Breast cancer screening programme in The Netherlands: an interim review.
The year 1988 saw the introduction of a nationwide breast cancer screening programme in the Netherlands. Some 10 years on, a network that extended throughout the country was in place and all women in the target group had been invited to undergo screening at least once. Of the original target group, which was made up of women aged 50-70, around 80% responded to this invitation, an indication of their trust. Is that trust justified? and do the results justify the enormous effort of all those who make the screening programme possible, the cooperation of the nearly 4 million women who by now have taken part and the 17 million pound sterling that are currently needed per year to fund the programme? The time has come to assess and review. The ultimate test is obviously whether women participating in the screening programme have a lower chance of dying of breast cancer than women not participating in the programme. Yet, as the breast cancer screening programme is one of the best documented facilities in the Dutch health care system, a reliable mid-term review can be made. The article by Fracheboud that appears in this issue provides a number of important results. Important elements of this programme are considered in this review.


Clinical effect of acute complete acromioclavicular joint dislocation treated with micro-movable and anatomical acromioclavicular plate.
We evaluated the long-term clinical results of acute complete acromioclavicular dislocations treated with micro-movable and anatomical acromioclavicular plate. Open reduction and internal fixation was performed using the MAAP in 16 patients (10 males, 6 females; mean age 36 years; range 16 to 63 years) with acute complete acromioclavicular joint dislocation. Radiographic evaluations were routinely conducted every 3 weeks until 3 months postoperatively. The MAAP were removed under local anesthesia after 3 months postoperatively. We evaluated the functional results by using the constant scoring system and radiological results in the last follow-up time. The mean follow up was 26 months (range 16 to 38 months). The mean Constant score was 94 (range, 78 to 100). The results were excellent in 12 patients (75.0%), good in 3 patients (18.8%) and satisfactory in 1 patient (6.2%). Three patients with scores of 80 to 90 had mild pain during activity, but have not affected the shoulder range of motion. One patient has both some pain and limited range of motion of shoulder joint. All patients but one have returned to their preoperative work without any limitations. Compared to the contralateral side, radiography showed anatomical reposition in the vertical plane in 14 cases, slight loss of reduction in 2 older patients. We recommend the MAAP fixation for surgical treatment of acute complete acromioclavicular joint dislocation as it could provide satisfactory shoulder functions and clinical results, with lower complication rate. However, it is necessary to continue to observe the clinical effects of this fixation technique.


Morphometry of the fibular collateral ligament: anatomic study with comprehensive review of the literature.
There is no consensus in the literature related to the morphology, bony attachments, and variations of the fibular collateral ligament (FCL) of the knee. Our purpose was to investigate FCL morphology and to review reports in the literature regarding this structure. Seventy knees from formalin-fixed, adult cadavers were dissected and a digital caliper was used to measure FCL length, width, distance from proximal attachment to articular surface (PA→AS), and distance from distal attachment to articular surface (DA→AS). The mean (SE) length and width of all FCLs was 48.3 (1.1) mm and 4 (0.16) mm, respectively. The mean (SE) PA→AS and DA→AS of all FCLs was 22 (0.8) mm and 24.8 (1) mm, respectively. We found a direct relationship between the PA→AS and DA→AS distances (Spearman rho = 0.527, P = 0.002) and this association was independent of age and sex. Two FCL variations were found: a bifurcate ligament with two distal bands and a trifurcate ligament with three distal bands, all of which attached to the fibular head. Our literature review revealed that only 2/10 cadaveric FCL morphology studies reported variations similar to the current study. Further, there was variability in the reported location of the FCL proximal attachment. When combining our data with these studies, 105/219 FCLs directly attached to the apex of the lateral epicondyle (LE), 10/219 to a fovea posterior to the LE, and 104/219 posterior and proximal to the LE. These data may have implications related to FCL injury and repair.


Immunisation with foamy virus Bet fusion proteins as novel strategy for HIV-1 epitope delivery.
The induction of 2F5- and 4E10-like antibodies broadly neutralising HIV-1 and targeting the membrane external proximal region (MPER) of the transmembrane envelope protein gp41 would be a major advancement for the development of a preventive HIV-1 vaccine, but successful attempts remain rare. Recent studies demonstrated that broadly reactive antibodies develop relatively late during infection and after intensive affinity maturation. Therefore, a prolonged antigen delivery might be beneficial to induce them. Replicating foamy viruses which are characterised by apathogenic but persistent infection could represent suitable carrier viruses for this purpose. In order to develop such a system, we modified the accessory foamy virus Bet protein to contain the MPER of gp41, or the MPER linked to the stabilising fusion peptide proximal region of gp41 and analysed here the antigenic and immunogenic properties of such hybrid proteins. The antigens, expressed and purified to homogeneity, were recognised by the monoclonal antibodies 2F5 and 4E10 with nanomolar affinities and induced high levels of antibodies specific to gp41 after immunisation of rats. The antisera also bound to virus particles attached to infected cells, and peptide-based epitope mapping showed that they recognised the 2F5 epitope. Although no HIV-1 neutralising activity was observed, the presented data demonstrate that using the foamy virus Bet for HIV-1 epitope delivery is successfully applicable. Together with the attractive potential for sustained antigen expression after transfer to replicating virus, these results should therefore provide a first basis for the development of chimeric foamy viruses as novel HIV-1 vaccine vectors.


Cytokine profile in cervical mucosa of Japanese patients with cervical intraepithelial neoplasia.
Immune responses in the uterine cervix are considered to play an important role in persistent human papillomavirus (HPV) infection and carcinogenesis, but many aspects of the mechanism are still unclear. The goal of this study was to measure cytokines to analyze immune responses in patients with cervical intraepithelial neoplasia (CIN). The levels of 17 cytokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, G-CSF, GM-CSF, INF-γ, MCP-1, MIP-1β, and TNFα) in cervical mucus were simultaneously measured using a multiplex immunoassay in 52 high-grade squamous intraepithelial lesion (HSIL) cases and overproduction of IL-1β, IL-8, and MIP-1β was identified. The levels of these 3 cytokines were measured in 130 patients with or without CIN lesions using enzyme-linked immunosorbent assay. The associations of the cytokine levels with the cytology, infecting HPV type, and status of cigarette smoking were investigated. IL-1β and IL-8 levels were associated with the cytology, and these levels were higher in HSIL cases than in NILM (negative for intraepithelial lesion and malignancy) and LSIL (low-grade squamous intraepithelial lesion) cases (P = 0.005, P = 0.001, respectively). The MIP-1β level was significantly lower in smokers (P = 0.018) and high-risk (HR)-HPV-infected patients (P = 0.021). Enhanced expression of IL-1β and IL-8 indicates that Th2 inflammatory responses become stronger in the local uterine cervical region with the progression of CIN lesions, and a decrease in the MIP-1β level may be advantageous for immunoescape of HPV. Cigarette smoking may further facilitate persistent HPV infection.


Caesarean section in uninsured women in the USA: systematic review and meta-analysis.
The aim of this study is to assess the odds of caesarean section (CS) for uninsured women in the USA and understand the underlying mechanisms as well as consequences of lower use. Systematic review and meta-analysis. PubMed, Embase, the Cochrane Library and CINAHL from the first year of records to April 2018. We included studies that reported data to allow the calculation of ORs of CS of uninsured as compared with insured women. The prespecified primary outcome was the adjusted OR of deliveries by CS of uninsured women as compared with privately or publicly insured women. The prespecified secondary outcome was the crude OR of deliveries by CS of uninsured women as compared with insured women. 12 articles describing 16 separate studies involving more than 8.8 million women were included in this study. We found: 0.70 times lower odds of CS in uninsured as compared with privately insured women (95% CI 0.63 to 0.78), with no relevant heterogeneity between studies (τ2=0.01); and 0.92 times lower odds for CS in uninsured as compared with publicly insured women (95% CI 0.80 to 1.07), with no relevant heterogeneity between studies (τ2=0.02). We found 0.70 times lower odds in uninsured as compared with privately and publicly insured women (95% CI 0.69 to 0.72). CSs are less likely to be performed in uninsured women as compared with insured women. While the higher rates for CS among privately insured women can be explained with financial incentives associated with private insurance, the lower odds among uninsured women draw attention at barriers to access for delivery care. In many regions, the rates for uninsured women are above, close or below the benchmarks for appropriate CS rates and could imply both, underuse and overuse.


Role of keratinocytes in human recurrent herpetic lesions. Ability to present herpes simplex virus antigen and act as targets for T lymphocyte cytotoxicity in vitro.
In human recurrent herpetic lesions epidermal keratinocytes are induced to express HLA class II (DR) antigens. Keratinocytes derived from human split skin and cultured in vitro were induced to express HLA-DR but not -DQ antigens with IFN gamma preparations. These stimulated keratinocytes presented herpes simplex antigen directly to autologous blood-derived T lymphocytes in four of four subjects (stimulation indices: 1.5-2.7), suggesting that keratinocytes may have an accessory herpes simplex virus (HSV) antigen-presenting role in addition to the Langerhans cells and macrophages in herpetic skin lesions. Blood mononuclear cells from eight herpes simplex seropositive subjects which were activated in vitro by HSV antigen for 6 d showed cytotoxicity specific for HSV in infected autologous keratinocytes. This was significantly increased by prestimulation with IFN gamma (51-56% to 83-85%). In four of eight patients some cytotoxicity also occurred against uninfected, IFN gamma-stimulated keratinocytes. Lymphocyte subset analysis showed that cytotoxicity against HSV-infected, IFN gamma-stimulated keratinocyte targets was mediated by both CD3+ T lymphocytes and Leu 11b+ natural killer cells. T lymphocyte cytotoxicity was mediated by both CD4+ and CD8+ T lymphocytes, suggesting a cytotoxic role for the activated CD4+ lymphocytes that initially predominate in herpetic lesions.


LC-MALDI-TOF imaging MS: a new approach in combining chromatography and mass spectrometry of copolymers.
A new approach that utilizes MALDI-TOF imaging mass spectrometry as a new detector for polymer chromatography is presented. For the first time, the individual retention behavior of single structural units of polyethylene oxide (PEO)/polypropylene oxide (PPO) copolymers and changes of the copolymer composition could be monitored. Composition specific calibration curves could be easily obtained by displaying the copolymer ion intensity data. This approach provides completely new insights in the chromatographic principle of copolymer separation and could be used to easily modify and adapt conditions for separation. In combination with electrospray deposition, homogeneous sample/matrix traces of surprisingly high spatial resolution could be obtained.


Intracellular two-phase Ca2+ release and apoptosis controlled by TRP-ML1 channel activity in coronary arterial myocytes.
Activation of the death receptor Fas has been reported to produce a two-phase intracellular Ca(2+) release response in coronary arterial myocytes (CAMs), which consists of local Ca(2+) bursts via lysosomal transient potential receptor-mucolipin 1 (TRP-ML1) channels and consequent Ca(2+) release from the sarcoplasmic reticulum (SR). The present study was designed to explore the molecular mechanism by which lysosomal Ca(2+) bursts are coupled with SR Ca(2+) release in mouse CAMs and to determine the functional relevance of this lysosome-associated two-phase Ca(2+) release to apoptosis, a common action of Fas activation with Fas ligand (FasL). By confocal microscopy, we found that transfection of CAMs with TRP-ML1 small interfering (si)RNA substantially inhibited FasL (10 ng/ml)-induced lysosome Ca(2+) bursts and consequent SR Ca(2+) release. In contrast, transfection of CAMs with plasmids containing a full-length TRP-ML1 gene enhanced FasL-induced two-phase Ca(2+) release. We further demonstrated that FasL significantly increased the colocalization of the lysosomal marker Lamp1 with ryanodine receptor 3 and enhanced a dynamic trafficking of lysosomes to the SR. When CAMs were treated with TRP-ML1 siRNA, FasL-induced interactions between the lysosomes and SR were substantially blocked. Functionally, FasL-induced apoptosis and activation of calpain and calcineurin, the Ca(2+) sensitive proteins that mediate apoptosis, were significantly attenuated by silencing TRP-ML1 gene but enhanced by overexpression of TRP-ML1 gene. These results suggest that TRP-ML1 channel-mediated lysosomal Ca(2+) bursts upon FasL stimulation promote lysosome trafficking and interactions with the SR, leading to apoptosis of CAMs via a Ca(2+)-dependent mechanism.


Fast assessment of stroke and transient ischaemic attack to prevent early recurrence (FASTER): a randomised controlled pilot trial.
Patients with transient ischaemic attack (TIA) or minor stroke are at high immediate risk of stroke. The optimum early treatment options for these patients are not known. Within 24 h of symptom onset, we randomly assigned, in a factorial design, 392 patients with TIA or minor stroke to clopidogrel (300 mg loading dose then 75 mg daily; 198 patients) or placebo (194 patients), and simvastatin (40 mg daily; 199 patients) or placebo (193 patients). All patients were also given aspirin and were followed for 90 days. Descriptive analyses were done by intention to treat. The primary outcome was total stroke (ischaemic and haemorrhagic) within 90 days. Safety outcomes included haemorrhage related to clopidogrel and myositis related to simvastatin. This study is registered as an International Standard Randomised Controlled Trial (number 35624812) and with ClinicalTrials.gov (NCT00109382). The median time to stroke outcome was 1 day (range 0-62 days). The trial was stopped early due to a failure to recruit patients at the prespecified minimum enrolment rate because of increased use of statins. 14 (7.1%) patients on clopidogrel had a stroke within 90 days compared with 21 (10.8%) patients on placebo (risk ratio 0.7 [95% CI 0.3-1.2]; absolute risk reduction -3.8% [95% CI -9.4 to 1.9]; p=0.19). 21 (10.6%) patients on simvastatin had a stroke within 90 days compared with 14 (7.3%) patients on placebo (risk ratio 1.3 [0.7-2.4]; absolute risk increase 3.3% [-2.3 to 8.9]; p=0.25). The interaction between clopidogrel and simvastatin was not significant (p=0.64). Two patients on clopidogrel had intracranial haemorrhage compared with none on placebo (absolute risk increase 1.0% [-0.4 to 2.4]; p=0.5). There was no difference between groups for the simvastatin safety outcomes. Immediately after TIA or minor stroke, patients are at high risk of stroke, which might be reduced by using clopidogrel in addition to aspirin. The haemorrhagic risks of the combination of aspirin and clopidogrel do not seem to offset this potential benefit. We were unable to provide evidence of benefit of simvastatin in this setting. This aggressive prevention approach merits further study.


Appreciation of osteoporosis among men with hyperparathyroidism.
To define the scope of bone disease among men referred for parathyroidectomy and to document bone density screening trends in this high-risk population. Clinical data were analyzed from a prospectively maintained database of 1000 patients undergoing parathyroidectomy for sporadic primary hyperparathyroidism at the Cleveland Clinic between 2000 and 2006. Information collected included demographics, reason for referral, body mass index, intraoperative findings, preoperative and postoperative laboratory values (serum calcium, phosphorus, parathyroid hormone, 25-hydroxyvitamin D, and 24-hour urinary calcium excretion), and preoperative and postoperative dual-energy x-ray absorptiometry (DXA) findings. Of the 1000 patients, 243 (24%) were men. As with women, bone health-related issues were the most common reason for referral (32%). In 2000, only 12% of men had preoperative DXA scans; this rose to 42% by 2005. The frequency of prereferral DXA screening increased throughout the study, but even by 2006, referring physicians did not screen most of their male patients. In 2000, the prevalence of bone disease (osteoporosis or osteopenia) in men was 8%, but with improved screening, this increased to 26%, approaching the 34% rate in women. Preoperative and postoperative levels of calcium, parathyroid hormone, phosphorus, 24-hour urinary calcium, and 25-hydroxyvitamin D were the same among men with and without bone disease. Analysis of postoperative DXA scans revealed that gains in postparathyroidectomy bone density were significantly greater in men than in women at all anatomic testing sites. In men, lowest T scores improved by a mean +/- SEM of +0.35 +/- 0.09 compared with +0.098 +/- 0.035 in women (P = .009). Men were 4 times less likely than women to have continued bone loss after parathyroidectomy. Clinical and laboratory parameters cannot identify men with hyperparathyroidism who have bone loss, illustrating the need for routine DXA screening. Despite this, DXA remains underused. Improved screening practices will favorably effect men's health.


Levodopa-induced dyskinesia in Parkinson disease: A population-based cohort study.
To assess dyskinesia frequency in a population-based cohort of patients with Parkinson disease (PD). Dyskinesia complicates levodopa treatment and affects quality of life. Utilizing the 1991-2010 population-based, parkinsonism-incident cohort of Olmsted County, MN (n = 669), accessed via the Rochester Epidemiology Project, we identified patients with PD and abstracted levodopa-related dyskinesia information. Of 309 patients with PD (46.2% with parkinsonisms), 279 (90.3%) received levodopa. Most (230/279; 82.4%) had been treated by a Mayo Clinic neurologist. Median age of the 309 patients with PD at the time of diagnosis was 74.1 years (range 33.1-97.8 years). Median-age levodopa initiation in this cohort was 75 years (range 37-98 years), and median-duration levodopa treatment was 6 years (range 2 months to 19.8 years). Dyskinesia was documented in 84 of 279 patients (30.1%). Median time from levodopa initiation to dyskinesia onset was 4 years (range 2 months to 20 years); those with dyskinesia (65.5%; 55/84) developed it within 5 years of levodopa initiation (9 within the first year). Dyskinesia was mild in 57/84 (67.9%), moderate in 16/84 (19.1%), and severe in 9/84 (10.7%); severity was not reported in 2 cases. Dyskinesia severity led to levodopa adjustments or amantadine initiation in 60.7% (51/84 of those with dyskinesia), with improvement in 23/51 (45.1%). Thirteen patients with dyskinesia underwent deep brain stimulation, reporting marked improvement. Postmortem examination confirmed Lewy body disease in 7 autopsied cases. Levodopa-induced dyskinesia affected 30% of the patients with PD in our cohort. Mayo neurologists favoring levodopa dosage optimization treated most patients. Dyskinesia was severe in 3.2% of all levodopa-treated patients with PD (10.7% of all patients with dyskinesia) with marked improvement among those treated with deep brain stimulation.


Micronucleus Assay: The State of Art, and Future Directions.
During almost 40 years of use, the micronucleus assay (MN) has become one of the most popular methods to assess genotoxicity of different chemical and physical factors, including ionizing radiation-induced DNA damage. In this minireview, we focus on the position of MN among the other genotoxicity tests, its usefulness in different applications and visibility by international organizations, such as International Atomic Energy Agency, Organization for Economic Co-operation and Development and International Organization for Standardization. In addition, the mechanism of micronuclei formation is discussed. Finally, foreseen directions of the MN development are pointed, such as automation, buccal cells MN and chromothripsis phenomenon.


Suppression of laser-induced choroidal neovascularization by a CCR3 antagonist.
To evaluate the efficacy of a novel CCR3 antagonist for laser injury-induced choroidal neovascularization (CNV) in mice. We evaluated YM-344031, a novel and selective small-molecule CCR3 antagonist. CNV was induced by laser injury in C57BL/6J mice, and its volume was measured after 7 days by confocal microscopy. Leakage from the CNV was also measured after 7 days by fluorescein angiography. The CCR3 antagonist was administered by gavage at 1 hour before and 1 day after the laser injury, or intravitreous injection immediately after the laser injury. After the laser injury, ELISA, Western blot analysis, and real-time RT-PCR for VEGF-A expression in the RPE/choroid, and immunohistochemistry for CCR3, CCL11, Ki67, and Rac1 was performed. Both oral administration and intravitreous injection of YM-344031 significantly suppressed the CNV volume (P < 0.0001 and P < 0.01, respectively). Pathologically significant leakage was significantly less common in YM-344031-injected mice (P < 0.0001). The mean VEGF protein level was significantly increased in vehicle-injected eyes after the laser injury (P < 0.05). Although the YM-344031-injected eyes did not show VEGF-A suppression after the laser injury, VEGF164 mRNA upregulation was significantly suppressed in YM-344031-injected mice (P < 0.05), and intravitreous injection of YM-344031 appeared to suppress CCR3, CCL11 (eotaxin), Ki67, and Rac1 expression after the laser injury. The present data suggest that the CCR3 antagonist YM-344031 can suppress CNV, via suppression of the upregulation of VEGF164 mRNA in VEGF isoform after the laser injury. Although our findings may warrant further investigation, YM-344031 may have potential as a new therapy for age-related macular degeneration.


Lifetime data analysis of disease and aging by the Weibull probability distribution.
We applied the Weibull distribution to the life-table and age-patterns of diseases in Japan. The life-table follows a composite Weibull distribution composed of initial failure and two stage wear-out failure periods. The extension of lifespan during the past century is manifested as increases in the scale parameters in all three periods and the shape parameters in the wear-out periods with female predominancy. The shape parameters of diseases show time-independent sex-dependent specific values. When consistent changes are observed, such as increases in the shape parameters of tuberculosis and pneumonia, legitimate causes such as prevention and repair at the societal and medical level are present. Cancer and arteriosclerosis share a common range of shape parameters suggesting analogous underlying biological processes. An analysis of the epidemiology of human aging and disease by the Weibull distribution discloses intrinsic properties of man as a biological organization as well as a societal presence.


In vitro inhibition of matrix metalloproteinases, invasion and growth of Fanconi anemia human FANCA and FANCC lymphoblasts by nutrient mixture.
Fanconi anemia is a rare genetic disorder with high propensity for development of cancers, such as aplastic anemia, leukemia and head and neck cancers. Collagen digesting matrix metalloproteinase (MMP) enzymes have been implicated in for their role in various malignancies and to promote metastasis. Biological agents that prevent extracellular matrix digestion by the MMPs have been shown to be promising therapeutic approaches to cancer. In this study, we investigated effects of a nutrient mixture (NM) containing, ascorbic acid, lysine, proline and green tea extract, on human FANCA and FANCC lymphoblasts for viability, MMP secretion and invasion. Human FANCA lymphoblasts GM13022 and HCS536 were challenged with NM at concentration range within 10-1000 µg/ml. Cell toxicity was assessed by Trypan blue dye exclusion test. Invasion was evaluated through Matrigel and gelatinase zymography for MMP activity. NM was toxic in dose dependent mode to HCS536 cells but not to GM13022 cells. GM13022 cells but not HCS536 cells exhibited MMP-9 secretion, which was inhibited by NM. Matrigel invasion was inhibited in HCS536 cells at 100 and 500 µg/ml by 27% and 93%, respectively. In GM13022 cells, the NM showed completely blocked Matrigel invasion at 500 µg/ml. NM inhibited MMP secretion and Matrigel invasion in FANCA and inhibited invasion and induced toxicity in FANCC lymphoblasts. These results suggest that the NM may have therapeutic potential in Fanconi anemia associated neoplasia.


[Prenatal patient cards and quality of prenatal care in public health services in Greater Metropolitan Vitória, Espírito Santo State, Brazil].
This study aimed to assess the completeness of prenatal care information on the patients' prenatal care cards, according to coverage by various public health services: Family Health Strategy (FHS), Community-Based Health Workers' Program (CBHWP), and traditional Primary Care Units (PCU) in Greater Metropolitan Vitória, Espírito Santo State, Brazil. In a cross-sectional study, 1,006 prenatal cards were randomly selected from postpartum women at maternity hospitals in the metropolitan area. Completeness of the cards was assessed according to the criteria proposed by Romero & Cunha, which measure the quality on a scale from excellent (< 5% incomplete cards) to very bad (> 50% incomplete cards). In general, completion of information on the cards was bad (> 20% incomplete), but cards were filled out better in the FHS than in the CBHWP and PCU, especially for tetanus vaccination (p = 0.016) and gestational weight (p = 0.039). In conclusion, the quality of prenatal care in the public health system in Greater Metropolitan Vitória fails to meet the Brazilian national guidelines for maternal and child health.


Recovery of steroid induced adrenal insufficiency.
Secondary adrenal insufficiency can result from insufficient stimulation of the adrenal glands due to inadequate secretion or synthesis of adrenocorticotropic hormone (ACTH). This can be caused by hypopituitarism, central nervous system injury (tumors, radiation, and surgery) or long-term glucocorticoid therapy. Glucocorticoids were introduced in the 1950s, and have been used for their anti-inflammatory and other pharmacological effects, and also as replacement therapy for adrenal insufficiency. However, chronic glucocorticoid use may lead to suppression of the hypothalamic pituitary adrenal axis through negative feedback. This may lead to secondary adrenal insufficiency. Typically, the hypothalamic pituitary adrenal axis recovers after cessation of glucocorticoids, but the timing of recovery can be variable and can take anywhere from 6-12 months. Understanding the effect of exogenous glucocorticoids on the hypothalamic pituitary adrenal axis, recovery of the axis, and tests used to assess the recovery, are crucial to avoid prescribing unnecessary steroid replacement or missing a critical diagnosis with detrimental consequences.


Enhancement of non-invasive trans-membrane drug delivery using ultrasound and microbubbles during physiologically relevant flow.
Sonoporation has been associated with drug delivery across cell membranes and into target cells, yet several limitations have prohibited further advancement of this technology. Higher delivery rates were associated with increased cellular death, thus implying a safety-efficacy trade-off. Meanwhile, there has been no reported study of safe in vitro sonoporation in a physiologically relevant flow environment. The objective of our study was not only to evaluate sonoporation under physiologically relevant flow conditions, such as fluid velocity, shear stress and temperature, but also to design ultrasound parameters that exploit the presence of flow to maximize sonoporation efficacy while minimizing or avoiding cellular damage. Human umbilical vein endothelial cells (EA.hy926) were seeded in flow chambers as a monolayer to mimic the endothelium. A peristaltic pump maintained a constant fluid velocity of 12.5 cm/s. A focused 0.5 MHz transducer was used to sonicate the cells, while an inserted focused 7.5 MHz passive cavitation detector monitored microbubble-seeded cavitation emissions. Under these conditions, propidium iodide, which is normally impermeable to the cell membrane, was traced to determine whether it could enter cells after sonication. Meanwhile, calcein-AM was used as a cell viability marker. A range of focused ultrasound parameters was explored, with several unique bioeffects observed: cell detachment, preservation of cell viability with no membrane penetration, cell death and preservation of cell viability with sonoporation. The parameters were then modified further to produce safe sonoporation with minimal cell death. To increase the number of favourable cavitation events, we lowered the ultrasound exposure pressure to 40 kPapk-neg and increased the number of cavitation nuclei by 50 times to produce a trans-membrane delivery rate of 62.6% ± 4.3% with a cell viability of 95% ± 4.2%. Furthermore, acoustic cavitation analysis showed that the low pressure sonication produced stable and non-inertial cavitation throughout the pulse sequence. To our knowledge, this is the first study to demonstrate a high drug delivery rate coupled with high cell viability in a physiologically relevant in vitro flow system.


A pilot study of IL-2Rα blockade during lymphopenia depletes regulatory T-cells and correlates with enhanced immunity in patients with glioblastoma.
Preclinical studies in mice have demonstrated that the prophylactic depletion of immunosuppressive regulatory T-cells (T(Regs)) through targeting the high affinity interleukin-2 (IL-2) receptor (IL-2Rα/CD25) can enhance anti-tumor immunotherapy. However, therapeutic approaches are complicated by the inadvertent inhibition of IL-2Rα expressing anti-tumor effector T-cells. To determine if changes in the cytokine milieu during lymphopenia may engender differential signaling requirements that would enable unarmed anti-IL-2Rα monoclonal antibody (MAbs) to selectively deplete T(Regs) while permitting vaccine-stimulated immune responses. A randomized placebo-controlled pilot study was undertaken to examine the ability of the anti-IL-2Rα MAb daclizumab, given at the time of epidermal growth factor receptor variant III (EGFRvIII) targeted peptide vaccination, to safely and selectively deplete T(Regs) in patients with glioblastoma (GBM) treated with lymphodepleting temozolomide (TMZ). Daclizumab treatment (n = 3) was well-tolerated with no symptoms of autoimmune toxicity and resulted in a significant reduction in the frequency of circulating CD4+Foxp3+ TRegs in comparison to saline controls (n = 3)( p = 0.0464). A significant (p<0.0001) inverse correlation between the frequency of TRegs and the level of EGFRvIII specific humoral responses suggests the depletion of TRegs may be linked to increased vaccine-stimulated humoral immunity. These data suggest this approach deserves further study. ClinicalTrials.gov NCT00626015.


Interfacial synthesis of a three-dimensional hierarchical MoS2-NS@Ag-NP nanocomposite as a SERS nanosensor for ultrasensitive thiram detection.
Interfacial self-assembly of ordered nanostructures at oil-water interfaces towards the fabrication of nanofilms has attracted the interest of plenty of scientists, since its discovery in 2004. Herein, further developments have been achieved, and we report a new strategy for the synthesis of a three-dimensional (3D) hierarchical nanostructure, through an interfacial synthesis driven microemulsion process. Thus, the synthesis route has been simplified, with the rigorous experimental conditions of traditional compositing technology. Combined with a two-step seed-mediated growth method for preparing uniform Ag-NPs, a plasmonic 3D MoS2-NS@Ag-NP nanostructure was successfully developed as a Surface-Enhanced Raman Scattering (SERS) active substrate, with plenty of surface hot spots, leading to an enhancement factor (EF) of 1.2 × 108 derived from both electromagnetic mechanism (EM) and chemical mechanism (CM) effects. The 3D MoS2-NS@Ag-NP nanostructure can be applied to detect trace thiram in apple juice and local lake water, with a detection limit as low as 10 ppb (42 nM), which is much lower than the maximal residue limit (MRL) of 7 ppm in fruit prescribed by the U.S. Environmental Protection Agency (EPA). Furthermore, quantitative analysis was achieved in the range of 10 ppb-1 ppm with good homogeneity and selectivity.


Histogenetic and morphological in vivo and in vitro data concerning androgen-estrogen-induced scent gland tumor in the Syrian hamster.
The scent gland tumor of the Syrian hamster is induced by exogenous androgen and estrogen. Microscopic nodules are induced normally in old males by endogenous androgen. The histogenesis of the scent gland tumor is complex and not completely understood. In this study microscopic preneoplastic nodules and macroscopic tumors were studied by light and electron microscopy, and the macroscopic tumors were grown in tissue culture on collagen-coated coverslips and on sponge foam matrices by the organ culture method. The cultures were fed with an unfiltered fetal calf serum-bovine serum ultrafiltrate medium, which contained endogenous androgen-estrogen, 110-100 pg and could maintain growth without additional androgen-estrogen. Exogenous androgen-estrogen was also added to some cultures. Scent gland tumors grown in organ culture contained cells of two shapes, spindle and ovoid arranged in cords. Cultures on coverslips showed radiating outgrowths of spindle cells suggesting either mesenchymal or Schwann cells. By electron microscopy, both in vivo and in vitro preneoplastic and tumor samples contained cells with segments of basal lamina, micropinocytotic vesicles, and junctional complexes. These features were similar to those of poorly differentiated experimental malignant rat schwannomas maintained in similar in vitro systems. Tumors grown in vivo and in vitro were associated with collagen fibrils with a periodicity ranging from 400 to 1075 A. The evidence reported in this paper suggests that one component of the scent gland tumor is an androgen-estrogen-induced poorly differentiated schwannoma.


Barriers against required nurse estimation models applying in Iran hospitals from health system experts' point of view.
One of the strategies for accessing effective nursing care is to design and implement a nursing estimation model. The purpose of this research was to determine barriers in applying models or norms for estimating the size of a hospital's nursing team. This study was conducted from November 2015 to March 2016 among three levels of managers at the Ministry of Health, medical universities, and hospitals in Iran. We carried out a qualitative study using a Colaizzi method. We used semistructured and in-depth interviews by purposive, quota, and snowball sampling of 32 participants (10 informed experts in the area of policymaking in human resources in the Ministry of Health, 10 decision makers in employment and distribution of human resources in treatment and administrative chancellors of Medical Universities, and 12 nursing managers in hospitals). The data were analyzed by Atlas.ti software version 6.0.15. The following 14 subthemes emerged from data analysis: Lack of specific steward, weakness in attracting stakeholder contributions, lack of authorities trust to the models, lack of mutual interests between stakeholders, shortage of nurses, financial deficit, non-native models, designing models by people unfamiliar with nursing process, lack of attention to the nature of work in each ward, lack of attention to hospital classification, lack of transparency in defining models, reduced nurses available time, increased indirect activity of nurses, and outdated norms. The main themes were inappropriate planning and policymaking in high levels, resource constraints, and poor design of models and lack of updating the model. The results of present study indicate that many barriers exist in applying models for estimating the size of a hospital's nursing team. Therefore, for designing an appropriate nursing staff estimation model and implementing it, in addition to considering the present barriers, identifying the norm required features may positively impact on norm acceptance and implementation.


Validation of the Childhood Health Assessment Questionnaire in the juvenile idiopathic myopathies. Juvenile Dermatomyositis Disease Activity Collaborative Study Group.
To examine the validity of the Childhood Health Assessment Questionnaire (CHAQ) in patients with juvenile idiopathic inflammatory myopathy (IIM). One hundred fifteen patients were enrolled in a multicenter collaborative study, during which subjects were assessed twice, 7-9 months apart. Physical function was measured using the CHAQ. Internal reliability was assessed using adjusted item-total correlations and item endorsement rates. Construct validity was assessed by comparing predicted and actual correlations of the CHAQ with other measures of physical function and disease activity. Responsiveness was assessed by calculating effect size (ES) and standardized response mean (SRM) in a group of a priori defined "improvers." Item-total correlations were high (rs range = 0.35-0.81), suggesting all items were related to overall physical function. Manual muscle testing and the Childhood Myositis Assessment Scale correlated moderate to strongly with the CHAQ (r = -0.64 and -0.75, both p < 0.001). Moderate correlations were also seen with the physician global assessment of disease activity (rs = 0.58, p < 0.001), parent global assessment of overall health (rs = -0.65, p < 0.001), Steinbrocker function class (rs = 0.69, p < 0.001), and global skin activity (rs = 0.40, p < 0.001), while global disease damage and skin damage had low correlations (rs = 0.13 and 0.07, p > or =0.17). Responsiveness of the CHAQ was high, with ES = 1.05 and SRM = 1.20. In this large cohort of patients with juvenile IIM, the CHAQ exhibited internal reliability, construct validity, and strong responsiveness. We conclude that the CHAQ is a valid measure of physical function in juvenile IIM, appropriate for use in therapeutic trials, and potentially in the clinical care of these patients.


Performance metrics for an application-driven selection and optimization of psychophysical sampling procedures.
When estimating psychometric functions with sampling procedures, psychophysical assessments should be precise and accurate while being as efficient as possible to reduce assessment duration. The estimation performance of sampling procedures is commonly evaluated in computer simulations for single psychometric functions and reported using metrics as a function of number of trials. However, the estimation performance of a sampling procedure may vary for different psychometric functions. Therefore, the results of these type of evaluations may not be generalizable to a heterogeneous population of interest. In addition, the maximum number of trials is often imposed by time restrictions, especially in clinical applications, making trial-based metrics suboptimal. Hence, the benefit of these simulations to select and tune an ideal sampling procedure for a specific application is limited. We suggest to evaluate the estimation performance of sampling procedures in simulations covering the entire range of psychometric functions found in a population of interest, and propose a comprehensive set of performance metrics for a detailed analysis. To illustrate the information gained from these metrics in an application example, six sampling procedures were evaluated in a computer simulation based on prior knowledge on the population distribution and requirements from proprioceptive assessments. The metrics revealed limitations of the sampling procedures, such as inhomogeneous or systematically decreasing performance depending on the psychometric functions, which can inform the tuning process of a sampling procedure. More advanced metrics allowed directly comparing overall performances of different sampling procedures and select the best-suited sampling procedure for the example application. The proposed analysis metrics can be used for any sampling procedure and the estimation of any parameter of a psychometric function, independent of the shape of the psychometric function and of how such a parameter was estimated. This framework should help to accelerate the development process of psychophysical assessments.


The effects of inverting prisms on the horizontal-vertical illusion: a systematic effect of downward gaze.
The purpose of this work is to compare the relative contributions from the extraocular and sensory systems on the magnitude of the horizontal-vertical illusion (HVI). The visual HVI refers to the general tendency to overestimate vertical extensions of small-scale lines on a picture plane relative to the horizontal by 4-16% depending on the method of measurement. The HVI line stimuli consisted of luminous vertical and horizontal lines forming "L-profiles" located in the frontoparallel plane at a 45 cm viewing distance, collinearly with a binocular gaze. The home position of gaze was aligned to the center of the screen with the ear-eye angle concordant with the environmental horizontal. Illusion strength was quantified when subjects fixated the HVI line stimuli in four quadrants of the visual field. The HVI was also viewed through prism lenses that inverted the retinal images by 180 degrees, thereby dissociating the sensory "up-down" direction from the oculomotor up-down frame of reference. The results revealed a systematically lower magnitude of the HVI in the bottom visual field regardless of whether subjects fixated the HVI with the distorting prisms or without. Taken together, these results suggest that the HVI is sensitive to small-angle gaze shifts. In agreement with several recent findings, these results are interpreted as implying that the brain imposes an enhanced analytic structure on the ascending sensory information during downward gaze.


Multi-Component Exercise with High-Intensity, Free-Weight, Functional Resistance Training in Pre-Frail Females: A Quasi-Experimental, Pilot Study.
No study has performed an exercise intervention that included high-intensity, free-weight, functional resistance training, and assessed frailty status as an inclusion criteria and outcome measure via original, standardized tools, in pre-frail females. Determine if the intervention strategy is not only feasible and safe, but can also improve frailty status, functional task performance, and muscle strength. Pilot, quasi-experimental. Community. 20 older-adults with pre-frailty characteristics. 12-weeks (3 days/week, 45-60 minutes/session) of multi-component exercise, inclusive of aerobic, resistance, balance and flexibility exercises. The crux of the program was balance and resistance exercises, the latter utilized high-intensity, free-weight, functional resistance training. The control group maintained their usual care. 1) Feasibility and safety (dropout, adherence, and adverse event); 2) Frailty (Frailty Phenotype, Clinical Frailty Scale, and gait speed); 3) Functional task performance (grip strength and sit-to-stand time); and 4) Isometric and isotonic strength of the knee extensors and elbow flexors. No participants dropped out of the intervention or experienced an adverse event, and adherence averaged 88.3%. The exercise group became less frail, whereas the control group became more frail. There was a significant within-group improvement in exercise participants gait speed (p ≤ 0.01, +0.24 m/sec), grip strength (p ≤ 0.01, +3.9 kg), and sit-to-stand time (p ≤ 0.01, -5.0 sec). There was a significant within-group improvement in exercise participants knee extension isometric torque (p ≤ 0.05, +7.4 Nm) and isotonic velocity (p = ≤ 0.01, +37.5 ˚/sec). Elbow flexion isotonic velocity significantly declined within the control group (p ≤ 0.01, -20.2 ˚/sec) and demonstrated a significant between-group difference (p ≤ 0.05, 40.73 ˚/sec) post-intervention. The intervention strategy appears to be feasible and safe, and may also improve frailty status, functional task performance, and muscle strength. These results help calculate effect size for a future randomized controlled trial.


The 2002 HBR list. Breakthrough ideas for today's business agenda.
In the last year, war broke out, the economy took a nosedive, and an alarming number of businesses went belly up. Thus it is with some urgency that we bring you this year's list of the seven best new business ideas to help you find your way through these complex times. History Returns. Many thought that the fall of the Berlin Wall marked the beginning of a new world order, one in which history didn't matter. But September 11, 2001, put an end to that theory. And it raised serious questions about globalization, security, and strategy. Enter the Everyday Leader--at Last. CEOs are used to getting all the glory, but leaders outside the limelight--middle managers and tempered radicals--are beginning to receive the attention they deserve. The Internet Is Not About You. The real value of the Internet may lie less in connecting individuals than in connecting databases, servers, and devices--a web not of people but of machines. Mind Your Behavior. Behavioral scientists are beginning to be able to accurately predict the ways individuals and crowds will respond to stimuli, and the implications for business are profound. Don't Delight Your Customers Away. Companies have wooed and coddled customers for too long. The truth is, people delight in being teased and are repelled by those who try too hard to befriend them. Games Are for Losers. Financial game playing and verbal politicking are seriously damaging businesses. Honesty is more than an admirable virtue; it's the foundation of every lasting enterprise. Three Cheers for Creativity (Sometimes). Creativity and best-practice replication are fundamentally different undertakings. Managers who mix the two achieve creativity that is merely incremental and replication that is sadly inept.


[Tarlov's cyst: definition, etiopathogenesis, propaedeutic and treatment].
Tarlov's cyst or perineurial cyst is disease on portion of the posterior nerve root in lumbo-sacral region. The lack of knowledge of physicians around the world about Tarlov's cyst as to their nature, significance and treatment also with differential diagnostics to radiculopathy in legs. With review of literature discuss about definition, etiopathogenesis, diagnostic investigation and treatment clinic and/or surgery. The pathogenesis Tarlov's cyst remains unclear; several cases have history of the trauma, old hemorrhage, congenital and iatrogenic. Cysts provoke low back pain, sacral radiculopathy, dyspareunia, urinary incontence. The magnetic resonance imaging is now the gold standard to diagnose cysts. The treatment is clinic or surgery depending neurologics finding and neuroimage.


Kinetochores capture astral microtubules during chromosome attachment to the mitotic spindle: direct visualization in live newt lung cells.
When viewed by light microscopy the mitotic spindle in newt pneumocytes assembles in an optically clear area of cytoplasm, virtually devoid of mitochondria and other organelles, which can be much larger than the forming spindle. This unique optical property has allowed us to examine the behavior of individual microtubules, at the periphery of asters in highly flattened living prometaphase cells, by video-enhanced differential interference-contrast light microscopy and digital image processing. As in interphase newt pneumocytes (Cassimeris, L., N. K. Pryer, and E. D. Salmon. 1988. J. Cell Biol. 107:2223-2231), centrosomal (i.e., astral) microtubules in prometaphase cells appear to exhibit dynamic instability, elongating at a mean rate of 14.3 +/- 5.1 microns/min (N = 19) and shortening at approximately 16 microns/min. Under favorable conditions the initial interaction between a kinetochore and the forming spindle can be directly observed. During this process the unattached chromosome is repeatedly probed by microtubules projecting from one of the polar regions. When one of these microtubules contacts the primary constriction the chromosome rapidly undergoes poleward translocation. Our observations on living mitotic cells directly demonstrate, for the first time, that chromosome attachment results from an interaction between astral microtubules and the kinetochore.


Plasmid DNA- and messenger RNA-based anti-cancer vaccination.
Tumor cells (over-) express specific antigens which allow them to be recognized and destroyed by the immune system. Triggering anti-tumor immunity in cancer patients by specific vaccination is foreseen as a safe and versatile method to control cancer. As a source of antigen, whole tumor cells, nucleic acids, proteins or derived peptides have been used. This review focuses on the utilization of vaccines based on plasmid DNA (pDNA) and messenger RNA (mRNA) coding for tumor associated antigens. Both vectors (pDNA and mRNA) are grouped under the designation "minimal nucleic acid vector" or MNAV. The current knowledge on anti-tumor vaccination based on MNAV-encoded tumor antigens, methods of delivery, principles of production and optimization is discussed. Furthermore, an up-to-date summary of published clinical trials using MNAV for the vaccination against solid tumors is given. Recent preclinical and early phase clinical trials demonstrate promising synergies between vaccination and other treatments such as chemotherapy or non-specific immune enhancement regimens. Combining optimized MNAV formulations and parallel adjuvant treatments could allow to turn MNAV-based vaccines into efficient anti-tumor immunotherapies in humans.


CARD15/NOD2 functions as an antibacterial factor in human intestinal epithelial cells.
Mutations in the CARD15/NOD2 gene, a putative intracellular pattern recognition receptor, have been linked to the risk for Crohn's disease. Because intestinal epithelial cells play a role as the barrier to luminal microorganisms, we investigated the expression and function of CARD15/NOD2 in intestinal epithelial cells. Expression of CARD15/NOD2 messenger RNA (mRNA) in intestinal epithelial cell lines and primary intestinal epithelial cells was assessed by reverse-transcription polymerase chain reaction (RT-PCR). Regulation of expression of CARD15/NOD2 by cytokines was determined by Northern blot using the SW480 cell line. Active CARD15/NOD2 protein in SW480 cells was assessed by the combination of immunoprecipitation and immunoblotting using anti-CARD15/NOD2 antisera. To identify the functional role of CARD15/NOD2 in intestinal epithelial cells, gentamicin protection assays of Salmonella typhimurium were performed using Caco2 cells stably transfected with either wild-type CARD15/NOD2 or the 3020insC mutant associated with Crohn's disease. CARD15/NOD2 mRNA was expressed in both intestinal epithelial cell lines and primary intestinal epithelial cells. CARD15/NOD2 mRNA and protein were up-regulated by tumor necrosis factor alpha (TNFalpha) in SW480 cells. The number of viable internalized S. typhimurium in Caco2 cells stably transfected with CARD15/NOD2 expression plasmid was lower than untransfected Caco2 cells or MOCK transfectant. In contrast, expression of a variant associated with Crohn's disease was unable to constrain bacterial survival. CARD15/NOD2 is expressed in intestinal epithelial cells and may serve as a key component of innate mucosal responses to luminal bacteria as an antibacterial factor. Failure in this activity may contribute to the development of Crohn's disease.


A Clinical Study of Acute Kidney Injury in Tropical Acute Febrile Illness.
Tropical Acute Febrile Illness (TAFI) is one of the most common causes of morbidity within the community. Acute Kidney Injury (AKI) due to infective and non infective causes is a major complication. Presence of AKI is a major cause of mortality among patients with TAFI. To study the spectrum of tropical acute febrile illness; the proportion, spectrum and staging of acute kidney injury; Renal Replacement Therapy (RRT) initiation and in-hospital mortality. A total of 600 TAFI patients were prospectively studied at a tertiary care centre in coastal Karnataka between September 2012 and September 2014 for the aetiology of TAFI; the development and staging of AKI based on Kidney disease: Improving global outcomes (KDIGO) guidelines; the initiation of RRT and in-hospital mortality. Data analysis was done using SPSS version 17.0 with statistical significance calculated using chi-square and Fisher's exact t-test for which p-value <0.05 was considered significant. The spectrum of TAFI, in decreasing order, was vivax malaria, leptospirosis, dengue fever, falciparum malaria, mixed malaria, enteric fever, scrub typhus and the most common aetiology was malaria. The proportion of AKI was 54%. The most common cause of AKI, its stages 2 and 3, RRT initiation and in-hospital mortality was leptospirosis; and AKI stage 1 was dengue fever. KDIGO AKI stage 1, 2 and 3 was seen in 46.9%, 31.2% and 21.9% of AKI patients, respectively. RRT initiation was required in 10.2% of AKI patients and in-hospital mortality was 3% among all patients. AKI, RRT initiationand in-hospital mortality were significantly associated with older age, fever duration and other presenting complaints, examination findings, renal function and other parameters, leptospirosis, dengue fever, falciparum malaria. The aetiology in about half of TAFI patients in coastal Karnataka was malaria. More than 50% develop AKI with greater than one-fifth of them progressing to AKI stage 3 and one-tenth requiring RRT. The most common cause of AKI, AKI stage 2, 3, RRT initiation and in-hospital mortality was leptospirosis. AKI was present in almost all patients with leptospirosis. Therefore leptospirosis was the most nephrotoxic acute febrile illness in the present study population. Dengue fever was the most common cause of AKI stage 1. Vivax malaria was the third most common cause of AKI. The factors like age, presenting complaints, examination findings, renal function and other parameters, aetiology and RRT initiation may be used to predict AKI and in-hospital mortality.


Mentoring in Medicine (MIM): Motivating and Enabling Disadvantaged Youth to Become the Next Generation of Minority Health Professionals.
Mentoring in Medicine, Inc (MIM) is a nonprofit health and science youth development organization based in the Bronx, NY. Founded in 2006 by three physicians and an engineer-trained entrepreneur, MIM's organizational goal is to expose socioeconomically disadvantaged students to the wide variety of health and science careers and to increase the health literacy of their communities. It is aligned with the outreach mission of the U.S. National Library of Medicine (NLM) whose former Director, Donald A.B. Lindberg M.D., fostered an enduring relationship. Technical assistance, evaluation, and financial support provided under his leadership helped MIM to become a nationally recognized organization leading the field to diversify health careers and to increase health literacy in often hard to reach populations. Through live and virtual programming, MIM has impacted nearly 58,000 students, parents, and educators in urban epicenters in the U.S. The MIM Team has helped 503 students who were discouraged to build a competitive application and matriculate in health professional school. MIM has 88 press features highlighting its work in the community.


Retrovirus-mediated expression of preprosomatostatin in rat pituitary GH3 cells: targeting of somatostatin to the regulated secretory pathway.
Somatostatin (SRIF) is a 14-amino acid peptide hormone that is synthesized as part of a larger precursor, prepro-SRIF, consisting of a signal peptide and a proregion of 80-90 amino acids; mature SRIF is located at the carboxyl-terminus of the precursor. We have used a recombinant retroviral expression vector encoding anglerfish prepor-SRIF-I to infect rat pituitary GH3 cells. The aim of these studies was to investigate the intracellular storage and secretion of the total pool of endogenous GH compared to that of SRIF. Several clonal lines of GH3 cells expressing high or low levels of SRIF were treated with TRH, forskolin, or depolarizing concentrations of potassium, and the levels of intracellular and secreted GH or SRIF were determined using highly sensitive RIAs. Approximately 65% of the total GH was secreted basally, whereas less than 20% of the SRIF-immunoreactive material was basally secreted. Forskolin treatment or potassium depolarization stimulated GH release, but only about 50% above basal levels. In contrast, SRIF secretion was stimulated approximately 5-fold in response to these secretagogues. Based on its lower basal rate of secretion compared to GH and its enhanced release in response to a variety of secretagogues, we conclude that the heterologously expressed SRIF is preferentially targeted to the regulated pathway in GH3 cells.


[The role of immune system in colon metastasis. Lymphangiogenesis or lymphedema in cancer tissue].
The extent of lymph node metastasis is a major determinant for the staging and the prognosis of most human malignancies. Although the clinical significance of lymph node involvement is well documented, molecular mechanisms that promote tumor spread into the lymphatic or blood vascular systems and widespread dissemination are not well understood. Although there is a large body of evidence that newly visualized lymphatics facilitate formation of metastases, it remains unclear whether these are "new" or simply pre-existing dilated vessels. High level of permeability of tumor blood capillaries brings about high tissue fluid and lymph formation. The physical forces but not the putative cancer-produced VEGF C may be responsible for more lymphatics seen around cancer than in normal tissue. The main question to be answered is: are there morphologic and functional differences between newly formed and pre-existing intra- or peri-tumoral lymphatics? In our experience specimens of gastric and colon cancer revealed presence of peri-tumoral but not intra-tumoral lymphatics. Tumor tissue contained numerous tissue fluid "lakes" communicating with lymphatics. We speculate that increased production of lymph in the tumor tissue caused by high blood capillary permeability brings about dilatation of the interstitial space and peri-tumoral lymphatics. Excessive lymph flow may drag tumor cells. This article is reviews current literature on the role of angiogenesis and lymphangiogenesis in cancer metastasis with respect to own research.


Comparing caregiver and clinician predictions of fitness to drive in people with Alzheimer's disease.
This observational study investigated family caregiver and clinician ratings of 75 drivers with Alzheimer's disease against scores on a standardized road test and a naturalistic driving evaluation. Clinician ratings by a physician specialized in dementia were significantly associated with road test error scores (r=.25, p=.03) but not naturalistic driving errors or global ratings of road test and naturalistic driving performance. Caregiver ratings were unrelated to either driving assessment, with two exceptions; adult child ratings of driving ability were correlated with road test error scores (r=.43, p=.02), and spousal ratings were inversely correlated with global ratings. Clinician ratings of driving competence were modestly correlated with road test performance, but caregiver ratings were more complex. Adult children may be more accurate reporters of driving ability than spouses, possibly because of less personal bias, but the reasons behind this discrepancy need further investigation.


Human IL-17: a novel cytokine derived from T cells.
A cDNA encoding human IL-17 (hIL-17) was cloned from a CD4+ T cell library. The predicted 155-amino acids sequence contains an N-terminal signal peptide and exhibits 72% amino acid identity with HVS13, an open reading frame from a T-lymphotropic Herpesvirus saimiri, and 63% with murine CTLA8. High levels of hIL-17 were induced from primary peripheral blood CD4+ T cells upon stimulation. When expressed in CV1/EBNA cells, recombinant hIL-17 was secreted in both glycosylated and nonglycosylated forms. A hIL-17.Fc fusion protein and supernatants from cells transfected with hIL-17 induced IL-6 and IL-8 production and enhanced the surface expression of the intracellular adhesion molecule-1 (ICAM-1) in human fibroblasts.


Soy Intake Modifies the Relation Between Urinary Bisphenol A Concentrations and Pregnancy Outcomes Among Women Undergoing Assisted Reproduction.
Experimental data in rodents suggest that the adverse reproductive health effects of bisphenol A (BPA) can be modified by intake of soy phytoestrogens. Whether the same is true in humans is not known. The purpose of this study was to evaluate whether soy consumption modifies the relation between urinary BPA levels and infertility treatment outcomes among women undergoing assisted reproduction. The study was conducted in a fertility center in a teaching hospital. We evaluated 239 women enrolled between 2007 and 2012 in the Environment and Reproductive Health (EARTH) Study, a prospective cohort study, who underwent 347 in vitro fertilization (IVF) cycles. Participants completed a baseline questionnaire and provided up to 2 urine samples in each treatment cycle before oocyte retrieval. IVF outcomes were abstracted from electronic medical records. We used generalized linear mixed models with interaction terms to evaluate whether the association between urinary BPA concentrations and IVF outcomes was modified by soy intake. Live birth rates per initiated treatment cycle were measured. Soy food consumption modified the association of urinary BPA concentration with live birth rates (P for interaction = .01). Among women who did not consume soy foods, the adjusted live birth rates per initiated cycle in increasing quartiles of cycle-specific urinary BPA concentrations were 54%, 35%, 31%, and 17% (P for trend = .03). The corresponding live birth rates among women reporting pretreatment consumption of soy foods were 38%, 42%, 47%, and 49% (P for trend = 0.35). A similar pattern was found for implantation (P for interaction = .02) and clinical pregnancy rates (P for interaction = .03) per initiated cycle, where urinary BPA was inversely related to these outcomes among women not consuming soy foods but unrelated to them among soy consumers. Soy food intake may protect against the adverse reproductive effects of BPA. As these findings represent the first report suggesting a potential interaction between soy and BPA in humans, they should be further evaluated in other populations.


Urachal carcinomas of the nonglandular type: salient features and considerations in pathologic diagnosis.
The vast majority of malignant urachal epithelial tumors have a glandular morphology (ie, adenocarcinoma), to which our principal understanding of urachal carcinoma and its prevailing set of diagnostic criteria are largely ascribed. The 2004 World Health Organization classification of genitourinary tumors recognizes other rarer histologic types of urachal carcinomas such as urothelial, squamous cell, and other carcinomas. However, the clinicopathologic data for these nonglandular groups of urachal carcinomas are very limited, being detailed only in sporadic case reports. Some of the criteria recommended for pathologic confirmation of urachal carcinomas were formulated almost exclusively for urachal adenocarcinoma and may not be relevant or applicable for nonglandular tumors. Here, we present 7 examples of pure (5) and mixed predominant (2) nonglandular urachal carcinomas. Patients were 45 to 85 years of age (mean, 64.1) with a male predominance (male-to-female ratio=6:1). Six tumors were related to the bladder [dome (3) or dome/supravesical (3)] and 1 was entirely supravesical. Histologically, 5 were urothelial carcinomas, of pure or mixed histology, all were high grade and invasive, and 2 were small cell carcinomas. Two urothelial carcinomas had focal (<5%) glandular differentiation and signet ring cell change, and 1 had admixed focal malignant squamous cells and high-grade dedifferentiated components. Four of 5 urachal urothelial carcinomas exhibited solid and partly cavitary or luminal growth with papillary structures and a variable amount of necrosis within the cavity. The 2 small cell carcinomas were pure, had classic undifferentiated neuroendocrine histology, were situated at the bladder dome, and were partly cavitary filled with necrotic debris. Urachal remnant was identified in 6 tumors mainly with dysplastic transitional cells in the urachal canal or rudimentary nests and tubules. All 6 bladder-related urachal tumors exhibited reverse invasive front from the surface, including 2 tumors that ulcerated the bladder mucosa. One tumor had concomitant in situ and noninvasive high-grade papillary urothelial carcinomas in the main bladder lumen. Sheldon stages at presentation were IIIA (2), IVA (3), and IVB (2). Follow-up in all 7 cases (<1 to 60 mo; median, 12.5 mo) showed that 6 patients had died of disease, including the 2 patients with small cell carcinoma. In conclusion, nonglandular urachal carcinoma may occur with pure histology or admixed with high-grade dedifferentiated morphologies and a minor adenocarcinoma component. These tumors may arise as deep-seated bladder-related or completely supravesical tumors along the urachal tract and may exhibit reverse invasive spread toward the bladder surface. Cavitary or luminal growth may occur that could be attributed to the intraurachal neoplastic proliferation. Urachal urothelial carcinomas in particular may contain papillary structures within the tumor and urachal cavity. Concomitant primary urothelial carcinoma outside of the urachus and tumor extension to bladder mucosa may occur, which should not negate diagnosis of an urachal primary. Behavior appears poor, as most tumors present with higher stage.


The relationship between sex hormones, the vaginal microbiome and immunity in HIV-1 susceptibility in women.
The role of sex hormones in regulating immune responses in the female genital tract has been recognized for decades. More recently, it has become increasingly clear that sex hormones regulate susceptibility to sexually transmitted infections through direct and indirect mechanisms involving inflammation and immune responses. The reproductive cycle can influence simian/human immunodeficiency virus (SHIV) infections in primates and HIV-1 infection in ex vivo cervical tissues from women. Exogenous hormones, such as those found in hormonal contraceptives, have come under intense scrutiny because of the increased susceptibility to sexually transmitted infections seen in women using medroxyprogesterone acetate, a synthetic progestin-based contraceptive. Recent meta-analyses concluded that medroxyprogesterone acetate enhanced HIV-1 susceptibility in women by 40%. In contrast, estradiol-containing hormonal contraceptives were not associated with increased susceptibility and some studies reported a protective effect of estrogen on HIV/SIV infection, although the underlying mechanisms remain incompletely understood. Recent studies describe a key role for the vaginal microbiota in determining susceptibility to sexually transmitted infections, including HIV-1. While Lactobacillus spp.-dominated vaginal microbiota is associated with decreased susceptibility, complex microbiota, such as those seen in bacterial vaginosis, correlates with increased susceptibility to HIV-1. Interestingly, sex hormones are inherently linked to microbiota regulation in the vaginal tract. Estrogen has been postulated to play a key role in establishing a Lactobacillus-dominated microenvironment, whereas medroxyprogesterone acetate is linked to hypo-estrogenic effects. The aim of this Review is to contribute to a better understanding of the sex-hormone-microbiome-immunity axis, which can provide key information on the determinants of HIV-1 susceptibility in the female genital tract and, consequently, inform HIV-1 prevention strategies.


Carrier-envelope phase dynamics and noise analysis in octave-spanning Ti:sapphire lasers.
We investigate the carrier-envelope phase dynamics of octave-spanning Ti:sapphire lasers and perform a complete noise analysis of the carrier-envelope phase stabilization. We model the effect of the laser dynamics on the residual carrier-envelope phase noise by deriving a transfer function representation of the octave-spanning frequency comb. The modelled phase noise and the experimental results show excellent agreement. This greatly enhances our capability of predicting the dependence of the residual carrier-envelope phase noise on the feedback loop filter, the carrier-envelope frequency control mechanism and the pump laser used.


Genetic structure of the ancestral population of modern humans.
Neutral DNA polymorphisms from an 8-kb segment of the dystrophin gene, previously ascertained in a worldwide sample (n = 250 chromosomes), were used to characterize the population ancestral to the present-day human groups. The ancestral state of each polymorphic site was determined by comparing human variants with their orthologous sites in the great apes. The "age before fixation" of the underlying mutations was estimated from the frequencies of the new alleles and analyzed in the context of these polymorphisms' distribution among 13 populations from Africa, Europe, Asia, New Guinea, and the Americas (n = 860 chromosomes in total). Seventeen polymorphisms older tan 100,000-200,000 years, which contributed approximately 90% to the overall nucleotide diversity, were common to all human groups. Polymorphisms endemic to human groups or continentally restricted were younger than 100,000-200,000 years. Africans (six populations) with 13 such sites stood out from the rest of the world (seven populations), where only 2 population-specific variants were observed. The similarity of the frequencies of the old polymorphisms in Africans and non-Africans suggested a similar profile of genetic variability in the population before the modern human's divergence. This ancestral population was characterized by an effective size of about 10,000 as estimated from the nucleotide diversity; this size may describe the number of breeding individuals over a long time during the Middle Pleistocene or reflect a speciation bottleneck from an initially larger population at the end of this period.


Obesity induced by a high-fat diet is associated with reduced brain insulin transport in dogs.
Insulin transported from plasma into the central nervous system (CNS) is hypothesized to contribute to the negative feedback regulation of body adiposity. Because CNS insulin uptake is likely mediated by insulin receptors, physiological interventions that impair insulin action in the periphery might also reduce the efficiency of CNS insulin uptake and predispose to weight gain. We hypothesized that high-fat feeding, which both reduces insulin sensitivity in peripheral tissues and favors weight gain, reduces the efficiency of insulin uptake from plasma into the CNS. To test this hypothesis, we estimated parameters for cerebrospinal fluid (CSF) insulin uptake and clearance during an intravenous insulin infusion using compartmental modeling in 10 dogs before and after 7 weeks of high-fat feeding. These parameters, together with 24-h plasma insulin levels measured during ad libitum feeding, also permitted estimates of relative CNS insulin concentrations. The percent changes of adiposity, body weight, and food intake after high-fat feeding were each inversely associated with the percent changes of the parameter k1k2, which reflects the efficiency of CNS insulin uptake from plasma (r = -0.74, -0.69, -0.63; P = 0.015, 0.03, and 0.05, respectively). These findings were supported by a non-model-based calculation of CNS insulin uptake: the CSF-to-plasma insulin ratio during the insulin infusion. This ratio changed in association with changes of k1k2 (r = 0.84, P = 0.002), body weight (r = -0.66, P = 0.04), and relative adiposity (r = -0.72, P = 0.02). By comparison, changes in insulin sensitivity, according to minimal model analysis, were not associated with changes in k1k2, suggesting that these parameters are not regulated in parallel. During high-fat feeding, there was a 60% reduction of the estimated CNS insulin level (P = 0.04), and this estimate was inversely associated with percent changes in body weight (r = -0.71, P = 0.03). These results demonstrate that increased food intake and weight gain during high-fat feeding are associated with and may be causally related to reduced insulin delivery into the CNS.


Time-correlated single-photon counting fluorescence lifetime confocal imaging of decayed and sound dental structures with a white-light supercontinuum source.
We report the demonstration of time-correlated single-photon counting (TCSPC) fluorescence lifetime imaging (FLIM) to ex vivo decayed and healthy dentinal tooth structures, using a white-light supercontinuum excitation source. By using a 100 fs-pulsed Ti:Sapphire laser with a low-frequency chirp to pump a 30-cm long section of photonic crystal fibre, a ps-pulsed white-light supercontinuum was created. Optical bandpass interference filters were then applied to this broad-bandwidth source to select the 488-nm excitation wavelength required to perform TCSPC FLIM of dental structures. Decayed dentine showed significantly shorter lifetimes, discriminating it from healthy tissue and hard, stained and thus affected but non-infected material. The white-light generation source provides a flexible method of producing variable-bandwidth visible and ps-pulsed light for TCSPC FLIM. The results from the dental tissue indicate a potential method of discriminating diseased tissue from sound, but stained tissue, which could be of crucial importance in limiting tissue resection during preparation for clinical restorations.


Selective protection of normal cells during chemotherapy by RY4 peptides.
Mitochondrial targeted Szeto-Schiller (SS) peptides have recently gained attention for their antioxidative stress ability; however, the functional variations between normal and cancer cells have not been determined. Here, we report the results of such experiments conducted with a newly designed class of peptide called RY4, which is based on SS peptide sequence characteristics. The RY4 peptide exhibits distinct differences in antioxidative stress response between normal and cancer cells when challenged with chemotherapeutics like the glycolytic inhibitor dichloroacetate (DCA), the platinating agent carboplatin, and the DNA damage inducer doxorubicin. Interestingly, only normal human cells were protected by the RY4 peptide and catalase (CAT) activity was significantly enhanced in normal but not tumor cells when incubated with RY4. Pull-down, coimmunoprecipitation, and LC/MS-MS proteomic analysis demonstrated that RY4 and catalase are capable of forming protein complexes. Finally, in vivo efficacy was evaluated by intraperitoneal administration of RY4 into a lung cancer xenograft model, which revealed significant myocardiocyte protection from doxorubicin-induced cardiotoxicity without diminishing doxorubicin's tumoricidal effects. Taken together, RY4 offers selective protection to normal cells from chemotherapy-induced toxicity by enhancing the activity of cellular antioxidant enzymes. RY4 peptides selectively reduce chemotherapeutic-induced oxidative stress and represent a new class of chemoprotective agents with clinical potential.


Water shortages and extreme events: a call for research.
Water shortages as a result of extreme weather events, such as flooding and severe cold, have the potential to affect significant numbers of people. Therefore, the need to build robust, coordinated plans based on scientific evidence is crucial. The literature review outlined in this short communication was conducted as part of a joint Drinking Water Inspectorate and Health Protection Agency (now Public Health England) report which aimed to review the scientific evidence base on extreme events, water shortages and the resulting health impacts. A systematic literature review was undertaken to identify published literature from both peer-reviewed and grey literature sources. The retrieved literature was then assessed using the Scottish Intercollegiate Guidelines Network quality assessment. The authors found very few scientific studies. However, a great deal of valuable grey literature was retrieved and used by the research team. In total, six main themes of importance that were identified by the review and discussed included health impacts, water quantity and quality, alternative supplies, vulnerable groups, communication with those affected and the emergency response. The authors conclude that more research needs to be conducted on health impacts and extreme events water shortages in order to build the future knowledge base and development of resilience.


Community-oriented primary care (COPC) in Barcelona, Spain: an urban COPC experience.
Community-oriented primary care (COPC) integrates comprehensive primary care with community health. Although it has had limited application in the United States, this model has been widely promoted among urban family physicians in Barcelona, Spain. This article describes the current status of COPC in four community clinics in Barcelona. Data were derived from a site visit that included direct observation and interviews with professionals involved in community health. The interviews explored how COPC has been implemented in each of the four primary care centers. We found that the degree to which COPC is practiced is quite varied and that it often coexists with other community health programs. A number of obstacles, including lack of support and funding from the government and lack of motivation and participation among health professionals, make practicing COPC in Barcelona a challenge. Despite these obstacles, COPC is flourishing in Barcelona. This experience may offer guidance for COPC implementation in the United States and other countries.


The impact of psychopathology, race, and environmental context on violent offending in a male adolescent sample.
Research has identified a multitude of demographic, psychological, and contextual factors that are associated with violent offending among youth. However, much of the previous research has focused on single factors, and little is known about the effects of these various factors in tandem. The present study examines whether certain community factors impact the effects of race and psychopathology on violent crime. Multivariate analyses were used to investigate race-psychopathology combinations and the moderating influence of exposure to community violence in a sample of youth (N = 1,116). Youth without antisocial personality disorder (APD) or psychopathy were less violent than the other diagnostic groups, and Black youth were less likely to have psychopathy compared with Whites and Hispanics. However, Black youth with APD and psychopathy were twice as likely to exhibit violent crime versatility. Furthermore, Black and Hispanic youth demonstrating aggressive conduct problems committed a greater number of violent crime types than Whites. This relationship was further qualified, such that Black and Hispanic youth with APD and psychopathy, who were more exposed to community violence, committed a greater number of violent crime types compared with Whites. These results suggest that prevention and intervention strategies should consider individual and community-level factors. (PsycINFO Database Record


Grating visual acuity using the preferential-looking method in elderly nursing home residents.
To assess the performance of two approaches to visual acuity testing in a group of nursing home residents with cognitive impairment. The study was a cross-sectional comparison of the effectiveness of two tests of visual acuity. Nursing home residents participating in a clinical trial were tested with both recognition acuity charts and grating acuity cards (Teller) by masked observers. Of the nursing home residents (n = 656) who participated in the study, 86% could respond to visual acuity testing in at least one eye. Eighty-four percent were testable using Teller cards versus 73% who were testable by Early-Treatment Diabetic Retinopathy Study (ETDRS) charts or Lea symbol charts. Forty-one percent of individuals with MiniMental Status Examination (MMSE) scores lower than 10 were testable by recognition acuity, whereas 61% were testable with grating acuity cards. Grating acuity correlated well with recognition acuity (R = 0.79; 95% CI, 0.75-0.98, intraclass correlation coefficient [ICC]). The correlation was slightly lower in individuals with decreased MMSE scores. Although grating acuity was one line better than recognition acuity on average and median acuities were the same, 24% of individuals had results that differed by three or more lines. Teller acuity cards can effectively test the vision in cognitively impaired individuals who are not testable by conventional means. Grating acuity results correlated well with those of recognition acuity, although differences of three or more lines were not uncommon. Wider use of grating acuity testing allows a more complete assessment of visual function in the cognitively impaired elderly.


Identification of service development needs for incarcerated adults with autism spectrum disorders in an Australian prison system.
The purpose of this paper is to identify and deliberate the service development needs required for the improvement of service provision for incarcerated adults with autism spectrum disorder (ASD) in NSW, Australia. Consultation groups were conducted to bring together n=5 key stakeholders from heath and correctional-based services in the prison system. A facilitated asynchronous e-mail-based discussion occurred amongst group members between consultation group meetings. Two main themes were identified: detecting persons with ASD and providing appropriate care. Participants discussed current service gaps with regard to the identification of people with ASD at the point of contact with the prison service, and the difficulties associated with diagnosing prisoners with ASD. The need for effective alert systems to detect persons with ASD in custody was identified. The current absence of ASD-specific support services in prison was highlighted, and recommendations for improvement suggested. Current health and correctional-based service provision failed to adequately support incarcerated adults with ASD. Improvements in prison-entry screening processes, alert systems and diagnostic practices are required. Multidisciplinary collaboration between prison-based and external service providers is required for the development of a model of care based on individualised case management to adequately support incarcerated adults with ASD in prison. Given the lack of reported service provision for incarcerated adults with ASD internationally, other prison-based services are likely to experience similar service development needs and see the relevance of the recommendations made directly from the study findings.


A neutralizing monoclonal antibody targeting the acid-sensitive region in chikungunya virus E2 protects from disease.
The mosquito-borne alphavirus, chikungunya virus (CHIKV), has recently reemerged, producing the largest epidemic ever recorded for this virus, with up to 6.5 million cases of acute and chronic rheumatic disease. There are currently no licensed vaccines for CHIKV and current anti-inflammatory drug treatment is often inadequate. Here we describe the isolation and characterization of two human monoclonal antibodies, C9 and E8, from CHIKV infected and recovered individuals. C9 was determined to be a potent virus neutralizing antibody and a biosensor antibody binding study demonstrated it recognized residues on intact CHIKV VLPs. Shotgun mutagenesis alanine scanning of 98 percent of the residues in the E1 and E2 glycoproteins of CHIKV envelope showed that the epitope bound by C9 included amino-acid 162 in the acid-sensitive region (ASR) of the CHIKV E2 glycoprotein. The ASR is critical for the rearrangement of CHIKV E2 during fusion and viral entry into host cells, and we predict that C9 prevents these events from occurring. When used prophylactically in a CHIKV mouse model, C9 completely protected against CHIKV viremia and arthritis. We also observed that when administered therapeutically at 8 or 18 hours post-CHIKV challenge, C9 gave 100% protection in a pathogenic mouse model. Given that targeting this novel neutralizing epitope in E2 can potently protect both in vitro and in vivo, it is likely to be an important region both for future antibody and vaccine-based interventions against CHIKV.


1009 nm continuous-wave ytterbium-doped fiber amplifier emitting 146 W.
In this Letter, we demonstrate a single-mode continuous-wave fiber laser amplifier emitting 146 W of average output power at a wavelength of 1009 nm. The wavelength and bandwidth of the seed oscillator are defined by a pair of fiber Bragg gratings. The seed is amplified in a two-stage ytterbium-doped rod-type amplifier to 146 W with a high slope efficiency of 64%, showing excellent beam quality and stability throughout the experiment. The ASE suppression is as high as 63 dB.


Molecular clocks in pharmacology.
Circadian rhythms regulate a vast array of biological processes and play a fundamental role in mammalian physiology. As a result, considerable diurnal variation in the pharmacokinetics, efficacy, and side effect profiles of many therapeutics has been described. This variation has subsequently been tied to diurnal rhythms in absorption, distribution, metabolism, and excretion, as well as in pharmacodynamic variables, such as target expression. More recently, the molecular basis of circadian rhythmicity has been elucidated with the identification of clock genes, which oscillate in a circadian manner in most cells and tissues and regulate transcription of large sets of genes. Ongoing research efforts are beginning to reveal the critical role of circadian clock genes in the regulation of pharmacologic parameters, as well as the reciprocal impact of drugs on circadian clock function. This chapter will review the role of circadian clocks in the pharmacokinetics and pharmacodynamics of drug response and provide several examples of the complex regulation of pharmacologic systems by components of the molecular circadian clock.


Innovative approaches to clinical development and trial design.
Pharmaceutical innovation is increasingly risky, costly and at times inefficient, which has led to a decline in industry productivity. Despite the increased investment in R&D by the industry, the number of new molecular entities achieving marketing authorization is not increasing. Novel approaches to clinical development and trial design could have a key role in overcoming some of these challenges by improving efficiency and reducing attrition rates. The effectiveness of clinical development can be improved by adopting a more integrated model that increases flexibility and maximizes the use of accumulated knowledge. Central to this model of drug development are novel tools, including modelling and simulation, Bayesian methodologies, and adaptive designs, such as seamless adaptive designs and sample-size re-estimation methods. Applications of these methodologies to early- and late-stage drug development are described with some specific examples, along with advantages, challenges, and barriers to implementation. Because they are so flexible, these new trial designs require significant statistical analyses, simulations and logistical considerations to verify their operating characteristics, and therefore tend to require more time for the planning and protocol development phase. Greater awareness of the distinct advantages of innovative designs by regulators and sponsors are crucial to increasing the adoption of these modern tools.


Effects of low-fat or full-fat fermented and non-fermented dairy foods on selected cardiovascular biomarkers in overweight adults.
The association between consumption of full-fat dairy foods and CVD may depend partly on the nature of products and may not apply to low-fat dairy foods. Increased circulating levels of inflammatory biomarkers after consumption of dairy product-rich meals suggest an association with CVD. In the present study, we tested the effects of low-fat and full-fat dairy diets on biomarkers associated with inflammation, oxidative stress or atherogenesis and on plasma lipid classes. Within full-fat dairy diets, we also compared fermented v. non-fermented products. In a randomised cross-over study, twelve overweight/obese subjects consumed during two 3-week periods two full-fat dairy diets containing either yogurt plus cheese (fermented) or butter, cream and ice cream (non-fermented) or a low-fat milk plus yogurt diet, with the latter being consumed between and at the end of the full-fat dairy dietary periods. The concentrations of six inflammatory and two atherogenic biomarkers known to be raised in CVD were measured as well as those of plasma F2-isoprostanes and lipid classes. The concentrations of six of the eight biomarkers tended to be higher on consumption of the low-fat dairy diet than on that of the fermented dairy diet and the concentrations of two plasmalogen lipid classes reported to be associated with increased oxidisability were also higher on consumption of the low-fat dairy diet than on that of the fermented dairy diet (P< 0.001), although plasma F2-isoprostane concentrations did not differ on consumption of any of the diets. On the other hand, the concentrations of plasma sphingomyelin and IL-6 were significantly higher on consumption of the non-fermented dairy diet than on that of the low-fat dairy diet (P< 0.02). In conclusion, short-term diets containing low-fat dairy products did not lead to a more favourable biomarker profile associated with CVD risk compared with the full-fat dairy products, suggesting that full-fat fermented dairy products may be the more favourable.


Why do farmers and veterinarians not report all bovine abortions, as requested by the clinical brucellosis surveillance system in France?
Since 2005, France has been officially free of brucellosis, an infectious disease that causes abortion in cattle and can be transmitted from cattle to humans. Recent animal and human cases have drawn attention to the need to prevent infection of humans and animals from any primary outbreaks. In order to detect any new outbreaks as soon as possible, a clinical surveillance system requires farmers and veterinarians to report each abortion and to test the aborting cow for brucellosis. However, under-reporting limits the sensitivity of this system. Our objective was to identify the barriers and motivations influencing field actors in their decision to report or not to report bovine abortions. We used a qualitative approach with semi-structured interviews of 12 cattle farmers and their eight veterinarians. Our analysis showed that four main themes influence the decision-making process of farmers and veterinarians: 1) the perceived risk of brucellosis and other abortive diseases; 2) the definition of a suspected case of brucellosis and other abortive diseases adopted by field actors, which is less sensitive than the mandatory definition; 3) the cost-benefit analysis conducted by actors, taking into account regulatory and health aspects, economic and financial losses, technical and practical factors; 4) the level of cooperation within the socio-technical network. We discussed how early detection may be improved by revising the definition of abortion, extending the time frame for notification and generalising the differential diagnosis of the causes of abortion. In contrast to quantitative approaches, qualitative studies can identify the factors (including unknown factors) influencing the decision-making process of field actors and reveal why they take those factors into consideration. Our qualitative study sheds light on the factors underlying the poor sensitivity of clinical brucellosis surveillance system for cattle in France, and suggests that early detection may be improved by considering actors' perceptions. We believe our findings may provide further insight into ways of improving other clinical surveillance systems and thus reduce the risk of disease.


Effects of texture properties of semi-solid food on the sensory test for pharyngeal swallowing effort in the older adults.
Increasing viscosity can reduce the risk of aspiration into the airway, but excessively thickened food may require more force and effort. We assumed that semi-solid foods with similar viscosities will behave differently in the oropharynx and there might exist the possibility that properties other than viscosity may have clinical relevance. This study aimed to find out the texture of semi-solid foods that affects the effort of pharyngeal swallow in the older adults. Nine kinds of semi-solid foods not requiring mastication were selected for texture profile analysis (TPA), and included whipped cream, mayonnaise, soft tofu, mango pudding, boiled mashed pumpkin, boiled mashed potatoes, boiled mashed sweet potatoes, red bean paste, and peanut butter. Hardness, adhesiveness and cohesiveness of each food were measured three times by using the rheometer. A blinded sensory test using a 9-point hedonic scale was also conducted in eighteen older adults people to investigate how much effort was required to swallow food, and how much of the food remained in the pharynx after swallowing. The correlation between texture and sensory outcome was statistically analyzed. Foods that belonged to the same viscosity category showed different texture values, and the participants also rated different scores respectively. Only adhesiveness among three properties was significantly correlated with the sensory test. (r = 0.882, p = 0.002 for difficult to swallow, r = 0.879, p = 0.002 for sense of residue). Adhesiveness was the most important property of the semi-solid foods, requiring most efforts in pharyngeal swallow in the older adults. If we select and provide food having low adhesiveness value in the same viscosity category, there might be the possibility to make it easier to swallow in older adults.


Interaction of Vaccination and Reduction of Antibiotic Use Drives Unexpected Increase of Pneumococcal Meningitis.
Antibiotic-use policies may affect pneumococcal conjugate-vaccine effectiveness. The reported increase of pneumococcal meningitis from 2001 to 2009 in France, where a national campaign to reduce antibiotic use was implemented in parallel to the introduction of the 7-valent conjugate vaccine, provides unique data to assess these effects. We constructed a mechanistic pneumococcal transmission model and used likelihood to assess the ability of competing hypotheses to explain that increase. We find that a model integrating a fitness cost of penicillin resistance successfully explains the overall and age-stratified pattern of serotype replacement. By simulating counterfactual scenarios of public health interventions in France, we propose that this fitness cost caused a gradual and pernicious interaction between the two interventions by increasing the spread of nonvaccine, penicillin-susceptible strains. More generally, our results indicate that reductions of antibiotic use may counteract the benefits of conjugate vaccines introduced into countries with low vaccine-serotype coverages and high-resistance frequencies. Our findings highlight the key role of antibiotic use in vaccine-induced serotype replacement and suggest the need for more integrated approaches to control pneumococcal infections.