Primates in peril: the significance of Brazil, Madagascar, Indonesia and the Democratic Republic of the Congo for global primate conservation.
Primates occur in 90 countries, but four-Brazil, Madagascar, Indonesia, and the Democratic Republic of the Congo (DRC)-harbor 65% of the world's primate species (439) and 60% of these primates are Threatened, Endangered, or Critically Endangered (IUCN Red List of Threatened Species 2017-3). Considering their importance for global primate conservation, we examine the anthropogenic pressures each country is facing that place their primate populations at risk. Habitat loss and fragmentation are main threats to primates in Brazil, Madagascar, and Indonesia. However, in DRC hunting for the commercial bushmeat trade is the primary threat. Encroachment on primate habitats driven by local and global market demands for food and non-food commodities hunting, illegal trade, the proliferation of invasive species, and human and domestic-animal borne infectious diseases cause habitat loss, population declines, and extirpation. Modeling agricultural expansion in the 21st century for the four countries under a worst-case-scenario, showed a primate range contraction of 78% for Brazil, 72% for Indonesia, 62% for Madagascar, and 32% for DRC. These pressures unfold in the context of expanding human populations with low levels of development. Weak governance across these four countries may limit effective primate conservation planning. We examine landscape and local approaches to effective primate conservation policies and assess the distribution of protected areas and primates in each country. Primates in Brazil and Madagascar have 38% of their range inside protected areas, 17% in Indonesia and 14% in DRC, suggesting that the great majority of primate populations remain vulnerable. We list the key challenges faced by the four countries to avert primate extinctions now and in the future. In the short term, effective law enforcement to stop illegal hunting and illegal forest destruction is absolutely key. Long-term success can only be achieved by focusing local and global public awareness, and actively engaging with international organizations, multinational businesses and consumer nations to reduce unsustainable demands on the environment. Finally, the four primate range countries need to ensure that integrated, sustainable land-use planning for economic development includes the maintenance of biodiversity and intact, functional natural ecosystems.


Descartes before the horse: I clone, therefore I am: the hepatitis C virus in current perspective.
In an unprecedented approach to viral discovery, the hepatitis C virus (HCV) was cloned before it was established by conventional methods of viral detection or by genomic characterization. Hepatitis C virus is a small (10-kb), single-stranded RNA virus with a genomic organization that places it in the family Flaviviridae. The virus is global in distribution, with a prevalence between 0.3% and 1.5%. The same agent causes parenterally acquired and sporadic non-A, non-B hepatitis. Nonparenteral modes of spread are poorly defined, but low-level sexual transmission is probable. There is a strong association between the presence of antibody to HCV (anti-HCV) and hepatocellular carcinoma; a causal role for HCV is suspected but has not been proved. Hepatitis C virus accounts for at least 85% of the cases of transfusion-associated hepatitis; an anti-HCV-reactive donor was retrospectively identified in nearly 90% of cases. Among donors confirmed by recombinant immunoblot assay (RIBA) to be anti-HCV positive, 80% to 90% are infectious. Hepatitis C virus RNA can be detected within 1 to 2 weeks of exposure and persists throughout the course of infection. Generally, the presence of anti-HCV cannot be confirmed until 9 to 20 weeks after exposure, creating a window period of seronegativity and potential infectivity. It is anticipated that the anti-HCV assay will reduce the number of cases of transfusion-associated hepatitis by 50% in the United States; a 70% reduction has been documented in Spain.


Pediatric emergency medical care in Yerevan, Armenia: a knowledge and attitudes survey of out-of-hospital emergency physicians.
Out-of-hospital emergency care is at an early stage of development in Armenia, with the current emergency medical services (EMS) system having emergency physicians (EPs) work on ambulances along with nurses. While efforts are underway by the Ministry of Health and other organizations to reform the EMS system, little data exists on the status of pediatric emergency care (PEC) in the country. We designed this study to evaluate the knowledge and attitudes of out-of-hospital emergency physicians in pediatric rapid assessment and resuscitation, and identify areas for PEC improvement. We distributed an anonymous, self-administered Knowledge and Attitudes survey to a convenience sample of out-of-hospital EPs in the capital, Yerevan, from August to September 2012. With a response rate of 80%, the majority (89.7%) of respondents failed a 10-question knowledge test (with a pre-defined passing score of ≥7) with a mean score of 4.17 ± 1.99 SD. Answers regarding the relationship between pediatric cardiac arrest and respiratory issues, compression-to-ventilation ratio in neonates, definition of hypotension, and recognition of shock were most frequently incorrect. None of the participants had attended pediatric-specific continuing medical education (CME) activities within the preceding 5 years. χ2 analysis demonstrated no statistically significant association between physician age, length of EMS experience, type of ambulance (general vs. resuscitation/critical care), or CME attendance and pass/fail status. The majority of participants agreed that PEC education in Armenia needs improvement (98%), that there is a need for pediatric-specific CME (98%), and that national out-of-hospital PEC guidelines would increase PEC safety, efficiency, and effectiveness (96%). Out-of-hospital emergency physicians in Yerevan, Armenia are deficient in pediatric-specific emergency assessment and resuscitation knowledge and training, but express a clear desire for improvement. There is a need to support additional PEC training and CME within the EMS system in Armenia.


Action on the social determinants of health: views from inside the policy process.
It is now well documented that many of the key drivers of health reside in our everyday living conditions. In the last two decades, public health has urged political action on these critical social determinants of health (SDH). As noted by the World Health Organisation, encouraging action in this area is challenging. Recent research has argued that public health researchers need to gain a deeper understanding of the complex and changing rationalities of policymaking. This, it seems, is the crucial next step for social determinants of health research. In this paper, we turn our attention to the practitioners of 'the art of government', in order to gain insight into how to secure upstream change for the SDH. Through interviews with policy actors (including politicians, senior government advisors, senior public servants and experienced policy lobbyists) the research sought to understand the nature of government and policymaking, as it pertains to action on the SDH. Through exploring the policy process, we examine how SDH discourses, evidence and strategies align with existing policy processes in the Australian context. Participants indicated that approaches to securing change that are based on linear conceptualisations of the policy process (as often found in public health) may be seen as 'out of touch' with the messy reality of policymaking. Rather, a more dialogic approach that embraces philosophical and moral reasoning (alongside evidence) may be more effective. Based on our findings, we recommend that SDH advocates develop a deeper awareness of the political and policy structures and the discursive conventions they seek to influence within specific settings.


A combination of the immunohistochemical markers CK7 and SATB2 is highly sensitive and specific for distinguishing primary ovarian mucinous tumors from colorectal and appendiceal metastases.
Primary ovarian mucinous tumors can be difficult to distinguish from metastatic gastrointestinal neoplasms by histology alone. The expected immunoprofile of a suspected metastatic lower gastrointestinal tumor is CK7-/CK20+/CDX2+/PAX8-. This study assesses the addition of a novel marker SATB2, to improve the diagnostic algorithm. A test cohort included 155 ovarian mucinous tumors (105 carcinomas and 50 borderline tumors) and 230 primary lower gastrointestinal neoplasms (123 colorectal adenocarcinomas and 107 appendiceal neoplasms). All cases were assessed for SATB2, PAX8 CK7, CK20, and CDX2 expression on tissue microarrays. Expression was scored in a 3-tier system as absent, focal (1-50% of tumor cells) and diffuse ( >50% of tumor cells) and then categorized into either absent/present or nondiffuse/diffuse. SATB2 and PAX8 expression was further evaluated in ovarian tumors from an international cohort of 2876 patients (expansion cohort, including 159 mucinous carcinomas and 46 borderline mucinous tumors). The highest accuracy of an individual marker in distinguishing lower gastrointestinal from ovarian mucinous tumors was CK7 (91.7%, nondiffuse/diffuse cut-off) followed by SATB2 (88.8%, present/absent cut-off). The most effective combination was CK7 and SATB2 with accuracy of 95.3% using the 3-tier interpretation, absent/focal/diffuse. This combination outperformed the standard clinical set of CK7, CK20 and CDX2 (87.5%). Re-evaluation of outlier cases confirmed ovarian origin for all but one case. The accuracy of SATB2 was confirmed in the expansion cohort (91.5%). SATB2 expression was also detected in 15% of ovarian endometrioid carcinoma but less than 5% of other ovarian histotypes. A simple two marker combination of CK7 and SATB2 can distinguish lower gastrointestinal from ovarian primary mucinous tumors with greater than 95% accuracy. PAX8 and CDX2 have value as second-line markers. The utility of CK20 in this setting is low and this warrants replacement of this marker with SATB2 in clinical practice.


Bacteriotherapy in children with recurrent upper respiratory tract infections.
Children with recurrent upper-airway infections (UI) represent a social issue for their economic burden and negative impact on families. Bacteriotherapy is a new therapeutic strategy that could potentially prevent infections. The current study tested the hypothesis that recurrent UI may be prevented by bacteriotherapy. This open study was conducted in an outpatient clinic, enrolling 80 children (40 males, mean age 5.26±2.52 years) suffering from recurrent UI. Children were treated with a nasal spray containing Streptococcus salivarius 24SMB and Streptococcus oralis 89a, 2 puffs per nostril twice a day for a week; this course was repeated for 3 months. The evaluated parameters were: number of UI and number of school and work absences; these outcomes were compared with those recorded in the past year. The mean number of UI significantly diminished: from 5.98 (2.30) in the past year to 2.75 (2.43) after treatment (p<0.0001). The number of school and work absences significantly diminished (from 4.50±2.81 to 2.80±3.42 and from 2.33±2.36 to 1.48±2.16 respectively; p<0.0001 for both). This preliminary experiment suggests that bacteriotherapy using Streptococcus salivarius 24SMB and Streptococcus oralis89a nasal spray could prevent recurrent UI in children.


Analysis of socioeconomic and environmental impacts of waste stabilization pond and unrestricted wastewater irrigation: interface with maintenance.
The effluent from the waste stabilization ponds (WSPs) of the University of Nigeria, Nsukka Campus, is used for irrigation by poor rural farmers. There has been fear that the poorly maintained WSPs and the reuse practices are contributing to environmental degradation and health hazards. In this study the environmental and socioeconomic impacts of the WSPs and reuse were evaluated based on data collected from questionnaires and the literature. The engineering and agricultural properties of soil in the irrigated and nonirrigated areas were compared. Comparison of the health status of the farmers and nonfarmers, of consumers of crops irrigated with wastewater and nonconsumers was performed using Student's t test and the z-score test. The occurrences of diarrhea, typhoid fever, and malaria among the various groups were used as indices. Analyses show that the health status of the farmers and consumers is poorer than those of nonfarmers and nonconsumers at the 5% level of significance. Vegetable cultivation using WSP effluent is a means of sustenance to the farmers and provides an affordable means of satisfying their nutritional deficiencies. However, the poorly maintained WSPs create odor and mosquito nuisances, trap and destroy livestock, and flood nearby compounds with waste debris. At both 1% and 5% levels of significance, communities around the ponds (< 300 m) suffer malaria more frequently than those who live far away (> or = 300 m). Cost-benefit analysis argues in favor of improvement of WSP management and irrigation reuse of wastewater. Dredging of the ponds, training workers and farmers, and adopting appropriate maintenance and monitoring strategies will greatly enhance the socioeconomic status of the urban poor farmers.


Upregulated expression of integrin α1 in mesangial cells and integrin α3 and vimentin in podocytes of Col4a3-null (Alport) mice.
Alport disease in humans, which usually results in proteinuria and kidney failure, is caused by mutations to the COL4A3, COL4A4, or COL4A5 genes, and absence of collagen α3α4α5(IV) networks found in mature kidney glomerular basement membrane (GBM). The Alport mouse harbors a deletion of the Col4a3 gene, which also results in the lack of GBM collagen α3α4α5(IV). This animal model shares many features with human Alport patients, including the retention of collagen α1α2α1(IV) in GBMs, effacement of podocyte foot processes, gradual loss of glomerular barrier properties, and progression to renal failure. To learn more about the pathogenesis of Alport disease, we undertook a discovery proteomics approach to identify proteins that were differentially expressed in glomeruli purified from Alport and wild-type mouse kidneys. Pairs of cy3- and cy5-labeled extracts from 5-week old Alport and wild-type glomeruli, respectively, underwent 2-dimensional difference gel electrophoresis. Differentially expressed proteins were digested with trypsin and prepared for mass spectrometry, peptide ion mapping/fingerprinting, and protein identification through database searching. The intermediate filament protein, vimentin, was upregulated ∼2.5 fold in Alport glomeruli compared to wild-type. Upregulation was confirmed by quantitative real time RT-PCR of isolated Alport glomeruli (5.4 fold over wild-type), and quantitative confocal immunofluorescence microscopy localized over-expressed vimentin specifically to Alport podocytes. We next hypothesized that increases in vimentin abundance might affect the basement membrane protein receptors, integrins, and screened Alport and wild-type glomeruli for expression of integrins likely to be the main receptors for GBM type IV collagen and laminin. Quantitative immunofluorescence showed an increase in integrin α1 expression in Alport mesangial cells and an increase in integrin α3 in Alport podocytes. We conclude that overexpression of mesangial integrin α1 and podocyte vimentin and integrin α3 may be important features of glomerular Alport disease, possibly affecting cell-signaling, cell shape and cellular adhesion to the GBM.


Dyslexia: the Role of Vision and Visual Attention.
Dyslexia is more than just difficulty with translating letters into sounds. Many dyslexics have problems with clearly seeing letters and their order. These difficulties may be caused by abnormal development of their visual "magnocellular" (M) nerve cells; these mediate the ability to rapidly identify letters and their order because they control visual guidance of attention and of eye fixations. Evidence for M cell impairment has been demonstrated at all levels of the visual system: in the retina, in the lateral geniculate nucleus, in the primary visual cortex and throughout the dorsal visuomotor "where" pathway forward from the visual cortex to the posterior parietal and prefrontal cortices. This abnormality destabilises visual perception; hence, its severity in individuals correlates with their reading deficit. Treatments that facilitate M function, such as viewing text through yellow or blue filters, can greatly increase reading progress in children with visual reading problems. M weakness may be caused by genetic vulnerability, which can disturb orderly migration of cortical neurones during development or possibly reduce uptake of omega-3 fatty acids, which are usually obtained from fish oils in the diet. For example, M cell membranes require replenishment of the omega-3 docosahexaenoic acid to maintain their rapid responses. Hence, supplementing some dyslexics' diets with DHA can greatly improve their M function and their reading.


Twenty-five years of the international Bled course for teachers of family medicine in Europe: Glancing back and looking forward.
The international Bled course for teacher training has played a central role in faculty development in family medicine for the past 25 years. The course was originally designed to promote faculty development for family medicine teachers in the new academic discipline of family medicine in Slovenia in 1990 and to introduce new topics into the family medicine curriculum. In this background paper, we perform a SCOT analysis (strengths, challenges, opportunities, and threats) of the current course, evaluating participant feedback and reviewing past topics and their impact on local and international teaching programmes. We also review the place of the course in the context of other teacher-training programmes in family medicine in Europe. We found that the structure and learning aims of the Bled course have remained stable over 25 years. It provides a safe, well-structured learning environment for the participants even though the course topic is different every year. The course has had a significant impact on curriculum development and teacher training in Slovenia as well as in many other countries in Europe and beyond. Because of the positive impact of the course and the high degree of satisfaction of the participants and course directors, it seems worthwhile to continue this endeavour. New directions for the course will depend on the learning needs of the participants and the evolving medical curricula in the countries they represent.


Women in the European Stroke Organisation: One, two, many… - A Top Down and Bottom Up approach.
An increasing proportion of physicians are women, yet they still face challenges with career advancement. In 2014, the European Stroke Organisation established the goal of increasing the number and participation of women within the society using a Top Down and Bottom Up approach. The 'Women's Initiative for Stroke in Europe' was created the same year by a group of women active within the organisation. We aimed to assess the current status of women in European Stroke Organisation, and to explore the change in sex differences after the introduction of focused approaches to address disparities in 2014. Using organisational records, we collected data on sex differences in core activities from 2008 up to 2017 including membership, participation in conferences, courses and in the official journal of the society, and positions of seniority and leadership. We estimated sex distribution differences in each of the activities from 2014 to date. In 2017, the proportion of female members was 40%, while 24% of fellows, 22% of the executive board and 19% of the editorial board in the official journal of the society were women. From 2014 to 2017, there was a significant increase in the proportion of female members (p = 0.0002) and in women participating in the annual conference as faculty (p = 0.001). There was no significant change in the sex distribution among the faculty members in junior educational activities (≤27%) or fellows. In 2017, the proportion of women holding positions of seniority and leadership is still significantly lower to the proportion of women attending educational activities. Transparent data on sex distribution will assist implementing tailored programmes to achieve progress against sex-based barriers.


Effects of a 6-Week Period of Polarized or Threshold Training on Performance and Fatigue in Elite Swimmers.
To quantify the impact of a polarized distribution of training intensity on performance and fatigue in elite swimmers. Twenty-two elite junior swimmers (12 males, age = 17 [3] y, and 10 females, age = 17 [3] y) participated in a crossover intervention study over 28 wk involving 2- × 6-wk training periods separated by 6 wk. Swimmers were randomly assigned to a training group for the first period: polarized (81% in zone 1, blood lactate concentration, [La]b ≤ 2 mmol·L-1; 4% in zone 2, 2 mmol·L-1 < [La]b ≤ 4 mmol·L-1; and 15% in zone 3, [La]b > 4 mmol·L-1) or threshold (65%/25%/10%). Before and after each period, they performed a 100-m maximal swimming test to determine performance, maximal [La]b, and oxygen consumption and an incremental swimming test to determine speed corresponding to [La]b = 4 mmol·L-1 (V4 mmol·L-1). Self-reported indices of well-being were collected with a daily questionnaire. Polarized training elicited small to moderately greater improvement than threshold training on 100-m performance (within-group change ± 90% confidence interval: 0.97% ± 1.02% vs 0.09% ± 0.94%, respectively) with less fatigue and better quality of recovery. There was no substantial gender effect. No clear differences were observed in physiological adaptations between groups. In elite junior swimmers, a 6-wk period of polarized training induced small improvements in 100-m time-trial performance and, in combination with less perceived fatigue, forms a viable option for coaches preparing such cohorts of swimmers for competition.


Prenatal prediction of osteogenesis imperfecta (OI type IV): exclusion of inheritance using a collagen gene probe.
Autosomal dominant osteogenesis imperfecta is caused by mutations in the COL1A2 and COL1A1 genes of type I collagen. In a family with OI type IV genetically linked to the COL1A2 gene, we attempted prenatal diagnosis in a pregnancy at risk by genotyping the DNA of the fetus for a COL1A2 gene associated RFLP. Our results showed that the fetus inherited the normal COL1A2 allele from her affected parent. Linkage analysis can thus be used in the prenatal diagnosis of dominantly inherited osteogenesis imperfecta.


Flare Rate in Patients with Rheumatoid Arthritis in Low Disease Activity or Remission When Tapering or Stopping Synthetic or Biologic DMARD: A Systematic Review.
To evaluate the risk of having a disease flare in patients with rheumatoid arthritis (RA) with low disease activity (LDA) or in remission when deescalating (tapering or stopping) disease-modifying antirheumatic drug (DMARD) therapy. A search in medical databases including publications from January 1950 to February 2015 was performed. Included were trials and observational studies in adults with RA who were in LDA or remission, evaluating ≥ 20 patients tapering or stopping DMARD. Flare rates had to have been reported. A metaanalysis was performed on studies deescalating tumor necrosis factor (TNF) blockers. Four studies evaluated synthetic DMARD. Flare rates ranged from 8% at 24 weeks to 63% at 4 months after deescalation. Fifteen studies reported on TNF blockers. Estimated flare rates by metaanalysis on studies tapering or stopping TNF blockers were 0.26 (95% CI 0.17-0.39) and 0.49 (95% CI 0.27-0.73) for good-quality and moderate-quality studies, respectively. Flare rates in 3 studies stopping tocilizumab were 41% after 6 months, 55% at 1 year, and 87% at 1 year. Flare rates in 3 studies deescalating abatacept were 34% at 1 year, 41% at 1 year, and 72% at 6 months. Five studies evaluating radiographic progression in patients deescalating treatment all found limited to no progression. Results suggest that more than one-third of patients with RA with LDA or in remission may taper or stop DMARD treatment without experiencing a disease flare within the first year. Dose reduction of TNF blockers results in lower flare rates than stopping and may be noninferior to continuing full dose. Radiological progression after treatment deescalation remains low, but may increase slightly.


Nanocytology of rectal colonocytes to assess risk of colon cancer based on field cancerization.
Developing a minimally invasive and cost-effective prescreening strategy for colon cancer is critical because of the impossibility of conducting colonoscopy on the entire at-risk population. The concept of field carcinogenesis, in which normal-appearing tissue away from a tumor has molecular and, consequently, nano-architectural abnormalities, offers one attractive approach to identify high-risk patients. In this study, we investigated whether the novel imaging technique partial wave spectroscopic (PWS) microscopy could risk-stratify patients harboring precancerous lesions of the colon, using an optically measured biomarker (L(d)) obtained from microscopically normal but nanoscopically altered cells. Rectal epithelial cells were examined from 146 patients, including 72 control patients, 14 patients with diminutive adenomas, 20 patients with nondiminutive/nonadvanced adenomas, 15 patients with advanced adenomas/high-grade dysplasia, 12 patients with genetic mutation leading to Lynch syndrome, and 13 patients with cancer. We found that the L(d) obtained from rectal colonocytes was well correlated with colon tumorigenicity in our patient cohort and in an independent validation set of 39 additional patients. Therefore, our findings suggest that PWS-measured L(d) is an accurate marker of field carcinogenesis. This approach provides a potential prescreening strategy for risk stratification before colonoscopy.


[Pharmacological preconditioning in carotid endarterectomy].
The study was aimed at examining efficacy of preoperative preparation (pharmacological preconditioning) for carotid endarterectomy in patients with chronic cerebrovascular insufficiency. For this purpose, we analysed the outcomes of surgical treatment in a total of 80 patients presenting with haemodynamically significant unilateral and bilateral lesions of carotid arteries. Of these, 40 patients were operated on immediately and a further 40 patients underwent surgery after pharmacological preconditioning with Actovegin taken at a daily dose of 1,200 mg for 1.5 months. It was demonstrated that preoperative preparation prior to surgery increases cerebral perfusion which is determined by means of single-photon emission computed tomography, thus substantially improving the outcomes of surgical treatment. Statistically significant differences in cognitive function of these groups of patients were revealed 7 days and 6 months after the operation. Improvement of cognitive functions was associated with fewer symptom-free postoperative cerebral ischaemic foci in various regions of the brain. A conclusion was made on a positive role of pharmacological preconditioning with Actovegin in surgical management of cerebrovascular insufficiency, first of all in relation to more complete restoration of cognitive functions.


Localization of Kv2.2 protein in Xenopus laevis embryos and tadpoles.
Voltage-gated potassium (Kv) channels sculpt neuronal excitability and play important developmental roles. Kv channels consist of pore-forming alpha- and auxiliary subunits. For many Kv alpha-subunits, existing mRNA probes and antibodies have allowed analysis of expression patterns, typically during adult stages. Here, we focus on the Kv2.2 alpha-subunit, for which the mRNA shows broad expression in the embryo and adult. A lack of suitable antibodies, however, has hindered detailed analysis of Kv2.2 protein localization, especially during development. We developed an antibody that specifically recognizes Kv2.2 protein in Xenopus laevis, a vertebrate well suited for study of early developmental stages. The Kv2.2 antibody recognized heterologously expressed Kv2.2 but not the closely related Kv2.1 protein. Immunodetection of the protein showed its presence at St 32 in ventrolateral regions of the hindbrain and spinal cord. At later stages, several sensory tissues (retina, otic, and olfactory epithelia) also expressed Kv2.2 protein. As development progressed in the central nervous system, Kv2.2 protein distribution expanded in close association with the cytoskeletal marker alpha-tubulin, consistent with growth of neuronal tracts. We analyzed the subcellular distribution of Kv2.2 protein within single cultured neurons. In addition to a surface membrane presence, Kv2.2 protein also resided intracellularly closely associated with alpha-tubulin, as in vivo. Furthermore, in contrast to Kv2.1, Kv2.2 protein localized to long, axonal-like processes, consistent with its in vivo location in tracts. Despite their primary sequence similarity, the contrasting localizations of Kv2.1 and Kv2.2 support different roles for the two during development and neuronal signaling.


Potentials evoked by chirp-modulated tones: a new technique to evaluate oscillatory activity in the auditory pathway.
Steady-state potentials are oscillatory responses generated by a rhythmic stimulation of a sensory pathway. The frequency of the response, which follows the frequency of stimulation, is maximal at a stimulus rate of 40 Hz for auditory stimuli. The exact cause of these maximal responses is not known, although some authors have suggested that they might be related to the 'working frequency' of the auditory cortex. Testing of the responses to different frequencies of stimulation may be lengthy if a single frequency is studied at a time. Our aim was to develop a fast technique to explore the oscillatory response to auditory stimuli, using a tone modulated in amplitude by a sinusoid whose frequency increases linearly in frequency ('chirp') from 1 to 120 Hz. Time-frequency transforms were used for the analysis of the evoked responses in 10 subjects. Also, we analyzed whether the peaks in these responses were due to increases of amplitude or to phase-locking phenomena, using single-sweep time-frequency transforms and inter-trial phase analysis. The pattern observed in the time-frequency transform of the chirp-evoked potential was very similar in all subjects: a diagonal band of energy was observed, corresponding to the frequency of modulation at each time instant. Two components were present in the band, one around 45 Hz (30-60 Hz) and a smaller one between 80 and 120 Hz. Inter-trial phase analysis showed that these components were mainly due to phase locking phenomena. A simultaneous testing of the amplitude-modulation-following oscillatory responses to auditory stimulation is feasible using a tone modulated in amplitude at increasing frequencies. The maximal energies found at stimulation frequencies around 40 Hz are probably due to increased phase-locking of the individual responses.


Differences in glutamate receptors and inflammatory cell numbers are associated with the resolution of pain in human rotator cuff tendinopathy.
The relationship between peripheral tissue characteristics and pain symptoms in soft tissue inflammation is poorly understood. The primary aim of this study was to determine immunohistochemical differences in tissue obtained from patients with persistent pain and patients who had become pain-free after surgical treatment for rotator cuff tendinopathy. The secondary aim was to investigate whether there would be differences in glutaminergic and inflammatory gene expression between disease-derived and healthy control cells in vitro. Supraspinatus tendon biopsies were obtained from nine patients with tendon pain before shoulder surgery and from nine further patients whose pain had resolved completely following shoulder surgery. Histological markers relating to the basic tendon characteristics, inflammation and glutaminergic signalling were quantified by immunohistochemical analysis. Gene expression of glutaminergic and inflammatory markers was determined in tenocyte explants derived from painful rotator cuff tendon tears in a separate cohort of patients and compared to that of explants from healthy control tendons. Dual labelling was performed to identify cell types expressing nociceptive neuromodulators. Tendon samples from patients with persistent pain demonstrated increased levels of metabotropic glutamate receptor 2 (mGluR2), kainate receptor 1 (KA1), protein gene product 9.5 (PGP9.5), CD206 (macrophage marker) and CD45 (pan-leucocyte marker) versus pain-free controls (p <0.05). NMDAR1 co-localised with CD206-positive cells, whereas PGP9.5 and glutamate were predominantly expressed by resident tendon cells. These results were validated by in vitro increases in the expression of mGluR2, N-methyl-D-aspartate receptor (NMDAR1), KA1, CD45, CD206 and tumour necrosis factor alpha (TNF-α) genes (p <0.05) in disease-derived versus control cells. We conclude that differences in glutamate receptors and inflammatory cell numbers are associated with the resolution of shoulder pain in rotator cuff tendinopathy, and that disease-derived cells exhibit a distinctly different neuro-inflammatory gene expression profile to healthy control cells.


Obstructive sleep apnea in 2-6 year old children referred for adenotonsillectomy.
Adenotonsillectomy is one of the most common surgical procedures performed in children. The indications for surgery are either frequent recurrent throat infections or hypertrophy of the tonsils/adenoid vegetation, which can cause obstructive sleep apnea (OSA). There is disagreement regarding the need for sleep studies before adenotonsillectomy to confirm a diagnosis of OSA. Several studies have evaluated questionnaires and physical examination as tools to identify OSA, with conflicting results. The aim of this study was to evaluate the prevalence of OSA among children referred for adenotonsillectomy and whether questionnaires or physical examination can help identify OSA. This is a prospective cohort study of children aged 2-6 years, referred for adenotonsillectomy. Polysomnography and an otorhinological examination were performed. Tonsillar size and the oral cavity were graded using Friedman's classification and Mallampati score, respectively. The Pediatric Sleep Questionnaire (PSQ) and OSA-18 were also completed. 100 children were included. The prevalence of OSA was 87%, with 52% having moderate to severe OSA. The usefulness of the PSQ and OSA-18 for detecting OSA was evaluated using multiple cutoff points, but none yielded acceptable values for both sensitivity and specificity. In logistic regression analyses predicting different levels of OSA severity, age, Friedman tonsillar size and Mallampati score were weakly associated with OSA. The prevalence of OSA is high among children referred for adenotonsillectomy and questionnaires and clinical characteristics are not sensitive enough to detect the presence or severity of OSA.


A dynamic framework for the study of optimal birth intervals reveals the importance of sibling competition and mortality risks.
Human reproductive patterns have been well studied, but the mechanisms by which physiology, ecology and existing kin interact to affect the life history need quantification. Here, we create a model to investigate how age-specific interbirth intervals adapt to environmental and intrinsic mortality, and how birth patterns can be shaped by competition and help between siblings. The model provides a flexible framework for studying the processes underlying human reproductive scheduling. We developed a state-based optimality model to determine age-dependent and family-dependent sets of reproductive strategies, including the state of the mother and her offspring. We parameterized the model with realistic mortality curves derived from five human populations. Overall, optimal birth intervals increase until the age of 30 after which they remain relatively constant until the end of the reproductive lifespan. Offspring helping each other does not have much effect on birth intervals. Increasing infant and senescent mortality in different populations decreases interbirth intervals. We show that sibling competition and infant mortality interact to lengthen interbirth intervals. In lower-mortality populations, intense sibling competition pushes births further apart. Varying the adult risk of mortality alone has no effect on birth intervals between populations; competition between offspring drives the differences in birth intervals only when infant mortality is low. These results are relevant to understanding the demographic transition, because our model predicts that sibling competition becomes an important determinant of optimal interbirth intervals only when mortality is low, as in post-transition societies. We do not predict that these effects alone can select for menopause.


Salivary Telomere Length and Lung Function in Adolescents Born Very Preterm: A Prospective Multicenter Study.
Preterm birth is associated with abnormal respiratory functions throughout life. The mechanisms underlying these long-term consequences are still unclear. Shortening of telomeres was associated with many conditions, such as chronic obstructive pulmonary disease. We aimed to search for an association between telomere length and lung function in adolescents born preterm. Lung function and telomere length were measured in 236 adolescents born preterm and 38 born full-term from the longitudinal EPIPAGE cohort. Associations between telomere length and spirometric indices were tested in univariate and multivariate models accounting for confounding factors in the study population. Airflows were significantly lower in adolescents born preterm than controls; forced expiratory volume in one second was 12% lower in the extremely preterm born group than controls (p<0.001). Lower birth weight, bronchopulmonary dysplasia and postnatal sepsis were significantly associated with lower airflow values. Gender was the only factor that was significantly associated with telomere length. Telomere length correlated with forced expiratory flow 25-75 in the extremely preterm adolescent group in univariate and multivariate analyses (p = 0.01 and p = 0.02, respectively). We evidenced an association between telomere length and abnormal airflow in a population of adolescents born extremely preterm. There was no evident association with perinatal events. This suggests other involved factors, such as a continuing airway oxidative stress leading to persistent inflammation and altered lung function, ultimately increasing susceptibility to chronic obstructive pulmonary disease.


Are autistic traits measured equivalently in individuals with and without an autism spectrum disorder? An invariance analysis of the Autism Spectrum Quotient Short Form.
It is common to administer measures of autistic traits to those without autism spectrum disorders (ASDs) with, for example, the aim of understanding autistic personality characteristics in non-autistic individuals. Little research has examined the extent to which measures of autistic traits actually measure the same traits in the same way across those with and without an ASD. We addressed this question using a multi-group confirmatory factor invariance analysis of the Autism Quotient Short Form (AQ-S: Hoekstra et al. in J Autism Dev Disord 41(5):589-596, 2011) across those with (n = 148) and without (n = 168) ASD. Metric variance (equality of factor loadings), but not scalar invariance (equality of thresholds), held suggesting that the AQ-S measures the same latent traits in both groups, but with a bias in the manner in which trait levels are estimated. We, therefore, argue that the AQ-S can be used to investigate possible causes and consequences of autistic traits in both groups separately, but caution is due when combining or comparing levels of autistic traits across the two groups.


Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease.
The CRISPR-associated protein Cas9 is an RNA-guided endonuclease that cleaves double-stranded DNA bearing sequences complementary to a 20-nucleotide segment in the guide RNA. Cas9 has emerged as a versatile molecular tool for genome editing and gene expression control. RNA-guided DNA recognition and cleavage strictly require the presence of a protospacer adjacent motif (PAM) in the target DNA. Here we report a crystal structure of Streptococcus pyogenes Cas9 in complex with a single-molecule guide RNA and a target DNA containing a canonical 5'-NGG-3' PAM. The structure reveals that the PAM motif resides in a base-paired DNA duplex. The non-complementary strand GG dinucleotide is read out via major-groove interactions with conserved arginine residues from the carboxy-terminal domain of Cas9. Interactions with the minor groove of the PAM duplex and the phosphodiester group at the +1 position in the target DNA strand contribute to local strand separation immediately upstream of the PAM. These observations suggest a mechanism for PAM-dependent target DNA melting and RNA-DNA hybrid formation. Furthermore, this study establishes a framework for the rational engineering of Cas9 enzymes with novel PAM specificities.


What aspects of social network are protective for dementia? Not the quantity but the quality of social interactions is protective up to 15 years later.
To test the association between several social networks variables reflecting both structural characteristics and quality of relationships with the risk of dementia and Alzheimer's disease 5 and up to 15 years later. The study sample is gathered from the Paquid cohort, a French population-based study of 3,777 elderly people evaluated at baseline and regularly revisited during a 15-year interval. The sample consisted of 2,089 subjects who completed the social network questionnaire and were free of dementia at the time of enrollment and also at the next two follow-ups to minimize the problem of reverse causality. The questionnaire collected at baseline included marital status, number of ties, nature of social network, satisfaction, perception of being understood/misunderstood, and reciprocity in relationships. The incident cases of dementia considered were those diagnosed at 5-year and subsequent follow-ups, resulting in 461 dementia and 373 Alzheimer's disease cases. The multivariate Cox model, including the six social network variables and adjusted for numerous potential confounders, showed significant associations with satisfaction and reciprocity in relationships. Participants who felt satisfied with their relations had a 23% reduced dementia risk. Participants who reported that they received more support than they gave over their lifetime had a 55% and 53% reduced risk for dementia and Alzheimer's disease, respectively. The only variables associated with subsequent dementia or Alzheimer's disease were those reflecting the quality of relationships. The delay between social network assessment and dementia diagnosis was from 5 up to 15 years, thus minimizing the problem of reverse causality.


Psychophysical evidence for the number sense.
It is now clear that most animals, including humans, possess an ability to rapidly estimate number. Some have questioned whether this ability arises from dedicated numerosity mechanisms, or is derived indirectly from judgements of density or other attributes. We describe a series of psychophysical experiments, largely using adaptation techniques, which demonstrate clearly the existence of a number sense in humans. The number sense is truly general, extending over space, time and sensory modality, and is closely linked with action. We further show that when multiple cues are present, numerosity emerges as the natural dimension for discrimination. However, when element density increases past a certain level, the elements become too crowded to parse, and the scene is perceived as a texture rather than array of elements. The two different regimes are psychophysically discriminable in that they follow distinct psychophysical laws, and show different dependencies on eccentricity, luminance levels and effects of perceptual grouping. The distinction is important, as the ability to discriminate numerosity, but not texture, correlates with formal maths skills.This article is part of the discussion meeting issue 'The origins of numerical abilities'.


Lithium-Oxygen Batteries and Related Systems: Potential, Status, and Future.
The goal of limiting global warming to 1.5 °C requires a drastic reduction in CO2 emissions across many sectors of the world economy. Batteries are vital to this endeavor, whether used in electric vehicles, to store renewable electricity, or in aviation. Present lithium-ion technologies are preparing the public for this inevitable change, but their maximum theoretical specific capacity presents a limitation. Their high cost is another concern for commercial viability. Metal-air batteries have the highest theoretical energy density of all possible secondary battery technologies and could yield step changes in energy storage, if their practical difficulties could be overcome. The scope of this review is to provide an objective, comprehensive, and authoritative assessment of the intensive work invested in nonaqueous rechargeable metal-air batteries over the past few years, which identified the key problems and guides directions to solve them. We focus primarily on the challenges and outlook for Li-O2 cells but include Na-O2, K-O2, and Mg-O2 cells for comparison. Our review highlights the interdisciplinary nature of this field that involves a combination of materials chemistry, electrochemistry, computation, microscopy, spectroscopy, and surface science. The mechanisms of O2 reduction and evolution are considered in the light of recent findings, along with developments in positive and negative electrodes, electrolytes, electrocatalysis on surfaces and in solution, and the degradative effect of singlet oxygen, which is typically formed in Li-O2 cells.


Expansion and Compression of Time Correlate with Information Processing in an Enumeration Task.
Perception of temporal duration is subjective and is influenced by factors such as attention and context. For example, unexpected or emotional events are often experienced as if time subjectively expands, suggesting that the amount of information processed in a unit of time can be increased. Time dilation effects have been measured with an oddball paradigm in which an infrequent stimulus is perceived to last longer than standard stimuli in the rest of the sequence. Likewise, time compression for the oddball occurs when the duration of the standard items is relatively brief. Here, we investigated whether the amount of information processing changes when time is perceived as distorted. On each trial, an oddball stimulus of varying numerosity (1-14 items) and duration was presented along with standard items that were either short (70 ms) or long (1050 ms). Observers were instructed to count the number of dots within the oddball stimulus and to judge its relative duration with respect to the standards on that trial. Consistent with previous results, oddballs were reliably perceived as temporally distorted: expanded for longer standard stimuli blocks and compressed for shorter standards. The occurrence of these distortions of time perception correlated with perceptual processing; i.e. enumeration accuracy increased when time was perceived as expanded and decreased with temporal compression. These results suggest that subjective time distortions are not epiphenomenal, but reflect real changes in sensory processing. Such short-term plasticity in information processing rate could be evolutionarily advantageous in optimizing perception and action during critical moments.


Treatment strategy for type 2 diabetes from the perspective of systemic vascular protection and insulin resistance.
This paper provides an update on the mechanisms of vascular impairment associated with insulin resistance and the pathogenesis of diabetic nephropathy and peripheral artery disease (PAD). It also considers the optimal treatment strategies for systemic vascular protection in light of recent findings. This area is of major clinical importance given the ongoing global epidemic of type 2 diabetes and the pivotal role played by insulin resistance in the mechanism of vascular impairment that manifests as macroangiopathy and microangiopathy. Timely diagnosis and intervention is critical in patients with systemic arteriosclerotic disease. Therefore, treatment strategies are aimed not only at targeting the presenting pathology, but also at reducing the risk of cardiovascular events. These efforts can help reduce the risk of both cardiovascular events and mortality. Treatment for PAD includes pharmacotherapy, endovascular treatment, and vascular reconstruction, along with exercise therapy. Because PAD can cause ischemia in the lower extremities, typical drug approaches include use of vasodilators and antiplatelet agents. Beraprost sodium and cilostazol are common choices in Japan, and their risks and benefits are discussed. Of note, beraprost has several therapeutic properties, including vascular endothelial protection, and antiplatelet and anti-inflammatory effects, in addition to vasodilatory activity. In patients with PAD, these activities improve the pathological process in the lower extremities and reduce the incidence of systemic vascular events. Recent preclinical findings indicate that beraprost improves not only ischemic extremities through its vasodilatory properties, but also reduces the insulin resistance which affects vascular endothelium. In this way, beraprost may contribute to an overall systemic vascular protective action. The use of agents, such as beraprost, which are capable of improving insulin resistance and resulting vascular endothelial function at an earlier disease stage, may ultimately contribute to increasing the life expectancy of patients with PAD.


Effect of soil temperature on optical frequency transfer through unidirectional dense-wavelength-division-multiplexing fiber-optic links.
Results of optical frequency transfer over a carrier-grade dense-wavelength-division-multiplexing (DWDM) optical fiber network are presented. The relation between soil temperature changes on a buried optical fiber and frequency changes of an optical carrier through the fiber is modeled. Soil temperatures, measured at various depths by the Royal Netherlands Meteorology Institute (KNMI) are compared with observed frequency variations through this model. A comparison of a nine-day record of optical frequency measurements through the 2×298 km fiber link with soil temperature data shows qualitative agreement. A soil temperature model is used to predict the link stability over longer periods (days-months-years). We show that optical frequency dissemination is sufficiently stable to distribute and compare, e.g., rubidium frequency standards over standard DWDM optical fiber networks using unidirectional fibers.


SUBTHRESHOLD LASER TREATMENT FOR SEROUS RETINAL DETACHMENT IN DOME-SHAPED MACULA ASSOCIATED WITH PATHOLOGIC MYOPIA.
To evaluate the effects of the subthreshold laser treatment as a therapeutic option in the treatment of serous retinal detachment secondary to dome-shaped macula. Prospective pilot study with 12-month follow-up. Each patient underwent a complete ophthalmologic examination, including best-corrected visual acuity, fluorescein angiography, indocyanine green angiography, and spectral domain optical coherence tomography. Eyes with persistent serous retinal detachment lasting for a minimum of 12 months and absence of best-corrected visual acuity improvement underwent subthreshold laser treatment in a single session. The laser spots were delivered in a contiguous mode, covering the area of hyperfluorescence recognized on indocyanine green angiography. The primary outcome measure was the best-corrected visual acuity changes at the 12-month examination. Secondary outcomes included changes in central foveal thickness (CFT) and number of eyes achieving a complete serous retinal detachment resolution. Twelve eyes (8 patients) were included in the study. Best-corrected visual acuity changed from a baseline value of 0.8 ± 0.2 (±SD, logarithm of the minimum angle of resolution [Snellen equivalent: 20/125]) to 0.48 ± 0.1 (Snellen equivalent: 20/60, P = 0.001). Central foveal thickness decreased from 320 ± 52 μm to a final value of 266 ± 41 μm (P = 0.001). Serous retinal detachment reduced in all cases; however, just one eye (8.3%) showed a complete resolution at the 12-month examination. Fluorescein angiography and indocyanine green angiography showed that the hyperfluorescence had disappeared after subthreshold laser treatment. No adverse event was registered, including enlargement of atrophic alterations and choroidal neovascularization. Subthreshold laser treatment can lead to improvements in visual acuity and central foveal thickness in myopic eyes with serous retinal detachment secondary to dome-shaped macula. Further studies are required to confirm our preliminary results.


Management of Calvarial Tumors: A Retrospective Analysis and Literature Review.
Tumors of various organs that metastasize to bone do not neglect calvarium as a target. The aim of this study was to characterize the calvarial tumors. We retrospectively reviewed 45 consecutive patients operated for calvarial masses from January 2002 till May 2012 at our hospital. Skull base tumors and patients ≤18 years were excluded. Three groups of lesions were found - calvarial metastases (15/45), primary tumors (5/45) and tumor-like lesions (25/45). Malignant lesions were equitable by gender distribution, higher age of onset (median age of primary =55; secondary = 60 years) and benign lesions by younger age (median = 35) and female bias (18/25). Calvarial metastases mostly presented with local swelling (10/15), local pain (6/15) and rarely neurologic deficit. There was associated dural sinus thrombosis (4/20 of malignant; 1/25 of benign lesions) and osteolysis (3/5 primary malignant, 13/15 secondary and 18/25 of benign lesions). Complete surgical excision was possible with minimal morbidity in all except one patient and nil mortality. Nearly half (20/45) of the calvarial lesions tend to be malignant with most of them presenting as silent painless masses. Surgical excision should be considered only after suitable investigation and appropriate neurosurgical set-up.


Morphometric Characteristics of the Skull in Horses and Donkeys-A Pilot Study.
Horses and donkeys belong to the genus Equus, but important differences exist between the species, many of which affect their management and welfare. This study compared skull morphology between horses and donkeys. Horse (n = 14) and donkey (n = 16) heads were obtained post-mortem, sectioned sagittally close to the midline, and photographed for subsequent measurement of various skull structures. Skull, cranial, nasal, and profile indices were calculated for topographical comparisons between the species. The olfactory bulb area (OBA), OB pitch (the angle between the hard palate and the OB axis), and whorl location (WL) were also measured. A General Linear Model determined the main effect of species with Sidak's multiple comparisons of species' differences among the various measurements. There was no species difference in cranial or nasal indices (p > 0.13), but donkeys had a larger cranial profile than horses (p < 0.04). Donkeys had a smaller OBA (p < 0.05) and a steeper OB pitch (p < 0.02) than horses. The WL corresponded to the level of the OB in horses but was extremely rostral in donkeys (p < 0.0001). These results show clear differentiation in skull morphology between horses and donkeys. This may be useful in validating other physiological and behavioural differences between horses and donkeys.


Vocal development in a Waddington landscape.
Vocal development is the adaptive coordination of the vocal apparatus, muscles, the nervous system, and social interaction. Here, we use a quantitative framework based on optimal control theory and Waddington's landscape metaphor to provide an integrated view of this process. With a biomechanical model of the marmoset monkey vocal apparatus and behavioral developmental data, we show that only the combination of the developing vocal tract, vocal apparatus muscles and nervous system can fully account for the patterns of vocal development. Together, these elements influence the shape of the monkeys' vocal developmental landscape, tilting, rotating or shifting it in different ways. We can thus use this framework to make quantitative predictions regarding how interfering factors or experimental perturbations can change the landscape within a species, or to explain comparative differences in vocal development across species.


Initial results of hypoxia imaging using 1-alpha-D: -(5-deoxy-5-[18F]-fluoroarabinofuranosyl)-2-nitroimidazole ( 18F-FAZA).
Tumour hypoxia is thought to play a significant role in the outcome of solid tumour therapy. Positron emission tomography (PET) is the best-validated noninvasive technique able to demonstrate the presence of hypoxia in vivo. The locally developed PET tracer for imaging hypoxia, 1-alpha-D: -(5-deoxy-5-[(18)F]-fluoroarabinofuranosyl)-2-nitroimidazole ((18)F-FAZA), has been shown to accumulate in experimental models of tumour hypoxia and to clear rapidly from the circulation and nonhypoxic tissues. The safety and general biodistribution patterns of this radiopharmaceutical in patients with squamous cell carcinoma of the head and neck (HNSCC), small-cell lung cancer (SCLC) or non-small-cell lung cancer (NSCLC), malignant lymphoma, and high-grade gliomas, were demonstrated in this study. Patients with known primary or suspected metastatic HNSCC, SCLC or NSCLC, malignant lymphoma or high-grade gliomas were dosed with 5.2 MBq/kg of (18)F-FAZA, then scanned 2-3 h after injection using a PET or PET/CT scanner. Images were interpreted by three experienced nuclear medicine physicians. The location and relative uptake scores (graded 0 to 4) of normal and abnormal (18)F-FAZA biodistribution patterns, the calculated tumour-to-background (T/B) ratio, and the maximum standardized uptake value were recorded. Included in the study were 50 patients (32 men, 18 women). All seven patients with high-grade gliomas showed very high uptake of (18)F-FAZA in the primary tumour. In six out of nine patients with HNSCC, clear uptake of (18)F-FAZA was observed in the primary tumour and/or the lymph nodes in the neck. Of the 21 lymphoma patients (15 with non-Hodgkin's lymphoma and 6 with Hodgkin's disease), 3 demonstrated moderate lymphoma-related uptake. Of the 13 lung cancer patients (12 NSCLC, 1 SCLC), 7 had increased (18)F-FAZA uptake in the primary lung tumour. No side effects of the administration of (18)F-FAZA were observed. This study suggests that (18)F-FAZA may be a very useful radiopharmaceutical to image hypoxia in the tumour types selected. Especially the high uptake by gliomas was encouraging. Given the good imaging properties, including acceptable T/B ratios in the tumour categories studied, (18)F-FAZA could be considered as a very promising agent for assessing the hypoxic fraction of these tumour types.


Therapy of human papillomavirus-related disease.
This chapter reviews the current treatment of chronic and neoplastic human papillomavirus (HPV)-associated conditions and the development of novel therapeutic approaches. Surgical excision of HPV-associated lower genital tract neoplasia is very successful but largely depends on secondary prevention programmes for identification of disease. Only high-risk HPV-driven chronic, pre-neoplastic lesions and some very early cancers cannot be successfully treated by surgical procedures alone. Chemoradiation therapy of cervical cancer contributes to the 66-79% cervical cancer survival at 5 years. Outlook for those patients with persistent or recurrent cervical cancer following treatment is very poor. Topical agents such as imiquimod (immune response modifier), cidofovir (inhibition of viral replication; induction apoptosis) or photodynamic therapy (direct damage of tumour and augmentation of anti-tumour immunity) have all shown some useful efficacy (~50-60%) in treatment of high grade vulvar intraepithelial neoplasia (VIN). Provider administered treatments of genital warts include cryotherapy, trichloracetic acid, or surgical removal which has the highest primary clearance rate. Patient applied therapies include podophyllotoxin and imiquimod. Recurrence after "successful" treatment is 30-40%. Further improvements could derive from a rational combination of current therapy with new drugs targeting molecular pathways mediated by HPV in cancer. Small molecule inhibitors targeting the DNA binding activities of HPV E1/E2 or the anti-apoptotic consequences of E6/E7 oncogenes are in preclinical development. Proteasome and histone deacetylase inhibitors, which can enhance apoptosis in HPV positive tumour cells, are being tested in early clinical trials. Chronic high-risk HPV infection/neoplasia is characterised by systemic and/or local immune suppressive regulatory or escape factors. Recently two E6/E7 vaccines have shown some clinical efficacy in high grade VIN patients and this correlated with strong and broad systemic HPV-specific T cell response and modulation of key local immune factors. Treatments that can shift the balance of immune effectors locally in combination with vaccination are now being tested. This article forms part of a special supplement entitled "Comprehensive Control of HPV Infections and Related Diseases" Vaccine Volume 30, Supplement 5, 2012.


Maternal exposure to predator scents: offspring phenotypic adjustment and dispersal.
Predation is a strong selective pressure generating morphological, physiological and behavioural responses in organisms. As predation risk is often higher during juvenile stages, antipredator defences expressed early in life are paramount to survival. Maternal effects are an efficient pathway to produce such defences. We investigated whether maternal exposure to predator cues during gestation affected juvenile morphology, behaviour and dispersal in common lizards (Zootoca vivipara). We exposed 21 gravid females to saurophagous snake cues for one month while 21 females remained unexposed (i.e. control). We measured body size, preferred temperature and activity level for each neonate, and released them into semi-natural enclosures connected to corridors in order to measure dispersal. Offspring from exposed mothers grew longer tails, selected lower temperatures and dispersed thrice more than offspring from unexposed mothers. Because both tail autotomy and altered thermoregulatory behaviour are common antipredator tactics in lizards, these results suggest that mothers adjusted offspring phenotype to risky natal environments (tail length) or increased risk avoidance (dispersal). Although maternal effects can be passive consequences of maternal stress, our results strongly militate for them to be an adaptive antipredator response that may increase offspring survival prospects.


Child testimonies during an outbreak of witch hysteria: Sweden 1670-1671.
Eight hundred and nine testimonies given by children between the ages of 1 and 16 to the priests of the parish of Rättvik, and to the Royal commission of inquiry, during an outbreak of witch hysteria in 1670-71 are examined. The result implies that the capacity to separate reality from fantasy as well as the tendency to give stereotyped testimonies are related to age, social influence from other children and sex. The results also suggest that the testimonies were influenced by the person investigating the child.


Insomnia and suicidal ideation in nonaffective psychosis.
Insomnia is a common symptom in the clinical course of schizophrenia. There is a robust association between insomnia and suicidality in other psychiatric disorders. Two previous studies found associations between insomnia and suicide attempt or completed suicide in patients with schizophrenia. We hypothesized that greater insomnia would be associated with greater levels of suicidal ideation in patients with schizophrenia and other nonaffective psychoses. We recruited 108 inpatients and outpatients age 18-65 between July 2010 and July 2016 with DSM-IV nonaffective psychosis (schizophrenia, schizoaffective disorder, or schizophreniform disorder). We investigated relationships between current insomnia (Insomnia Severity Index [ISI]), suicidal ideation over the past week, and lifetime history of suicide attempt (Beck Scale for Suicide Ideation [BSS]) in regression analyses. After controlling for potential confounders, insomnia was a significant indicator of suicidal ideation (β = 0.27, p = 0.032). Insomnia was also a significant indicator of a high BSS score (≥16; OR = 1.14, 95% CI: 1.01-1.28, p = 0.029). Furthermore, participants with severe insomnia were almost 15 times more likely to have a lifetime history suicide attempt than participants without current insomnia (OR = 14.8, 95% CI: 1.4-157, p = 0.025). Insomnia was also an indicator of greater PANSS total (β = 0.33, p = 0.001), positive subscale (β = 0.32, p = 0.002), and general subscale (β = 0.40, p < 0.001) scores. Insomnia is associated with suicidal ideation, lifetime suicide attempt, and greater psychopathology in patients with schizophrenia. Our findings suggest that formal assessment of insomnia may be germane to the clinical care of patients with schizophrenia as a marker of suicide risk and symptom severity.


The Poisoning Information Database Covers a Large Proportion of Real Poisoning Cases in Korea.
The poisoning information database (PIDB) provides clinical toxicological information on commonly encountered toxic substances in Korea. The aim of this study was to estimate the coverage rate of the PIDB by comparing the database with the distribution of toxic substances that real poisoning patients presented to 20 emergency departments. Development of the PIDB started in 2007, and the number of toxic substances increased annually from 50 to 470 substances in 2014. We retrospectively reviewed the medical records of patients with toxic exposure who visited 20 emergency departments in Korea from January to December 2013. Identified toxic substances were classified as prescription drug, agricultural chemical, household product, animal or plant, herbal drug, or other. We calculated the coverage rate of the PIDB for both the number of poisoning cases and the kinds of toxic substances. A total of 10,887 cases of intoxication among 8,145 patients was collected. The 470 substances registered in the PIDB covered 89.3% of 8,891 identified cases related to poisoning, while the same substances only covered 45.3% of the 671 kinds of identified toxic substances. According to category, 211 prescription drugs, 58 agricultural chemicals, 28 household products, and 32 animals or plants were not covered by the PIDB. This study suggested that the PIDB covered a large proportion of real poisoning cases in Korea. However, the database should be continuously extended to provide information for even rare toxic substances.


Monoclonal antibodies to epithelium-specific components of the human milk fat globule membrane: production and reaction with cells in culture.
Three hybridomas producing monoclonal antibodies (IgG), reacting with components of the human mammary milk fat globule have been isolated. When tested for binding to a wide range of human cell lines and strains, all three antibodies show negative reactions with fibroblasts, lymphoblastoid cells, and a large number of epithelial cell lines of non-breast origin. Two of the antibodies (1.10.F3 and 3.14.A3) reacted with seven out of eight breast cancer lines tested, and with epithelial cells cultured from human milk. The other antibody (3.15.C3) reacted with only two of the breast cancer cell lines.


Defective secretion of islet hormones in chromogranin-B deficient mice.
Granins are major constituents of dense-core secretory granules in neuroendocrine cells, but their function is still a matter of debate. Work in cell lines has suggested that the most abundant and ubiquitously expressed granins, chromogranin A and B (CgA and CgB), are involved in granulogenesis and protein sorting. Here we report the generation and characterization of mice lacking chromogranin B (CgB-ko), which were viable and fertile. Unlike neuroendocrine tissues, pancreatic islets of these animals lacked compensatory changes in other granins and were therefore analyzed in detail. Stimulated secretion of insulin, glucagon and somatostatin was reduced in CgB-ko islets, in parallel with somewhat impaired glucose clearance and reduced insulin release, but normal insulin sensitivity in vivo. CgB-ko islets lacked specifically the rapid initial phase of stimulated secretion, had elevated basal insulin release, and stored and released twice as much proinsulin as wildtype (wt) islets. Stimulated release of glucagon and somatostatin was reduced as well. Surprisingly, biogenesis, morphology and function of insulin granules were normal, and no differences were found with regard to beta-cell stimulus-secretion coupling. We conclude that CgB is not required for normal insulin granule biogenesis or maintenance in vivo, but is essential for adequate secretion of islet hormones. Consequentially CgB-ko animals display some, but not all, hallmarks of human type-2 diabetes. However, the molecular mechanisms underlying this defect remain to be determined.


Anonymization of longitudinal electronic medical records.
Electronic medical record (EMR) systems have enabled healthcare providers to collect detailed patient information from the primary care domain. At the same time, longitudinal data from EMRs are increasingly combined with biorepositories to generate personalized clinical decision support protocols. Emerging policies encourage investigators to disseminate such data in a deidentified form for reuse and collaboration, but organizations are hesitant to do so because they fear such actions will jeopardize patient privacy. In particular, there are concerns that residual demographic and clinical features could be exploited for reidentification purposes. Various approaches have been developed to anonymize clinical data, but they neglect temporal information and are, thus, insufficient for emerging biomedical research paradigms. This paper proposes a novel approach to share patient-specific longitudinal data that offers robust privacy guarantees, while preserving data utility for many biomedical investigations. Our approach aggregates temporal and diagnostic information using heuristics inspired from sequence alignment and clustering methods. We demonstrate that the proposed approach can generate anonymized data that permit effective biomedical analysis using several patient cohorts derived from the EMR system of the Vanderbilt University Medical Center.


A candidate gene investigation of methylphenidate response in adult attention-deficit/hyperactivity disorder patients: results from a naturalistic study.
Attention-deficit/hyperactivity disorder (ADHD) is a common childhood onset neuropsychiatric disorder with a complex and heterogeneous symptomatology. Persistence of ADHD symptoms into adulthood is common. Methylphenidate (MPH) is a widely prescribed stimulant compound that may be effective against ADHD symptoms in children and adults. However, MPH does not exert satisfactory effect in all patients. Several genetic variants have been proposed to predict either treatment response or adverse effects of stimulants. We conducted a literature search to identify previously reported variants associated with MPH response and additional variants that were biologically plausible candidates for MPH response. The response to MPH was assessed by the treating clinicians in 564 adult ADHD patients and 20 genetic variants were successfully genotyped. Logistic regression was used to test for association between these polymorphisms and treatment response. Nominal associations (p < 0.05) were meta-analysed with published data from previous comparable studies. In our analyses, rs1800544 in the ADRA2A gene was associated with MPH response at a nominal significance level (OR 0.560, 95 % CI 0.329-0.953, p = 0.033). However, this finding was not affirmed in the meta-analysis. No genetic variants revealed significant associations after correction for multiple testing (p < 0.00125). Our results suggest that none of the studied variants are strong predictors of MPH response in adult ADHD as judged by clinician ratings, potentially except for rs1800544. Consequently, pharmacogenetic testing in routine clinical care is not supported by our analyses. Further studies on the pharmacogenetics of adult ADHD are warranted.


A comparison of the effects of controlled-release levodopa (Madopar CR) with conventional levodopa in late Parkinson's disease.
In this multicentre study a controlled-release formulation of levodopa and the decarboxylase inhibitor benserazide (Madopar CR) was evaluated in patients with Parkinson's disease exhibiting dose-related fluctuations in motor performance in response to conventional levodopa preparations. The effect of Madopar CR, with or without conventional levodopa/benserazide, on the proportion of time spent "on", "off" or "intermediate" was compared with that of previous conventional levodopa/decarboxylase inhibitor therapy. Evaluation of the two periods of optimum therapy was based on both patient diary data and investigator opinion. Forty seven patients completed the study but full patient diaries were available for only 37. The mean optimum total daily dosage of conventional Madopar was 820 mg taken in a mean of 6.4 doses, compared with a mean optimum daily dosage of combined Madopar CR and conventional Madopar of 1088 mg, taken in a mean of 5.2 doses. Conventional Madopar was taken in addition to Madopar CR in all but eight patients. Madopar CR was felt to be advantageous in 83% and disadvantageous in 11% of patients completing the study. Considering the 37 patients for whom diary data were available, Madopar CR therapy resulted in an increase in the mean time spent "on" (p = 0.016) and a decrease in the mean time spent "off" (p = 0.029) compared with conventional Madopar alone. Individually 25 out of 37 had an increase in "on" time and 19 out of 37 experienced a decrease in "off" time. Thus Madopar CR was found to be beneficial in a significant proportion of patients experiencing fluctuations in response to conventional levodopa.


The effects of selective electrical stimulation of the rat cochlea on hippocampal field potentials.
The hippocampus is a brain structure well known for its importance to spatial learning and memory. As such it is assumed to use multisensory integration in order to generate mathematical maps of the spatial environment. Auditory inputs to the hippocampus have been described to a limited extent. Although it has been demonstrated that natural auditory stimulation can evoke local field potentials (LFPs) and single neuron responses in the hippocampus, and that noise trauma and tinnitus can cause maladaptive hippocampal plasticity, to the best of our knowledge no one has selectively, electrically stimulated the cochlea and recorded electrophysiological responses in the hippocampus. Here, we used unilateral electrical cochlear stimulation in rats while recording LFPs in the hippocampus bilaterally, with a multi-electrode array. We observed triphasic LFP responses bilaterally from unilateral stimulation, but with larger amplitudes on the contralateral side, especially for Phase 1 and 3 of the LFP, for which the latencies were also longer. The latencies under urethane anaesthesia were consistent with auditory information from the cochlea travelling across approximately 6-7 synapses. These data demonstrate that the cochlea contributes a powerful auditory input to the hippocampus, which is also lateralized, and may have implications for spatial cognition in cochlear implant patients.


HLA-C is necessary for optimal human immunodeficiency virus type 1 infection of human peripheral blood CD4 lymphocytes.
The hypothesis that open conformers of HLA-C on target cells might directly exert an effect on their infectability by human immunodeficiency virus (HIV) has been suggested previously. This was tested by exploiting the peculiar specificity of monoclonal antibody (mAb) L31 for HLA-C open conformers to show that normal levels of Env-driven fusion were restored in HLA-C transfectants of a major histocompatibility complex-deleted (fusion-incompetent) cell line. The physiological relevance of this finding is now confirmed in this report, where small interfering RNA (siRNA) technology was used to silence HLA-C expression in peripheral blood lymphocytes (PBLs) from 11 healthy donors. Infectability by HIV (strains IIIB and Bal and primary isolates) was significantly reduced (P=0.016) in silenced cells compared with cells that maintained HLA-C expression in 10 of the 11 PBL donors. Normal infectability was resumed, together with HLA-C expression, when the effect of siRNA interference waned after several days in culture. Additional confirmation of the HLA-C effect was obtained in several assays employing HLA-C-positive and -negative cell lines, a number of HIV strains and also pseudoviruses. In particular, viruses pseudotyped with env genes from HIV strains AC10 and QH0692.42 were assayed on siRNA-silenced lymphocytes from three healthy donors: the differences in infection with pseudoviruses were even higher than those observed in infections with normal viruses.


Cue reactivity in converted and native intermittent smokers.
Nondaily, or intermittent smokers (ITS), who constitute a substantial fraction of U.S. smokers, are thought to smoke in response to cues. Previous cue reactivity research showed no difference between ITS and daily smokers in response to cues. This report examines whether "converted" ITS (CITS) with a history of past daily smoking differ from "native" ITS (NITS) in craving and smoking in response to cues. A total of 146 CITS (who previously smoked daily for at least 6 months) and 73 NITS participated. Participants were exposed to 5 active cues (smoking, alcohol, negative affect, positive affect, and smoking prohibitions) and a control neutral cue, in separate sessions. Changes in craving were assessed pre-post cue exposure. Smoking behavior (smoking [y/n], smoking latency, number of cigarettes, number of puffs, and increase in carbon monoxide [CO]) was observed. Analyses contrasted response to each active cue compared to the neutral cue and controlled for order effects and for time since last cigarette, which differed between groups. Regardless of cues, CITS reported higher craving and greater change in craving, were more likely to smoke, tended to progress faster to smoking, and showed greater increases in CO when they did smoke. NITS and CITS showed similar cue reactivity on most measures, though NITS took more puffs after viewing smoking cues (compared to neutral) than did CITS. Though CITS show some remnants of their history of daily smoking, CITS and NITS demonstrate similar cue reactivity, suggesting that they would not require different behavioral approaches to help them quit.


Water: neglected, unappreciated and under researched.
Water, an essential nutrient, is often ignored in reports of dietary surveys and nutrition. Although it is ubiquitous in foods and beverages, the attention is often focused on the minerals or calorific values of the fluids imbibed rather than the water per se. Water is often taken for granted by many in Western countries due to its abundant availability through water systems. In developing countries, however, water and sanitation raise significant problems. This review overviews (i) the global perspective of the potable water supply, (ii) human rights and water, (iii) dietary guidelines and sources of water and (iv) the physiology of water balance. Gaps in knowledge and understanding around hydration and water requirements are also discussed. Nutritionists are urged to look at the bigger picture of the global water supply and to use good judgement and common sense when advising on water requirements.


Analysis of the gut microbiota in the old order Amish and its relation to the metabolic syndrome.
Obesity has been linked to the human gut microbiota; however, the contribution of gut bacterial species to the obese phenotype remains controversial because of conflicting results from studies in different populations. To explore the possible dysbiosis of gut microbiota in obesity and its metabolic complications, we studied men and women over a range of body mass indices from the Old Order Amish sect, a culturally homogeneous Caucasian population of Central European ancestry. We characterized the gut microbiota in 310 subjects by deep pyrosequencing of bar-coded PCR amplicons from the V1-V3 region of the 16S rRNA gene. Three communities of interacting bacteria were identified in the gut microbiota, analogous to previously identified gut enterotypes. Neither BMI nor any metabolic syndrome trait was associated with a particular gut community. Network analysis identified twenty-two bacterial species and four OTUs that were either positively or inversely correlated with metabolic syndrome traits, suggesting that certain members of the gut microbiota may play a role in these metabolic derangements.


Topological portraits of multiscale coordination dynamics.
Living systems exhibit complex yet organized behavior on multiple spatiotemporal scales. To investigate the nature of multiscale coordination in living systems, one needs a meaningful and systematic way to quantify the complex dynamics, a challenge in both theoretical and empirical realms. The present work shows how integrating approaches from computational algebraic topology and dynamical systems may help us meet this challenge. In particular, we focus on the application of multiscale topological analysis to coordinated rhythmic processes. First, theoretical arguments are introduced as to why certain topological features and their scale-dependency are highly relevant to understanding complex collective dynamics. Second, we propose a method to capture such dynamically relevant topological information using persistent homology, which allows us to effectively construct a multiscale topological portrait of rhythmic coordination. Finally, the method is put to test in detecting transitions in real data from an experiment of rhythmic coordination in ensembles of interacting humans. The recurrence plots of topological portraits highlight collective transitions in coordination patterns that were elusive to more traditional methods. This sensitivity to collective transitions would be lost if the behavioral dynamics of individuals were treated as separate degrees of freedom instead of constituents of the topology that they collectively forge. Such multiscale topological portraits highlight collective aspects of coordination patterns that are irreducible to properties of individual parts. The present work demonstrates how the analysis of multiscale coordination dynamics can benefit from topological methods, thereby paving the way for further systematic quantification of complex, high-dimensional dynamics in living systems.


Longitudinal Assessment of Medical Student Emotional Intelligence Over Preclinical Training.
Given the importance of emotional intelligence (EI) in physician leadership and success, the possible decline of EI over the course of training is a concern in medical education. To assess the EI of osteopathic medical students as they progress through the preclinical years of medical school, to discover the course of EI over time, and to identify specific core EI competencies to be enhanced or coached. This exploratory, longitudinal study used survey data to assess the course and competencies of EI in the incoming 2019 class of medical students at a college of osteopathic medicine. Changes in EI overall scores and the 8 core competency scale scores were assessed, and additional demographic data were collected for use in analysis. Methods of analysis included paired samples t test, independent samples t test, Pearson product moment correlation, and multiple regression analysis. Statistical significance was defined as P<.05. Sixty-two participants completed the Six Seconds Emotional Intelligence Assessment survey at orientation and again after their second year. Overall EI scores declined over the course of the preclinical training for men and women (t61=4.24, P<.001), although no differences were noted by gender when independent-samples t tests were run. However, 2 of the 8 competency scales of the composite EI scores did not decline over time-Enhanced Emotional Literacy and Pursue Noble Goals. A weak negative correlation was found between Medical College Admission Test scores and the scale of Apply Consequential Thinking (r=-0.3, P<.05). A weak positive correlation was found between grade point average and intrinsic motivation (r=0.3, P<.05) and optimism (r=0.3, P<.05), and a moderate positive correlation existed between preboard examination scores and intrinsic motivation (r=0.5, P<.001) and optimism (r=0.4, P<.01). None of the chosen independent variables (academic grades, test scores, age, or gender) contributed to overall EI scores. During the preclinical training, osteopathic medical students' overall EI scores, as well as 6 of the 8 core competency scales, declined. Therefore, EI coaching is warranted to address specific core competencies needed to build, enhance, and prevent the decline of EI through preclinical training.


Newer endovascular tools: a review of experimental and clinical aspects.
The history of treatment of intracranial aneurysms dates back to the late 18th century. These early physicians largely based their crude techniques around "wire insertion alone, galvanopuncture (electrothrombosis), and fili-galvanopuncture (wire insertion together with electrothrombosis)," albeit with overwhelmingly unfavorable outcomes. By the end of the 20th century, treatment options progressed to include two highly effective, and safe, procedures: surgical clipping and endovascular coiling. These methods have been found to be effective treatments for a large portion of aneurysms, but there still exists a subset of patients that do not respond well to these therapies. While much progress has been made in stent-assisted coiling including the development of newer stents aimed at keep the coil ball from protruding into the parent vessel, the introduction of flow diverters has characterized a new phase in the endovascular treatment of intracranial aneurysms. This treatment paradigm is rapidly becoming the treatment of choice for large and complex aneurysms internal carotid artery. Intrasaccular flow diverters such as the Woven EndoBridge device (WEB) and Luna device are showing promise in the treatment of wide neck bifurcation aneurysms. Other newer devices including the pCONus Bifurcating Aneurysm Implant and Endovascular Clip Systems (eCLIPs) are showing promise in small clinical and preclinical studies. As technology improves, newer devices with ingenious designs are constantly being introduced into the clinical arena. Most of these devices try to address the limitations of traditional endovascular methods in regard to providing a safe and effective treatment of wide-necked bifurcation aneurysms. Several large prospective studies are underway and once completed, the role of these newer devices will be better defined. It is easy to anticipate that with advances in 3D techniques and printing, a future in which customized devices are designed based on the individual anatomy and morphology of the aneurysm in a given patient is within reach.


Relationship of the expression of the multidrug resistance gene product (P-glycoprotein) in human colon carcinoma to local tumor aggressiveness and lymph node metastasis.
P-glycoprotein mediates classic multidrug resistance by functioning as an efflux pump that excretes lipophilic chemotherapeutic drugs from cancer cells. We now report an association of P-glycoprotein in colon carcinomas with another tumor property, i.e., enhancement of local tumor aggressiveness. P-glycoprotein was detected with monoclonal antibody immunohistochemistry in 65 of 95 primary colon adenocarcinomas, which were stage B1 or greater. In all but 1 of the 95 cases, solitary invading carcinoma cells were present at the leading edge of the tumor. This subpopulation of invasive carcinoma cells expressed P-glycoprotein (P-Gp+) in 47 of the 95 surgically resected colon specimens. Cases were grouped on the basis of the presence (Group 1, 47 cases) or absence (Group 2, 48 cases) of P-Gp+ invasive carcinoma cells. There was a significantly greater incidence of vessel invasion (P less than 0.001) and lymph node metastases (P less than 0.01) in Group 1 cases. Groups 1 and 2 did not differ with respect to tumor size, depth of invasion of the bowel wall, histological grade, maximum tumor size, mitotic index, mucin production, or presence of perineural invasion (P greater than 0.1). Our findings indicate that P-Gp+ invasive colon cancer cells may have an increased potential for dissemination, suggesting that P-glycoprotein may influence cell behavior.


The impact of state community benefit laws on the community health orientation and health promotion services of hospitals.
This study examined the effect of state community benefit laws and guidelines on the community health orientation and the provision of hospital-based health promotion services in hospitals. The sample included all not-for-profit and investor-owned acute-care hospitals in the United States during the year 2000. Multiple regression procedures were used to test the effect of community benefit laws and type of ownership while controlling for organizational and environmental variables. The results of these procedures indicated that, on average, not-for-profit hospitals in the ten states with community benefit laws/guidelines reported significantly more community health orientation activities than did not-for-profit hospitals in the forty other states. The results of the multiple regression procedures also indicated that, on average, the investor-owned hospitals in the ten states with laws/guidelines reported significantly more community health orientation activities than did the investor-owned hospitals in the forty other states. The study found that community benefit laws had the effect of decreasing ownership-related differences in reported community health orientation activities. Further, Levene's test of equality of variance showed that the not-for-profit hospitals in community benefit states exhibited significantly lower variance in the community health orientation activities when compared with the not-for-profit hospitals in non-community benefit states. However, none of the statistical tests supported the hypotheses that community benefit laws compelled or induced hospitals to offer significantly more health promotion services. The study concluded that coercive measures such as community benefit laws were effective in compelling not-for-profit hospitals to report increased community orientation activities, and it also concluded that the mimetic pressures associated with these laws were effective in inducing investor-owned hospitals to report increased community orientation activities.


High rates of oxygen reduction over a vapor phase-polymerized PEDOT electrode.
The air electrode, which reduces oxygen (O2), is a critical component in energy generation and storage applications such as fuel cells and metal/air batteries. The highest current densities are achieved with platinum (Pt), but in addition to its cost and scarcity, Pt particles in composite electrodes tend to be inactivated by contact with carbon monoxide (CO) or by agglomeration. We describe an air electrode based on a porous material coated with poly(3,4-ethylenedioxythiophene) (PEDOT), which acts as an O2 reduction catalyst. Continuous operation for 1500 hours was demonstrated without material degradation or deterioration in performance. O2 conversion rates were comparable with those of Pt-catalyzed electrodes of the same geometry, and the electrode was not sensitive to CO. Operation was demonstrated as an air electrode and as a dissolved O2 electrode in aqueous solution.


MicroRNA-21 abrogates palmitate-induced cardiomyocyte apoptosis through caspase-3/NF-κB signal pathways.
The aim of the study was to investigate the role of microRNA-21 (miR-21) in cardiomyocyte apoptosis and to determine a possible mechanism. H9c2 embryonic rat heart-derived cells were used in the study. Cell viability was determined using the 3-(4.5-dimethyl-2-thiazolyl)-2,5- diphenyl-2-H-tetrazolium bromide (MTT) assay, and flow cytometry was used to evaluate cell apoptosis. Reverse transcription-polymerase chain reaction and western blot assays were used to detect mRNA and protein expression of the apoptosis-related proteins and miR-21. ELISA was used to detect reactive oxygen species (ROS). Palmitate exposure greatly reduced miR-21 expression in cardiomyocytes. Apoptosis increased when miR-21 was inhibited with or without palmitate exposure. Consistently, reduced apoptosis was observed when miR-21 was overexpressed in cardiomyocytes. Caspase-3 activity was reduced after palmitate exposure. Bcl-2 protein expression was increased in H9c2 cells when transfected with the miR-21 mimic. MiR-21 overexpression alone did not induce ROS or DNA fragmentation; however, in conjunction with palmitate exposure, miR-21 mimic reduced ROS and DNA fragmentation. Moreover, palmitate administration overcame the antioxidant effect of 3 mM N-acetylcysteine to significantly inhibit apoptosis, DNA fragmentation, and caspase-3 activity. The exposure to palmitate greatly reduced p65 and p-p38 expression in the nucleus. A p38 inhibitor had no effect on the expression of Bcl-2 and cleaved caspase-3 in H9c2 cells alone; however, when combined with exposure to palmitate the p38 inhibitor induced Bcl-2 expression and inhibited caspase-3 activity. The p38 inhibitor by itself did not induce apoptosis, ROS production, or DNA fragmentation in H9c2 cells, but when palmitate was included with the p38 inhibitor, apoptosis, ROS production, and DNA fragmentation were reduced. miR-21 protects cardiomyocytes from apoptosis that is induced by palmitate through the caspase-3/NF-κB signal pathways.


Optimal pressure regulation of the pneumatic ventricular assist device with bellows-type driver.
The bellows-type pneumatic ventricular assist device (VAD) generates pneumatic pressure with compression of bellows instead of using an air compressor. This VAD driver has a small volume that is suitable for portable devices. However, improper pneumatic pressure setup can not only cause a lack of adequate flow generation, but also cause durability problems. In this study, a pneumatic pressure regulation system for optimal operation of the bellows-type VAD has been developed. The optimal pneumatic pressure conditions according to various afterload conditions aiming for optimal flow rates were investigated, and an afterload estimation algorithm was developed. The developed regulation system, which consists of a pressure sensor and a two-way solenoid valve, estimates the current afterload and regulates the pneumatic pressure to the optimal point for the current afterload condition. Experiments were performed in a mock circulation system. The afterload estimation algorithm showed sufficient performance with the standard deviation of error, 8.8 mm Hg. The flow rate could be stably regulated with a developed system under various afterload conditions. The shortcoming of a bellows-type VAD could be handled with this simple pressure regulation system.


PCR detection of Pneumocystis carinii in bronchoalveolar lavage specimens: analysis of sensitivity and specificity.
Although PCR detection of Pneumocystis carinii DNA has been described, little is known about the sensitivity or specificity of the assay in routine laboratory practice. We had the unique opportunity to use a bronchoalveolar lavage (BAL) specimen bank with samples for which the direct examination results for P. carinii were known. DNA purified from 129 selected specimens was amplified by using the primers described previously (A. E. Wakefield, F. J. Pixley, S. Banerji, K. Sinclair, R. F. Miller, E. R. Moton, and J. M. Hopkin, Mol. Biochem. Parasitol. 43:69-76, 1990). Of the 129 specimens, 37 were positive for P. carinii by direct examination. All 37 specimens were positive for P. carinii by PCR, yielding a 100% sensitivity and 100% negative predictive value for the assay. An additional 23 specimens were repeatedly positive for P. carinii by PCR but were not positive by direct examination. Review of the patient charts for these specimens with discordant results demonstrated that five of the patients were actually positive for P. carinii, as determined by either biopsy or examination of repeat or prior BAL specimens. A response to empiric therapy for P. carinii pneumonia was seen in an additional two patients. Of the remaining specimens, 8 produced no significant isolates other than P. carinii, while 12 contained culture-confirmed significant respiratory pathogens in addition to P. carinii (two fungal, nine bacterial, and one viral pathogen). Cytomegalovirus, which was of unknown significance, was isolated from 16 additional specimens. Overall, the specificity of the PCR assay was 79.3% compared to the results of direct examination. We hypothesized that the apparently poor specificity of the PCR assay was due to the increased sensitivity of the assay compared to that of direct examination. The sensitivity of the PCR assay was therefore assessed with BAL specimens containing P. carinii cysts. Serial dilutions of this preparation were evaluated by direct examination and PCR. PCR was found to be 100-fold more sensitive than direct examination, which detected one to two cysts per amplification. No false-positive results were detected in controls containing no DNA or by using target DNA from various fungal, viral, or bacterial respiratory pathogens. We conclude that PCR detection of P. carinii in BAL specimens is very sensitive and should be considered for patients whose specimens do not yield a diagnosis. The increased sensitivity of the PCR assay may help to identify those patients with low-titer infections who might benefit from directed antibiotic therapy for P. carinii and would otherwise be missed by direct examination alone.