of squamous cell carcinoma were correlated with tumor vascularity. Twenty-two surgically treated cases were examined for the histological effects of BAI. Five of 6 cases (83.3%) of squamous cell carcinoma showed IIb effects by Shimosato's criteria. These results showed that the therapeutic effect of BAI was excellent in cases of squamous cell carcinoma in comparison with cases of adenocarcinoma. Serious side effects including esophago-bronchial fistula, massive hemoptysis and esophageal ulcer were observed in 4 cases.


Occurrence of malignant neoplasms in patients with chronic lymphatic leukaemia.
In a group of 280 patients with chronic lymphatic leukaemia, a coexistence of malignant tumours diagnosed simultaneously with the leukaemia was found in 14 cases or 5.0%. No significant difference was observed between the development of the second malignant neoplasm in the patients studied and the expected incidence in the general population, taking into account also the sex and age of patients and their place of residence. No statistically significant relation was found between the duration of CLL and the occurrence of second malignancies. No significant correlation was found between the total dose of cytostatic drugs, corticosteroids or radiotherapy and the incidence of cancer.


Pioglitazone prevents acute liver injury induced by ethanol and lipopolysaccharide through the suppression of tumor necrosis factor-alpha.
Peroxisome proliferator-activated receptor-gamma (PPAR-gamma), which is a ligand-dependent transcriptional factor, forms a heterodimer with retinoid X receptor (RXR) and controls many genes that are relevant to the regulation of lipid metabolism and insulin sensitization. Recent studies have shown that stimulation of PPAR-gamma inhibits the production of inflammatory cytokines in monocytes and macrophages. Alcohol and lipopolysaccharide (LPS) have already been shown to induce liver injury through the activation of many inflammatory cytokines. Thus, the activation of PPAR-gamma by its ligand may represent a potential effect causing liver injury. In this study, we investigated the effects of pioglitazone, a ligand for PPAR-gamma, on acute liver injury induced by ethanol and LPS. Female Sprague-Dawley rats that weighed 300 g were given ethanol (5 g/kg body weight) intragastrically and received an intraperitoneal injection of LPS 24 hr later. Subsequently, pioglitazone (1 mg/kg body weight) or vehicle alone was injected intraperitoneally 10 min and 24 hr after ethanol administration. Plasma levels of aspartate transaminase and alanine aminotransferase were measured by spectrophotometer. Plasma levels of tumor necrosis factor-alpha (TNF-alpha) were also determined using an enzyme-linked immunosorbent assay. Plasma and hepatic levels of lipid peroxide were measured, and the histologic findings of the liver were examined. Reverse transcription-polymerase reaction analysis of TNF-alpha, PPAR-gamma, RXR-alpha, and beta-actin mRNA was performed. Western blot analysis using the p65 subunit of NF-kappaB was also performed. Pioglitazone prevented increase in plasma aspartate transaminase, alanine aminotransferase, and TNF-alpha levels but had no effect on plasma and hepatic levels of lipid peroxide. Pioglitazone also prevented hepatic inflammation and necrosis induced by ethanol and LPS. Ethanol and LPS induction of TNF-alpha mRNA in the liver was blunted by pioglitazone; however, RXR-alpha mRNA was not affected. PPAR-gamma mRNA levels were suppressed by ethanol and LPS but recaptured by pioglitazone. Western blot analysis showed that pioglitazone did not inhibit translocation of NF-kappaB to nuclei. These results suggest that pioglitazone may prevent liver injury induced by ethanol and LPS through the suppression of TNF-alpha.


Automatic detection of obstructive sleep apnea using speech signals.
Obstructive sleep apnea (OSA) is a common disorder associated with anatomical abnormalities of the upper airways that affects 5% of the population. Acoustic parameters may be influenced by the vocal tract structure and soft tissue properties. We hypothesize that speech signal properties of OSA patients will be different than those of control subjects not having OSA. Using speech signal processing techniques, we explored acoustic speech features of 93 subjects who were recorded using a text-dependent speech protocol and a digital audio recorder immediately prior to polysomnography study. Following analysis of the study, subjects were divided into OSA (n=67) and non-OSA (n=26) groups. A Gaussian mixture model-based system was developed to model and classify between the groups; discriminative features such as vocal tract length and linear prediction coefficients were selected using feature selection technique. Specificity and sensitivity of 83% and 79% were achieved for the male OSA and 86% and 84% for the female OSA patients, respectively. We conclude that acoustic features from speech signals during wakefulness can detect OSA patients with good specificity and sensitivity. Such a system can be used as a basis for future development of a tool for OSA screening.


Modulation of microglia can attenuate neuropathic pain symptoms and enhance morphine effectiveness.
Microglia play a crucial role in the maintenance of neuronal homeostasis in the central nervous system, and microglia production of immune factors is believed to play an important role in nociceptive transmission. There is increasing evidence that uncontrolled activation of microglial cells under neuropathic pain conditions induces the release of proinflammatory cytokines (interleukin - IL-1beta, IL-6, tumor necrosis factor - TNF-alpha), complement components (C1q, C3, C4, C5, C5a) and other substances that facilitate pain transmission. Additionally, microglia activation can lead to altered activity of opioid systems and neuropathic pain is characterized by resistance to morphine. Pharmacological attenuation of glial activation represents a novel approach for controlling neuropathic pain. It has been found that propentofylline, pentoxifylline, fluorocitrate and minocycline decrease microglial activation and inhibit proinflammatory cytokines, thereby suppressing the development of neuropathic pain. The results of many studies support the idea that modulation of glial and neuroimmune activation may be a potential therapeutic mechanism for enhancement of morphine analgesia. Researchers and pharmacological companies have embarked on a new approach to the control of microglial activity, which is to search for substances that activate anti-inflammatory cytokines like IL-10. IL-10 is very interesting since it reduces allodynia and hyperalgesia by suppressing the production and activity of TNF-alpha, IL-1beta and IL-6. Some glial inhibitors, which are safe and clinically well tolerated, are potential useful agents for treatment of neuropathic pain and for the prevention of tolerance to morphine analgesia. Targeting glial activation is a clinically promising method for treatment of neuropathic pain.


Facilitation of aggressive and sexual behaviors by saccharin deprivation in rats.
The 'deprivation effect' (DE) phenomenon is expressed as an increase in the level of free choice consumption of drugs, alcohol, or saccharin following a period of forced abstinence in humans and in several species of laboratory animals. The DE may reflect relapse-like drinking and be relevant for modeling addictive behaviors. In humans, drug or alcohol abstinence is commonly associated with the increased physical and sexual abuse. The present study aimed to study whether aggressive and sexual behaviors are affected by the conditions known to result in the DE. Adult male Wistar rats had unlimited free choice access to water and saccharin-containing solution (0.1%, w/v). After the preference for saccharin was established, animals underwent repeated 7-day-long episodes of saccharin deprivation. It was found that (i) aggressive and sexual behaviors were facilitated after 7 days of saccharin deprivation, and (ii) DE was significantly reduced in animals that were allowed to interact with the nonaggressive male or receptive female conspecifics during the last day of deprivation exposure. These results suggest that the saccharin deprivation exposures may facilitate aggressive and sexual behaviors. Alternative behavioral strategy may serve as a mechanism of coping with the deprivation state.


Post-translational modification of a chimeric EPO-Fc hormone is more important than its molecular size in defining its in vivo hematopoietic activity.
Recombinant erythropoietin (EPO) has been marketed as biopharmaceutical for anemia and chronic renal failure. Long-acting EPO variants that aimed at achieving less frequent dosing have been generated, either by the addition of glycosylation sites or increasing its molecular weight. The hEPO cDNA linked to the human IgG Fc fragment was cloned as a single codifying gene on the pAdtrack-CMV vector, yielding the recombinant adenoviral genome. For in vitro and in vivo expression assays cervical cancer cell line (SiHa) and nulliparous goats were used, respectively. The hematopoietic activity of EPO-Fc, expressed as the differential increment of hematocrit was evaluated in B6D2F1 mice. NP-HPLC of the 2AB-labeled N-glycan was carried out to profile analysis. The direct transduction of mammary secretory cells with adenoviral vector is a robust methodology to obtain high levels of EPO of up to 3.5mg/mL in goat's milk. SiHa-derived EPO-Fc showed significant improvement in hematopoietic activity compared to the commercial hEPO counterpart or with the homologous milk-derived EPO-Fc. The role of the molecular weight seemed to be important in enhancing the hematopoietic activity of SiHa-derived EPO-Fc. However, the lack of sialylated multi-antennary glycosylation profile in milk-derived EPO-Fc resulted in lower biological activity. The low content of tri- or tetra-antennary sialylated N-glycans linked to the chimeric EPO-Fc hormone, expressed in the goat mammary gland epithelial cells, defined its in vivo hematopoietic activity. The sialylated N-glycan content plays a more significant role in the in vivo biological activity of hEPO than its increased molecular weight.


A prospective study of health-related quality-of-life outcomes for patients with low-risk prostate cancer managed by active surveillance or radiation therapy.
Patients with low-risk prostate cancer (PCa) often have excellent oncologic outcomes. However, treatment with curative intent can lead to decrements in health-related quality of life (HRQoL). Patients treated with radical prostatectomy have been shown to suffer declines in urinary and sexual HRQoL as compared to those managed with active surveillance (AS). Similarly, patients treated with external-beam radiation therapy (EBRT) are hypothesized to experience greater declines in bowel HRQoL. As health-related quality-of-life (HRQoL) concerns are paramount when selecting among treatment options for low-risk PCa, this study examined HRQoL outcomes in men undergoing EBRT as compared to AS in a prospective, racially diverse cohort. A prospective study of HRQoL in patients with PCa enrolled in the Center for Prostate Disease Research (CPDR) Multicenter National Database was initiated in 2007. The current study included patients diagnosed through April 2014. HRQoL was assessed with the Expanded Prostate Cancer Index Composite (EPIC) and the Medical Outcomes Study Short Form (SF-36). Temporal changes in HRQoL were compared for patients with low-risk PCa managed on AS vs. EBRT at baseline, 1-, 2-, and 3 years post-PCa diagnosis. Longitudinal patterns were modeled using linear regression models fitted with generalized estimating equations (GEE), adjusting for baseline HRQoL, demographic, and clinical patient characteristics. Of the 499 eligible patients with low-risk PCa, 103 (21%) selected AS and 60 (12%) were treated with EBRT. Demographic characteristics of the treatment groups were similar, though a greater proportion of patients in the EBRT group were African American (P = 0.0003). At baseline, both treatment groups reported comparable HRQoL. EBRT patients experienced significantly worse bowel function and bother at 1 year (adjusted mean score: 87 vs. 95, P = 0.001 and 89 vs. 95, P = 0.008, respectively) and 2 years (87 vs. 93, P = 0.007 and 87 vs. 96, P = 0.002, respectively) compared to patients managed on AS. In contrast to those on AS, more than half the number of patients who received EBRT experienced a decline in bowel function (52% vs. 17%, p=0.003) and bother (52% vs. 15%, P = 0.002) from baseline to 1 year. Patients who received EBRT were significantly more likely to experience a decrease in more than one functional domain (urinary, sexual, bowel, or hormonal) at 1 year when compared with those on AS (60% vs. 28%, P = 0.004). Patients receiving EBRT for low-risk prostate cancer suffer declines in bowel HRQoL. These declines are not experienced by patients on AS, suggesting that management of low-risk prostate cancer with AS may offer a means for preserving HRQoL following prostate cancer diagnosis.


Why do certain primary health care teams respond better to intimate partner violence than others? A multiple case study.
To analyse how team level conditions influenced health care professionals' responses to intimate partner violence. We used a multiple embedded case study. The cases were four primary health care teams located in a southern region of Spain; two of them considered "good" and two s "average". The two teams considered good had scored highest in practice issues for intimate partner violence, measured via a questionnaire (PREMIS - Physicians Readiness to Respond to Intimate Partner Violence Survey) applied to professionals working in the four primary health care teams. In each case quantitative and qualitative data were collected using a social network questionnaire, interviews and observations. The two "good" cases showed dynamics and structures that promoted team working and team learning on intimate partner violence, had committed social workers and an enabling environment for their work, and had put into practice explicit strategies to implement a women-centred approach. Better individual responses to intimate partner violence were implemented in the teams which: 1) had social workers who were knowledgeable and motivated to engage with others; 2) sustained a structure of regular meetings during which issues of violence were discussed; 3) encouraged a friendly team climate; and 4) implemented concrete actions towards women-centred care.


Correlated fine structure branching ratios arising from state-selected predissociation of ClO (A 2Pi 3/2).
We have extended our investigation of the v'-dependent predissociation dynamics of the ClO A(2)Pi(3/2) state using velocity-map ion imaging. Correlated fine-structure branching ratios are reported for v' = 0-5. The measured branching ratios are non-statistical and are qualitatively inconsistent with adiabatic dissociation dynamics. The coupling constants between the A(2)Pi(3/2) state and several dissociative excited state potentials have been optimized, as have the locations of the crossing points, based on comparison to previously reported v'-dependent predissociation rates. Using these optimized potentials we have modeled the branching ratios in the diabatic limit but the lack of agreement with experiments suggests the importance of exit channel coupling. Coupled channel calculations including 9 coupled potentials provide modest improvement with experiment but differences remain.


Differentiation between senescence (M1) and crisis (M2) in human fibroblast cultures.
Normal human fibroblasts undergo only a limited number of divisions in culture and eventually enter a nonreplicative state designated senescence or mortality stage 1 (M1). Expression of certain viral oncogenes, such as the SV40 large T antigen (SV40 T-Ag), can elicit a significant extension of replicative life span, but these cultures eventually also cease dividing. This proliferative decline has been designated crisis or mortality stage 2 (M2). BrdU incorporation assays are commonly used to distinguish between senescence (<5% labeling index) and crisis (>30% labeling index). It has not been possible, however, to ascertain whether the high labeling index, indicative of ongoing DNA replication, was caused by the presence of T-Ag. We used gene targeting to knock out both copies of the p21(CIP1/WAF1) gene in presenescent human fibroblasts. p21 -/- cells displayed an extended life span but eventually entered a nonproliferative state. In their terminally nonproliferative state both p21 +/+ and p21 -/- cultures were positive for the senescence-associated beta-galactosidase (SA-beta-gal) activity; in contrast, the labeling index of p21 +/+ cells was low (<5%) whereas the labeling index of p21 -/- cells was high (>30%). The observation that p21 -/- and SV40 T-Ag-expressing cells behave identically with respect to life span extension as well as the high labeling index in the terminally nonproliferative state indicates that crisis is not a phenomenon induced solely by viral oncogenes, but a physiological state resulting from the bypass of normal senescence mechanisms. The widely used biomarker for senescence, SA-beta-gal, cannot distinguish between senescence and crisis. We propose that all SA-beta-gal-positive cultures should be further examined for their BrdU labeling index.


Nationwide survey of the role of travel medicine in primary care in Germany.
With more persons traveling to (sub)tropical destinations, travel medicine is an increasing challenge for the health care system in many countries. This study investigated the role and scope of travel medicine provided by general practitioners (GPs) in primary health care, assessed the determinants of specific prophylactic recommendations, and identified the needs of GPs regarding training and cooperation in travel medicine. A national survey was conducted among randomly selected GPs in Germany. Information on the type and extent of activities in travel medicine, the number and characteristics of patients, the use of specific information systems, and training and cooperation, was obtained with a standardized postal questionnaire. Representativeness was assessed by a computer-assisted telephone survey among nonrespondents. Univariate and multivariate statistical analysis was performed. Of all GPs (n=1,320), 95% provided medical pretravel advice or post-travel counseling (13.2 patients/month). The most common topics in pretravel consultation were immunizations (95%), malaria chemoprophylaxis (94%), advice on exposure prophylaxis (41%), and advice on protection from sexually transmitted infections (STIs) (43%). GPs frequently advised certain patient groups (e.g., cardiovascular patients 68%, pregnant women 20%). Travelers' diarrhea was the most common topic in post-travel consultation (89%), followed by fever (38%). Of the GPs, 25% cooperated regularly with institutions with special expertise in travel medicine or tropical diseases. Sixty-four percent had received some training in travel medicine, and 86% stated a need for additional training. In the multivariate analysis, independent determinants for advice on exposure prophylaxis and STI risks included special training in travel medicine, use of country-specific information sources, and location of the practice in the former West Germany and urban areas. Travel medicine is of importance in primary health care in Germany. There is a need for improved counseling on exposure and STI prophylaxis. GPs' training in travel medicine and cooperation with institutions such as institutes, clinics and laboratories for tropical medicine should be improved.