Assessment of sperm nuclear quality after in vitro maturation of fresh or frozen/thawed mouse pre-pubertal testes.
Is nuclear quality of in vitro generated spermatozoa from fresh or frozen/thawed pre-pubertal mouse testes similar to that of their in vivo counterparts? The production of spermatozoa with aneuploidy, DNA fragmentation or chromatin condensation defects was not significantly increased in organotypic cultures compared to in vivo controls. Although murine spermatozoa have been produced in vitro from pre-pubertal testes, their nuclear DNA integrity has never been investigated. Fresh and frozen/thawed testicular fragments from 6 to 7 days postpartum (dpp) mice were cultured for 30 days. Testicular tissues were frozen by controlled slow freezing (CSF) or solid surface vitrification (SSV). In total, 30 fresh, 30 CSF, 30 SSV testes were used for in vitro maturation and 6 testes from 36 to 37 dpp mice were used as in vivo controls. Murine spermatozoa were extracted from pooled in vitro cultured testicular fragments and from in vivo controls. Sperm aneuploidy was analyzed by fluorescence in situ hybridization (FISH), DNA fragmentation by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling, chromatin condensation by aniline blue staining, telomere length and number by quantitative FISH, DNA oxidation by immunocytochemical detection of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Because of the low spermatogenic yield in cultures, a hundred spermatozoa extracted from pooled tissues were examined and compared to their in vivo counterparts. Most of spermatozoa generated in vitro and in vivo were haploid, contained unfragmented DNA and normally condensed chromatin. A similar proportion of spermatozoa with aneuploidy, DNA fragmentation or chromatin condensation defects was found in cultures and in vivo. No significant difference in telomere length was found within the nuclei of in vitro and in vivo generated spermatozoa. However, the number of telomere spots was lower in gametes obtained from cultures of fresh, CSF and SSV testes than in their natural counterparts (P < 0.01). Moreover, the proportion of spermatozoa containing 8-OHdG was significantly increased in frozen/thawed tissues in comparison to fresh tissues and in vivo controls (P < 0.05). None. Further studies will be needed to enhance the production of spermatozoa in organotypic cultures while preserving their quality, to investigate epigenetic modifications and embryonic development. This is the first study comparing the nuclear quality of in vitro and in vivo generated murine spermatozoa. The organotypic culture system will have to be adapted for human tissue and extensive analyses of human gamete quality will have to be performed before potential clinical applications can be envisaged. This work was supported by Rouen University Hospital, Ligue contre le Cancer, Agence de la Biomédecine, Association Laurette Fugain, France Lymphome Espoir, and co-supported by European Union and Région Normandie. Europe gets involved in Normandie with European Régional Development Fund (ERDF). The authors declare that they have no conflict of interest.


Structure--activity relationships of the didemnins.
Bioactivities of 42 didemnin congeners, either isolated from the marine tunicates Trididemnun solidum and Aplidium albicans or prepared synthetically and semisynthetically, have been compared. The growth inhibition of various murine and human tumor cells and plaque reduction of HSV-1 and VSV grown on cultured mammalian cells were used to assess cytotoxicity and antiviral activity. Biochemical assays for macromolecular synthesis (protein, DNA, and RNA) and enzyme inhibition (dihydrofolate reductase, thymidylate synthase, DNA polymerase, RNA polymerase, and topoisomerases I and II) were also performed to specify the mechanisms of action of each analogue. Immunosuppressive activity of the didemnins was determined using a mixed lymphocyte reaction (MLR) assay. These assays revealed that the native cyclic depsipeptide core is an essential structural requirement for most of the bioactivites of the didemnins, especially for cytotoxicities and antiviral activities. The linear side-chain portion of the peptide can be altered with a gain, in some cases, of bioactivities. In particular, dehydrodidemnin B, tested against several types of tumor cells and in in vivo studies in mice, as well as didemnin M, tested for the mixed lymphocyte reaction and graft vs host reaction in murine systems, showed remarkable gains in their in vitro and in vivo activities compared to didemnin B.


The Prognostic Implications of Two-Dimensional Speckle Tracking Echocardiography in Hypertrophic Cardiomyopathy: Current and Future Perspectives.
Two-dimensional speckle tracking echocardiography represents a novel, simple, and reproducible technique for the estimation of left ventricular myocardial deformation (strain) and the evaluation of left ventricular twist mechanics. During the last few years, its clinical and prognostic implications in cardiomyopathies and hypertrophic cardiomyopathy (HCM), in particular, have been rapidly increasing. Reduced global longitudinal strain is associated with more severe disease and confers an increased risk for major cardiac events, independently of other clinical and echocardiographic risk factors. Left ventricular dyssynchrony also seems promising as a risk factor for sudden cardiac events. With respect to left atrial mechanics, left atrial reservoir, conduit, and contractile strain may also confer an increased prognostic value for atrial fibrillation, major cardiac events, and even sudden death. Although right ventricular global longitudinal strain is impaired in HCM compared with healthy controls and individuals with physiological hypertrophy, its prognostic significance is currently unknown. Conclusively, 2-dimensional speckle tracking imaging seems promising for HCM. However, future studies are needed to incorporate this new imaging technique in the standard evaluation of an HCM individual.


Ethical, Legal and Social Issues Surrounding Research on Genetic Contributions to Anti-Social Behavior.
Scientific study of genetic contributions to chronic antisocial behavior has stemmed from many lines of research in recent years. Genetic research involving twin, family, and adoption studies have traditionally been used to compare the health and behavior outcomes of individuals who share the same environment or hereditary lineage; several of these studies have concluded that heredity plays some role in the formation of chronic antisocial behavior, including various forms of aggression and chronic norm-defiance. However, the ethical, social, and legal environment surrounding research on the biological contributions to antisocial behavior in the United States is contentious. Although there has been some discussion in the last few decades regarding the ethical, social, and legal concerns around this type of research within academic and policy circles, analysis and discussion of these concerns rarely appear together. This paper explores the main themes that interact to form the basis of much of the resistance to positing biological contributions to antisocial behavior.


The Effects of Social Skills Training vs. Psychoeducation on Negative Attitudes of Mothers of Persons with Schizophrenia: A Pilot Study.
We compared the effects of two brief psychoeducation programs and social skills training on the negative attitudes of mothers with a son who has schizophrenia. 15 mothers with strong negative feelings towards a sons with schizophrenia were assigned by convenience to participate in one of three brief (5 session) group programs at an outpatient clinic: lecture-based psychoeducation, video-based psychoeducation, or social skills training. Assessments using the Patient Rejection Scale were conducted with the mothers at post-treatment, and 3-, 6-, and 9-months later. Mothers in the three groups demonstrated significantly different patterns of changes in their negative attitudes following treatment. Whereas the mothers who received the two psychoeducation interventions showed reductions in rejecting attitudes immediately following the program, their scores gradually increased at the subsequent follow-up assessments. In contrast, the mothers in the social skills training group showed reductions in negative attitudes that were sustained across all of the follow-up assessments. Brief social skills training may be more effective than psychoeducation in reducing negative attitudes of parents who have an offspring with schizophrenia.


Lived experiences of pediatric oncology nurses in Iran.
Caring is a valuable task. The staff in any profession that involves patients' fear, anxiety, pain, and suffering may experience similar feelings. As a professional group, oncology nurses deal with patients and their relatives and caregivers under very stressful conditions. They encounter pain, suffering, and death as a part of their daily life. A number of studies have evaluated the experiences of pediatric oncology nurses in other countries. Therefore, conducting a survey about the experiences of Iranian nurses of caring for children with cancer can reveal their demands, stress, and limitations. In a qualitative research, in-depth, unstructured individual interviews with open-ended questions were conducted to evaluate the experiences of pediatric oncology nurses in a hospital in a metropolitan city of Iran. The subjects all consented to participate and had at least one year of working experience in the ward. Content analysis was performed to analyze the data. The lived experiences of pediatric oncology nurses were categorized in five main themes. These themes included attachment, supportive care, trying to repress feelings, feeling of helplessness, and the need to be supported. According to these results, nurses who provide care for children with cancer require support. This research also highlighted the roles, limitations, and needs of nurses in pediatric oncology wards.


The human body may buffer small differences in meal size and timing during a 24-h wake period provided energy balance is maintained.
Because approximately 20% of the work force in the industrialized world have irregular working hours, it is pertinent to study the consequences of eating at irregular, especially nighttime hours. We studied the postprandial responses during nocturnal fasting vs. eating throughout a 24-h wake period. Seven healthy males were studied twice in a crossover design. After a 6-d diet adjustment period [high fat diet, 45 energy percent (en%) fat, 40 en% carbohydrates)] with sleep from 2300 to 0700 h, the men were kept awake for 24 h at the metabolic ward and given either 6 isoenergetic meals, i.e., every 4 h (N-eat) or 4 isoenergetic meals from 0800 to 2000 h followed by a nocturnal fast (N-fast), with the same 24-h energy intake. Energy expenditure, substrate utilization, activity, heat release, body temperature and blood variables were measured over 24 h. Energy expenditure and blood glucose, triacylglycerol, insulin and glucagon concentrations were lower and nonesterified fatty acids concentrations were higher during the nocturnal fast than during nocturnal eating (P < 0.05); however, no 24-h differences between the protocols were apparent. Nocturnal fasting slightly altered the secretory patterns of the thyroid hormones and cortisol (P < 0.05). We found no clear indication that it would be more favorable to ingest few larger daytime meals than smaller meals throughout the 24-h period. The body seems to be able to buffer small differences in meal size and timing provided energy balance is maintained.


Oral health status and treatment need among institutionalised hearing-impaired and blind children and young adults in Udaipur, India. A comparative study.
The aim of this study was to assess and compare the oral health status and the treatment needs of the institutionalised hearing-impaired and blind children and young adults in the city of Udaipur, Rajasthan, India. A descriptive cross-sectional study was conducted among 498 institutionalised hearing-impaired and blind people, aged 4 to 23 years, in the city of Udaipur, Rajasthan. The World Health Organization oral health assessment basic methods and form (1997) were used for data collection. Clinical examinations were carried out in the institute's medical room or classroom by single examiner with the aid of a mouth mirror, explorer and Community Periodontal Index (CPI) probe under adequate natural light (Type III examination). The resulting data were entered into statistical software and analysed by applying the chi-square test, ANOVA, t-test and stepwise multiple linear regression analysis. The total mean DMFT (decayed-missing-filled teeth) and mean dft scores were 1.77 and 0.27 respectively. The largest component of DMFT was the D, with a mean of 1.49. The F component of 0.08 was very low. Mean DMFT/dft was greater among hearing-impaired than among blind subjects. Overall, 159 (32%) were periodontally healthy (CPI=0), 162 (32%) had shallow pockets (CPI=3) and 36 (7%) had deeper pockets (CPI=4). A higher percentage of the blind (87; 43%) than the hearing-impaired (72; 24%) subjects were periodontally healthy (CPI score=0). One-surface fillings were the most commonly provided form of past treatment. The findings in this study highlight the lack of dental treatment for this group. Overall oral health status was poorer in the hearing-impaired than in the blind subjects.


Incidence and outcomes of major trauma patients managed in the Australian Capital Territory.
To determine the incidence and characteristics of major traumatic injury treated in the Australian Capital Territory (ACT) over a 5-year period. A retrospective analysis of ACT Level 1 Trauma Centre registry data from July 2007 to June 2012 was conducted. Inclusion criteria were: major trauma (Injury Severity Score (ISS) >15), intensive care unit admission, hospital stay >3 days or penetrating injury. There were 931 patients with an ISS >15, a 12.5% increase over the 5-year period (P = 0.003). Unadjusted mortality rates in the ISS >15 group remained stable at 9.9%. About half of all injuries occurred outside the ACT. The largest age group represented was 16-24 years. Road trauma was the most prevalent injury mechanism, but its incidence reduced over the 5-year period (52.2-40.3% [P = 0.005]). There was a 3.1% increase in admissions following violence. The largest demographic affected by major trauma in the ACT and surrounds is young people. Injury prevention should remain focused on road trauma but also target violence and high-risk recreation activities. Further investigation around the circumstances of major traumatic injury in young people is required. Funding and cross-border agreements should be reviewed to minimize financial disadvantage to the ACT.


Immunologic reconstitution in the DiGeorge syndrome by fetal thymic transplant.
An infant with idiopathic hypoparathyroidism was found to have no evidence of thymus by radiologic examinations, including pneumomediastinum. Immunoglobulin concentrations were normal but there was clear evidence of inadequate cellular immunity including gradually developing lymphopenia. The infant suffered recurrent diarrhea and pneumonia, chronic rhinorrhea and moniliasis. At age 7 months fetal thymic tissue was implanted in the rectus abdominis muscle. There was immediate increase in lymphocyte counts which have remained normal on most determinations. Other aspects of immunologic function were restored and remain intact. The child, now 6 years of age, although mentally retarded, has no unusual infections. Immunologic reconstitution was thus achieved by transplant of fetal thymus.


Radial keratotomy. A retrospective study of cases observed at the Kresge Eye Institute for six months.
The conclusions alter the review of 25 eyes six months following radial keratotomy are as follows: (1) The improvement in uncorrected visual acuity is dependent on multiple variables that substantially alter the results achieved. The degree to which each of these variables influences the outcome has yet to be determined; however, radial keratotomy does reduce myopia. (2) The time required for stabilization of the corneal curvature was about three months, although gradual reduction in the correction achieved occurred in a few cases between three and six months. (3) Postoperative glare at night and/or fluctuating vision was an important complaint in over one third of the patients. (4) Proper patient selection and informed consent are essential. The possibility of getting rid of glasses is not likely except for the patient with less than 3 to 4 diopters of myopia. Most patients who underwent the procedure were pleased with the improvement in unaided visual acuity, although it may have been less than they expected.


Proximal Hamstring Tendinopathy: Clinical Aspects of Assessment and Management.
Synopsis Proximal hamstring tendinopathy (PHT) typically manifests as deep buttock pain at the hamstring common origin. Both athletic and nonathletic populations are affected by PHT. Pain and dysfunction are often long-standing and limit sporting and daily functions. There is limited evidence regarding diagnosis, assessment, and management; for example, there are no randomized controlled trials investigating rehabilitation of PHT. Some of the principles of management established in, for example, Achilles and patellar tendinopathy would appear to apply to PHT but are not as well documented. This narrative review and commentary will highlight clinical aspects of assessment and management of PHT, drawing on the available evidence and current principles of managing painful tendinopathy. The management outline presented aims to guide clinicians as well as future research. J Orthop Sports Phys Ther 2016;46(6):483-493. Epub 15 Apr 2016. doi:10.2519/jospt.2016.5986.


An experimental study on the anti-Ehrlich ascites carcinoma effect of purified toad venom extract.
The objective of this paper was to study the anti-Ehrlich ascites carcinoma effect of purified toad venom extract and its mechanism. Mouse model of Ehrlich ascites carcinoma was established with cisplatin as the control to observe the inhibitory effect of purified toad venom extract on malignant peritoneal effusion in mice. The results showed that compared with the control group, ascites volume, number of tumour cells and tumour cell viability decreased and ascites inhibition rate reached over 50% in each treatment group, and with the increase of the dose, incidence of ascites showed a downward trend. The number of tumour cells in ascites and tumour cell viability in the purified toad venom high-dose group were lower than those of the cisplatin group. Compared with the model group, survival time was prolonged in varying degrees in the purified toad venom groups and cisplatin group. The study concluded that purified extract of toad venom has an anti-Ehrlich ascites carcinoma effect.


Characterizing Lysine Acetylation of Glucokinase.
Glucokinase (GK) catalyzes the phosphorylation of glucose to form glucose-6-phosphate as the substrate of glycolysis for energy production. Acetylation of lysine residues in Escherichia coli GK has been identified at multiple sites by a series of proteomic studies, but the impact of acetylation on GK functions remains largely unknown. In this study, we applied the genetic code expansion strategy to produce site-specifically acetylated GK variants which naturally exist in cells. Enzyme assays and kinetic analyses showed that lysine acetylation decreases the GK activity, mostly resulting from acetylation of K214 and K216 at the entrance of the active site, which impairs the binding of substrates. We also compared results obtained from the glutamine substitution method and the genetic acetyllysine incorporation approach, showing that glutamine substitution is not always effective for mimicking acetylated lysine. Further genetic studies as well as in vitro acetylation and deacetylation assays were performed to determine acetylation and deacetylation mechanisms, which showed that E. coli GK could be acetylated by acetyl-phosphate without enzymes and deacetylated by CobB deacetylase. This article is protected by copyright. All rights reserved.


Inefficacy of metronidazole in experimental infections of Leishmania donovani, L. mexicana, and Trypanosoma brucei brucei.
Metronidazole has been claimed in several earlier reports to be active in human cases of leishmaniasis and trypanosomiasis. Its efficacy against the protozoa causing these diseases was tested in hamsters infected with Leishmania mexicana or L. donovani, and in mice infected with Trypanosoma brucei brucei. In separate experiments, hamsters were either inoculated intradermally into the nose with 5 million amastigotes of L. mexicana or intracardially with 10-30 million amastigotes of L. donovani, and mice were infected intraperitoneally with 30 million T. b. brucei. Metronidazole was administered in four oral doses on alternate days for a total of 375 mg/kg to hamsters and 500 mg/kg to mice. Sodium stibogluconate (Pentostam) served as a positive control. In hamsters the extent of infection was assessed by the appearance of flagellates in blood agar cultures of nose and spleen, by counting amastigotes in nose and liver impression smears, and by measuring the size of nose lesions. Ultrastructure of nose lesions before and after treatment with metronidazole or Pentostam was also evaluated. Infection in mice was assessed by the extent of parasitemia and/or survival to 30 days. In no case did metronidazole-treated animals differ from untreated controls. Metronidazole shows no activity against experimental infections of leishmaniasis or trypanosomiasis in these animal models.


Disseminated mycobacteriosis in a cat (abstract).
A 12-year-old, FIV-positive domestic cat from rural Northland presented with diffuse and slowly progressive swellings of the vulva, tail base, head, and distal limbs. Fine needle aspirate biopsy of the subcutis revealed large numbers of pleomorphic acid fast bacteria. At necropsy, the dermis and subcutis in the affected areas were expanded by a pale yellow opaque infiltrate. Large ulcerated cutaneous swellings were present in the left maxillary area and bilaterally at the point of the jaw corresponding to the submandibular lymph nodes. Cutaneous nodules were present in the pre-auricular area. The skin of the carpi and tarsi was thickened and there was extensive hair loss, also evident over the tail base. The internal organs were normal in appearance. Histopathologically, the cutaneous swellings consisted of sheets of large foamy macrophages, macrophages, containing yellow/brown pigment and large numbers of acid-fast bacilli, that extended from the superficial dermis to the subcutis and fascia. Infiltrates of acid fast-laden macrophages were present in the liver, lung, spleen, and lymph nodes. Infection by a novel saprophytic Mycobacterium spp was considered most likely.


Suicide risk assessment in the depressed elderly patient with cancer.
Although several studies have identified different factors that increase the risk for suicide in the elderly, depression, especially in cancer patients, puts people at even greater risk. In geriatric patients with co-morbid illnesses, depression is often diagnosed or overlooked because of the assumption that it is to be expected. When patients with cancer are diagnosed with depression, it is imperative that clinicians have a clear-cut tool for assessing and identifying signs of suicidality in order to develop a treatment plan to manage the patient. This article delineates the steps involved in conducting a thorough and systematic suicide risk assessment in depressed elderly patients with cancer.


Antiviral therapy of hepatitis C in chronic kidney diseases: meta-analysis of controlled clinical trials.
Hepatitis C virus (HCV) infection remains frequent in patients with chronic kidney disease and the detrimental role of HCV on survival is well-established in this population. Several authors have reported on efficacy and safety of antiviral therapy for hepatitis C in this polulation but there is no clear consensus on management. To evaluate efficacy and safety of antiviral therapy for hepatitis C in patients with chronic kidney disease, we performed a systematic review of the published medical literature and completed a meta-analysis of controlled clinical trials. The primary outcome was sustained virological response (as a measure of efficacy); the secondary outcome was drop-out rate (as a measure of tolerability). We used the random effects model of Der Simonian and Laird, with heterogeneity and sensitivity analyses. We identified 13 studies including 539 unique patients; 10 (76.9%) concerned patients on maintenance dialysis. Only prospective, controlled clinical trials were included. Pooling of study results showed a significant increase of viral response in study (patients treated with antiviral therapy) than control patients (patients who did not receive therapy), the pooled odds ratio (OR) of failure to obtain a sustained viral response was 0.081 [95% confidence intervals (CI), 0.029-0.230], P = 0.0001. The pooled OR of drop-out rate was significantly increased in study vs control patients, OR = 0.389 (95% CI, 0.155-0.957), P = 0.04. The studies were heterogeneous with regard to viral response and drop-out rate. In the subset of clinical trials (n = 6) involving only dialysis patients receiving interferon (IFN) monotherapy for chronic HCV, there was a significant difference in the risk of failure to obtain a sustained viral response (study vs control patients), OR = 0.054 (95% CI, 0.019; 0.150), P = 0.0001 (random-effects model). No significant (NS) heterogeneity was found (Q = 14.604, P = 1.0). No difference in the drop-out rate between study and control patients was shown, OR = 0.920 (95% CI, 0.367; 2.311), NS. This result being homogeneous (Q = 3.639, P = 0.388). Our meta-analysis showed that the viral response was greater in patients with chronic kidney disease who received antiviral therapy than controls. No difference in the drop-out rate between study and control patients occurred in the subgroup of dialysis patients on IFN monotherapy. These results support IFN-based therapy for hepatitis C in patients on maintenance dialysis.


Subcutaneous hydatid cyst in the popliteal fossa at the site of a previous wasp sting.
We report an uncommon case of a primary Echinococcus cyst that developed in the subcutaneous tissue of the right popliteal fossa, at the spot of a previous wasp sting, suggesting the possibility of an unusual transmission of the eggs by insects. This unusual presentation was initially considered as a Baker cyst until parasytological results verified Echinococcus hydatidosus, the larval form of Echinococcus granulosus, as diagnosis. However, the most common path of Echinococcus granulosus infection is through contact with a definitive host or by ingestion of ova through contaminated water or food. Transmission by insects should also be reconsidered in endemic areas.


Comparative efficacy of risperidone and clozapine in the treatment of patients with refractory schizophrenia or schizoaffective disorder: a retrospective analysis.
Clozapine is effective in up to 60% of patients with refractory schizophrenia, whereas the efficacy of risperidone remains unknown. This retrospective study examined the relative efficacy of these drugs in chronically institutionalized patients refractory to conventional antipsychotic agents. A total of 24 patients who at different time periods had received adequate trials of both clozapine and risperidone and met our inclusion criteria for minimum dose and duration of each trial were included; for clozapine, a minimum dose of 300 mg/day had to be maintained for at least 12 weeks, and for risperidone, a minimum dose of 6 mg/day for at least 6 weeks. Information obtained from systematic retrospective chart review was blindly rated by 2 psychiatrists using the 7-point Clinical Global Impressions-Improvement (CGI-I) scale on overall clinical state and along specific symptom domains of positive symptoms, negative symptoms, and aggressive behavior. The mean +/- SD dose was 520+/-94 mg/day for clozapine and 7.5+/-2.2 mg/day for risperidone. Fourteen patients (58%) were classified as responders to clozapine, while 6 (25%) responded to risperidone (CGI-I score of 1 or 2); on specific symptom domains, response rates to clozapine were 38% (9/24) on positive symptoms, 29% (7/24) on negative symptoms, and 71% (12/17) on aggressive behavior. For risperidone, response rates were 17% (4/24) on positive symptoms, 8% (2/24) on negative symptoms, and 41% (7/17) on aggressive behavior. The results of this study support the utility of first giving a risperidone trial in a treatment algorithm for refractory patients because of its better risk/benefit profile compared with clozapine. Clozapine, however, remains our gold standard in the management of these patients.


Perioperative Infusion of Glucagon-Like Peptide-1 Prevents Insulin Resistance After Surgical Trauma in Female Pigs.
Insulin resistance is an independent negative predictor of outcome after elective surgery and increases mortality among surgical patients in intensive care. The incretin hormone glucagon-like peptide-1 (GLP-1) potentiates glucose-induced insulin release from the pancreas but may also increase insulin sensitivity in skeletal muscle and directly suppress hepatic glucose release. Here, we investigated whether a perioperative infusion of GLP-1 could counteract the development of insulin resistance after surgery. Pigs were randomly assigned to three groups; surgery/control, surgery/GLP-1, and sham/GLP-1. Both surgery groups underwent major abdominal surgery. Whole-body glucose disposal (WGD) and endogenous glucose release (EGR) were assessed preoperatively and postoperatively using D-[6,6-2H2]-glucose infusion in combination with hyperinsulinemic euglycemic step-clamping. In the surgery/control group, peripheral insulin sensitivity (i.e., WGD) was reduced by 44% relative to preoperative conditions, whereas the corresponding decline was only 9% for surgery/GLP-1 (P < 0.05). Hepatic insulin sensitivity (i.e., EGR) remained unchanged in the surgery/control group but was enhanced after GLP-1 infusion in both surgery and sham animals (40% and 104%, respectively, both P < 0.05). Intraoperative plasma glucose increased in surgery/control (∼20%) but remained unchanged in both groups receiving GLP-1 (P < 0.05). GLP-1 diminished an increase in postoperative glucagon levels but did not affect skeletal muscle glycogen or insulin signaling proteins after surgery. We show that GLP-1 improves intraoperative glycemic control, diminishes peripheral insulin resistance after surgery, and suppresses EGR. This study supports the use of GLP-1 to prevent development of postoperative insulin resistance.


Ganoderma lucidum suppresses growth of breast cancer cells through the inhibition of Akt/NF-kappaB signaling.
Ganoderma lucidum (Reishi, Lingzhi) is a popular Asian mushroom that has been used for more than 2 millennia for the general promotion of health and was therefore called the "Mushroom of Immortality." Ganoderma lucidum was also used in traditional Chinese medicine to prevent or treat a variety of diseases, including cancer. We previously demonstrated that Ganoderma lucidum suppresses the invasive behavior of breast cancer cells by inhibiting the transcription factor NF-kappaB. However, the molecular mechanisms responsible for the inhibitory effects of Ganoderma lucidum on the growth of highly invasive and metastatic breast cancer cells has not been fully elucidated. Here, we show that Ganoderma lucidum inhibits proliferation of breast cancer MDA-MB-231 cells by downregulating Akt/NF-kappaB signaling. Ganoderma lucidum suppresses phosphorylation of Akt on Ser473 and downregulates the expression of Akt, which results in the inhibition of NF-kappaB activity in MDA-MB-231 cells. The biological effect of Ganoderma lucidum was demonstrated by cell cycle arrest at G0/G1, which was the result of the downregulation of expression of NF-kappaB-regulated cyclin D1, followed by the inhibition of cdk4. Our results suggest that Ganoderma lucidum inhibits the growth of MDA-MB-231 breast cancer cells by modulating Akt/NF-kappaB signaling and could have potential therapeutic use for the treatment of breast cancer.


Gambogic acid inhibits STAT3 phosphorylation through activation of protein tyrosine phosphatase SHP-1: potential role in proliferation and apoptosis.
The transcription factor, STAT3, is associated with proliferation, survival, and metastasis of cancer cells. We investigated whether gambogic acid (GA), a xanthone derived from the resin of traditional Chinese medicine, Garcinia hanburyi (mangosteen), can regulate the STAT3 pathway, leading to suppression of growth and sensitization of cancer cells. We found that GA induced apoptosis in human multiple myeloma cells that correlated with the inhibition of both constitutive and inducible STAT3 activation. STAT3 phosphorylation at both tyrosine residue 705 and serine residue 727 was inhibited by GA. STAT3 suppression was mediated through the inhibition of activation of the protein tyrosine kinases Janus-activated kinase 1 (JAK1) and JAK2. Treatment with the protein tyrosine phosphatase (PTP) inhibitor pervanadate reversed the GA-induced downregulation of STAT3, suggesting the involvement of a PTP. We also found that GA induced the expression of the PTP SHP-1. Deletion of the SHP-1 gene by siRNA suppressed the ability of GA to inhibit STAT3 activation and to induce apoptosis, suggesting the critical role of SHP-1 in its action. Moreover, GA downregulated the expression of STAT3-regulated antiapoptotic (Bcl-2, Bcl-xL, and Mcl-1), proliferative (cyclin D1), and angiogenic (VEGF) proteins, and this correlated with suppression of proliferation and induction of apoptosis. Overall, these results suggest that GA blocks STAT3 activation, leading to suppression of tumor cell proliferation and induction of apoptosis.


Characterization of Ultra-fine Grained and Nanocrystalline Materials Using Transmission Kikuchi Diffraction.
One of the challenges in microstructure analysis nowadays resides in the reliable and accurate characterization of ultra-fine grained (UFG) and nanocrystalline materials. The traditional techniques associated with scanning electron microscopy (SEM), such as electron backscatter diffraction (EBSD), do not possess the required spatial resolution due to the large interaction volume between the electrons from the beam and the atoms of the material. Transmission electron microscopy (TEM) has the required spatial resolution. However, due to a lack of automation in the analysis system, the rate of data acquisition is slow which limits the area of the specimen that can be characterized. This paper presents a new characterization technique, Transmission Kikuchi Diffraction (TKD), which enables the analysis of the microstructure of UFG and nanocrystalline materials using an SEM equipped with a standard EBSD system. The spatial resolution of this technique can reach 2 nm. This technique can be applied to a large range of materials that would be difficult to analyze using traditional EBSD. After presenting the experimental set up and describing the different steps necessary to realize a TKD analysis, examples of its use on metal alloys and minerals are shown to illustrate the resolution of the technique and its flexibility in term of material to be characterized.


Electrocatalytic CO2 Reduction to Formate at Low Overpotentials on Electrodeposited Pd Films: Stabilized Performance by Suppression of CO Formation.
Palladium nanoparticles are effective for catalytic CO2 reduction. However, CO, one of the most important products in the CO2 reduction sequence, has strong affinity for the Pd surface and poisons the catalytic sites rapidly. In this research, an electrodeposited Pd film exhibits high activity for CO2 reduction to formate with the suppression of CO formation at low overpotentials. The substrates, electrodeposition process and the post-treatment of the Pd films affect the CO2 reduction pathway significantly. The cyclic voltammetry deposition produces films that exhibit more porous morphologies and have higher current efficiencies for formate than those of films produced at constant potential. These films show stable CO2 reduction performance at low overpotentials and have high current efficiencies (≈50-60 % depending on the substrate) for formate formation at a potential of -0.4 V versus the reversible hydrogen electrode without any detectable CO formation. It seems that the Pd surface generated by the new electrodeposition process described here produces a nanostructure that can promote formate formation and suppress CO formation.


Wnt5a signaling controls cytokinesis by correctly positioning ESCRT-III at the midbody.
Wnts activate at least two signaling pathways, the β-catenin-dependent and -independent pathways. Although the β-catenin-dependent pathway is known to contribute to G1-S transition, involvement of the β-catenin-independent pathway in cell cycle regulation remains unclear. Here, we show that Wnt5a signaling, which activates the β-catenin-independent pathway, is required for cytokinesis. Dishevelled 2 (Dvl2), a mediator of Wnt signaling pathways, was localized to the midbody during cytokinesis. Beside the localization of Dvl2, Fz2, a Wnt receptor, was detected in the midbody with the endosomal sorting complex required for transport III (ESCRT-III) subunit, CHMP4B. Depletion of Wnt5a, its receptors, and Dvl increased multinucleation. The phenotype observed in Wnt5a-depleted cells was rescued by the addition of purified Wnt5a but not Wnt3a, which is a ligand for the β-catenin-dependent pathway. Moreover, depletion of Wnt5a signaling caused loss of stabilized microtubules and mislocalization of CHMP4B at the midbody, which affected abscission. Inhibition of the stabilization of microtubules at the midbody led to the mislocalization of CHMP4B, while depletion of CHMP4B did not affect the stabilization of microtubules, suggesting that the correct localization of CHMP4B depends on microtubules. Fz2 was localized to the midbody in a Rab11-dependent manner, probably along stabilized microtubules. Fz2 formed a complex with CHMP4B upon Wnt5a stimulation and was required for proper localization of CHMP4B at the midbody, while CHMP4B was not necessary for the localization of Fz2. These results suggest that Wnt5a signaling positions ESCRT-III in the midbody properly for abscission by stabilizing midbody microtubules.


Current trends in the understanding and treatment of social phobia in youth.
Social phobia is a common psychiatric problem in children and adolescents that has recently gained serious attention in clinical practice and research. This article describes the clinical presentation of social phobia and reviews several emergent areas of related research to social phobia in children and adolescents. We begin with a summary of research regarding the developmental pathways of childhood social anxiety into social phobia, highlighting normative developmental factors, behavioral inhibition, pathophysiology, genetics, and parenting/environmental factors. This is followed by a critical review of findings from studies providing evidence for and against the existence of subtypes of social phobia. Common comorbid disorders and sequelae of social phobia in children and adolescents are discussed, bringing to attention the critical need for effective treatments, the current status of which is reviewed in the final section. In sum, this review demonstrates that through research in multiple disciplines, scientists are now developing a better understanding of the risk and protective factors of social phobia in children and adolescents, ultimately leading to improvements in primary prevention and intervention efforts for children at serious risk for long-term problems with education, employment, social relationships, and independent adult functioning.


[The calcium channel: electrophysiological findings in cardiac cells].
General properties of the calcium channel are analyzed in the myocardium under voltage clamp conditions both in multicellular properties and in single isolated cells. More recently the patch-clamp has allowed us to study single channels. In normal conditions, the selectivity of the calcium channel to Ca2+ ions is very high; however, in the absence of calcium many divalent cations and even Na ions can go through this channel. Kinetic analysis shows: calcium channel inactivation depends on Ca2+ entry rather than on membrane potential, opening of this channel requires at least two transitions of closed states before the open state. Many works refer to pharmacology of the calcium channel in heart tissue. beta-adrenergic stimulation induces a large increase in current amplitude related to the increase in maximal conductance without variation in the unitary conductance. Two interpretations are available: an increase in the opening probability and/or an increase in the number of available channels, both are consecutives to phosphorylations of the channel. Cholinergic stimulation seems to have little effect. Studies of calcium antagonists have revealed that all these substances have, at various levels, use-dependent and voltage-dependent inhibitory effect. Moreover, some dihydropyridine derivatives can even, activate the channel. Antiarrhythmic as well as general anaesthetic agents have an inhibitory action on the calcium channel besides their effects on the sodium channel or the Na-Ca exchange. Very recently, the existence of another calcium conductance was demonstrated. It is characterized by a low threshold, a pure voltage-dependent inactivation, a relatively weak sensitivity to anticalcic agents and neurotransmettors.


The HSP70 co-chaperone DNAJC14 targets misfolded pendrin for unconventional protein secretion.
Mutations in SLC26A4, which encodes pendrin, are responsible for hearing loss with an enlarged vestibular aqueduct and Pendred syndrome. The most prevalent mutation in East Asia is p.H723R (His723Arg), which leads to defects in protein folding and cell-surface expression. Here we show that H723R-pendrin can be rescued to the cell surface by an HSP70 co-chaperone DNAJC14-dependent unconventional trafficking pathway. Blockade of ER-to-Golgi transport or activation of ER stress signals induced Golgi-independent cell-surface expression of H723R-pendrin and restored its cell-surface Cl(-)/HCO3(-) exchange activity. Proteomic and short interfering RNA screenings with subsequent molecular analyses showed that Hsc70 and DNAJC14 are required for the unconventional trafficking of H723R-pendrin. Moreover, DNAJC14 upregulation was able to induce the unconventional cell-surface expression of H723R-pendrin. These results indicate that Hsc70 and DNAJC14 play central roles in ER stress-associated unconventional protein secretion and are potential therapeutic targets for diseases such as Pendred syndrome, which arise from transport defects of misfolded proteins.


A critical review on the pathways of carbamazepine transformation products in oxidative wastewater treatment processes.
Carbamazepine (CBZ) is an anticonvulsant drug, released in domestic and hospital wastewater, and one of the drugs most commonly detected in surface water. Conventional secondary processes do a very poor job of removing it (<25 %), but its concentrations are significantly reduced by polishing oxidation processes. However, there are still many unknowns regarding the transformation products generated and their fate. This review first presents the journey of CBZ and its transformation products (TPs) in wastewater, from human consumption to discharge in water bodies. It then goes on to detail the diversity of mechanisms responsible for CBZ degradation and the generation of multiple TPs, laying the emphasis on the different types of advanced oxidation processes (AOP). 135 TPs were reported and a map describing their formation/degradation pathways was drawn up. This work highlights the wide range of physicochemical properties and toxicity effects of TPs on aquatic organisms and provides information about TPs of interest for future research. Finally, this review concludes on the importance of quantifying TPs and of determining kinetic characteristics to produce more accurate reaction schemes and computer-based fate predictions.


Model Based or Model Free? Comparing Adaptive Methods for Estimating Thresholds in Neuroscience.
The quantification of human perception through the study of psychometric functions Ψ is one of the pillars of experimental psychophysics. In particular, the evaluation of the threshold is at the heart of many neuroscience and cognitive psychology studies, and a wide range of adaptive procedures has been developed to improve its estimation. However, these procedures are often implicitly based on different mathematical assumptions on the psychometric function, and unfortunately, these assumptions cannot always be validated prior to data collection. This raises questions about the accuracy of the estimator produced using the different procedures. In the study we examine in this letter, we compare five adaptive procedures commonly used in psychophysics to estimate the threshold: Dichotomous Optimistic Search (DOS), Staircase, PsiMethod, Gaussian Processes, and QuestPlus. These procedures range from model-based methods, such as the PsiMethod, which relies on strong assumptions regarding the shape of Ψ, to model-free methods, such as DOS, for which assumptions are minimal. The comparisons are performed using simulations of multiple experiments, with psychometric functions of various complexity. The results show that while model-based methods perform well when Ψ is an ideal psychometric function, model-free methods rapidly outshine them when Ψ deviates from this model, as, for instance, when Ψ is a beta cumulative distribution function. Our results highlight the importance of carefully choosing the most appropriate method depending on the context.


Validation of the GerdQ questionnaire for the management of gastro-oesophageal reflux disease in Japan.
The GerdQ scoring system may be a useful tool for managing gastro-oesophageal reflux disease. However, GerdQ has not been fully validated in Asian countries. To validate the Japanese version of GerdQ and to compare this version to the Carlsson-Dent questionnaire (CDQ) in both general and hospital-based populations. The questionnaires, including the Japanese versions of GerdQ and CDQ, and questions designed to collect demographic information, were sent to a general population via the web, and to a hospital-based population via conventional mail. The optimal cutoff GerdQ score and the differences in the characteristics between GerdQ and CDQ were assessed. The answers from 863 web-responders and 303 conventional-mail responders were analysed. When a GerdQ cutoff score was set at 8, GerdQ significantly predicted the presence of reflux oesophagitis. Although the GerdQ scores were correlated with the CDQ scores, the concordance rates were poor. Multivariate analysis results indicated that, the additional use of over-the-counter medications was associated with GerdQ score ≥ 8, but not with CDQ score ≥ 6. The GerdQ cutoff score of 8 was appropriate for the Japanese population. Compared with CDQ, GerdQ was more useful for evaluating treatment efficacy and detecting patients' unmet medical needs.


Recent Advances on Multi-Parameter Flow Cytometry to Characterize Antimicrobial Treatments.
The investigation on antimicrobial mechanisms is a challenging and crucial issue in the fields of food or clinical microbiology, as it constitutes a prerequisite to the development of new antimicrobial processes or compounds, as well as to anticipate phenomenon of microbial resistance. Nowadays it is accepted that a cells population exposed to a stress can cause the appearance of different cell populations and in particular sub-lethally compromised cells which could be defined as viable but non-culturable (VBNC). Recent advances on flow cytometry (FCM) and especially on multi-parameter flow cytometry (MP-FCM) provide the opportunity to obtain high-speed information at real time on damage at single-cell level. This review gathers MP-FCM methodologies based on individual and simultaneous staining of microbial cells employed to investigate their physiological state following different physical and chemical antimicrobial treatments. Special attention will be paid to recent studies exploiting the possibility to corroborate MP-FCM results with additional techniques (plate counting, microscopy, spectroscopy, molecular biology techniques, membrane modeling) in order to elucidate the antimicrobial mechanism of action of a given antimicrobial treatment or compound. The combination of MP-FCM methodologies with these additional methods is namely a promising and increasingly used approach to give further insight in differences in microbial sub-population evolutions in response to antimicrobial treatments.


Human error and patient-controlled analgesia pumps.
Contrary to the prevailing attitude that error is a source of blame and punishment, errors can be an opportunity to discover a problem and institute activities to correct the problem to reduce the likelihood of recurrence. Often the source of error may be the system in which it occurred, not the person associated with it. Error in any domain, including healthcare, is difficult to identify and address because persons are reluctant to report errors for fear of self-incrimination. The discipline of human factors addresses issues related to human performance including use error. Human factors analysis provides insight into the etiology of use errors and how they can be reduced. Patient-controlled analgesia (PCA) pumps were developed to allow the patient or care-giver more control over pain relief. The PCA pumps can be programmed to deliver pain medication on a continuous basis, intermittently, or as a bolus. Selected adverse incidents involving PCA pumps that were due to use error and reported to the U.S. Food and Drug Administration are described. Finally, implications of those findings and the potential for reducing use error by applying considerations of the discipline of human factors are discussed.


A randomized double-blinded study assessing the dose-response of ropivacaine with dexmedetomidine for maintenance of labor with epidural analgesia in nulliparous parturients.
Background: The combination of ropivacaine and dexmedetomidine has been used as an epidural analgesic for inducing labor. However, there is limited data regarding the administration of epidural analgesia for labor maintenance, hence, this study aimed to determine the optimum concentration through dose-response curves of ropivacaine plus dexmedetomidine, which could be used along with the Programmed Intermittent Epidural Bolus (PIEB) technique. Methods: One hundred parturients were randomized into 4 groups who were administered four different doses of ropivacaine (dexmedetomidine at 0.4 μg mL-1): 0.04%, 0.06%, 0.08%, and 0.1%. The primary outcome that was determined included the proportion of patients experiencing breakthrough pain during their 1st stage of labor. Breakthrough pain was described as a visual analog scale [VAS] score of >30 mm, requiring supplemental epidural analgesia after the administration of at least one patient-controlled bolus. The effective concentration of analgesia that was used for labor maintenance in 50% (EC50) and 90% (EC90) of patients were calculated with the help of probit regression. Secondary outcomes included epidural block characteristics, side effects, neonatal outcomes, and patient satisfaction. Results: The results indicated that the proportion of patients without breakthrough pain was 45% (10/22), 55% (12/22), 67% (16/24), and 87% (20/23) for 0.04%, 0.06%, 0.08%, and 0.10% doses of the analgesic that were administered, respectively. The EC50 value was 0.051% (95% confidence interval [CI], 0.011%-0.065%) while the EC90 value was recorded to be 0.117% (95% CI, 0.094%-0.212%). Side effects were similar among groups. Conclusion: A ropivacaine dose of 0.117% can be used as epidural analgesia for maintaining the 1st stage of labor when it was combined with dexmedetomidine (0.4 μg mL-1) and the PIEB technique. Clinical Trial Register: https://www.chictr.org.cn/index.aspx, identifier ChiCTR2200059557.


Effect of Three Colostrum Diets on Passive Transfer of Immunity and Preweaning Health in Calves on a California Dairy following Colostrum Management Training.
Following colostrum management training, a randomized field trial was conducted on a California dairy to determine the effect of supplementing pooled colostrum with either colostrum-derived replacer (CDR) or second-milking colostrum (transition milk) on failure of passive transfer (FPT) and preweaning morbidity risks. A total of 166 calves were randomly assigned to 4L first-milking pooled colostrum (treatment 1), 2L first-milking pooled colostrum and 2L of CDR (treatment 2), or 2L first-milking pooled colostrum and 2L second-milking pooled colostrum (treatment 3). Mean 24-hour serum TP and IgG for treatments 2 (TP 5.2 g/dL, IgG 15.9 g/L) and 3 (TP 5.4 g/dL, IgG 18.3 g/L) did not statistically differ but were significantly lower than for treatment 1 (TP 5.9 g/dL, IgG 24.6 g/L). Risk of FPT did not differ for treatments 1, 2, and 3 (0.0%, 9.3%, and 1.9%, resp.). Similarly, the preweaning risk of diarrhea (81.0%, 92.5%, and 87.0%, resp.) or pneumonia (6.9%, 13.2%, and 18.5%, resp.) did not differ between treatments. Feeding 4L first-milking pooled colostrum resulted in adequate passive transfer. When first-milking pooled colostrum quantity is inadequate, CDR or second-milking pooled colostrum can be used to supplement the required colostrum volume and IgG mass without adversely affecting the risks of FPT or preweaning diarrhea and pneumonia.


Climate and demography of the planktivorous Cassin's auklet Ptychoramphus aleuticus off northern California: implications for population change.
1. We performed demographic analyses on Cassin's auklet Ptychoramphus aleuticus, a zooplanktivorous seabird inhabiting the variable California Current System, to understand how temporal environmental variability influences population dynamics. 2. We used capture-recapture data from 1986 to 2002 to rank models of interannual variation in survival, breeding propensity, breeding success, and recruitment. 3. All demographic parameters exhibited temporal variability. Interannual variation in survival was best modelled as a nonlinear function of the winter Southern Oscillation Index (SOI). Breeding propensity was best modelled as a threshold function of local sea surface temperature. Breeding success and recruitment were best modelled with year-dependent annual variation. 4. Changes in the SOI force El Niño/La Niña events, which in turn alter prey availability to seabirds in this system. Demographic responses varied during El Niños/La Niñas. Survival diminished substantially during the 1997-98 El Niño event, while breeding propensity was affected during both the 1992 and 1998 El Niños. Breeding success was reduced during the 1992, 1993, and 1998 El Niños, but was unusually high in 2002. Recruitment was higher at the beginning and end of this time-series. 5. While demographic responses varied interannually, parameter values covaried in a positive fashion, a situation conducive to rapid population change. During the 11 years study period, the Farallon auklet breeding population declined at 6.05 +/- 0.80% (SE) per year, a cumulative decline of 49.7%. This study demonstrates how climate variability has influenced key demographic processes for this diminished marine bird population.


A randomized phase II trial of mitoxantrone, estramustine and vinorelbine or bcl-2 modulation with 13-cis retinoic acid, interferon and paclitaxel in patients with metastatic castrate-resistant prostate cancer: ECOG 3899.
To test the hypothesis that modulation of Bcl-2 with 13-cis retinoic acid (CRA)/interferon-alpha2b (IFN) with paclitaxel (TAX), or mitoxantrone, estramustine and vinorelbine (MEV) will have clinical activity in men with metastatic castrate-resistant prostate cancer (CRPC). 70 patients were treated with either MEV (Arm A) in a 3-week cycle or CRA/IFN/TAX with an 8-week cycle (Arm B). Patients were assessed for response, toxicity, quality of life (QOL), and the effect of treatment on Bcl-2 levels in peripheral blood mononuclear cells (PBMC). The PSA response rates were 50% and 23%, measurable disease response rates (CR+PR) 14% and 15%, and median overall survival 19.4 months and 13.9 months on Arm A and Arm B respectively. Transient grade 4 neutropenia occurred in 18 and 2 patients, and grade 3 to 4 thrombosis in 7 patients and 1 patient in Arm A and Arm B respectively. Patients on Arm B reported a clinically significant decline in QOL between baseline and week 9/10 (.71 s.d.), and a significantly lower level of QOL than Arm A (p = 0.01). As hypothesized, Bcl-2 levels decreased with CRA/IFN therapy only in Arm B (p = 0.03). Treatment with MEV was well tolerated and demonstrated clinical activity in patients with CRPC. Given the adverse effect of CRA/IFN/TAX on QOL, the study of other novel agents that target Bcl-2 family proteins is warranted. The feasibility of measuring Bcl-2 protein in a cooperative group setting is hypothesis generating and supports further study as a marker for Bcl-2 targeted therapy. CDR0000067865.


Emerging role of integrase inhibitors in the management of treatment-experienced patients with HIV infection.
Integrase is an essential HIV-1-specific enzyme that is an active target for antiretroviral drug development. Recently, a new class of drugs that specifically inhibits strand transfer, one of the three steps of HIV integration into the host DNA, has been developed. Two drugs in this class have reached late stages of development for use in HIV-1 infected individuals: raltegravir, which has just been approved for use in treatment-experienced patients, and elvitegravir, currently in phase III trials. Both are potent with an IC(50) in the 30 nM range and active in vitro against wild type as well as in strains highly resistant to all other existing classes of drugs. Clinical trials in both treatment-naïve and -experienced patients have demonstrated raltegravir to be highly effective with an excellent tolerability profile and no specific clinical or metabolic side effects. Longer follow up is necessary to ensure this early safety profile is sustained. The rapid rate of viral decay observed with raltegravir challenges the current understanding of HIV-1 turnover and may open new strategies for long term treatment and management of infected patients.


Measurement of Flow-Mediated Vasodilatation.
The FMDJ study, a multicenter prospective observational study conducted in Japan, demonstrated the acceptable reliability of measurement of flow-mediated vasodilatation (FMD) using a semi-automatic device in individual institutions. However, in about 10% of Japanese subjects, adequate scans to determine the brachial arterial diameter failed to be obtained. The prevalence of inadequate scans was higher in women than in men, while obesity had no influence on the inadequate scan rate. The FMDJ study also proposed that attending periodic refresher courses on the measurement of FMD is needed for maintaining competency. Finally, the FMDJ study proposed reference values for FMD. Thus, FMD measurement may be categorized as a clinically applicable tool on the basis of class IIb (exploratory cohort study with good reference standards) evidence.


Bioenergy production from diluted poultry manure and microbial consortium inside Anaerobic Sludge Bed Reactor at sub-mesophilic conditions.
In this study, anaerobic treatability of diluted chicken manure (with an influent feed ratio of 1 kg of fresh chicken manure to 6 L of tap water) was investigated in a lab-scale anaerobic sludge bed (ASB) reactor inoculated with granular seed sludge. The ASB reactor was operated at ambient temperature (17-25°C) in order to avoid the need of external heating up to higher operating temperatures (e.g., up to 35°C for mesophilic digestion). Since heat requirement for raising the temperature of incoming feed for digestion is eliminated, energy recovery from anaerobic treatment of chicken manure could be realized with less operating costs. Average biogas production rates were calculated ca. 210 and 242 L per kg of organic matter removed from the ASB reactor at average hydraulic retention times (HRTs) of 13 and 8.6 days, respectively. Moreover, average chemical oxygen demand (COD) removal of ca. 89% was observed with suspended solids removal more than 97% from the effluent of the ASB reactor. Influent ammonia, on the other hand, did not indicate any free ammonia inhibition due to dilution of the raw manure while pH and alkalinity results showed stability during the study. Microbial quantification results indicated that as the number of bacterial community decreased, the amount of Archaea increased through the effective digestion volume of the ASB reactor. Moreover, the number of methanogens displayed an uptrend like archaeal community and a strong correlation (-0.645) was found between methanogenic community and volatile fatty acid (VFA) concentration especially acetate.


[The design of a family of ultrasonic diagnostic units based on up-to-date computer technologies].
Small ultrasonic devices are now being used in medical practice. However, the vast majority of them represent designingly completed systems which cannot include additional soft- and hardwares to accumulate measurements and to set up databases, to make additional postprocessing, to transmit measurements along networks and telephone lines, etc. On the other hand, in our and foreign countries, work is under way in designing the systems of archiving and treating ultrasonic images, working places for a physician engaged in radiation diagnosis. But at the same time direct joining of ultrasonic equipment with standard computer facilities is impossible in many practical cases. To design ultrasonic devices by using Multimedia IBM PC technology which may integrate various informational media, such as display, sonic information, and textual data is one of the ways of solving this problem. With this, the hardwares of ultrasonic devices are realized in the construction of standard IBM PC modules. Sonomed ultrasonic devices may be considered to be alternatives of Multimedia IBM PC-based devices.


Heterotrimeric G alpha q/G alpha 11 proteins function upstream of vascular endothelial growth factor (VEGF) receptor-2 (KDR) phosphorylation in vascular permeability factor/VEGF signaling.
Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) functions by activating two receptor-tyrosine kinases, Flt-1 (VEGF receptor (VEGFR)-1) and KDR (VEGFR-2), both of which are selectively expressed on primary vascular endothelium. KDR is responsible for VPF/VEGF-stimulated endothelial cell proliferation and migration, whereas Flt-1 down-modulates KDR-mediated endothelial cell proliferation. Our most recent works show that pertussis toxin-sensitive G proteins and Gbetagamma subunits are required for Flt-1-mediated down-regulation of human umbilical vein endothelial cell (HUVEC) proliferation and that Gq/11 proteins are required for KDR-mediated RhoA activation and HUVEC migration. In this study, we demonstrate that Gq/11 proteins are also required for VPF/VEGF-stimulated HUVEC proliferation. Our results further indicate that Gq/11 proteins specifically mediate KDR signaling such as intracellular Ca2+ mobilization rather than Flt-1-induced CDC42 activation and that a Gq/11 antisense oligonucleotide completely inhibits MAPK phosphorylation induced by KDR but has no effect on Flt-1-induced MAPK activation. More importantly, we demonstrate that Gq/11 proteins interact with KDR in vivo, and the interaction of Gq/11 proteins with KDR does not require KDR tyrosine phosphorylation. Surprisingly, the Gq/11 antisense oligonucleotide completely inhibits VPF/VEGF-stimulated KDR phosphorylation. Expression of a constitutively active mutant of G11 but not Gq can cause phosphorylation of KDR and MAPK. In addition, a Gbetagamma minigene, hbetaARK1(495), inhibits VPF/VEGF-stimulated HUVEC proliferation, MAPK phosphorylation, and intracellular Ca2+ mobilization but has no effect on KDR phosphorylation. Taken together, this study demonstrates that Gq/11 proteins mediate KDR tyrosine phosphorylation and KDR-mediated HUVEC proliferation through interaction with KDR.


Dynamics of non-cohabiting sex partnering in sub-Saharan Africa: a modelling study with implications for HIV transmission.
To develop an analytical understanding of non-cohabiting sex partnering in sub-Saharan Africa (SSA) using nationally representative sexual behaviour data. A non-homogenous Poisson stochastic process model was used to describe the dynamics of non-cohabiting sex. The model was applied to 25 countries in SSA and was fitted to Demographic and Health Survey data. The country-specific mean values and variances of the distributions of number of non-cohabiting partners were estimated. The model yielded overall robust fits to the empirical distributions stratified by marital status and sex. The median across all country-specific mean values was highest for unmarried men at 0.574 non-cohabiting partners over the last 12 months, followed by that of unmarried women at 0.337, married men at 0.192 and married women at 0.038. The median of variances was highest for unmarried men at 0.127, followed by married men at 0.057, unmarried women at 0.003 and married women at 0.000. The largest variability in mean values across countries was for unmarried men (0.103-1.206), and the largest variability in variances was among unmarried women (0.000-1.994). Non-cohabiting sex appears to be a random 'opportunistic' phenomenon linked to situations that may facilitate it. The mean values and variances of number of partners in SSA show wide variation by country, marital status and sex. Unmarried individuals have larger mean values than their married counterparts, and men have larger mean values than women. Unmarried individuals appear to play a disproportionate role in driving heterogeneity in sexual networks and possibly epidemiology of sexually transmitted infections.


Engineering ultrasmall metal nanoclusters for photocatalytic and electrocatalytic applications.
In view of many of the fundamental properties of ultrasmall noble metal nanoclusters progressively being uncovered, it has become increasingly clear that this class of materials has enormous potential for photocatalytic and electrocatalytic applications due to their unique electronic and optical properties. In this Minireview, we highlight the key electronic and optical properties of metal nanoclusters which are essential to photocatalysis and electrocatalysis. We further use these properties as the basis for our discussion to map out directions or principles for the rational design of high performance photocatalysts and electrocatalysts, highlighting several successful attempts along this direction.


Four different hernias are encountered in the anterior part of the diaphragm.
A retrospective clinical study was performed to evaluate the hernias encountered in the anterior part of the diaphragm. Twenty patients (14 males, 6 females; aged 7 days-7 years) with hernias located in the anterior part of the diaphragm who were treated surgically formed the study group. The exact locations, contents and additional malformations were evaluated. The locations were parasternal in 14 and retrosternal in six. Parasternal locations were the right side in 11, left side in two and bilateral in one patient. Three patients had trisomy 21 syndrome. A sac was presented in all cases and included the colon in 12 patients. A patient with retrosternal location also had trisomy 21 syndrome. The patients with retrosternal hernias also presented with sacs, and the colon was the most commonly included viscus. In the presented series, no intrapericardial herniations or anteromedial defects were encountered. Comparison of previously reported patients and the present series suggests that the anterior part of the diaphragm hosts various hernias of congenital origin in its different locations. According to the exact location and the presence or absence of sacs, four different types of hernias occur in this area: retrosternal hernias with a sac, intrapericardial herniation, and parasternal and anteromedial hernias with either unilateral or bilateral involvements. Since four different hernias were distinguished, the term Morgagni hernia does not include or define all the hernias of the anterior part of the diaphragm. We believe they should, therefore be designated according to the location and presence or absence of a sac.


Using Continuous Glucose Monitoring Values for Bolus Size Calculation in Smart Multiple Daily Injection Systems: No Negative Impact on Post-bolus Glycemic Outcomes Found in Real-World Data.
Recent evidence shows that it may be safe to estimate bolus sizes based on continuous glucose monitoring (CGM) rather than blood glucose (BG) values using glycemic trend-adjusted bolus calculators. Users may already be doing this in the real world, though it is unclear whether this is safe or effective for calculators not employing trend adjustment. We assessed real-world data from a smart multiple daily injections (MDIs) device users with a CGM system, hypothesizing that four-hour post-bolus outcomes using CGM values are not inferior to those using BG values. Our data set included 184 users and spanned 18 months with 79 000 bolus observations. We tested differences using logistic regression predicting CGM or BG value usage based on outcomes and confirmed initial results using a mixed model regression accounting for within-subject correlations. Comparing four-hour outcomes for bolus events using CGM and BG values revealed no differences using our initial approach (P > .183). This finding was confirmed by our mixed model regression approach in all cases (P > .199), except for times below range outcomes. Higher times below range were predictive of lower odds of CGM-based bolus calculations (OR = 0.987, P < .0001 and OR = 0.987, P = .0276, for time below 70 and 54 mg/dL, respectively). We found no differences in four-hour post-bolus glycemic outcomes when using CGM or BG except for time below range, which showed evidence of being lower for CGM. Though preliminary, our results confirm prior findings showing non-inferiority of using CGM values for bolus calculation compared with BG usage in the real world.


[Effect of macrocyclic esters on mitochondrial and phosphatidylcholine membranes].
It has been found that some macrocyclic esters have no ionophoric properties, but can block valinomycin-induced potassium transport in mitochondrial membranes and lessen the potassium current induced by valinomycin in phosphatidylcholine bilayers. It has been also discovered that 36-member cyclic esters of succinic acid and propylene glycoles decrease fluorescence of puridine nucleotides in mitochondria and produce a modifying effect on bimolecular phosphatidylcholine membranes in the medium of litium, calcium and magnesium chlorides at unilateral injection. The results obtained suggest that cyclopolyesters under investigation form mixed complexes with the macromolecules in the composition of mitochondrial and phosphatidylcholine membranes with the participation of the integrated ion.


Genetic risk for schizophrenia and psychosis in Alzheimer disease.
Psychotic symptoms, defined as the occurrence of delusions or hallucinations, are frequent in Alzheimer disease (AD), affecting ~40 to 60% of individuals with AD (AD with psychosis (AD+P)). In comparison with AD subjects without psychosis, AD+P subjects have more rapid cognitive decline and poor outcomes. Prior studies have estimated the heritability of psychosis in AD at 61%, but the underlying genetic sources of this risk are not known. We evaluated a Discovery Cohort of 2876 AD subjects with (N=1761) or without psychosis (N=1115). All subjects were genotyped using a custom genotyping array designed to evaluate single-nucleotide polymorphisms (SNPs) with evidence of genetic association with AD+P and include SNPs affecting or putatively affecting risk for schizophrenia and AD. Results were replicated in an independent cohort of 2194 AD subjects with (N=734) or without psychosis (N=1460). We found that AD+P is associated with polygenic risk for a set of novel loci and inversely associated with polygenic risk for schizophrenia. Among the biologic pathways identified by the associations of schizophrenia SNPs with AD+P are endosomal trafficking, autophagy and calcium channel signaling. To the best of our knowledge, these findings provide the first clear demonstration that AD+P is associated with common genetic variation. In addition, they provide an unbiased link between polygenic risk for schizophrenia and a lower risk of psychosis in AD. This provides an opportunity to leverage progress made in identifying the biologic effects of schizophrenia alleles to identify novel mechanisms protecting against more rapid cognitive decline and psychosis risk in AD.


Synthesis of 1,4-anthracene-9,10-dione derivatives and their regulation of nitric oxide, IL-1β and TNF-α in activated RAW264.7 cells.
Mitoxantrone is an anthracenedione antineoplastic and immunosuppressive agent approved for multiple sclerosis treatment. Novel mono- and disubstituted anthraquinone derivatives, analogues of mitoxantrone, were synthesized through the addition of lipophilic amino alcohols and evaluated for their effect on IL-1β, TNF-α and nitric oxide production by LPS/IFN-γ-stimulated RAW264.7 cells. The disubstituted 1,4-anthracene-9,10-dione 10 showed significant inhibition of nitric oxide, TNF-α and IL-1β production at the concentration of 5 μg/mL, with a much lower cytotoxicity than mitoxantrone. The monosubstituted 3, 4, 11, 12 and 13 also displayed a moderate to good inhibitory capacity on IL-1β production. However, the methylated compounds 11, 12 and 13 failed to inhibit the TNF-α production, and compound 13 was the only one to decrease the production of nitric oxide. None of these derivatives was toxic at the tested concentrations. Compounds 10 and 13 had better inhibitory capacity of the inflammatory mediators analyzed, with reliable viability of the cells.


Autophagy activation in COL6 myopathic patients by a low-protein-diet pilot trial.
A pilot clinical trial based on nutritional modulation was designed to assess the efficacy of a one-year low-protein diet in activating autophagy in skeletal muscle of patients affected by COL6/collagen VI-related myopathies. Ullrich congenital muscular dystrophy and Bethlem myopathy are rare inherited muscle disorders caused by mutations of COL6 genes and for which no cure is yet available. Studies in col6 null mice revealed that myofiber degeneration involves autophagy defects and that forced activation of autophagy results in the amelioration of muscle pathology. Seven adult patients affected by COL6 myopathies underwent a controlled low-protein diet for 12 mo and we evaluated the presence of autophagosomes and the mRNA and protein levels for BECN1/Beclin 1 and MAP1LC3B/LC3B in muscle biopsies and blood leukocytes. Safety measures were assessed, including muscle strength, motor and respiratory function, and metabolic parameters. After one y of low-protein diet, autophagic markers were increased in skeletal muscle and blood leukocytes of patients. The treatment was safe as shown by preservation of lean:fat percentage of body composition, muscle strength and function. Moreover, the decreased incidence of myofiber apoptosis indicated benefits in muscle homeostasis, and the metabolic changes pointed at improved mitochondrial function. These data provide evidence that a low-protein diet is able to activate autophagy and is safe and tolerable in patients with COL6 myopathies, pointing at autophagy activation as a potential target for therapeutic applications. In addition, our findings indicate that blood leukocytes are a promising noninvasive tool for monitoring autophagy activation in patients.


CpGIMethPred: computational model for predicting methylation status of CpG islands in human genome.
DNA methylation is an inheritable chemical modification of cytosine, and represents one of the most important epigenetic events. Computational prediction of the DNA methylation status can be employed to speed up the genome-wide methylation profiling, and to identify the key features that are correlated with various methylation patterns. Here, we develop CpGIMethPred, the support vector machine-based models to predict the methylation status of the CpG islands in the human genome under normal conditions. The features for prediction include those that have been previously demonstrated effective (CpG island specific attributes, DNA sequence composition patterns, DNA structure patterns, distribution patterns of conserved transcription factor binding sites and conserved elements, and histone methylation status) as well as those that have not been extensively explored but are likely to contribute additional information from a biological point of view (nucleosome positioning propensities, gene functions, and histone acetylation status). Statistical tests are performed to identify the features that are significantly correlated with the methylation status of the CpG islands, and principal component analysis is then performed to decorrelate the selected features. Data from the Human Epigenome Project (HEP) are used to train, validate and test the predictive models. Specifically, the models are trained and validated by using the DNA methylation data obtained in the CD4 lymphocytes, and are then tested for generalizability using the DNA methylation data obtained in the other 11 normal tissues and cell types. Our experiments have shown that (1) an eight-dimensional feature space that is selected via the principal component analysis and that combines all categories of information is effective for predicting the CpG island methylation status, (2) by incorporating the information regarding the nucleosome positioning, gene functions, and histone acetylation, the models can achieve higher specificity and accuracy than the existing models while maintaining a comparable sensitivity measure, (3) the histone modification (methylation and acetylation) information contributes significantly to the prediction, without which the performance of the models deteriorate, and, (4) the predictive models generalize well to different tissues and cell types. The developed program CpGIMethPred is freely available at http://users.ece.gatech.edu/~hzheng7/CGIMetPred.zip.


Pericardial Adipose Tissue Thrombospondin-1 Associates With Antiangiogenesis in Ischemic Heart Disease.
Accumulation of ectopic pericardial adipose tissue has been associated with cardiovascular complications which, in part, may relate to adipose-derived factors that regulate vascular responses and angiogenesis. We sought to characterize adipose tissue microvascular angiogenic capacity in subjects who underwent elective cardiac surgeries including aortic, valvular, and coronary artery bypass grafting. Pericardial adipose tissue was collected intraoperatively and examined for angiogenic capacity. Capillary sprouting was significantly blunted (twofold, p <0.001) in subjects with coronary artery disease (CAD) (age 60 ± 9 years, body mass index [BMI] 32 ± 4 kg/m2, low-density lipoprotein cholesterol [LDL-C] 95 ± 46 mg/100 ml, n = 29) compared with age-, BMI-, and LDL-C matched subjects without angiographic obstructive CAD (age 59 ± 10 y, BMI 35 ± 9 kg/m2, LDL-C 101 ± 40 mg/100 ml, n = 12). For potential mechanistic insight, we performed mRNA expression analyses using quantitative real-time polymerase chain reaction and observed no significant differences in pericardial fat gene expression of proangiogenic mediators vascular endothelial growth factor-A (VEGF-A), fibroblast growth factor-2 (FGF-2), and angiopoietin-1 (angpt1), or anti-angiogenic factors soluble fms-like tyrosine kinase-1 (sFlt-1) and endostatin. In contrast, mRNA expression of anti-angiogenic thrombospondin-1 (TSP-1) was significantly upregulated (twofold, p = 0.008) in CAD compared with non-CAD subjects, which was confirmed by protein western-immunoblot analysis. TSP-1 gene knockdown using short hairpin RNA lentiviral delivery significantly improved angiogenic deficiency in CAD (p <0.05). In conclusion, pericardial fat in subjects with CAD may be associated with an antiangiogenic profile linked to functional defects in vascularization capacity. Local paracrine actions of TSP-1 in adipose depots surrounding the heart may play a role in mechanisms of ischemic heart disease.


Bone ultrasound velocity of appropriately grown for gestational age concordant twins.
In neonates, quantitative ultrasound has been shown to be predictive of bone breakability and an index of biophysical property of bone. There are no data on bone speed of sound (SOS) in appropriate for gestational age (AGA) twins. The purpose of this study was to test the hypothesis that AGA twins who had normal intrauterine growth have bone SOS similar to that of AGA singletons. We measured tibial midshaft bone SOS within the first 96 hours of life in 25 pairs of AGA twins, 27 to 40 weeks gestation, and compared them with our data of 73 singletons. There were no significant differences in bone SOS between twin and singleton infants in any of the parameters studied. We found no significant difference in bone SOS between twins of the same pair. AGA twins have bone SOS very similar to that of AGA singletons. We suggest that intrauterine weight sparing occurs also in terms of biophysical properties of bone.


The societal impact of a biologic treatment of ankylosing spondylitis: a case study based on secukinumab.
Aim: This study estimates the societal benefits of using biologics like secukinumab (SEC) instead of nonsteroidal anti-inflammatory drugs for treating patients with ankylosing spondylitis in Germany. Materials & methods: A Markov and a population model were used to predict the functional impairment of German ankylosing spondylitis patients using SEC or nonsteroidal anti-inflammatory drugs. This was translated into avoided productivity losses, which were valued according to gross value added. Results: The productivity impairment of SEC users was predicted to decrease by 20 percentage points, corresponding to 12.8 and 32.7 million hours in paid and unpaid work and a monetary value of €1.6 billion from 2016 to 2030. Accounting for economic spillover effects increases the societal value to € 3.3 billion. Conclusion: The improvements in functional impairment due to biologics could lead to sizable productivity effects.


Prospective associations among borderline personality disorder symptoms, interpersonal problems, and aggressive behaviors.
This study examined the prospective relationships among borderline personality disorder (BPD) symptoms, interpersonal problems, and types of aggressive behaviors (i.e., experiencing psychological and physical victimization and perpetrating psychological and physical aggression) in a psychiatric sample (N = 139) over the course of 2 years. We controlled for other PD symptoms and demographic variables. BPD symptoms at baseline were associated with interpersonal sensitivity, interpersonal ambivalence, interpersonal aggression, need for social approval, and lack of sociability 6 months later. In turn, interpersonal sensitivity predicted not experiencing physical aggression, interpersonal aggression predicted experiencing physical aggression and perpetrating both psychological and physical aggression, need for social approval predicted experiencing both psychological and physical aggression, and lack of sociability predicted perpetrating physical aggression 2 years later. Results demonstrated that interpersonal problems mediated the relationship between BPD and later violent behaviors. Our findings suggest the importance of distinguishing between these groups of aggressive behaviors in terms of etiological pathways, maintenance processes, and treatment interventions.


Localization of Chlamydia trachomatis hypothetical protein CT311 in host cell cytoplasm.
The chlamydia-specific hypothetical protein CT311 was detected both inside and outside of the chlamydial inclusions in Chlamydia trachomatis-infected cells. The extra-inclusion CT311 molecules were distributed in the host cell cytoplasm with a pattern similar to that of CPAF, a known Chlamydia-secreted protease. The detection of CT311 was specific since the anti-CT311 antibody labeling was only removed by absorption with CT311 but not CPAF fusion proteins. In addition, both anti-CT311 and anti-CPAF antibodies only detected their corresponding endogenous proteins without cross-reacting with each other or any other antigens in the whole cell lysates of C. trachomatis-infected cells. Although both CT311 and CPAF proteins were first detected 12 h after infection, localization of CT311 into host cell cytosol was delayed until 24 h while CPAF secretion into host cell cytosol was already obvious by 18 h after infection. The host cell cytosolic localization of CT311 was further confirmed in human primary cells. CT311 was predicted to contain an N-terminal secretion signal sequence and the CT311 signal sequence directed secretion of PhoA into bacterial periplasmic region in a heterologous assay system, suggesting that a sec-dependent pathway may play a role in the secretion of CT311 into host cell cytosol. This hypothesis is further supported by the observation that secretion of CT311 in Chlamydia-infected cells was blocked by a C16 compound known to inhibit signal peptidase I. These findings have provided important molecular information for further understanding the C. trachomatis pathogenic mechanisms.


Scope and design of the Following Rehabilitation, Economics and Everyday-Dialysis Outcome Measurements (FREEDOM) Study.
Conventional thrice-weekly hemodialysis (HD) has limited the ability to generate further improvements in patient quality of life, morbidity, and mortality. Daily HD (DHD) offers the promise of providing clinical and economic benefits. The objectives of the Following Rehabilitation, Economics and Everyday-Dialysis Outcome Measurements Study are to evaluate outcomes of DHD (6 times/wk) with the NxStage System One (NxStage Medical Inc, Lawrence, MA) device. Cohort study with matched control group. The DHD group will include up to 500 participants at 70 clinical sites, enrolling for 3 years with a minimum of 1-year follow-up. Study candidates include adult patients (age >or= 18 years) with end-stage renal disease who are considered suitable candidates for DHD with the NxStage System One device by the treating physician and who have Medicare as their primary insurance payer. The control group will consist of a matched thrice-weekly in-center HD cohort derived from the US Renal Data System database using a 10:1 ratio, totaling 5,000 patients. Treatment with DHD and "standard of care" thrice-weekly HD. The primary intent-to-treat analysis compares hospitalization days/patient-year between the DHD and thrice-weekly HD groups. Other outcomes recorded in both groups include non-treatment-related medical expenditures. In addition, in the DHD cohort, changes in quality-of-life measures (baseline, 4 and 12 months, and every 6 months thereafter); urea kinetics; parameters related to anemia, bone and mineral metabolism, and nutrition; vascular access interventions; and use of medications will be examined. This study has the potential to elucidate the health and economic benefits of DHD and complement results of current clinical trials.


Sentinel lymph node biopsy in melanoma: our 8-year clinical experience in a single French institute (2002-2009).
Since the introduction of sentinel lymph node biopsy (SLNB), its use as a standard of care for patients with clinically node-negative cutaneous melanoma remains controversial. We wished to evaluate our experience of SLNB for melanoma. A single center observational cohort of 203 melanoma patients with a primary cutaneous melanoma (tumour thickness > 1 mm) and without clinical evidence of metastasis was investigated from 2002 to 2009. Head and neck melanoma were excluded. SLN was identified following preoperative lymphoscintigraphy and intraoperative gamma probe interrogation. The SLN identification rate was 97%. The SLN was tumor positive in 44 patients (22%). Positive SLN was significantly associated with primary tumor thickness and microscopic ulceration. The median follow-up was 39.5 (5-97) months. Disease progression was significantly more frequent in SLN positive patients (32% vs 13%, p = 0.002). Five-year DFS and OS of the entire cohort were 79.6% and 84.6%, respectively, with a statistical significant difference between SLN positive (58.7% and 69.7%) and SLN negative (85% and 90.3%) patients (p = 0.0006 and p = 0.0096 respectively). Postoperative complications after SLNB were observed in 12% of patients. Our data confirm previous studies and support the clinical usefulness of SLNB as a reliable and accurate staging method in patients with cutaneous melanoma. However, the benefit of additional CLND in patients with positive SLN remains to be demonstrated.


Retention of the rural allied health workforce in New South Wales: a comparison of public and private practitioners.
Policy initiatives to improve retention of the rural health workforce have relied primarily on evidence for rural doctors, most of whom practice under a private business model. Much of the literature for rural allied health (AH) workforce focuses on the public sector. The AH professions are diverse, with mixed public, private or combined practice settings. This study explores sector differences in factors affecting retention of rural AH professionals. This study compared respondents from the 2008 Rural Allied Health Workforce (RAHW) survey recruiting all AH professionals in rural New South Wales. Comparisons between public (n = 833) and private (n = 756) groups were undertaken using Chi square analysis to measure association for demographics, job satisfaction and intention to leave. The final section of the RAHW survey comprised 33 questions relating to retention. A factor analysis was conducted for each cohort. Factor reliability was assessed and retained factors were included in a binary logistic regression analysis for each cohort predicting intention to leave. Six factors were identified: professional isolation, participation in community, clinical demand, taking time away from work, resources and 'specialist generalist' work. Factors differed slightly between groups. A seventh factor (management) was present only in the public cohort. Gender was not a significant predictor of intention to leave. Age group was the strongest predictor of intention to leave with younger and older groups being significantly more likely to leave than middle aged.In univariate logistic analysis (after adjusting for age group), the ability to get away from work did not predict intention to leave in either group. In multivariate analysis, high clinical demand predicted intention to leave in both the public (OR = 1.40, 95% CI = 1.08, 1.83) and private (OR = 1.61, 95% CI = 1.15, 2.25) cohorts. Professional isolation (OR = 1.39. 95% CI = 1.11, 1.75) and Participation in community (OR = 1.57, 95% CI = 1.13, 2.19) also contributed to the model in the public cohort. This paper demonstrates differences between those working in public versus private sectors and suggests that effectiveness of policy initiatives may be improved through better targeting.


Hydrogen-bond strengthening upon photoinduced electron transfer in ruthenium-anthraquinone dyads interacting with hexafluoroisopropanol or water.
Quinones play a key role as primary electron acceptors in natural photosynthesis, and their reduction is known to be facilitated by hydrogen-bond donors or protonation. In this study, the influence of hydrogen-bond donating solvents on the thermodynamics and kinetics of intramolecular electron transfer between Ru(bpy)(3)(2+) (bpy = 2,2'-bipyridine) and 9,10-anthraquinone redox partners linked together via one up to three p-xylene units was investigated. Addition of relatively small amounts of hexafluoroisopropanol to dichloromethane solutions of these rigid rodlike donor-bridge-acceptor molecules is found to accelerate intramolecular Ru(bpy)(3)(2+)-to-anthraquinone electron transfer substantially because anthraquinone reduction occurs more easily in the presence of the strong hydrogen-bond donor. Similarly, the rates for intramolecular electron transfer are significantly higher in acetonitrile/water mixtures than in dry acetonitrile. In dichloromethane, an increase in the association constant between hexafluoroisopropanol and anthraquinone by more than 1 order of magnitude following quinone reduction points to a significant strengthening of the hydrogen bonds between the hydroxyl group of hexafluoroisopropanol and the anthraquinone carbonyl functions. The photoinduced intramolecular long-range electron transfer process thus appears to be followed by proton motion; hence the overall photoinduced reaction may be considered a variant of stepwise proton-coupled electron transfer (PCET) in which substantial proton density (rather than a full proton) is transferred after the electron transfer has occurred.


A fluorescent reporter of caspase activity for live imaging.
There is a growing interest in the mechanisms that control the apoptosis cascade during development and adult life. To investigate the regulatory events that trigger apoptosis in whole tissues, we have devised a genetically encoded caspase sensor that can be detected in live and fixed tissue by standard confocal microscopy. The sensor comprises two fluorophores, mRFP, monomeric red fluorescent protein (mRFP) and enhanced green fluorescent protein (eGFP), that are linked by an efficient and specific caspase-sensitive site. Upon caspase activation, the sensor is cleaved and eGFP translocates to the nucleus, leaving mRFP at membranes. This is detected before other markers of apoptosis, including anti-cleaved caspase 3 immunoreactivity. Moreover, the sensor does not perturb normal developmental apoptosis and is specific, as cleavage does not occur in Drosophila embryos that are unable to activate the apoptotic cascade. Importantly, dying cells can be recognized in live embryos, thus opening the way for in vivo imaging. As expected from the high conservation of caspases, it is also cleaved in dying cells of chick embryos. It is therefore likely to be generally useful to track the spatiotemporal pattern of caspase activity in a variety of species.


Assembly of very low density lipoprotein: a two-step process of apolipoprotein B core lipidation.
The liver plays a primary role in lipid metabolism. Important functions include the synthesis and incorporation of hydrophobic lipids, triacylglycerols and cholesteryl esters into the core of water-miscible particles called lipoproteins and the secretion of these particles into the circulation for transport to distant tissues. In this article, we present a brief overview of one aspect of the assembly process of very low density lipoproteins, namely, possible mechanisms for combining core lipids with apolipoprotein B. This is a complex process in which apolipoprotein B interacts with core lipids to form very low density lipoproteins by a two-step process that can be dissociated biochemically.


Evaluation of different fragment sizes and cryoprotectants for cryopreservation of feline testicular tissues.
This study aimed to evaluate tissue damage of feline testicles sectioned in two different sizes (0.3 or 0.5 cm3 ) and submitted to different cryoprotectants (propanediol or glycerol). Testicles obtained from 12 domestic cats were sectioned in 0.3 and 0.5 cm3 sized pieces and immediately evaluated by TBARS and semi-quantitatively by histomorphology. The remaining fragments were placed in cryotubes with 1 ml Egg yolk Tris Equex STM extender containing 3% glycerol or 3% propanediol and cryopreserved by fast-freezing technique. Frozen-thawed fragments were also evaluated by TBARS and histomorphology. Statistical analysis was performed using one-way ANOVA with Student-Newmann-Keuls post hoc test, with p < .05. Fresh and cryopreserved tissues generally exhibited a similar morphology concerning detachment of cells from the basement membrane and observation of nucleoli, with a great proportion scored as 0 (no alteration). When present, alterations were slight and the morphology was considered to be good (most classified in scores 1). Pyknosis was the main anomaly observed as score 2 in 54.6% and 58.4% of 0.3-cm3 fragments cryopreserved in propanediol and glycerol, respectively (16.7% scored 2 in fresh tissue). In TBARS evaluation, 0.5-cm3 fragments cryopreserved in glycerol produced less free radical compared to the 0.3 cm3 cryopreserved in glycerol or propanediol. Our results showed that glycerol was more efficient than propanediol to cryopreserve 0.5-cm3 fragments; this might be attributed to the fact that glycerol molecular weight is larger than propanediol and so its perfusion in the testicular tissue is slower.