Do porous tantalum implants help preserve bone?: evaluation of tibial bone density surrounding tantalum tibial implants in TKA.
TKA with conventional metal-backed tibial implants subjects the tibial metaphysis to stress shielding, with resultant loss of bone density. We hypothesized tibial bone mineral density in patients with porous tantalum (trabecular metal) tibial baseplates would (1) more closely parallel tibial bone mineral density in the nonoperative control limb and (2) be better maintained than in conventional historical controls. We prospectively followed 41 patients (35 men, six women) 60 years of age or younger undergoing TKA with uncemented trabecular metal tibial components. Patients underwent dual-energy xray absorptiometry scans of both proximal tibiae preoperatively and at 2 months, 1 year, and 2 years postoperatively. We determined bone mineral density in three selected regions of interest (Zone 1, between the pegs; Zone 2, beneath the pegs; Zone 3, directly below entire baseplate). Precision analysis revealed a precision error of 4% or less for each region of interest, indicating adequate power to detect bone mineral density changes of 8% or greater. Bone mineral density percent change was different between the operative and nonoperative knees only in Zone 3 and only at 2 months. There was no change in bone mineral density in any zone in the nonoperative knee at any time. Only in Zone 3 did the bone mineral density decrease at 2 months in the operative knee. Trabecular metal implants appear to maintain tibial bone mineral density in a parallel fashion to the nonoperative limb in this population and better than historical controls.


A case series of 48 patients treated with thalidomide.
Thalidomide is approved by the Food and Drug Administration (FDA) for erythema nodosum leprosum, but has been used in many other dermatological conditions that are refractory to standard therapy. The medical records of 48 patients treated with thalidomide at Baylor College of Medicine (Houston, TX) were retrospectively reviewed to determine the conditions treated with thalidomide, dosing, efficacy, treatment duration, side effects, adverse events, and reason for discontinuing therapy. Forty-eight patients (men = 18, women = 30) with a mean age of 49.6 years (range: 20-79) were included in this study. Patients were treated for prurigo nodularis, discoid lupus erythematosus, tumid lupus erythematosus, subacute cutaneous lupus erythematosus, systemic lupus erythematosus, lichen planus, lichen planopilaris, cutaneous sarcoidosis, and prurigo nodularis. All conditions were refractory to standard therapy. Patients were treated for a mean of 7.5 months (range: 3 days to 70 months). In most of the disorders, a majority of patients experienced clinical improvement. The most common reason for discontinuation of therapy was side effects, the most frequent being peripheral neuropathy. This study was limited by being retrospective in nature. Thalidomide effectively treats some dermatologic conditions that are refractory to standard medications. There are inconveniences associated with obtaining the medication and it is expensive. Physicians must be vigilant for possible side effects, especially peripheral neuropathy.


Bioenergy and Breast Cancer: A Report on Tumor Growth and Metastasis.
As many as 80% of the 296,000 women and 2,240 men diagnosed with breast cancer in the United States will seek out complementary and alternative medicine (CAM) treatments. One such therapy is Healing Touch (HT), recognized by the National Center for Complementary and Integrative Health (NCCIH) as a treatment modality. Using a multiple experimental groups design, fifty-six six- to eight-week-old Balb/c mice were injected with 4T1 breast cancer tumor cells and randomly divided into intervention and positive control groups. Five days after tumor cell injection, mice in the intervention groups received HT either daily or every other day for 10 minutes by one HT practitioner. At 15 days after tumor cell injection, tumor size was measured, and metastasis was evaluated by a medical pathologist after necropsy. Tumor size did not differ significantly among the groups (F(3,52) = 0.75, p value = 0.53). The presence of metastasis did not differ across groups (chi-square(3) = 3.902, p = 0.272) or when compared within an organ (liver: chi-square(3) = 2.507, p = 0.474; lungs: chi-square(3) = 3.804, p = 0.283; spleen: chi-square(3) = 0.595, p = 0.898). However, these results did indicate a moderate, though insignificant, positive impact of HT and highlight the need for continued research into dose, length of treatment, and measurable outcomes (tumor size, metastasis) to provide evidence to suggest application for nursing care.


Downregulation of microRNAs 145-3p and 145-5p is a long-term predictor of postmenopausal breast cancer risk: The ORDET prospective study.
miRNAs have been implicated in the regulation of key metabolic, inflammatory, and malignant pathways; hence, they might be considered both predictors and players of cancer development. Using a case-control study design nested in the ORDET prospective cohort study, we addressed the possibility that specific mRNAs can serve as early predictors of breast cancer incidence in postmenopausal women. We compared leukocyte miRNA profiles of 133 incident postmenopausal breast cancer cases and profiles of 133 women who remained healthy over a follow-up period of 20 years. The analysis identified 20 differentially expressed miRNAs, 15 of which were downregulated. Of the 20 miRNAs, miR145-5p and miR145-3p, each derived from another arm of the respective pre-miRNA, were consistently and significantly downregulated in all the databases that we surveyed. For example, analysis of more than 1,500 patients (the UK Metabric cohort) indicated that high abundance of miR145-3p and miR145-5p was associated with longer, and for miR145-3p also statistically significant, survival. The experimental data attributed different roles to the identified miRNAs: Although the 5p isoform was associated with invasion and metastasis, the other isoform seems related to cell proliferation. These observations and the prospective design of our study lend support to the hypothesis that downregulation of specific miRNAs constitutes an early event in cancer development. This finding might be used for breast cancer prevention. The identification of the miRNAs as long-term biomarkers of breast cancer may have an impact on breast cancer prevention and early detection.


Factors Associated With Aggressiveness of End-of-Life Care for Lung Cancer Patients and Associated Costs of Care.
Results of previous studies demonstrated that high-intensity end-of-life (EOL) care improves neither cancer patients' survival nor quality of life. Our objective was to assess the incidence of and factors associated with aggressiveness of care during the last 30 days of life (DOL) of lung cancer (LC) patients and the impacts of aggressiveness of care in EOL-care costs. Using French national hospital database, all patients with LC who died between January 1, 2010, and December 31, 2011, or between January 1, 2015, and January 31, 2016, were included. EOL-care aggressiveness was assessed using the following criteria: chemotherapy administered within the last 14 DOL; more than one hospitalization within the last 30 DOL; admission to the intensive care unit within the last 30 DOL; and palliative care initiated < 3 days before death. Expenditures were limited to direct costs, from a health care payer's perspective. Among 79,746 adult LC patients identified; 57% had at least one indicator of EOL-care aggressiveness (49% repeated hospitalizations, 12% intensive care unit admissions, 9% chemotherapy, 5% palliative care). It increased significantly between the 2 periods (56% vs. 58%, P < .001). Young age, male sex, shorter time since diagnosis, comorbidities, no malnutrition, type of care facility other than general hospital, social deprivation, and low-density population were independently associated with having one or more indicator of aggressive EOL care. The mean EOL cost was €8152 ± 5117 per patient, but the cost was significantly higher for patients with at least one EOL-care aggressiveness criterion (€9480 vs. €6376, P < .001). In France, a majority of LC patients had at least one criterion of aggressive EOL care that had a major economic impact on the health care system.


Recurrent BRAF Gene Fusions in a Subset of Pediatric Spindle Cell Sarcomas: Expanding the Genetic Spectrum of Tumors With Overlapping Features With Infantile Fibrosarcoma.
Infantile fibrosarcomas (IFS) represent a distinct group of soft tissue tumors occurring in patients under 2 years of age and most commonly involving the extremities. Most IFS show recurrent ETV6-NTRK3 gene fusions, sensitivity to chemotherapy, and an overall favorable clinical outcome. However, outside these well-defined pathologic features, no studies have investigated IFS lacking ETV6-NTRK3 fusions, or tumors with the morphology resembling IFS in older children. This study was triggered by the identification of a novel SEPT7-BRAF fusion in an unclassified retroperitoneal spindle cell sarcoma in a 16-year-old female by targeted RNA sequencing. Fluorescence in situ hybridization screening of 9 additional tumors with similar phenotype and lacking ETV6-NTRK3 identified 4 additional cases with BRAF gene rearrangements in the pelvic cavity (n=2), paraspinal region (n=1), and thigh (n=1) of young children (0 to 3 y old). Histologically, 4 cases including the index case shared a fascicular growth of packed monomorphic spindle cells, with uniform nuclei and fine chromatin, and a dilated branching vasculature; while the remaining case was composed of compact cellular sheets of short spindle to ovoid cells. In addition, a minor small blue round cell component was present in 1 case. Mitotic activity ranged from 1 to 9/10 high power fields. Immunohistochemical stains were nonspecific, with only focal smooth muscle actin staining demonstrated in 3 cases tested. Of the remaining 5 BRAF negative cases, further RNA sequencing identified 1 case with EML4-NTRK3 in an 1-year-old boy with a foot IFS, and a second case with TPM3-NTRK1 fusion in a 7-week-old infant with a retroperitoneal lesion. Our findings of recurrent BRAF gene rearrangements in tumors showing morphologic overlap with IFS expand the genetic spectrum of fusion-positive spindle cell sarcomas, to include unusual presentations, such as older children and adolescents and predilection for axial location, thereby opening new opportunities for kinase-targeted therapeutic intervention.


Falling out of bed: a relatively benign occurrence.
To determine the likelihood of injuries occurring to infants and children who fall out of bed. A review of hospital records where children were documented to have fallen from a crib or bed and where the children were immediately assessed and examined following the fall. Two hundred seven children younger than 6 years of age. One hundred twenty-four falls were from cribs and 83 from beds. The heights of the falls were 25 inches from beds or 41 inches when the child climbed over the bed rails and then fell, and 32 inches from cribs or 54 inches for those who fell after climbing the crib rails. There were 29 superficial injuries such as contusions or minor lacerations. One simple skull fracture was noted incidentally on skull roentgenogram and there was one fractured clavicle. There were no serious, multiple, visceral, or life-threatening injuries. Calculation of the momentum of impact between the injured and noninjured showed no significant difference. Because falls from short distances are unlikely to produce serious injury, the reliability of the history should be questioned when a child has significant injuries said to have resulted from a short fall.


Cost-effectiveness analysis and the consistency of decision making: evidence from pharmaceutical reimbursement in australia (1991 to 1996).
The principle aim of this study was to generate a league table of drugs considered by the Australian Pharmaceutical Benefits Advisory Committee (PBAC) for reimbursement. The table was used to test the hypothesis that decisions made by the PBAC are consistent with the maxim of economic efficiency. In addition, we explored whether the past decisions by the PBAC revealed a threshold incremental cost-effectiveness ratio beyond which the PBAC is not prepared to recommend reimbursement of a drug. All 355 submissions made to the PBAC between January 1991 and June 1996 were reviewed. Submissions using cost per life-year gained (26 submissions) or the cost per quality adjusted life-year (QALY) gained (9 submissions) were ranked in a league table and compared with advice given by the PBAC about that drug. The confidentiality restrictions for the submissions require that the individual drug details cannot be revealed in this article. There was a statistically significant difference between the cost per life-year gained for drugs that were recommended for listing and those that were not, suggesting that the PBAC has been broadly consistent with the use of economic efficiency as a criterion for decision making. We did not find an explicit threshold beyond which the PBAC was unwilling to pay for additional life years gained. However, between 1992 and 1996 the PBAC appears to have been unlikely to recommend a drug for listing if the additional cost per life-year exceeded 76 000 Australian dollars [$AU] (1998/1999 values) and was unlikely to reject a drug for which the additional cost per life-year gained was less than $AU42 000. The cost-effectiveness ratio was not the only factor determining the reimbursement decision. The results of this preliminary study indicate that decisions to recommend a drug for listing by the PBAC in the last few years have, by and large, been consistent with the notion of economic efficiency.


Vector densities that potentiate dengue outbreaks in a Brazilian city.
To identify the critical vector density that potentiates dengue outbreaks in an endemic site and to identify obstacles to anti-dengue activities, we correlated a series of dengue outbreaks in a Brazilian city with the intensity of its anti-vector source-reduction activities. The proportion of houses infested by vector mosquitoes correlated inversely with intensity of anti-mosquito interventions, and the vector population developed independently of rainfall. Local periods of drought promoted vector abundance in two ways: residents stored water in which vector mosquitoes could breed, and cholera outbreaks due to contaminated water diverted local health workers from routine anti-vector activities. One dengue outbreak became apparent to authorities more than two months after it commenced but would have been identified almost immediately had dengue-like disease in indicator hospitals been monitored. Active surveillance, therefore, offers a window of opportunity for promptly executed anti-dengue interventions. Source-reduction measures that suppress vector infestations to less than 1% of houses effectively avert outbreaks of dengue.


Job-loss and weight gain in British adults: Evidence from two longitudinal studies.
Overweight and obesity have been associated with unemployment but less is known about changes in weight associated with changes in employment. We examined weight changes associated with job-loss, retirement and maintaining employment in two samples of working adults in the United Kingdom. This was a prospective study of 7201 adults in the European Prospective Investigation of Cancer (EPIC)-Norfolk study (aged 39-76 years) and 4539 adults in the British Household Panel Survey (BHPS) who were followed up over 43 months and 26 months, respectively. In both samples, changes in measured (EPIC) and self-reported (BHPS) weight were computed for each participant and assessed in relation to three employment transitions: maintaining paid employment, retirement and job-loss. Regression models adjusted for potential confounders. Further analyses evaluated the contribution of diet, physical activity and smoking to weight gain. In EPIC-Norfolk, weight change differed across the three employment transitions for women but not men. The mean (95% CI) annualised change in weight for women who became unemployed over the follow-up period was 0.70 (0.55, 0.85) kg/y while those who maintained employment gained 0.49 (0.43, 0.55) kg/y (P = 0.007). Accounting for changes in smoking, diet and physical activity did not substantially alter the difference in weight gain among groups. In BHPS, job-loss was associated with weight gain of 1.56 (0.89, 2.23) kg/y, while those who maintained employment 0.60 (0.53, 0.68) kg/y (P < 0.001). In both samples, weight changes associated with retirement were similar to those staying in work. In BHPS, job-loss was also associated with significant declines in self-reported well-being and increases in sleep-loss. Two UK-based samples of working adults reveal strong associations between job-loss and excess weight gain. The mediating behaviours are so far unclear but psychosocial mechanisms and sleep-loss may contribute to the excess weight gain among individuals who become unemployed.


Circulating Short-Chain Fatty Acids in Preterm Birth: A Pilot Case-Control Study.
Short-chain fatty acids (SCFAs) are by-products from microbial metabolism of fibers with anti-inflammatory effects. SCFAs may mitigate inflammatory conditions such as obesity through modulation of histone acetylation. Lipid metabolism and inflammation play critical roles during pregnancy. However, few studies have examined maternal SCFAs in relation to pregnancy outcomes. This pilot study examined plasma SCFAs in spontaneous preterm birth. This study used stored plasma from an existing cohort to measure seven (proponic acid, methanoic acid, butanoic acid, isovaleric acid, pentanoic acid, methylpropylbutanoic and methylbutanoic acids) SCFAs in 20 women with spontaneous preterm delivery (< 37 weeks gestation) and 30 women with a healthy term delivery (≥ 37 weeks gestation). All women had singleton pregnancies and provided serum at the time of admission to labor and delivery. SCFAs were measured by purge and trap gas chromatography/mass spectrometry. SCFAs were log transformed. Logistic regression with penalized likelihood approach examined associations between SCFAs and preterm birth, adjusting for age, BMI, and race. We also explored if SCFAs had a linear association with pre-pregnancy BMI. Propionic acid had a negative association with preterm birth [odds ratioadj: 0.56, 95% confidence interval 0.41-0.86). There was a negative association between propionic acid and BMI after adjustments (β = -0.14, p = 0.0011). No other associations were found. Lower levels of propionic acid were associated with preterm birth and correlated with higher BMI. Larger studies should explore if circulatory SCFAs protect against inflammatory pathways during pregnancy and are associated with adverse outcomes when measured earlier in pregnancy.


Anti-CyHV-3 Effect of Fluorescent, Tricyclic Derivative of Acyclovir 6-(4-MeOPh)-TACV in vitro.
Cyprinid herpesvirus 3 (CyHV-3) is a virus infecting carp with disease symptoms of gill necrosis, fish discoloration, sunken eyes, and mortality reaching 90%. Several research groups have examined how to potentially abate the consequences of viral activity. Recently we showed that acyclovir inhibits CyHV-3 replication in vitro and in the present study we examined the anti-CyHV-3 activity of the tricyclic derivative of acyclovir 6-(4-MeOPh)-TACV (T-ACV), a fluorescent molecule known for higher lipophilicity than acyclovir, and therefore potentially better candidate for application in vivo. CCB and KF1 cell lines were incubated with T-ACV at concentrations of 0, 66.67, and 133.33 μM for three days and toxicity examined with MTT and CV assays. To investigate the antiviral activity of T-ACV, the lines were infected with CyHV-3 or mock infected and incubated for three days with the drug at concentrations of 0 or 66.67 μM. The activity of T-ACV was evaluated by plaque assay and TaqMan qPCR. T-ACV at a concentration of 66.67 μM displayed low toxicity and inhibited CyHV-3 activity by 13-29%, varying by cell line and method. The low anti-CyHV-3 activity of T-ACV indicates that it would be reasonable to screen several tricyclic derivatives of acyclovir for such activity.


Standardisation of information submitted to an endpoint committee for cause of death assignment in a cancer screening trial – lessons learnt from CAP (Cluster randomised triAl of PSA testing for Prostate cancer).
In cancer screening trials where the primary outcome is target cancer-specific mortality, the unbiased determination of underlying cause of death (UCD) is crucial. To minimise bias, the UCD should be independently verified by expert reviewers, blinded to death certificate data and trial arm. We investigated whether standardising the information submitted for UCD assignment in a population-based randomised controlled trial of prostate-specific antigen (PSA) testing for prostate cancer reduced the reviewers' ability to correctly guess the trial arm. Over 550 General Practitioner (GP) practices (>415,000 men aged 50-69 years) were cluster-randomised to PSA testing (intervention arm) or the National Health Service (NHS) prostate cancer risk management programme (control arm) between 2001 and 2007. Assignment of UCD was by independent reviews of researcher-written clinical vignettes that masked trial arm and death certificate information. A period of time after the process began (the initial phase), we analysed whether the reviewers could correctly identify trial arm from the vignettes, and the reasons for their choice. This feedback led to further standardisation of information (second phase), after which we re-assessed the extent of correct identification of trial arm. 1099 assessments of 509 vignettes were completed by January 2014. In the initial phase (n = 510 assessments), reviewers were unsure of trial arm in 33% of intervention and 30% of control arm assessments and were influenced by symptoms at diagnosis, PSA test result and study-specific criteria. In the second phase (n = 589), the respective proportions of uncertainty were 45% and 48%. The percentage of cases whereby reviewers were unable to determine the trial arm was greater following the standardisation of information provided in the vignettes. The chances of a correct guess and an incorrect guess were equalised in each arm, following further standardisation. It is possible to mask trial arm from cause of death reviewers, by using their feedback to standardise the information submitted to them. ISRCTN92187251.


Association between Dietary Energy Density and Risk of Breast, Endometrial, Ovarian, and Colorectal Cancer among Canadian Women.
Background: Dietary energy density (DED) is strongly associated with cancer-associated metabolic disorders such as obesity and metabolic syndrome and may thus influence carcinogenesis. However, little is known about its association with cancer. Therefore, we investigated the association of DED with risk of breast, endometrial, ovarian, and colorectal cancers in the Canadian Study of Diet, Lifestyle, and Health.Methods: We conducted a case-cohort study that included an age-stratified subcohort of 3,120 of the 39,532 female participants who completed self-administered lifestyle and dietary questionnaires at baseline, and in whom, respectively, 922, 188, 104, and 269 incident breast, endometrial, ovarian, and colorectal cancer cases were diagnosed, respectively. We estimated HRs and 95% confidence intervals for the association of DED with risk of these cancers using Cox proportional hazards regression models modified for the case-cohort design.Results: There was no statistically significant association between DED and risk of breast, endometrial, ovarian, and colorectal cancers.Conclusions: Our study suggests that DED is not independently associated with risk of breast, endometrial, ovarian, and colorectal cancers among women.Impact: Further investigation of the association between DED and risk of these cancers in larger prospective studies is warranted, as demonstration of associations may have important implications for primary prevention of these cancers. Cancer Epidemiol Biomarkers Prev; 27(3); 338-41. ©2017 AACR.


Guepar total knee prosthesis. Experience at the Vancouver General Hospital.
Eighty-three Guepar valgus-hinge prostheses and 30 prostheses with collinear femoral and the tibial components were inserted in 97 patients at Vancouver General Hospital between March 1975 and May 1978. One hundred and nine arthroplasties were reviewed between January 1979 and April 1980, after an average follow-up of 19 months. It was found that the amount of bone resected made arthrodesis difficult, that there was a very high incidence of patellar instability and that the disintegration of the Silastic bumper created a severe chemical synovitis. The results were excellent in 32, good in 23, fair in 8, and poor in 30. Patellar subluxation occurred in 28 per cent of the valgus prostheses, and in 10 per cent with the straight prostheses; it did not occur with the straight prostheses in rheumatoid joints. The chemical synovitis led to failure with devastating complications necessitating further operations in some knees.


Quantifying avian avoidance of offshore wind turbines: Current evidence and key knowledge gaps.
The risk of collision between birds and turbines is seen as one of the key issues in the planning process for offshore wind farms. In some cases, predictions of collision risk have led to projects either being withdrawn from the planning process, or refused planning consent. Despite this, the evidence base on which collision risk is assessed is extremely limited and assessments rely on models which can be highly sensitive to assumptions, notably about bird collision avoidance behaviour. We present a synthesis of the current state of knowledge about collision risk and avoidance behaviour in seabirds. Evidence suggests species-specific responses to turbines and that in order to avoid collision, most birds adjust their flight paths at some distance from the turbines, rather than making last-second adjustments. We highlight the key gaps in knowledge and make recommendations for future data collection.


Effect of Community Health Worker Support on Clinical Outcomes of Low-Income Patients Across Primary Care Facilities: A Randomized Clinical Trial.
Addressing the social determinants of health has been difficult for health systems to operationalize. To assess a standardized intervention, Individualized Management for Patient-Centered Targets (IMPaCT), delivered by community health workers (CHWs) across 3 health systems. This 2-armed, single-blind, multicenter randomized clinical trial recruited patients from 3 primary care facilities in Philadelphia, Pennsylvania, between January 28, 2015, and March 28, 2016. Patients who resided in a high-poverty zip code, were uninsured or publicly insured, and who had a diagnosis for 2 or more chronic diseases were recruited, and patients were randomized to either the CHW intervention or the control arm (goal setting only). Follow-up assessments were conducted at 6 and 9 months after enrollment. Data were analyzed using an intention-to-treat approach from June 2017 to March 2018. Participants set a chronic disease management goal with their primary care physician; those randomized to the CHW intervention received 6 months of tailored support. The primary outcome was change in self-rated physical health. The secondary outcomes were self-rated mental health, chronic disease control, patient activation, patient-reported quality of primary care, and all-cause hospitalization. Of the 592 participants, 370 (62.5%) were female, with a mean (SD) age of 52.6 (11.1) years. Participants in both arms had similar improvements in self-rated physical health (mean [SD], 1.8 [11.2] vs 1.6 [9.9]; P = .89). Patients in the intervention group were more likely to report the highest quality of care (odds ratio [OR], 1.8; 95% CI, 1.4-2.4; risk difference [RD], 0.12; P < .001) and spent fewer total days in the hospital at 6 months (155 days vs 345 days; absolute event rate reduction, 69%) and 9 months (300 days vs 471 days; absolute event rate reduction, 65%). This reduction was driven by a shorter average length of stay (difference, -3.1 days; 95% CI, -6.33 to 0.22; P = .06) and a lower mean number of hospitalizations (difference, -0.3; 95% CI, -0.6 to 0.0; P = .07) among patients who were hospitalized. Patients in the intervention group had a lower odds of repeat hospitalizations (OR, 0.4; 95% CI, 0.2-0.9; RD, -0.24; P = .02), including 30-day readmissions (OR, 0.3; 95% CI, 0.1-0.9; RD, -0.17; P = .04). A standardized intervention did not improve self-rated health but did improve the patient-perceived quality of care while reducing hospitalizations, suggesting that health systems may use a standardized intervention to address the social determinants of health. ClinicalTrials.gov identifier: NCT02347787.


Prevalence of the extended-spectrum beta-lactamase and qnr genes in clinical isolates of Escherichia coli.
This study was performed to investigate the prevalence of qnr genes in clinical isolates of Escherichia coli from Korea that produce extended-spectrum beta-lactamases (ESBLs). During the period of May to June 2005, we collected clinical isolates of E. coli that were intermediate or resistant to ceftazidime and/or cefotaxime from 11 Korean hospitals. Antimicrobial susceptibility was determined by the disk diffusion and agar dilution methods. ESBL production was confirmed phenotypically by the double-disk synergy test. ESBL and qnr genes were searched for by PCR amplification, and the PCR products were then subjected to direct sequencing. Double-disk synergy tests were positive in 84.3% (118/140) of ceftazidime- and/or cefotaxime-nonsusceptible E. coli isolates. The most prevalent types of ESBL in E. coli isolates were CTX-M-14 (N=41) and CTX-M-15 (N=58). Other ESBLs were also identified, including CTX-M-3 (N=7), CTX-M-9 (N=8), CTX-M-12 (N=1), CTX-M-57 (N=1), SHV-2a (N=2), SHV-12 (N=17) and TEM-52 (N=4). The qnrA1 and qnrB4 genes were identified in 4 and 7 ESBL-producing isolates, respectively. CTX-M-type enzymes were the most common type of ESBL in E. coli isolates from Korea, and the qnr genes were not uncommon in ESBL-producing E. coli isolates. Dissemination of E. coli containing both ESBL and qnr genes could compromise the future usefulness of the expanded-spectrum antibiotics for the treatment of infections.


Serial changes in pulmonary haemodynamics during human pregnancy: a non-invasive study using Doppler echocardiography.
1. Serial pulmonary haemodynamic investigations were performed in 13 women before conception, at monthly intervals throughout pregnancy, and then at 6 months after delivery. 2. Mean pulmonary artery pressure was calculated from pulsed Doppler pulmonary velocities. Pulmonary flow was measured by Doppler and cross-sectional echocardiography. These two measurements were used to calculate pulmonary vascular resistance. 3. Mean non-pregnant pulmonary artery pressure was 13.8 mmHg and no significant change was demonstrated during pregnancy. 4. Pulmonary flow increased from 4.88 to 7.19 litres/min during pregnancy. 5. Pulmonary vascular resistance decreased from 2.85 resistance units before pregnancy to 2.17 resistance units at 8 weeks gestation. Thereafter there was no further significant change, values returning to pre-pregnant levels by 6 months after delivery.


Human Blood Autoantibodies in the Detection of Colorectal Cancer.
Colorectal cancer (CRC) is the second most common malignancy in the western world. Early detection and diagnosis of all cancer types is vital to improved prognosis by enabling early treatment when tumours should be both resectable and curable. Sera from 3 different cohorts; 42 sera (21 CRC and 21 matched controls) from New York, USA, 200 sera from Pittsburgh, USA (100 CRC and 100 controls) and 20 sera from Dundee, UK (10 CRC and 10 controls) were tested against a panel of multiple tumour-associated antigens (TAAs) using an optimised multiplex microarray system. TAA specific IgG responses were interpolated against the internal IgG standard curve for each sample. Individual TAA specific responses were examined in each cohort to determine cutoffs for a robust initial scoring method to establish sensitivity and specificity. Sensitivity and specificity of combinations of TAAs provided good discrimination between cancer-positive and normal serum. The overall sensitivity and specificity of the sample sets tested against a panel of 32 TAAs were 61.1% and 80.9% respectively for 6 antigens; p53, AFP, K RAS, Annexin, RAF1 and NY-CO16. Furthermore, the observed sensitivity in Pittsburgh sample set in different clinical stages of CRC; stage I (n = 19), stage II (n = 40), stage III (n = 34) and stage IV (n = 6) was similar (73.6%, 75.0%, 73.5% and 83.3%, respectively), with similar levels of sensitivity for right and left sided CRC. We identified an antigen panel of sufficient sensitivity and specificity for early detection of CRC, based upon serum profiling of autoantibody response using a robust multiplex antigen microarray technology. This opens the possibility of a blood test for screening and detection of early colorectal cancer. However this panel will require further validation studies before they can be proposed for clinical practice.


E-cigarette curiosity among U.S. middle and high school students: Findings from the 2014 national youth tobacco survey.
Curiosity is a potential risk factor for electronic cigarette (e-cigarette) use, which has increased considerably among US youth in recent years. We examined the relationship between curiosity about e-cigarettes and perceived harm, comparative addictiveness, and e-cigarette advertisement exposure. Data came from the 2014 National Youth Tobacco Survey, a nationally representative survey of U.S. middle and high school students. In 2014, 2.5% of middle school and 9.2% of high school students currently used cigarettes, while 3.9% of middle school and 13.4% of high school students reported current e-cigarette use. Among never e-cigarette users (n=17,286), descriptive statistics assessed curiosity about e-cigarettes by combustible tobacco use, sex, race/ethnicity, and school level. Associations between curiosity and perceived harm (absolute and comparative to cigarettes), comparative addictiveness, and e-cigarette advertising exposure were explored using multivariate models in 2015. Among youth who never used e-cigarettes, 25.8% reported curiosity about e-cigarettes. Higher levels of perceived absolute harm and comparative harm were associated with lower levels of curiosity, while no association was observed between comparative addictiveness and curiosity. Among never combustible tobacco users, the odds of high curiosity were greater among non-Hispanic blacks (odds ratio (OR): 1.39; 95% confidence interval (CI):1.02-1.88), Hispanics (OR=1.79; 95% CI:1.48-2.16), and non-Hispanic 'Other' (OR=1.47; 95% CI:1.15-1.89) race/ethnicities than non-Hispanic whites. One-quarter of middle and high school students who have never used e-cigarettes are curious about the products, with greater curiosity among those with lower perceptions of harm from these products. These findings may help inform future strategies aimed at reducing curiosity about e-cigarettes among youth.


Do wealth and inequality associate with health in a small-scale subsistence society?
In high-income countries, one's relative socio-economic position and economic inequality may affect health and well-being, arguably via psychosocial stress. We tested this in a small-scale subsistence society, the Tsimane, by associating relative household wealth (n = 871) and community-level wealth inequality (n = 40, Gini = 0.15-0.53) with a range of psychological variables, stressors, and health outcomes (depressive symptoms [n = 670], social conflicts [n = 401], non-social problems [n = 398], social support [n = 399], cortisol [n = 811], body mass index [n = 9,926], blood pressure [n = 3,195], self-rated health [n = 2523], morbidities [n = 1542]) controlling for community-average wealth, age, sex, household size, community size, and distance to markets. Wealthier people largely had better outcomes while inequality associated with more respiratory disease, a leading cause of mortality. Greater inequality and lower wealth were associated with higher blood pressure. Psychosocial factors did not mediate wealth-health associations. Thus, relative socio-economic position and inequality may affect health across diverse societies, though this is likely exacerbated in high-income countries.


Modeling Contagion Through Social Networks to Explain and Predict Gunshot Violence in Chicago, 2006 to 2014.
Every day in the United States, more than 200 people are murdered or assaulted with a firearm. Little research has considered the role of interpersonal ties in the pathways through which gun violence spreads. To evaluate the extent to which the people who will become subjects of gun violence can be predicted by modeling gun violence as an epidemic that is transmitted between individuals through social interactions. This study was an epidemiological analysis of a social network of individuals who were arrested during an 8-year period in Chicago, Illinois, with connections between people who were arrested together for the same offense. Modeling of the spread of gunshot violence over the network was assessed using a probabilistic contagion model that assumed individuals were subject to risks associated with being arrested together, in addition to demographic factors, such as age, sex, and neighborhood residence. Participants represented a network of 138 163 individuals who were arrested between January 1, 2006, and March 31, 2014 (29.9% of all individuals arrested in Chicago during this period), 9773 of whom were subjects of gun violence. Individuals were on average 27 years old at the midpoint of the study, predominantly male (82.0%) and black (75.6%), and often members of a gang (26.2%). Explanation and prediction of becoming a subject of gun violence (fatal or nonfatal) using epidemic models based on person-to-person transmission through a social network. Social contagion accounted for 63.1% of the 11 123 gunshot violence episodes; subjects of gun violence were shot on average 125 days after their infector (the person most responsible for exposing the subject to gunshot violence). Some subjects of gun violence were shot more than once. Models based on both social contagion and demographics performed best; when determining the 1.0% of people (n = 1382) considered at highest risk to be shot each day, the combined model identified 728 subjects of gun violence (6.5%) compared with 475 subjects of gun violence (4.3%) for the demographics model (53.3% increase) and 589 subjects of gun violence (5.3%) for the social contagion model (23.6% increase). Gunshot violence follows an epidemic-like process of social contagion that is transmitted through networks of people by social interactions. Violence prevention efforts that account for social contagion, in addition to demographics, have the potential to prevent more shootings than efforts that focus on only demographics.


Differential interactions of FGFs with heparan sulfate control gradient formation and branching morphogenesis.
The developmental activities of morphogens depend on the gradients that they form in the extracellular matrix. Here, we show that differences in the binding of fibroblast growth factor 7 (FGF7) and FGF10 to heparan sulfate (HS) underlie the formation of different gradients that dictate distinct activities during branching morphogenesis. Reducing the binding affinity of FGF10 for HS by mutating a single residue in its HS-binding pocket converted FGF10 into a functional mimic of FGF7 with respect to gradient formation and regulation of branching morphogenesis. In particular, the mutant form of FGF10 caused lacrimal and salivary gland epithelium buds to branch rather than to elongate. In contrast, mutations that reduced the affinity of the FGF10 for its receptor affected the extent, but not the nature, of the response. Our data may provide a general model for understanding how binding to HS regulates other morphogenetic gradients.


Active and inactive genes localize preferentially in the periphery of chromosome territories.
The intranuclear position of a set of genes was analyzed with respect to the territories occupied by the whole chromosomes in which these genes are localized. Genes and their respective chromosome territories were simultaneously visualized in three-dimensionally preserved nuclei applying dual color fluorescence in situ hybridization. Three coding (DMD, MYH7, and HBB) and two noncoding sequences (D1Z2 and an anonymous sequence) were analyzed in four different cell types, including cells where DMD and MYH7 are actively transcribed. Spatial analysis by confocal laser scanning microscopy revealed that the genes are preferentially located in the periphery of chromosome territories. This positioning was independent from the activity of the genes. In contrast, the non-expressed anonymous fragment was found randomly distributed or localized preferentially in the interior of the corresponding chromosome territory. Furthermore, the distribution of the analyzed genes within the territorial peripheries was found to be highly characteristic for each gene, and, again, independent from its expression. The impact of these findings with regard to the three-dimensional arrangement of the linear DNA string within chromosome territories, as well as with respect to a putative nuclear subcompartment confining gene expression, are discussed.


Fermented milk containing Bifidobacterium lactis DN-173 010 in childhood constipation: a randomized, double-blind, controlled trial.
Constipation is a frustrating symptom affecting 3% of children worldwide. A fermented dairy product containing Bifidobacterium lactis strain DN-173 010 was effective in increasing stool frequency in constipated women. Our aim was to assess the effects of this product in constipated children. In this prospective randomized, double-blind, controlled trial, 159 constipated children (defecation frequency < 3 times per week) were randomly allocated to receive either a fermented dairy product that contains B lactis DN-173 010 (n = 79) or a control product (n = 80) twice a day for 3 weeks. The primary endpoint was the change in stool frequency from baseline to after 3 weeks of product consumption. Analyses were by intention to treat. Eleven children did not return to any follow-up visit (5 in the probiotic group, 6 in the control group) and were therefore excluded from the final analysis. Thus, 74 children in each group were analyzed. The change in stool frequency from baseline to after 3 weeks of product consumption increased in both groups, but the difference was not statistically significant (2.9 ± 3.2 in probiotic group versus 2.6 ± 2.6 in control group, P = .35). There were no serious adverse events. In constipated children, the fermented dairy product containing B lactis strain DN-173 010 did increase stool frequency, but this increase was comparable in the control group. There is currently not sufficient evidence to recommend fermented dairy products containing B lactis strain DN-173 010 in this category of patients. Future studies should focus on whether a longer period of probiotic products is more effective in children who have a short history of constipation.


Inherited CARD9 deficiency in 2 unrelated patients with invasive Exophiala infection.
Exophiala species are mostly responsible for skin infections. Invasive Exophiala dermatitidis disease is a rare and frequently fatal infection, with 42 cases reported. About half of these cases had no known risk factors. Similarly, invasive Exophiala spinifera disease is extremely rare, with only 3 cases reported, all in patients with no known immunodeficiency. Autosomal recessive CARD9 deficiency has recently been reported in otherwise healthy patients with severe fungal diseases caused by Candida species, dermatophytes, or Phialophora verrucosa. We investigated an 8-year-old girl from a nonconsanguineous Angolan kindred, who was born in France and developed disseminated E. dermatitidis disease and a 26 year-old woman from an Iranian consaguineous kindred, who was living in Iran and developed disseminated E. spinifera disease. Both patients were otherwise healthy. We sequenced CARD9 and found both patients to be homozygous for loss-of-function mutations (R18W and E323del). The first patient had segmental uniparental disomy of chromosome 9, carrying 2 copies of the maternal CARD9 mutated allele. These are the first 2 patients with inherited CARD9 deficiency and invasive Exophiala disease to be described. CARD9 deficiency should thus be considered in patients with unexplained invasive Exophiala species disease, even in the absence of other infections.


Secretion of human serum albumin by Kluyveromyces lactis overexpressing KlPDI1 and KlERO1.
The control of protein conformation during translocation through the endoplasmic reticulum is often a bottleneck for heterologous protein production. The core pathway of the oxidative folding machinery includes two conserved proteins: Pdi1p and Ero1p. We increased the dosage of the genes encoding these proteins in the yeast Kluyveromyces lactis and evaluated the secretion of heterologous proteins. KlERO1, an orthologue of Saccharomyces cerevisiae ERO1, was cloned by functional complementation of the ts phenotype of an Scero1 mutant. The expression of KlERO1 was induced by treatment of the cells with dithiothreitol and by overexpression of human serum albumin (HSA), a disulfide bond-rich protein. Duplication of either PDI1 or ERO1 led to a similar increase in HSA yield. Duplication of both genes accelerated the secretion of HSA and improved cell growth rate and yield. Increasing the dosage of KlERO1 did not affect the production of human interleukin 1beta, a protein that has no disulfide bridges. The results confirm that the ERO1 genes of S. cerevisiae and K. lactis are functionally similar even though portions of their coding sequence are quite different and the phenotypes of mutants overexpressing the genes differ. The marked effects of KlERO1 copy number on the expression of heterologous proteins with a high number of disulfide bridges suggests that control of KlERO1 and KlPDI1 is important for the production of high levels of heterologous proteins of this type.


Sexy DEG/ENaC channels involved in gustatory detection of fruit fly pheromones.
Hydrocarbon pheromones on the cuticle of Drosophila melanogaster modulate the complex courtship behavior of males. Recently, three members of the degenerin/epithelial Na+ channel (DEG/ENaC) family of sodium channel subunits, Ppk25, Ppk23, and Ppk29 (also known as Nope), have been shown to function in gustatory perception of courtship-modulating contact pheromones. All three proteins are required for the activation of male courtship by female pheromones. Specific interactions between two of them have been demonstrated in cultured cells, suggesting that, in a subset of cells where they are coexpressed, these three subunits function within a common heterotrimeric DEG/ENaC channel. Such a DEG/ENaC channel may be gated by pheromones, either directly or indirectly, or alternatively may control the excitability of pheromone-sensing cells. In addition, these studies identify taste neurons that respond specifically to courtship-modulating pheromones and mediate their effects on male behavior. Two types of pheromone-sensing taste neurons, F and M cells, have been defined on the basis of their specific response to either female or male pheromones. These reports set the stage for the dissection of the molecular and cellular mechanisms that mediate gustatory detection of contact pheromones.


Effect of salinity on the quantity and quality of carotenoids accumulated by Dunaliella salina (strain CONC-007) and Dunaliella bardawil (strain ATCC 30861) Chlorophyta.
Dunaliella salina and D. bardawil are well-known microalgae accumulating high levels of beta-carotene under growth-limiting conditions. In both taxa, this pigment is primarily composed of the isomers 9-cis and all-trans. The 9-cis beta-carotene occurs only in natural sources and is the most attractive from a commercial point of view. The conditions that enhance the preferred accumulation of 9-cis beta-carotene in D. salina are controversial and they have not been well established yet. This study examined the effect of salinity on the quantity and quality of total carotenoids and beta-carotene isomers accumulated by D. salina (strain CONC-007) and D. bardawil (strain ATCC 30861) grown in two media with different nutritional compositions (PES and ART) and at salt concentrations of 1M, 2M and 3M NaCl. Total carotenoids were determined by spectrophotometry and beta-carotene isomers, by HPLC. The highest carotenoid contents per cell were obtained at 2M NaCl in both taxa. In both media, an increase of the 9-cis/all-trans beta-carotene ratio was observed in D. bardawil when the salt concentration increased, with a maximum value of 2.6 (in ART medium at 3M NaCl). In D. salina this ratio did not exhibit the same pattern, and the salt concentrations for maximal ratios were different in both media. The highest ratio obtained for this strain was 4.3 (in ART medium at 2M NaCl).


[Clinical experience of laparoscopic adrenalectomy: the National Defense Medical College experience].
Twenty-seven laparoscopic adrenalectomies (LapAdx) were performed at the National Defense Medical College between 1999 and 2004. We evaluated the results of LapAdx (group L) compared to the results of open adrenalectomy (group O). Twenty-six of the 27 LapAdx were successfully performed, but one patient with a large pheochromocytoma required open surgey because of arterial bleeding in the renal hilus. The mean operating time in group L (185 +/- 19 min) was not significantly different from that in group O (206 +/- 13 min). The mean estimated blood loss in group L (111 +/- 61 g) was significantly lower than that in group O (308 +/- 67 g). The starting time for oral feeding and for ambulation was significantly earlier in group L than in group O. There was a major complication (intraoperative bleeding) in which a group L patient required a blood transfusion. We also compared the surgical results of 26 patients in LapAdx divided chronologically into the first half and the last half to determine the surgical skill acquired. The operating time was significantly shorter and blood loss significantly less for patients in the last half. In addition, the operating time and blood loss for the first-time LapAdx operators were comparable with those of experienced surgeons. Our results support the efficacy and the minimal invasiveness of LapAdx. The accumulated experience and knowledge regarding laparoscopic surgery in our institute were important in improving surgical procedures and results.


Antireflective coatings for multijunction solar cells under wide-angle ray bundles.
Two important aspects must be considered when optimizing antireflection coatings (ARCs) for multijunction solar cells to be used in concentrators: the angular light distribution over the cell created by the particular concentration system and the wide spectral bandwidth the solar cell is sensitive to. In this article, a numerical optimization procedure and its results are presented. The potential efficiency enhancement by means of ARC optimization is calculated for several concentrating PV systems. In addition, two methods for ARCs direct characterization are presented. The results of these show that real ARCs slightly underperform theoretical predictions.


Separating stimulus-driven and response-related LRP components with Residue Iteration Decomposition (RIDE).
When the lateralized readiness potential (LRP) is recorded in stimulus-response compatibility (SRC) tasks, two processes may overlap in the LRP, stimulus-driven response priming and activation based on response selection rules. These overlapping processes are hard to disentangle with standard analytical tools. Here, we show that Residue Iteration Decomposition (RIDE), based on latency variability, separates the overlapping LRP components from a Simon task into stimulus-driven and response-related components. SRC affected LRP amplitudes only in the stimulus-driven component, whereas LRP onsets were affected only in the response-locked component. Importantly, the compatibility effect in reaction times was more similar to the effect in the onsets of the RIDE-derived response-locked LRP component than in the unseparated LRP. Thus, RIDE-separated LRP components are devoid of distortions inherent to standard LRPs.


EphA2 immunoconjugate as molecularly targeted chemotherapy for ovarian carcinoma.
EphA2 is overexpressed in many types of human cancer but is absent or expressed at low levels in normal epithelial tissues. We investigated whether a novel immunoconjugate containing an anti-EphA2 monoclonal antibody (1C1) linked to a chemotherapeutic agent (monomethyl auristatin phenylalanine [MMAF]) through a noncleavable linker maleimidocaproyl (mc) had antitumor activity against ovarian cancer cell lines and tumor models. Specificity of 1C1-mcMMAF was examined in EphA2-positive HeyA8 and EphA2-negative SKMel28 ovarian cancer cells by antibody binding and internalization assays. Controls were phosphate-buffered saline (PBS), 1C1, or control IgG-mcMMAF. Viability and apoptosis were investigated in ovarian cancer cell lines and tumor models (10 mice per group). Antitumor activities were tested in the HeyA8-luc and SKOV3ip1 orthotopic mouse models of ovarian cancer. Endothelial cells were identified by use of immunohistochemistry and anti-CD31 antibodies. All statistical tests were two-sided. The 1C1-mcMMAF immunoconjugate specifically bound to EphA2-positive HeyA8 cells but not to EphA2-negative cells and was internalized by HeyA8 cells. Treatment with 1C1-mcMMAF decreased the viability of HeyA8-luc cells in an EphA2-specific manner. In orthotopic mouse models, treatment with 1C1-mcMMAF inhibited tumor growth by 85%-98% compared with that in control mice (eg, for weight of HeyA8 tumors, 1C1-mcMMAF = 0.05 g and control = 1.03 g; difference = 0.98 g, 95% confidence interval [CI] = 0.40 to 1.58 g; P = .001). Even in bulkier disease models with HeyA8-luc cells, 1C1-mcMMAF treatment, compared with control treatment, caused regression of established tumors and increased survival of the mice (eg, 1C1-mcMMAF vs control, mean = 60.6 days vs 29.4 days; difference = 31.2 days, 95% CI = 27.6 to 31.2 days; P = .001). The antitumor effects of 1C1-mcMMAF therapy, in SKOV3ip1 tumors, for example, were statistically significantly related to decreased proliferation (eg, 1C1-mcMMAF vs control, mean = 44.1% vs 55.8% proliferating cells; difference = 11.7%, 95% CI = 2.45% to 20.9%; P = .01) and increased apoptosis of tumor cells (eg, 1C1-mcMMAF vs control, mean = 8.6% vs 0.9% apoptotic cells; difference = 7.7%, 95% CI = 3.8% to 11.7%; P < .001) and of mouse endothelial cells (eg, 1C1-mcMMAF vs control, mean 2.8% vs 0.4% apoptotic endothelial cells; difference = 2.4%, 95% CI = 1.4% to 4.6%; P = .034). The 1C1-mcMMAF immunoconjugate had antitumor activity in preclinical models of ovarian carcinoma.


Post-exposure prophylaxis against SARS-CoV-2 in close contacts of confirmed COVID-19 cases (CORIPREV): study protocol for a cluster-randomized trial.
Post-exposure prophylaxis (PEP) is a well-established strategy for the prevention of infectious diseases, in which recently exposed people take a short course of medication to prevent infection. The primary objective of the COVID-19 Ring-based Prevention Trial with lopinavir/ritonavir (CORIPREV-LR) is to evaluate the efficacy of a 14-day course of oral lopinavir/ritonavir as PEP against COVID-19 among individuals with a high-risk exposure to a confirmed case. This is an open-label, multicenter, 1:1 cluster-randomized trial of LPV/r 800/200 mg twice daily for 14 days (intervention arm) versus no intervention (control arm), using an adaptive approach to sample size calculation. Participants will be individuals aged > 6 months with a high-risk exposure to a confirmed COVID-19 case within the past 7 days. A combination of remote and in-person study visits at days 1, 7, 14, 35, and 90 includes comprehensive epidemiological, clinical, microbiologic, and serologic sampling. The primary outcome is microbiologically confirmed COVID-19 infection within 14 days after exposure, defined as a positive respiratory tract specimen for SARS-CoV-2 by polymerase chain reaction. Secondary outcomes include safety, symptomatic COVID-19, seropositivity, hospitalization, respiratory failure requiring ventilator support, mortality, psychological impact, and health-related quality of life. Additional analyses will examine the impact of LPV/r on these outcomes in the subset of participants who test positive for SARS-CoV-2 at baseline. To detect a relative risk reduction of 40% with 80% power at α = 0.05, assuming the secondary attack rate in ring members (p0) = 15%, 5 contacts per case and intra-class correlation coefficient (ICC) = 0.05, we require 110 clusters per arm, or 220 clusters overall and approximately 1220 enrollees after accounting for 10% loss-to-follow-up. We will modify the sample size target after 60 clusters, based on preliminary estimates of p0, ICC, and cluster size and consider switching to an alternative drug after interim analyses and as new data emerges. The primary analysis will be a generalized linear mixed model with logit link to estimate the effect of LPV/r on the probability of infection. Participants who test positive at baseline will be excluded from the primary analysis but will be maintained for additional analyses to examine the impact of LPV/r on early treatment. Harnessing safe, existing drugs such as LPV/r as PEP could provide an important tool for control of the COVID-19 pandemic. Novel aspects of our design include the ring-based prevention approach, and the incorporation of remote strategies for conducting study visits and biospecimen collection. This trial was registered at www.ClinicalTrials.gov ( NCT04321174 ) on March 25, 2020.


Management of obese patients with diabetes mellitus: comparison of advice education with group management.
The purpose of our study was to compare the effect on diabetes control of group management with the advice-educational technique traditionally used in managing obese outpatients with poorly controlled non-insulin-dependent diabetes mellitus (NIDDM). Forty-one patients were randomly assigned to these two treatment programs, and 32 patients completed the 6-mo study. Initially, patients were seen for 1-h sessions at 1- and 2-wk intervals and later at 1-mo intervals. Patients were asked to do home blood glucose monitoring, decrease caloric intake, increase exercise, and if they were taking insulin, to adjust the dose to attain approximate euglycemia and to stabilize food and exercise patterns. The combined groups reduced mean +/- SD glycohemoglobin from 10.9 +/- 3.1 to 9.4 +/- 2.4% (P less than .05). Internal Health Locus of Control Scale was negatively and significantly correlated with initial and subsequent glycohemoglobin values (the more internal, the lower the glycohemoglobin). At the end of the study the patients in the group management program had significantly lower blood glucose levels than those given advice and education, but no significant differences in glycohemoglobin values or percentage overweight were observed. One patient had a normal initial glycohemoglobin, and only 4 patients had values in the normal range of 4-6.8% at the end of the study. Better management programs need to be developed for treating obese outpatients with NIDDM.


Detection and identification of a novel lactic acid bacterial flora within the honey stomach of the honeybee Apis mellifera.
This investigation concerned the question of whether honeybees collect bacteria that are beneficial for humans from the flowers that contribute to formation of their honey. Bacteria originating from the types of flowers involved, and found in different anatomic parts of the bees, in larvae, and in honey of different types, were sampled during a 2-year period. 16S rRNA sequencing of isolates and clones was employed. A novel bacterial flora composed of lactic acid bacteria (LAB) of the genera Lactobacillus and Bifidobacterium, which originated in the honey stomach of the honeybee, was discovered. It varied with the sources of nectar and the presence of other bacterial genera within the honeybee and ended up eventually in the honey. It appeared that honeybees and the novel LAB flora may have evolved in mutual dependence on one another. It was suggested that honey be considered a fermented food product because of the LAB involved in honey production. The findings are seen as having clear implications for future research in the area, as providing a better understanding the health of honeybees and of their production and storage of honey, and as having clear relevance for future honeybee and human probiotics.


Time-resolved and temperature tuneable measurements of fluorescent intensity using a smartphone fluorimeter.
A smartphone fluorimeter capable of time-based fluorescence intensity measurements at various temperatures is reported. Excitation is provided by an integrated UV LED (λex = 370 nm) and detection obtained using the in-built CMOS camera. A Peltier is integrated to allow measurements of the intensity over T = 10 to 40 °C. All components are controlled using a smartphone battery powered Arduino microcontroller and a customised Android application that allows sequential fluorescence imaging and quantification every δt = 4 seconds. The temperature dependence of fluorescence intensity for four emitters (rhodamine B, rhodamine 6G, 5,10,15,20-tetraphenylporphyrin and 6-(1,4,8,11-tetraazacyclotetradecane)2-ethyl-naphthalimide) are characterised. The normalised fluorescence intensity over time of the latter chemosensor dye complex in the presence of Zn2+ is observed to accelerate with an increasing rate constant, k = 1.94 min-1 at T = 15 °C and k = 3.64 min-1 at T = 30 °C, approaching a factor of ∼2 with only a change in temperature of ΔT = 15 °C. Thermally tuning these twist and bend associated rates to optimise sensor approaches and device applications is proposed.


Regulation of protein kinase C-related protein kinase 2 (PRK2) by an intermolecular PRK2-PRK2 interaction mediated by Its N-terminal domain.
Protein kinase C-related protein kinases (PRKs) are effectors of the Rho family of small GTPases and play a role in the development of diseases such as prostate cancer and hepatitis C. Here we examined the mechanism underlying the regulation of PRK2 by its N-terminal region. We show that the N-terminal region of PRK2 prevents the interaction with its upstream kinase, the 3-phosphoinositide-dependent kinase 1 (PDK1), which phosphorylates the activation loop of PRK2. We confirm that the N-terminal region directly inhibits the kinase activity of PRK2. However, in contrast to previous models, our data indicate that this inhibition is mediated in trans through an intermolecular PRK2-PRK2 interaction. Our results also suggest that amino acids 487-501, located in the linker region between the N-terminal domains and the catalytic domain, contribute to the PRK2-PRK2 dimer formation. This dimerization is further supported by other N-terminal domains. Additionally, we provide evidence that the region C-terminal to the catalytic domain intramolecularly activates PRK2. Finally, we discovered that the catalytic domain mediates a cross-talk between the inhibitory N-terminal region and the activating C-terminal region. The results presented here describe a novel mechanism of regulation among AGC kinases and offer new insights into potential approaches to pharmacologically regulate PRK2.


Osteoporosis services in secondary care: a UK survey.
A 1994 survey indicated that only 13 health authorities in the UK were purchasing access to dual X-ray absorptiometry (DXA), the most accurate measure of osteoporosis risk. By 1998 the number of centres (including private facilities providing DXA) was 161. All these were sent a questionnaire concerning their activities. 124 (77%) responded, and the survey found that DXA machines operate, on average, for only 3.6 days a week. Funding of and access to diagnostic services for osteoporosis varies greatly. There is clear scope for greater efficiency in the use of existing DXA machines and more equitable access to diagnostic services is required for effective management of osteoporosis.


Nanodiamond as a New Hyperpolarizing Agent and Its (13)C MRS.
In this work, we have hyperpolarized carbonaceous nanoparticles (D ≈ 10 nm), that is, "nanodiamonds", with 1.1% (13)C (natural abundance) using dynamic nuclear polarization (DNP). The polarization buildup curve showed a signal enhancement with relative intensity up to 4700 at 1.4 K and 100 mW microwave power. (13)C magnetic resonance spectra (MRS) were obtained from the sample at 7 T, and the signal decayed with a T1 of 55 ± 3s. Notably, polarization was possible in the absence of added radical, consistent with previous results showing endogenous unpaired electrons in natural nanodiamonds. These likely contribute to the shorter T1's compared to those of highly pure diamond. Despite the relatively short T1, these observations suggest that natural nanodiamonds may be useful for in vivo applications.


Plasma Amyloid Beta Concentrations in Aged and Cognitively Impaired Pet Dogs.
Longevity-associated neurological disorders have been observed across human and canine aging populations. Alzheimer's disease (AD) and canine cognitive dysfunction syndrome (CDS) represent comparable diseases affecting both species as they age. Translational diagnostic and therapeutic research is needed for these incurable diseases. The amyloid β (Aβ) peptide family are AD-associated peptides with identical amino acid sequences between dogs and humans. Plasma Aβ42 concentration increases with age and decreases with AD in humans, and cerebrospinal fluid (CSF) concentration decreases in AD and correlates inversely with the amyloid load within the brain. Similarly, CSF Aβ42 concentrations decrease in dogs with CDS but there is limited and conflicting information on plasma Aβ42 concentrations in aging dogs and dogs with CDS. We measured plasma concentrations of Aβ42 and Aβ40 with an ultrasensitive single-molecule array assay (SIMOA) in a population of healthy aging dogs of different life stages (n = 36) and dogs affected with CDS (n = 11). In addition, the ratio of Aβ42/β40 was calculated. The mean plasma concentrations of Aβ42 and Aβ40 increased significantly with age (r2 = 0.27, p = 0.001; and r2 = 0.42, p < 0.001, respectively) and with life stage: puppy/junior group (0.43-2 years): 1.23 ± 0.95 and 38.26 ± 49.43 pg/mL; adult/mature group (2.1-9 years): 10.99 ± 5.45 and 131.05 ± 80.17 pg/mL; geriatric/senior group (9.3-14.5 years): 18.65 ± 16.65 and 192.88 ± 146.38 pg/mL, respectively. Concentrations of Aβ42 and Aβ40 in dogs with CDS (11.0-15.6 years) were significantly lower than age-matched healthy dogs at 11.61 ± 6.39 and 150.23 ± 98.2 pg/mL (p = 0.0048 and p = 0.001), respectively. Our findings suggest the dynamics of canine plasma amyloid concentrations are analogous to that found in aging humans with and without AD.


[Count Henri de Bonneval (1806-1882), practitioner of homeopathy, agronomist and philanthropist].
Count Henri de Bonneval, was born in Bordeaux in 1806, in the line of descent of one of the most ancient French families of noble rank. He was assistant manager of the Strasbourg stud farm in 1830, when Louis-Philippe, an Orleanist, ascended to the throne of the Bourbon Charles X. As several other legitimists, Count Henri refused to take an oath to the new king and prefered to resign his position. Interested in medicine, he was deeply impressed by Hahnemann's Organon der Heilkunst and decided to leave France for KOthen, in Saxony, in order to learn homeopathy directly from its founder Back to France, he defended in Montpellier the first French medical thesis devoted to homeopathy and then opened a consulting room in Bordeaux. He rapidly gained a solid reputation and a large audience as a practitioner of homeopathy. At the same time, Henri acquired the Chateau de Latresne and the 500 acres surrounding land. He renovated and brought up to date the agricultural and wine-producing activities of the estate. The medical doctor soon proved to be an expert agronomist, extending his competence to the famous vineyard Chateau Canon of St. Emilion. Throughout his life, the Count showed notable qualities of philanthropy, materialized at Latresne by the construction of a church and, adjacent to the chapel, a boarding school, two classrooms and shelters for poor or sick old people. At the end of his life, Henri de Bonneval wrote a comprehensive book, that includes the presentation and discussions of the homeopathic methods, some philosophical reflections and personal memories.


Mycobacteriophage D29 contains an integration system similar to that of the temperate mycobacteriophage L5.
A mycobacteriophage D29 DNA fragment cloned in pRM64, a shuttle plasmid that transforms Mycobacterium smegmatis, was sequenced. The determined sequence was 2592 nucleotides long and had a mean G+C content of 63.7 mol%, similar to that of mycobacterial DNA. Four ORFs were identified: one with strong homology to dCMP deaminase genes; one homologous to mycobacteriophage L5 gene 36, whose function is unknown; one encoding a possible excisase; and one encoding an integrase. The intergenic region between the putative excisase gene and the integrase gene had a lower than average G+C content and showed the presence of the same attP core sequence as mycobacteriophage L5. Transformation experiments using subclones of pRM64 indicated that the integrase gene and all the intergenic region were essential for stable transformation. A subclone containing the integrase gene and the core attP sequence was able to transform but recombinants were highly unstable. Southern analysis of total DNA from cells transformed with pRM64 and its derivatives showed that all the plasmids were integrated at one specific site of the bacterial chromosome. A recombinant exhibiting a high level of resistance to the selective drug kanamycin had two plasmids integrated at different sites. These results demonstrated that the D29 sequences contained in pRM64 were integrative, indicating that the generally hold view of D29 as a virulent phage must be reviewed.


Benefit of laparoscopic radical nephrectomy in patients with a high BMI.
The aim of the present study was to analyze all relevant data collected thus far at our hospital in order to compare the laparoscopic vs. open techniques in terms of the intraoperative results and to analyze whether patients with a higher BMI may derive greater benefit from laparoscopy. We reviewed the medical records of 93 patients treated with laparoscopic nephrectomy and 91 patients treated with the open method at our institution. We analyzed the data with the aim of determining what kind of factor would be associated with an increase in estimated blood loss and prolongation of operative time, and we compared the results for laparoscopic and open nephrectomy. A statistically significant correlation was observed between body mass index and operating time (P = 0.018) or estimated blood loss (P = 0.017) in laparoscopic nephrectomy. Multivariate analysis demonstrated that body mass index was an independent risk factor for prolonged operating time (P = 0.030, odds ratio = 1.174). In the open nephrectomy patients, body mass index also had a significant correlation with operating time (P < 0.001) and estimated blood loss (P < 0.001). Multivariate analysis also revealed that operating time depended on body mass index (P < 0.001, odds ratio = 1.348) and tumor size (P = 0.030, odds ratio = 1.320), and the odds ratio of body mass index was higher than that in laparoscopic nephrectomy. As body mass index increases, both laparoscopic nephrectomy and open nephrectomy become technically more difficult. However, our data showed that for patients with a high body mass index, the laparoscopic approach is comparatively safer and of greater benefit.


The importance of psychological and social factors in influencing the uptake and maintenance of physical activity after stroke: a structured review of the empirical literature.
Background. People with stroke are not maintaining adequate engagement in physical activity (PA) for health and functional benefit. This paper sought to describe any psychological and social factors that may influence physical activity engagement after stroke. Methods. A structured literature review of studies indexed in MEDLINE, CinAHL, P&BSC, and PsycINFO using search terms relevant to stroke, physical disabilities, and PA. Publications reporting empirical findings (quantitative or qualitative) regarding psychological and/or social factors were included. Results. Twenty studies from 19 publications (9 surveys, 1 RCT, and 10 qualitative studies) were included. Seventeen studies reported findings pertinent to psychological factors and fourteen findings pertinent to social factors. Conclusion. Self-efficacy, physical activity beliefs, and social support appear particularly relevant to physical activity behaviour after stroke and should be included in theoretically based physical interventions. The Transtheoretical Model and the Theory of Planned Behaviour are candidate behavioural models that may support intervention development.


Knowledge Across Reference Frames: Cognitive Maps and Image Spaces.
In human and non-human animals, conceptual knowledge is partially organized according to low-dimensional geometries that rely on brain structures and computations involved in spatial representations. Recently, two separate lines of research have investigated cognitive maps, that are associated with the hippocampal formation and are similar to world-centered representations of the environment, and image spaces, that are associated with the parietal cortex and are similar to self-centered spatial relationships. We review evidence supporting cognitive maps and image spaces, and we propose a hippocampal-parietal network that can account for the organization and retrieval of knowledge across multiple reference frames. We also suggest that cognitive maps and image spaces may be two manifestations of a more general propensity of the mind to create low-dimensional internal models.


The impact of cancer and its treatment on physical activity levels and behavior in Hong Kong Chinese childhood cancer survivors.
Research indicates that regular physical activity is associated with numerous physiological and psychological health benefits for childhood cancer survivors. A review of the literature reveals that no study has so far examined the physical activity levels and behavior of Hong Kong Chinese childhood cancer survivors, and how the cancer and its treatment affect the physical activity and other behavior of these children remains unclear. The aims of this study were to assess the physical activity levels of Hong Kong Chinese childhood cancer survivors and to explore the factors that affect their adherence to and maintenance of regular physical activity. A cross-sectional study was used. A total of 128 childhood cancer survivors (9-16-year-olds) who underwent medical follow-up in the outpatient clinic were invited to participate in the study. There was a significant decline in physical activity levels among childhood cancer survivors. Most of them did not take physical exercise regularly. Concern about academic performance, fatigue, and a decrease in physical strength and endurance after remission prevented them from engaging in regular physical activity. This study indicates that many childhood cancer survivors did not engage in regular physical activity and that they overlooked or underestimated its importance. It is essential for nurses to correct misconceptions about physical activity among childhood cancer survivors and their parents and, most importantly, to advocate the principle of regular physical activity for these children, with the aim of enhancing their physical and psychological well-being.


Clinical features and treatment outcomes of 41 dogs with sublingual ectopic thyroid neoplasia.
Thyroid neoplasia is common in dogs, but there are few reports of dogs with ectopic, sublingual thyroid tumors. To describe clinical features and outcomes of dogs with ectopic, sublingual thyroid neoplasia. Five hundred and forty-four dogs with thyroid neoplasia. This retrospective study reviewed the medical records of dogs referred for thyroid neoplasia between 1995 and 2013. Data extracted included signalment, extent of thyroid disease (eutopic or ectopic; metastasis), serum thyroxine (T4) concentration, treatment, and survival. Of 544 dogs with thyroid neoplasia, 41 (7.5%) dogs had ectopic sublingual thyroid tumors. The clinical features of these 41 dogs were similar to the cohort group of 503 dogs with eutopic or ectopic mediastinal thyroid tumors, but dogs with sublingual tumors were younger and less likely to have metastatic disease (15% versus 30%, P < .05). Of the 41 dogs, 28 received treatment: 21 with surgery (which included partial hyoidectomy in 13), 7 with radioiodine alone, and 13 with surgery followed by administration of radioiodine. Overall median survival was 562 days (range, 1-1,850 days). When compared with eutopic thyroid carcinomas, ectopic sublingual thyroid tumors generally have a less aggressive biologic behavior. Many dogs have prolonged survival, even without treatment, although death because of local tumor invasiveness or metastasis can develop in some dogs. Surgical thyroidectomy, including partial hyoidectomy, is generally effective for control of local disease. Administration of radioiodine, alone or in combination with surgical treatment, is recommended for multifocal disease or metastasis.


T cell-dendritic cell interaction in vivo: random encounters favor development of long-lasting ties.
Understanding the complexity of the functional communication between cells composing the immune system is central to improving our capacity to manipulate it and conceive better strategies to combat microbial pathogens. So far, these studies have been based on immunohistochemistry of fixed tissues and in vitro attempts to reproduce functional connections between cells. The application of two-photon laser microscopy to the observation of viable immune cells in their natural environment where foreign antigens are carried to trigger an immune response opens a new era for these studies. They reveal exceptional properties of the locomotion of T cells that facilitate encounters with dendritic cells and the receipt of information that promotes T cell survival, death, or initiation of immune responses. These studies also complement in vitro observations addressing the importance of time of stimulation in determining T cell fates.


Fecal transplant modifies urine chemistry risk factors for urinary stone disease.
Urinary stone disease (USD) is a major health concern. There is a need for new treatment modalities. Recently, our group provided evidence for an association between the GMB composition and USD. The accessibility of the Gut Microbiome (GMB) makes it an attractive target for investigation and therefore, in these studies we have evaluated the extent to which the whole gut microbial community in fecal transplants can affect urinary stone risk parameters in an animal model. Fresh fecal pellets were collected from Zucker lean rats, homogenized in PBS (100 mg/mL), filtered through a 70 μm strainer and then orally gavaged into C57BL/6NTac germ-free mice. Twenty-four hours urine collections and GMB analysis were performed over time for 1 month. Kidney and gut tissue were harvested from transplanted mice for western blot analysis of expression levels of the Slc26a6 transporter involved in oxalate balance. Urinary calcium decreased after fecal transplant by 55% (P < 0.001). Urinary oxalate levels were on average 24% lower than baseline levels (P < 0.001). Clostridiaceae family was negatively correlated with urinary oxalate at 4 weeks after transplant (r = -0.83, P < 0.01). There was a 0.6 unit average increase in urinary pH from a baseline of 5.85 (SE ± 0.028) to 6.49 (SE ± 0.04) (P < 0.001) after transplant. There was a concomitant 29% increase in gastrointestinal alkali absorption (P < 0.001) 4-weeks after fecal transplant. Slc26a6 expression increased by 90% in the cecum after transplant. Our results suggest that the gut microbiome may impact metabolism, alters urinary chemistry, and thereby may influence USD; the accessibility of the GMB can potentially be leveraged for therapeutic interventions.


Use of mass spectrometry to study signaling pathways.
Activation of cells by extracellular stimuli, such as growth factors, initiates a cascade of events involving posttranslational modifications, including phosphorylation, formation of protein complexes, and induction or repression of gene expression. Traditionally, genetic methods or specific biochemical assays have been used to identify molecules involved in signaling pathways. Lately, mass spectrometry, combined with elegant biochemical approaches, has become a powerful tool for identifying proteins and posttranslational modifications. With this protocol, we hope to bridge the gap between the biochemical and molecular aspects of signal transduction pathways and the mass spectrometric tools and techniques that are available to study them. We provide methods for large-scale cell culture and immunoprecipitation of tyrosine-phosphorylated proteins, silver staining of gels, trypsin digests, and protein identification by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and nanoelectrospray tandem mass spectrometry. We discuss the special requirements for the identification of phosphorylation sites in proteins by mass spectrometry. We describe enrichment of phosphopeptides from unseparated peptide mixtures by immobilized metal affinity column (IMAC) and the use of phosphatases to identify phosphorylated peptides. We also discuss specialized methods, such as precursor ion scanning in the negative mode and direct sequencing of phosphopeptides in the positive mode. Our goal is to provide detailed methods in use today for proteomic applications in general and for receptor-mediated signaling pathways in particular.


Wnt signaling and Dupuytren's disease.
Dupuytren's disease is a benign fibromatosis of the hands and fingers that leads to flexion contractures. We hypothesized that multiple genetic and environmental factors influence susceptibility to this disease and sought to identify susceptibility genes to better understand its pathogenesis. We conducted a genomewide association study of 960 Dutch persons with Dupuytren's disease and 3117 controls (the discovery set) to test for association between the disease and genetic markers. We tested the 35 single-nucleotide polymorphisms (SNPs) most strongly associated with Dupuytren's disease (P<1×10(-4)) in the discovery set in three additional, independent case series comprising a total of 1365 affected persons and 8445 controls from Germany, the United Kingdom, and The Netherlands. Initially, we observed a significant genomewide association between Dupuytren's disease and 8 SNPs at three loci. Tests of replication and joint analysis of all data from 2325 patients with Dupuytren's disease and 11,562 controls yielded an association with 11 SNPs from nine different loci (P<5.0×10(-8)). Six of these loci contain genes known to be involved in the Wnt-signaling pathway: WNT4 (rs7524102) (P=2.8×10(-9); odds ratio, 1.28), SFRP4 (rs16879765) (P=5.6×10(-39); odds ratio, 1.98), WNT2 (rs4730775) (P=3.0×10(-8); odds ratio, 0.83), RSPO2 (rs611744) (P=7.9×10(-15); odds ratio, 0.75), SULF1 (rs2912522) (P=2.0×10(-13); odds ratio, 0.72), and WNT7B (rs6519955) (P=3.2×10(-33); odds ratio, 1.54). This study implicates nine different loci involved in genetic susceptibility to Dupuytren's disease. The fact that six of these nine loci harbor genes encoding proteins in the Wnt-signaling pathway suggests that aberrations in this pathway are key to the process of fibromatosis in Dupuytren's disease.


Estimation of annual probabilities of changing disability levels in Australians with relapsing-remitting multiple sclerosis.
Transition probabilities are the engine within many health economics decision models. However, the probabilities of progression of disability due to multiple sclerosis (MS) have not previously been estimated in Australia. To estimate annual probabilities of changing disability levels in Australians with relapsing-remitting MS (RRMS). Combining data from Ausimmune/Ausimmune Longitudinal (2003-2011) and Tasmanian MS Longitudinal (2002-2005) studies (n = 330), annual transition probabilities were obtained between no/mild (Expanded Disability Status Scale (EDSS) levels 0-3.5), moderate (EDSS 4-6.0) and severe (EDSS 6.5-9.5) disability. From no/mild disability, 6.4% (95% confidence interval (CI): 4.7-8.4) and 0.1% (0.0-0.2) progressed to moderate and severe disability annually, respectively. From moderate disability, 6.9% (1.0-11.4) improved (to no/mild state) and 2.6% (1.1-4.5) worsened. From severe disability, 0.0% improved to moderate and no/mild disability. Male sex, age at onset, longer disease duration, not using immunotherapies greater than 3 months and a history of relapse were related to higher probabilities of worsening. We have estimated probabilities of changing disability levels in Australians with RRMS. Probabilities differed between various subgroups, but due to small sample sizes, results should be interpreted with caution. Our findings will be helpful in predicting long-term disease outcomes and in health economic evaluations of MS.


Acute renal failure in hemorrhagic hypotension: cellular energetics and renal function.
In male Wistar rats, renal adenosine triphosphate (ATP), inorganic phosphate (Pi) and intracellular pH were measured by 31phosphorus nuclear magnetic resonance (31P NMR) and correlated with renal function before, during, and for one hour after a period of 30 to 40 minutes hemorrhagic hypotension. In animals which suffered no change in these metabolites during hypotension, retransfusion immediately restored normal renal function. When metabolite changes were observed during hypotension, they occurred suddenly with severe ATP depletion, Pi accumulation, and intracellular acidosis occurring almost concurrently. Metabolic changes of this magnitude were always associated with renal dysfunction in the post-hypotensive period, which occurred even when the period of biochemical change was only 10 to 15 minutes. The abnormalities in post-hypotensive renal function resemble the pattern of change seen in human acute tubular necrosis (ATN): depressed glomerular filtration rate (GFR), urine output varying from polyuria to oliguria, decreased urine to plasma inulin ratio, increased urinary sodium concentration, increased fractional excretion of sodium, and increased fractional excretion of potassium. It is postulated that changes in renal cellular energy status during hemorrhagic hypotension distinguish pre-renal failure from early or incipient ATN.


Long-term effectiveness and predictors of success of low-intensity shockwave therapy in phosphodiesterase type 5 inhibitors non-responders.
Objective: To evaluate the long-term (18 months) effectiveness, safety, and factors that may predict the success of low-intensity shockwave lithotripsy (Li-SWT) in patients with erectile dysfunction (ED) who fail to respond to oral phosphodiesterase type 5 inhibitors (PDE5i). Patients and Methods: This prospective study included 52 patients with ED of vascular origin who failed to respond to oral PDE5i. The International Index of Erectile Function-Erectile Function domain (IIEF-EF) and Erection Hardness Score (EHS) questionnaires were used to evaluate EF. Patients under went two Li-SWT treatment sessions per week for 3 weeks, followed by a 3-week treatment-free period, and the cycle was repeated until each patient received 12 treatment sessions. Patients were followed-up after Li-SWT at 3, 6, 12, and 18 months. Results: At the 18-month follow-up, 33 patients (63.5%) were able to achieve an erection sufficient for penetration with or without PDE5i (22 were maintained on oral PDE5i). The remaining 19 patients (36.5%) had a poor response to Li-SWT and oral PDE5i. The initial response showed some decline in 50% of the initial responders. Younger men (aged <45 years), short ED duration (<2 years), and moderate ED severity responded better to Li-SWT. There were no adverse side-effects. Conclusion: In the present study, Li-SWT was a safe and effective treatment in 63.5% of men with ED who failed to respond to oral PDE5i. Factors such as age (<45 years), ED duration (<2 years), and ED severity can predict treatment outcome in such patients. Abbreviations: CDU: colour Doppler ultrasonography; ED: erectile dysfunction; EDV: end-diastolic velocity; EF: erectile function; EHS: Erection Hardness Score; FU: follow-up; IIEF-EF: International Index of Erectile Function-EF domain; Li-SWT: low-intensity shockwave lithotripsy; PDE5i: phosphodiesterase type 5 inhibitors; PGE1: prostaglandin E1; PSV: peak systolic velocity; RI: resistive index; VOD: veno-occlusive dysfunction.


Protective role of arginase in a mouse model of colitis.
Arginase is the endogenous inhibitor of inducible NO synthase (iNOS), because both enzymes use the same substrate, l-arginine (Arg). Importantly, arginase synthesizes ornithine, which is metabolized by the enzyme ornithine decarboxylase (ODC) to produce polyamines. We investigated the role of these enzymes in the Citrobacter rodentium model of colitis. Arginase I, iNOS, and ODC were induced in the colon during the infection, while arginase II was not up-regulated. l-Arg supplementation of wild-type mice or iNOS deletion significantly improved colitis, and l-Arg treatment of iNOS(-/-) mice led to an additive improvement. There was a significant induction of IFN-gamma, IL-1, and TNF-alpha mRNA expression in colitis tissues that was markedly attenuated with l-Arg treatment or iNOS deletion. Treatment with the arginase inhibitor S-(2-boronoethyl)-l-cysteine worsened colitis in both wild-type and iNOS(-/-) mice. Polyamine levels were increased in colitis tissues, and were further increased by l-Arg. In addition, in vivo inhibition of ODC with alpha-difluoromethylornithine also exacerbated the colitis. Taken together, these data indicate that arginase is protective in C. rodentium colitis by enhancing the generation of polyamines in addition to competitive inhibition of iNOS. Modulation of the balance of iNOS and arginase, and of the arginase-ODC metabolic pathway may represent a new strategy for regulating intestinal inflammation.


Scientific opinion on the tolerable upper intake level for vitamin B6.
Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver a scientific opinion on the tolerable upper intake level (UL) for vitamin B6. Systematic reviews of the literature were conducted by a contractor. The relationship between excess vitamin B6 intakes and the development of peripheral neuropathy is well established and is the critical effect on which the UL is based. A lowest-observed-effect-level (LOAEL) could not be established based on human data. A reference point (RP) of 50 mg/day is identified by the Panel from a case-control study, supported by data from case reports and vigilance data. An uncertainty factor (UF) of 4 is applied to the RP to account for the inverse relationship between dose and time to onset of symptoms and the limited data available. The latter covers uncertainties as to the level of intake that would represent a LOAEL. This leads to a UL of 12.5 mg/day. From a subchronic study in Beagle dogs, a LOAEL of 50 mg/kg body weight (bw) per day can be identified. Using an UF of 300, and a default bw of 70 kg, a UL of 11.7 mg/day can be calculated. From the midpoint of the range of these two ULs and rounding down, a UL of 12 mg/day is established by the Panel for vitamin B6 for adults (including pregnant and lactating women). ULs for infants and children are derived from the UL for adults using allometric scaling: 2.2-2.5 mg/day (4-11 months), 3.2-4.5 mg/day (1-6 years), 6.1-10.7 mg/day (7-17 years). Based on available intake data, EU populations are unlikely to exceed ULs, except for regular users of food supplements containing high doses of vitamin B6.


MO-D-213CD-01: Cartesian Methods for Rapid Time-Resolved MR Angiography.
The physics of the MR image formation fundamentally trades off spatial resolution with temporal resolution. Time spent in acquiring data for the second image of a time series can alternatively be spent in sampling higher spatial frequencies for the first image to improve its spatial resolution. Historically this tradeoff has been addressed by making the k-space sampling rate high, such as with very short repetition times, and with methods such as view sharing in which only a portion of k-space is updated from one image to the next in a time series. Over a decade ago the method of parallel acquisition was proposed in which the signals detected by the individual elements comprising a multi-element receiver coil are used to provide further spatial discrimination and reduce acquisition time. These approaches include those based in image space (SENSE) or in k- space (SMASH, GRAPPA). In the last decade these methods have been integrated in contrast-enhanced MRA (CE-MRA) to provide a radical improvement in performance. CE-MRA is an application particularly well suited to these methods. The general desire for MRA images to be three-dimensional allows the use of 2D implementation of parallel acquisition, generally much more robust than 1D implementation. Also, the SNR loss associated with parallel acquisition is tempered in CE-MRA because high, arterial-phase signal is sampled throughout the data acquisition. Cartesian MR data acquisition, performed along a rectilinear sampling pattern in k-space, offers specific advantages in relative ease of implementation of 2D parallel acquisition and in "freezing" the status of the time-varying object at a specific timepoint by use of centric view ordering. This presentation will provide a review of these methods and how they have been effectively developed and integrated within the last decade for improved time-resolved MRA. Cartesian k-space sampling patterns can now be quickly selected on a patient- and anatomy-specific basis for optimum acceleration. Receiver coil arrays have been adapted to allow up to 20× reduction in the number of k-space points sampled for a given spatial resolution. Reconstruction hardware now allows generation of 3D images within only hundreds of msec after data acquisition, permitting real-time generation of diagnostic quality images and their use in interactively guiding other processes. 1. Understand recently developed physics techniques which have allowed a 20x improvement in the speed of data acquisition for MR angiography 2. Understand how Cartesian sampling of k-space facilitates the practical and effective implementation of these techniques 3. Show how contemporary implementation of these physics techniques has provided a significant improvement in MRA image quality over the last decade.


Coumestrol modulates Akt and Wnt/β-catenin signaling during the attenuation of adipogenesis.
Coumestrol is a natural phytochemical present in plants such as red clover and soy, and has been reported to stimulate the estrogen receptor as a major phytoestrogen. While the molecular mechanisms responsible for the anti-adipogenic effects of phytoestrogens such as genistein and daidzein have been previously investigated, the effects of coumestrol on adipogenesis remain to be elucidated. We observed that coumestrol dose-dependently attenuates MDI (mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin)-induced lipid accumulation, consistent with an earlier study, while significantly inhibiting MDI-induced adipogenesis in the first 48 hours of differentiation, a critical time window for anti-adipogenic effects. Coumestrol treatment suppressed MDI-induced protein expression of PPARγ and C/EBPα in adipocytes, leading to the subsequent downregulation of FAS and aP2 expression. Akt and GSK3β were phosphorylated shortly after MDI stimulation, and these responses were inhibited by coumestrol treatment. Coumestrol also increased LRP6 protein expression, resulting in the recovery of β-catenin downregulation by MDI, while attenuating MDI-induced downregulation of Wnt10b. In addition, mRNA and protein expression of c-Myc and cyclin D1, target genes of β-catenin, were both recovered by coumestrol treatment. These results suggest that coumestrol inhibits adipocyte differentiation via regulation of Akt and Wnt/β-catenin signaling and may have potential for development as an agent to prevent adipogenesis.


Nutraceutical potential of monofloral honeys produced by the Sicilian black honeybees (Apis mellifera ssp. sicula).
In the light of the growing interest in food and food products obtained through organic and environmentally friendly techniques, the present work represents the first approach to the evaluation of the biological profile of some Sicilian honeys produced in purity by the local black honeybees. Samples exhibited up to 10 times more total phenolics and higher antioxidant capacity than what already reported for the same variety of honeys produced by other honeybee subspecies from Sicily, other Italian regions and abroad. Noteworthy, the gallic acid contents in medlar and almond honeys represented the highest level of single phenolic acid reported in honey so far. A broad antimicrobial spectrum was showed by all of the honey samples and a good correlation between their inhibition capacity and polyphenolic contents was measured. Experimental results highlighted samples among the honeys characterised by the highest nutraceutical added value and most excellent quality.