Offspring body size and metabolic profile - effects of lifestyle intervention in obese pregnant women.
Worldwide, the prevalence of obesity has reached epidemic proportions. In Denmark one third of all pregnant women are overweight and 12 % are obese. Perhaps even more concerning, a dramatic rise in the prevalence of childhood overweight and obesity has also been evident over recent decades. The obesity epidemic is not simply a consequence of poor diet or sedentary lifestyles. Obesity is a multifactorial condition in which environmental, biological and genetic factors all play essential roles. The Developmental Origins of Health and Disease (DoHaD) hypothesis has highlighted the link between prenatal, perinatal and early postnatal exposure to certain environmental factors and subsequent development of obesity and non-communicable diseases. Maternal obesity and excessive gestational weight gain, resulting in over-nutrition of the fetus, are major contributors to obesity and metabolic disturbances in the offspring. Pregnancy offers the opportunity to modify the intrauterine environment, and maternal lifestyle changes during gestation may confer health benefits to the child. The overall aim with this PhD thesis was to study the effects of maternal obesity on offspring body size and metabolic outcomes, with special emphasis on the effects of lifestyle intervention during pregnancy. The thesis is based on a literature review, description of own studies and three original papers/manuscripts (I, II and III). In paper I, we used data from the Danish Medical Birth Registry. The aim of this paper was to examine the impact of maternal pregestational Body Mass Index (BMI) and smoking on neonatal abdominal circumference (AC) and weight at birth and to define reference curves for birth AC and weight in offspring of healthy, non-smoking, normal weight women. Data on 366,886 singletons were extracted and analyzed using multivariate linear regressions. We found that birth AC and weight increased with increasing pregestational BMI and decreased with smoking. Reference curves were created for offspring of healthy, non-smoking mothers with normal pregestational BMI. Paper II and III are based on an offspring follow-up of a randomized controlled trial (RCT) with 360 obese pregnant women. The intervention during pregnancy consisted of two major components: dietary advice and physical activity. The intervention resulted in a small, but significant difference in gestational weight gain compared to the control group. A number of 301 completed the trial and were eligible for the follow-up. We managed to include 157 mother and child dyads in the follow-up, which was conducted in Odense University Hospital and Aarhus University Hospital, Skejby between February 2010 and November 2012. At that time the children were in the ages 2.5-3 years. In addition to the children from the RCT, a group of 97 children born to lean mothers were included as an external reference group. The follow-up consisted of a clinical examination with anthropometric measures, DEXA scans and fasting blood samples for evaluation of metabolic outcomes. In paper II the effect of the maternal intervention on offspring body composition and anthropometric outcomes was studied. The primary outcome was BMI Z-score and secondary outcomes were: body composition values by DEXA (fat mass, lean mass and fat percentage), BMI, percentage of overweight or obese children and skin fold thicknesses. We found no significant differences in offspring outcomes between randomized groups of the preceding RCT. Neither was any differences detected between offspring of the RCT or the external reference group born to lean mothers. Paper III focused on the metabolic outcomes in the offspring. We additionally studied the predictive values of birth weight (BW) and birth abdominal circumference (BAC) on metabolic risk factors. We found that both BAC and BW were significantly associated with several risk factors in early childhood. All metabolic measurements in RCT offspring were similar, and no differences were detected between the RCT offspring and the external reference group of offspring of lean mothers. Lifestyle intervention in obese pregnant women has the potential to modify the intrauterine environment and confer long-term benefits to the child. In this follow-up study, lifestyle intervention in pregnancy did not result in changes in offspring body composition or metabolic risk factors at 2.8 years. This might be due to a limited difference in gestational weight gain between follow-up attendees. When comparing offspring of obese women with offspring of normal weight mothers all outcomes were similar. We speculate that obese mothers entering a lifestyle intervention RCT regardless of the intervention have a high motivation to focus on healthy lifestyle during pregnancy, which makes it difficult to determine the effects of the randomized lifestyle intervention compared to an unselected control group of obese women. Our studies (paper I and III) on birth abdominal circumference show that abdominal size at birth is a good predictor of later adverse metabolic profile. Abdominal circumference at birth may reflect visceral adiposity and this measurement together with birth weight are strongly associated to later adverse metabolic outcome. Future studies should be performed in other populations to confirm this.


In vitro formation of a c-type cytochrome.
C-type cytochromes are essential for almost all organisms; they are characterized by the covalent attachment of heme to protein through two thioether bonds to a Cys-Xaa-Xaa-Cys-His peptide motif. Here we show, contrary to opinion of 30 years standing, that a c-type cytochrome can form from heme and apoprotein in vitro under mild conditions and in the absence of any biosynthesis apparatus. This reaction occurs provided formation of a disulfide bond within the Cys-Xaa-Xaa-Cys-His motif is avoided. There are important implications for understanding in vivo cytochrome c assembly.


Recommendations for imaging tumor response in neurofibromatosis clinical trials.
Neurofibromatosis (NF)-related benign tumors such as plexiform neurofibromas (PN) and vestibular schwannomas (VS) can cause substantial morbidity. Clinical trials directed at these tumors have become available. Due to differences in disease manifestations and the natural history of NF-related tumors, response criteria used for solid cancers (1-dimensional/RECIST [Response Evaluation Criteria in Solid Tumors] and bidimensional/World Health Organization) have limited applicability. No standardized response criteria for benign NF tumors exist. The goal of the Tumor Measurement Working Group of the REiNS (Response Evaluation in Neurofibromatosis and Schwannomatosis) committee is to propose consensus guidelines for the evaluation of imaging response in clinical trials for NF tumors. Currently used imaging endpoints, designs of NF clinical trials, and knowledge of the natural history of NF-related tumors, in particular PN and VS, were reviewed. Consensus recommendations for response evaluation for future studies were developed based on this review and the expertise of group members. MRI with volumetric analysis is recommended to sensitively and reproducibly evaluate changes in tumor size in clinical trials. Volumetric analysis requires adherence to specific imaging recommendations. A 20% volume change was chosen to indicate a decrease or increase in tumor size. Use of these criteria in future trials will enable meaningful comparison of results across studies. The proposed imaging response evaluation guidelines, along with validated clinical outcome measures, will maximize the ability to identify potentially active agents for patients with NF and benign tumors.


Prevalence of peri-implantitis in patients with implant-supported fixed prostheses.
The purpose of this study was to evaluate periimplantitis prevalence in patients using implant-supported fixed prostheses that did not have any routine maintenance care. A total of 161 implants (27 patients) were evaluated in patients using implant-supported fixed prostheses. Collected data included information related to patient general health and local factors such as characteristics of implants, time in function, type of loading, positioning, Modified Bleeding Index, bacterial plaque, bleeding on probing (BOP), marginal recession, probing depth (PD), keratinized mucosa, and radiographic bone loss (BL). Factors related to the prostheses were also evaluated. The exclusion criteria were patients that have had any follow-up visit for plaque control of the prosthesis and/or the implants. From a total of 161 implants, 116 (72%) presented without peri-implantitis (PD > 4 mm + BOP + BL > 2 mm) while 45 (28%) had some sign of the disease. Implants placed in the maxilla were 2.98 times more likely to develop the disease (P < .05). Moreover, patients aged ≤ 60 years old were 3.24 times more likely to develop peri-implantitis (P < .05). Another analysis with statistical relevance (P < .05) was that implants with less than 3 mm interimplant distance were three times more likely to have peri-implantitis. There was no statistical relevance considering other analyses. It can be concluded that patients aged ≤ 60 years have a greater chance of presenting periimplantitis, as well as for implants positioned in the maxilla and those placed with an interimplant distance < 3 mm.


Is Endoscopic Therapy Effective for Angioectasia in Obscure Gastrointestinal Bleeding?: A Systematic Review of the Literature.
We aimed to summarize pooled rebleeding rates of angioectasia after therapeutic endoscopy, and compare these to historical control (no intervention) rates. Obscure gastrointestinal bleeding continues to be challenging to diagnose and treat; in America, small bowel angioectasias are the most common cause. Technology advances led to higher diagnostic yield for these lesions; however, therapeutic impact of endoscopy remains unclear. A PubMed search (June 1, 2006 to September 19, 2013) with 2 independent reviews sought articles reporting rebleeding rates of symptomatic angioectasia without therapy (natural history) and after endoscopic treatment. This study list was added to studies in the 2007 American Gastroenterological Association systematic review. Data on number of patients who underwent endoscopic therapy, type of therapy used, number of patients who experienced rebleeding, and follow-up time were extracted. Rebleeding data were pooled and weighted averages were reported with 95% confidence intervals (CI). Twenty-four articles (n=490 patients) with data on endoscopic therapy for angioectasia and 6 natural history cohorts (n=130) receiving no therapy for angioectasia were eligible. Of the endoscopic therapy patients, 121 at push enteroscopy and 427 at balloon-assisted enteroscopy; 209/490 (42.7%; 95% CI, 38%-47%) rebled. Of the control (no therapy) patients, 64/130 (49.2%; 95% CI, 40%-58%) rebled. Number needed to treat is estimated at 15 to 16. Rebleeding rate after endoscopic therapy for symptomatic small bowel angioectasia may be comparable to that expected without therapy. Endoscopic therapy may be ineffective; if effective, the needed to treat is estimated to be high. Controlled studies, with intervention-stratified and etiology-stratified outcomes are needed.


The thrombotic events in polycythemia vera patients may be related to increased oxidative stress.
This study was designed to compare the oxidative stress parameters of patients with polycythemia vera (PV) to those of healthy volunteers and to investigate the probable relationship between vascular events and parameters of oxidative status such as total oxidative status (TOS), total antioxidant status, oxidative stress index (OSI) and malondialdehyde (MDA) in PV patients. Thirty-five PV patients (20 males and 15 females) and 20 healthy volunteers (11 males and 9 females) were enrolled. The oxidative status parameters of the patients were measured by spectrophotometric analyses at the time of diagnosis and at 6 months after treatment which consisted of phlebotomy and 100 mg/day acetyl salicylic acid with or without hydroxyurea for the high- and low-risk disease group, respectively. These parameters were compared both to healthy controls and to each other, in order to obtain the values before and after treatment. In addition, during diagnosis, the oxidative status parameters of patients with PV and a history of a vascular event were compared with those of patients with no history of a vascular event. The TOS, OSI and MDA values were significantly higher in the patients than in the control group at the time of diagnosis. At 6 months after phlebotomy and 100 mg/day acetyl salicylic acid therapy, the TOS, OSI and MDA values were significantly lower in the patients when compared to the pretreatment values. The TOS and OSI levels were notably higher in the patients with a vascular-event history than in those without this history. Oxidative stress parameters were increased in PV patients.


Explanations for medically unexplained symptoms: a qualitative study on GPs in daily practice consultations.
General practice is the centre of care for patients with medically unexplained symptoms (MUS). Providing explanations for MUS, i.e. making sense of symptoms, is considered to be an important part of care for MUS patients. However, little is known how general practitioners (GPs) do this in daily practice. This study aimed to explore how GPs explain MUS to their patients during daily general practice consultations. A thematic content analysis was performed of how GPs explained MUS to their patients based on 39 general practice consultations involving patients with MUS. GP provided explanations in nearly all consultations with MUS patients. Seven categories of explanation components emerged from the data: defining symptoms, stating causality, mentioning contributing factors, describing mechanisms, excluding explanations, discussing the severity of symptoms and normalizing symptoms. No pattern of how GPs constructed explanations with the various categories was observed. In general, explanations were communicated as a possibility and in a patient-specific way; however, they were not very detailed. Although explanations for MUS are provided in most MUS consultations, there seems room for improving the explanations given in these consultations. Further studies on the effectiveness of explanations and on the interaction between patients and GP in constructing these explanations are required in order to make MUS explanations more suitable in daily primary care practice.


Postoperative pneumoperitoneum as detected by CT: prevalence, duration, and relevant factors affecting its possible significance.
To determine the prevalence and duration of postoperative pneumoperitoneum as detected on computed tomography (CT) and to evaluate factors that influence postoperative pneumoperitoneum. One hundred three CT examinations of 89 patients performed after abdominal surgery for various indications were prospectively collected and reviewed. The presence and volume of free air were noted and correlated with patients' sex, age, and habitus, with the time interval between surgery and CT, with the type of surgery, and with the presence of drains. Pneumoperitoneum was seen in 44% of examinations performed in the first 3 days after surgery and in 30% between the 4th and 18th postoperative days. The prevalence and volume of free air decreased with the time interval between surgery and CT. It was not detected in any of the 11 examinations performed beyond the 18th postoperative day. The volume of free air in the majority of examinations did not exceed 10 mL and ranged from 0.2 to 10 mL in 19 patients (66% of patients with free air). Larger volumes of free air, ranging from 10 to 20 mL and from 20 to 40 mL, were observed in 5 (17%) and 4 (14%) patients, respectively, and only one patient had more than 40 mL of free air. Free air was found significantly more often in male and in asthenic patients; age had no significant effect. Free air was more prevalent in the presence of drains. The type of surgery did not significantly influence the prevalence of postoperative pneumoperitoneum, although it was noted slightly less often after laparoscopic surgery than after open laparotomy. Pneumoperitoneum is a common phenomenon after abdominal surgery, decreasing in frequency with time. The air is most often residual and not a sign of disruption of the gastrointestinal tract. Obesity, female sex, and occurrence of free air several weeks after surgery are factors suggestive of a leak, but the significance of a postoperative pneumoperitoneum on CT should be determined mainly by the clinical setting.


The impact of work time control on physicians' sleep and well-being.
Physicians' work schedules are an important determinant of their own wellbeing and that of their patients. This study considers whether allowing physicians control over their work hours ameliorates the effects of demanding work schedules. A questionnaire was completed by hospital physicians regarding their work hours (exposure to long shifts, short inter-shift intervals, weekend duties, night duties, unpaid overtime; and work time control), sleep (quantity and disturbance) and wellbeing (burnout, stress and fatigue). Work time control moderated the negative impact that frequent night working had upon sleep quantity and sleep disturbance. For participants who never worked long shifts, work time control was associated with fewer short sleeps, but this was not the case for those who did work long shifts. Optimizing the balance between schedule flexibility and patient needs could enhance physicians' sleep when working the night shift, thereby reducing their levels of fatigue and enhancing patient care.


Utilization of prostate cancer screening according to dietary patterns and other demographic variables. The adventist health study-2.
Prostate-specific antigen test and digital rectal examination are considered important screening methods for early detection of prostate cancer. However, the utilization of prostate cancer screening varies widely and there is limited knowledge of the predictors of utilization. Self-reported prostate cancer screening utilization within the last 2 years was investigated among 11,162 black and non-black North American Seventh-day Adventist men, aged 50-75 years, with different dietary patterns and lifestyle characteristics. Blacks were more likely to screen for prostate cancer than non-blacks (Odds Ratio (OR)=1.38 (95% confidence interval (CI): 1.20-1.57). Those with a vegetarian diet, especially vegans, were less likely to follow screening guidelines, particularly among non-Blacks: vegans (OR=0.47, 0.39-0.58), lacto-ovo-vegetarians (OR=0.75, 0.66-0.86), and pesco-vegetarians (OR=0.74, 0.60-0.91) compared to non-vegetarians after adjusting for age, BMI, marital status, education, income, and family history of cancer. Trends for dietary patterns remained unchanged after stratification on age, family history of cancer, education, personal income, marital status, and BMI. Among black men, diet patterns showed no significant associations with utilization of prostate cancer screening, although vegans tended to underutilize screening compared to non-vegetarians (OR=0.70, 0.44-1.10). Vegetarians, especially non-black vegans, are less likely to follow recommended prostate cancer screening guidelines. The effect of diet was attenuated, and not statistically significant, among black men. Since only about 60% of US men follow prostate cancer screening guidelines, it is important to study reasons for non-compliance in order to increase utilization of preventive measures against prostate cancer.


Microbial fuel cell biosensor for rapid assessment of assimilable organic carbon under marine conditions.
The development of an assimilable organic carbon (AOC) detecting marine microbial fuel cell (MFC) biosensor inoculated with microorganisms from marine sediment was successful within 36 days. This established marine MFC was tested as an AOC biosensor and reproducible microbiologically produced electrical signals in response to defined acetate concentration were achieved. The dependency of the biosensor sensitivity on the potential of the electron-accepting electrode (anode) was investigated. A linear correlation (R(2) > 0.98) between electrochemical signals (change in anodic potential and peak current) and acetate concentration ranging from 0 to 150 μM (0-3600 μg/L of AOC) was achieved. However, the present biosensor indicated a different-linear relation at somewhat elevated acetate concentration ranging from 150 to 450 μM (3600-10,800 μg/L of AOC). This high concentration of acetate addition could be measured by coulombic measurement (cumulative charges) with a linear correlation. For the acetate concentration detected in this study, the sensor recovery time could be controlled within 100 min.


Type 2 diabetic mice demonstrate slender long bones with increased fragility secondary to increased osteoclastogenesis.
Type 2 diabetics often demonstrate normal or increased bone mineral density, yet are at increased risk for bone fracture. Furthermore, the anti-diabetic oral thiazolidinediones (PPARgamma agonists) have recently been shown to increase bone fractures. To investigate the etiology of possible structural and/or material quality defects, we have utilized a well-described mouse model of Type 2 diabetes (MKR). MKR mice exhibit muscle hypoplasia from birth with reduced mass by the pre-diabetic age of 3 weeks. A compensatory hyperplasia ensues during early (5 weeks) development; by 6-8 weeks muscle is normal in structure and function. Adult whole-bone mechanical properties were determined by 4-point bending to test susceptibility to fracture. Micro-computed tomography and cortical bone histomorphometry were utilized to assess static and dynamic indices of structure, bone formation and resorption. Osteoclastogenesis assays were performed from bone marrow-derived non-adherent cells. The 8-week and 16-week, but not 3-week, male MKR had slender (i.e., narrow relative to length) femurs that were 20% weaker (p<0.05) relative to WT control femurs. Tissue-level mineral density was not affected. Impaired periosteal expansion during early diabetes resulted from 250% more, and 40% less of the cortical bone surface undergoing resorption and formation, respectively (p<0.05). Greater resorption persisted in adult MKR on both periosteal and endosteal surfaces. Differences were not limited to cortical bone as the distal femur metaphysis of 16 week MKR contained less trabecular bone and trabecular separation was greater than in WT by 60% (p<0.05). At all ages, MKR marrow-derived cultures demonstrated the ability for enhanced osteoclast differentiation in response to M-CSF and RANK-L. Taken together, the MKR mouse model suggests that skeletal fragility in Type 2 diabetes may arise from reduced transverse bone accrual and increased osteoclastogenesis during growth that is accelerated by the diabetic/hyperinsulinemic milieu. Further, these results emphasize the importance of evaluating diabetic bone based on morphology in addition to bone mass.


Seasonal effects on the occurrence of nocturnal leg cramps: a prospective cohort study.
It has been anecdotally reported that nocturnal leg cramps in pregnant women are worse in summer. We analyzed population-level data to determine whether the symptom burden of nocturnal leg cramps is seasonal in the general population. We examined time-series data for 2 independent measures of the symptom burden of leg cramps: (a) new quinine prescriptions (reflecting new or escalating treatment of leg cramps) from December 2001 to October 2007 among adults aged 50 years and older, which were obtained from linked health care databases that contain the prescribing information for the 4.2 million residents of British Columbia, Canada; and (b) the Internet search volume from February 2004 to March 2012 for the term "leg cramps" (reflecting public interest), which we obtained from Google Trends data and geographically limited to the United States and Australia. We assessed seasonality by determining how well a least-squares sinusoidal model predicted variability in the outcomes. New quinine prescriptions and Internet searches related to leg cramps were both seasonal, with highs in mid-summer and lows in mid-winter, and a peak-to-peak variability that was about two-thirds of the mean. Seasonality accounted for 88% of the observed monthly variability in new quinine prescriptions (p < 0.001) and 70% of the observed variability in Internet searches related to leg cramps (p < 0.001). New quinine prescriptions and Internet searches related to leg cramps were seasonal and roughly doubled between the winter lows and summer highs. Why a disorder of peripheral motor neurons displays such strong seasonality warrants exploration.


An improved microassay for Plasmodium falciparum cytoadherence using stable transformants of Chinese hamster ovary cells expressing CD36 or intercellular adhesion molecule-1.
Stable transformants of Chinese hamster ovary (CHO) cell lines expressing high levels of human CD36 or intercellular adhesion molecule-1 (ICAM-1) have been produced as target cells for cytoadherence of Plasmodium falciparum-infected erythrocytes. An improved adherence microassay has been designed using small sample volumes and allowing convenient and reliable measurements on a large number of samples. The assay can be used both with purified proteins spotted on plastic and with the stably transformed CHO cell lines. The same assay plate can be evaluated either microscopically or by scintillation counting after use of 3H-hypoxanthine-labeled parasites. Using the microassay, functional expression of the transfected receptor molecules on CHO-CD36 and CHO-ICAM was confirmed using parasites with different cytoadherence phenotypes and cytoadherence inhibition experiments with a panel of anti-CD36 antibodies. The use of isolates from The Gambia confirmed the applicability of these assays for laboratory studies of these isolates.


Providing Extrinsic Reward for Test Performance Undermines Long-Term Memory Acquisition.
Based on numerous studies showing that testing studied material can improve long-term retention more than restudying the same material, it is often suggested that the number of tests in education should be increased to enhance knowledge acquisition. However, testing in real-life educational settings often entails a high degree of extrinsic motivation of learners due to the common practice of placing important consequences on the outcome of a test. Such an effect on the motivation of learners may undermine the beneficial effects of testing on long-term memory because it has been shown that extrinsic motivation can reduce the quality of learning. To examine this issue, participants learned foreign language vocabulary words, followed by an immediate test in which one-third of the words were tested and one-third restudied. To manipulate extrinsic motivation during immediate testing, participants received either monetary reward contingent on test performance or no reward. After 1 week, memory for all words was tested. In the immediate test, reward reduced correct recall and increased commission errors, indicating that reward reduced the number of items that can benefit from successful retrieval. The results in the delayed test revealed that reward additionally reduced the gain received from successful retrieval because memory for initially successfully retrieved words was lower in the reward condition. However, testing was still more effective than restudying under reward conditions because reward undermined long-term memory for concurrently restudied material as well. These findings indicate that providing performance-contingent reward in a test can undermine long-term knowledge acquisition.


Strategic appraisal of environmental risks: a contrast between the United Kingdom's Stern Review on the Economics of Climate Change and its Committee on Radioactive Waste Management.
In this article, we compare two high-profile strategic policy reviews undertaken for the U.K. government on environmental risks: radioactive waste management and climate change. These reviews took very different forms, both in terms of analytic approach and deliberation strategy. The Stern Review on the Economics of Climate Change was largely an exercise in expert modeling, building, within a cost-benefit framework, an argument for immediate reductions in carbon emissions. The Committee on Radioactive Waste Management, on the other hand, followed a much more explicitly deliberative and participative process, using multicriteria decision analysis to bring together scientific evidence and stakeholder and public values. In this article, we ask why the two reviews were different, and whether the differences are justified. We conclude that the differences were mainly due to political context, rather than the underpinning science, and as a consequence that, while in our view "fit for purpose," they would both have been stronger had they been less different. Stern's grappling with ethical issues could have been strengthened by a greater degree of public and stakeholder engagement, and the Committee on Radioactive Waste Management's handling of issues of uncertainty could have been strengthened by the explicitly probabilistic framework of Stern.


The UPR preserves mature oligodendrocyte viability and function in adults by regulating autophagy of PLP.
Maintaining cellular proteostasis is essential for oligodendrocyte viability and function; however, its underlying mechanisms remain unexplored. Unfolded protein response (UPR), which comprises 3 parallel branches, inositol requiring enzyme 1 (IRE1), pancreatic ER kinase (PERK), and activating transcription factor 6α (ATF6α), is a major mechanism that maintains cellular proteostasis by facilitating protein folding, attenuating protein translation, and enhancing autophagy and ER-associated degradation. Here we report that impaired UPR in oligodendrocytes via deletion of PERK and ATF6α did not affect developmental myelination but caused late-onset mature oligodendrocyte dysfunction and death in young adult mice. The detrimental effects of the impaired UPR on mature oligodendrocytes were accompanied by autophagy impairment and intracellular proteolipid protein (PLP) accumulation and were rescued by PLP deletion. Data indicate that PLP was degraded by autophagy and that intracellular PLP accumulation was cytotoxic to oligodendrocytes. Thus, these findings imply that the UPR is required for maintaining cellular proteostasis and the viability and function of mature oligodendrocytes in adults by regulating autophagy of PLP.


High prevalence of overweight and obesity among 6-year-old children in Finnmark County, North Norway.
The aim was to determine the prevalence of overweight and obesity among 6-year-old children in Finnmark, the northernmost county of Norway. This is a survey of 1774 children born during 1999 and 2000 from 18 of 19 child healthcare centres in Finnmark. Body mass index data extracted retrospectively in 2007 from health records at the age of 6 years were compared with international definitions of over- and underweight. The prevalence figures were further compared with socio-demographic figures on municipality level. Overall, 19% of the children were classified as overweight or obese; 5% were classified as obese. The prevalence of overweight and obesity was higher among girls (22%) than among boys (16%) (p < 0.01). The prevalence of underweight was 8% among both girls and boys. Despite large variations in the prevalence of overweight and obesity between municipalities (9-35%), no association was found with municipality figures on socio-demographic factors. In the northernmost county Finnmark, the prevalence of overweight including obesity among 6-year-old children was somewhat higher than in previous surveys from Norway, especially among girls.


Jump Shrug Height and Landing Forces Across Various Loads.
The purpose of this study was to examine the effect that load has on the mechanics of the jump shrug. Fifteen track and field and club/intramural athletes (age 21.7 ± 1.3 y, height 180.9 ± 6.6 cm, body mass 84.7 ± 13.2 kg, 1-repetition-maximum (1RM) hang power clean 109.1 ± 17.2 kg) performed repetitions of the jump shrug at 30%, 45%, 65%, and 80% of their 1RM hang power clean. Jump height, peak landing force, and potential energy of the system at jump-shrug apex were compared between loads using a series of 1-way repeated-measures ANOVAs. Statistical differences in jump height (P < .001), peak landing force (P = .012), and potential energy of the system (P < .001) existed; however, there were no statistically significant pairwise comparisons in peak landing force between loads (P > .05). The greatest magnitudes of jump height, peak landing force, and potential energy of the system at the apex of the jump shrug occurred at 30% 1RM hang power clean and decreased as the external load increased from 45% to 80% 1RM hang power clean. Relationships between peak landing force and potential energy of the system at jump-shrug apex indicate that the landing forces produced during the jump shrug may be due to the landing strategy used by the athletes, especially at lighter loads. Practitioners may prescribe heavier loads during the jump-shrug exercise without viewing landing force as a potential limitation.


Building reproductive health research and audit capacity and activity in the Pacific Islands (BRRACAP) study: methods, rationale and baseline results.
Clinical research and audit in reproductive health is essential to improve reproductive health outcomes and to address the Millennium Development Goals 4 and 5. Research training, mentoring and a supportive participatory research environment have been shown to increase research activity and capacity in low to middle income countries (LMIC). This paper details the methods, rationale and baseline findings of a research program aimed at increasing clinical research activity and audit in the six Pacific Islands of Fiji, Samoa, Tonga, Vanuatu, Cook Islands and the Solomon Islands. Twenty-eight clinician participants were selected by the five Ministries of Health and the Fiji National University to undergo a research capacity building program which includes a research workshop and mentoring support to perform research and audit as teams in their country. Data on the participants' characteristics, knowledge and experiences were collected from structured interviews, questionnaires, focus groups, and an online survey. The interviews and the two focus groups were audio-recorded and all replies were analysed in a thematic framework. The 28 participants included 9 nurses/midwives, 17 medical doctors of whom 8 were specialists in reproductive health and 2 other health workers. Most (24, 86%) were required to perform research as part of their employment and yet 17 (61%) were not confident in writing a research proposal, 13 (46%) could not use an electronic spreadsheet and the same number had not analysed quantitative data. The limited environmental enablers contributed to poor capacity with only 11 (46%) having access to a library, 10 (42%) receiving management support and 6 (25%) having access to an experienced researcher. Barriers to research that affected more than 70% of the participants were time constraints, poor coordination, no funding and a lack of skills. Building a research capacity program appropriate for the diversity of Pacific clinicians required research evidence and collaborative effort of key stakeholders in the Pacific Islands and the region. The participants had limited research knowledge, skills and experience and would require individualized training and continuous intensive mentorship to realize their potential as clinician researchers for their services in the Pacific.


MTHFR gene C677T and A1298C polymorphisms and homocysteine levels in primary open angle and primary closed angle glaucoma.
To investigate the methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C genotypes and plasma concentrations of total homocysteine (tHcy) in Pakistani patients with primary open angle glaucoma (POAG) and primary closed angle glaucoma (PCAG). This was a prospective case-control study. A total of 295 patients (173 POAG, 122 PCAG) and 143 age- and sex-matched controls were subdivided into two ethnic groups, Punjabis (Punjab province, central Pakistan) and Pathans (North-West Frontier Province, northern Pakistan). Genotypes of the MTHFR C677T and A1298C polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). An enzyme-linked immunosorbent assay was used to determine the total serum homocysteine (tHcy) levels. Associations were determined by logistic regression analysis. Frequency distributions of genotypes and combined genotypes as well as homocysteine levels were obtained. The overall distribution of the C677T genotype was found to be significantly associated with PCAG (CC 69%, CT 21%, TT 10%; p=0.001, chi(2)=12.6), but not with POAG (CC 71%, CT 28%, TT 1%; p=0.98, chi(2)=0.02) as compared to the controls (CC 71%, CT 29%, TT 1%). The Pathan cohorts revealed no association with the disease; however, the Punjabis demonstrated a significant association with PCAG (CC 75%, CT 11%, TT 13%; p<0.001, chi(2)=17.2). PCAG in the Punjabi subjects was also significantly associated with the A1298C polymorphism (AA 43%, AC 54%, CC 3%; p<0.001, chi(2)=33.9) as compared to the controls. Combined genotype data showed no association with POAG; however, a significant association with all combined genotypes was observed in the overall PCAG subjects (p<0.05, chi(2)=20.1). This difference was particularly apparent in the TTAA and TTAC combinations that were completely absent in the control groups (p<0.05. chi(2)=49.6). Mean serum tHcy levels were found to be significantly increased in the POAG (15.2+/-1.28 micromol/l, p<0.001) and PCAG (20.8+/-4.8 micromol/l) groups as compared to the controls (10.0+/-0.97 micromol/l). The tHcy levels in the TT and AC genotype were significantly elevated in the PCAG group (67+/-12.39 micromol/l, p<0.001; 23+/-5.94 micromol/l, p=0.027) as compared to the controls. The TT and AC genotypes of MTHFR C677T and A1298C polymorphisms and the combined genotype TTAC were associated with PCAG in Punjabi subjects of Pakistani origin and correlated with the high serum tHcy levels seen in these patients.


Steroid therapy for bacterial meningitis.
Routine dexamethasone therapy for bacterial meningitis in pediatric patients is controversial. Two experts debated this topic at the 1993 meeting of the Infectious Diseases Society of America. Both experts agreed that for management of Haemophilus influenzae meningitis, dexamethasone significantly reduced sensorineural hearing loss and probably reduced other long-term sequelae. Because relatively few patients with pneumococcal and meningococcal meningitis have been studied, no conclusions could be reached regarding the effectiveness of dexamethasone. Dr. Urs Schaad emphasized the impressive anti-inflammatory effects of dexamethasone in experimental pneumococcal meningitis and the lack of any adverse events when given to children for 2 or 4 days. He recommended routine use of dexamethasone in treating pediatric patients with bacterial meningitis. Dr. Sheldon Kaplan expressed concern regarding the effectiveness of steroids in treating pneumococcal meningitis, especially when penicillin-resistant and cephalosporin-resistant isolates are present, and he addressed the question of the long-term effects of administration of dexamethasone in children with viral meningitis. He advised against the routine use of dexamethasone for non-H. influenzae meningitis.


Methodologic issues in the use of bleeding as an outcome in transfusion medicine studies.
Prophylactic platelet transfusions are given to thrombocytopenic patients to prevent bleeding. The benefit of platelet transfusions has frequently been assessed by measuring the count increment; however, more recently, an assessment of bleeding has been used because it is a more clinically relevant outcome measure. The purpose of this study was to identify platelet transfusion trigger studies that used bleeding as an outcome measure, compare and contrast methods used to document bleeding and analyze bleeding outcomes, and identify and discuss methodologic issues to consider when bleeding is used as a study outcome. A systematic search to identify platelet transfusion trigger studies was performed. Relevant articles were reviewed to identify how bleeding data was captured and analyzed, and methodologic considerations were identified. Seven articles meeting the predefined entry criteria were identified. Methods used to document bleeding included chart review and clinical assessment. The frequency of assessment and the type of personnel performing the assessment were variable. Four approaches to analysis were identified: descriptive; comparison of the proportions of patients having at least one bleed; comparison of patient days with bleeding expressed as a proportion of the total days at risk of bleeding; and time-to-event (first bleed) analysis. Methodologic issues for consideration when designing a clinical study with bleeding as the outcome measure included approaches to minimize bias in the documentation and classification of bleeding and selection of an analysis approach that is appropriate to the question being asked. The need for development of a valid and reliable bleeding scale was also identified.


A framework for experimental scenarios of global change in marine systems using coral reefs as a case study.
Understanding the consequences of rising CO2 and warming on marine ecosystems is a pressing issue in ecology. Manipulative experiments that assess responses of biota to future ocean warming and acidification conditions form a necessary basis for expectations on how marine taxa may respond. Although designing experiments in the context of local variability is most appropriate, local temperature and CO2 characteristics are often unknown as such measures necessitate significant resources, and even less is known about local future scenarios. To help address these issues, we summarize current uncertainties in CO2 emission trajectories and climate sensitivity, examine region-specific changes in the ocean, and present a straightforward global framework to guide experimental designs. We advocate for the inclusion of multiple plausible future scenarios of predicted levels of ocean warming and acidification in forthcoming experimental research. Growing a robust experimental base is crucial to understanding the prospect form and function of marine ecosystems in the Anthropocene.


Fabrication of micro DOE using micro tools shaped with focused ion beam.
A novel method is proposed to fabricate micro Diffractive Optical Elements (DOE) using micro cutting tools shaped with focused ion beam (FIB) milling. Micro tools with nanometric cutting edges and complicated shapes are fabricated by controlling the tool facet's orientation relative to the FIB. The tool edge radius of less than 30 nm is achieved for the nano removal of the work materials. Semi-circular micro tools and DOE-shaped micro tools are developed to fabricate micro-DOE and sinusoidal modulation templates. Experiments show that the proposed method can be a high efficient way in fabricating micro-DOE with nanoscale surface finishes.


Evaluation of immunodiagnostic potential of ESAT-6 synthetic peptides mixture in Egyptian pulmonary tuberculosis patients.
Identification of M. tuberculosis ESAT-6 proteins has enhanced research for developing new specific and reliable in vitro diagnostic tests for pulmonary tuberculosis. The aim of this study was to assess the diagnostic potential of ESAT-6 synthetic overlapping peptides mixture (Pepmix) for diagnosis of pulmonary M. tuberculosis infection in Egypt. IFN-gammaand IL-17 production was assessed in whole-blood culture assay after overnight stimulation with M. tuberculosis crude culture filtrate (CF), purified 38 kDa antigen, and ESAT-6 Pepmix. Concentration of IFN-gamma and IL-17 was evaluated in the culture supernatant by ELISA. Patients with active pulmonary TB patients (n = 37), Tuberculin Skin Test (TST) positive healthy subjects (n = 16) and TST negative healthy subjects (n = 16) were included. Results indicated that IFN-gammaproduction following stimulation with crude CF, and 38 kDa purified antigen was not significantly different between patients and TST+ healthy controls. In contrast, Pepmix clearly distinguished between patients and TST+ healthy subjects with sensitivity of 70.3 % and specificity 96.9 %. The concentration of secreted IL-17 was significantly lower than IFN-gamma, and no statistically significant difference was found between groups. It is concluded that IFN-gamma assay using ESAT-6 Pepmix, as antigenic stimulus, has the potential to rapidly diagnose pulmonary TB infection.


PfSETvs methylation of histone H3K36 represses virulence genes in Plasmodium falciparum.
The variant antigen Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), which is expressed on the surface of P. falciparum-infected red blood cells, is a critical virulence factor for malaria. Each parasite has 60 antigenically distinct var genes that each code for a different PfEMP1 protein. During infection the clonal parasite population expresses only one gene at a time before switching to the expression of a new variant antigen as an immune-evasion mechanism to avoid the host antibody response. The mechanism by which 59 of the 60 var genes are silenced remains largely unknown. Here we show that knocking out the P. falciparum variant-silencing SET gene (here termed PfSETvs), which encodes an orthologue of Drosophila melanogaster ASH1 and controls histone H3 lysine 36 trimethylation (H3K36me3) on var genes, results in the transcription of virtually all var genes in the single parasite nuclei and their expression as proteins on the surface of individual infected red blood cells. PfSETvs-dependent H3K36me3 is present along the entire gene body, including the transcription start site, to silence var genes. With low occupancy of PfSETvs at both the transcription start site of var genes and the intronic promoter, expression of var genes coincides with transcription of their corresponding antisense long noncoding RNA. These results uncover a previously unknown role of PfSETvs-dependent H3K36me3 in silencing var genes in P. falciparum that might provide a general mechanism by which orthologues of PfSETvs repress gene expression in other eukaryotes. PfSETvs knockout parasites expressing all PfEMP1 proteins may also be applied to the development of a malaria vaccine.


Immunogenicity and efficacy of Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan peptide mimotope-protein conjugates in human immunoglobulin transgenic mice.
Peptide mimotopes of capsular polysaccharides have been proposed as antigens for vaccines against encapsulated pathogens. In this study, we determined the antibody response to and efficacy of P13, a peptide mimetic of the Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan (GXM), in mice that produce human antibodies. P13 was conjugated to tetanus toxoid (TT) or diphtheria toxoid (DT) and administered subcutaneously in Alhydrogel with or without CpG to mice transgenic for human immunoglobulin loci (XenoMouse mice) and expressing either immunoglobulin G2 (IgG2) (G2 mice) or IgG4 (G4 mice). Mice were vaccinated and revaccinated two or three times. The serum antibody responses of the mice to GXM and P13 and antibody idiotype expression were analyzed by an enzyme-linked immunosorbent assay. The results showed that both P13-TT and P13-DT were antigenic, inducing a mimetic response to P13 in both G2 and G4 mice, and immunogenic, inducing a mimotope response including VH3 (idiotype)-positive antibodies to GXM in G2 but not G4 mice. CpG led to higher titers of IgG to P13 and GXM in P13-TT-vaccinated G2 mice. C. neoformans challenge of P13-protein conjugate-vaccinated and control G2 mice induced anamnestic IgG- and VH3-positive responses to GXM and was associated with a significantly decreased risk of death and a prolongation of survival in P13-DT-vaccinated mice compared to phosphate-buffered saline-treated or protein carrier-vaccinated mice. These findings reveal that P13 elicited a human antibody response with VH3 expression in human immunoglobulin transgenic mice that has been observed for human antibodies to GXM and support the concept that peptide mimotope-based vaccines may hold promise for the treatment of C. neoformans infections.


Hand-held and integrated single-cell pipettes.
Successful single-cell isolation is a primary step for subsequent chemical and biological analyses of single cells. Conventional single-cell isolation methods often encounter operational complexity, limited efficiency, deterioration of cell viability, incompetence in the isolation of a single-cell into nanoliter liquid, and/or inability to select single adherent cells with specific phenotypes. Here, we develop a hand-held single-cell pipet (hSCP) that is rapid, operationally simple, highly efficient, and inexpensive for unbiased isolation of single viable suspended cells directly from submicroliter cell suspensions into nanoliter droplets without the assistance of any additional equipment. An integrated SCP (iSCP) has also been developed for selective isolation of single suspended and adherent cells according to the fluorescence imaging and morphological features. The isolated single cells can be conveniently transferred into standard 96-/384-well plates, Petri dishes, or vials for cloning, PCR, and other single-cell biochemical assays.


Erythropoietin Increases GABAA Currents in Human Cortex from TLE Patients.
Erythropoietin (EPO) is a hematopoietic growth factor that has an important role in the erythropoiesis. EPO and its receptor (EPO-R) are expressed all over in the mammalian brain. Furthermore, it has been reported that EPO may exert neuroprotective effect in animal models of brain disorders as ischemia and epilepsy. Here, we investigate whether EPO could modulate the GABA-evoked currents (IGABA) in both human epileptic and non-epileptic control brain tissues. Therefore, we transplanted in Xenopus oocytes cell membranes obtained from autoptic and surgical brain tissues (cortex) of seven temporal lope epilepsy (TLE) patients and of five control patients. Two microelectrodes voltage-clamp technique has been used to record IGABA. Moreover, qRT-PCR assay was performed in the same human tissues to quantify the relative gene expression levels of EPO/EPO-R. To further confirm experiments in oocytes, we performed additional experiments using patch-clamp recording in slices obtained from rat cerebellum. We show that exposure to EPO significantly increased the amplitude of the IGABA in all the patients analyzed. No differences in the expression of EPO and EPO-R in both TLE and control patients have been found. Notably, the increase of IGABA has been recorded also in rat cerebellar slices. Our findings show a new modulatory action of EPO on GABAA receptors (GABAA-Rs). This effect could be relevant to balance the GABAergic dysfunction in human TLE. This article is part of a Special Issue entitled: Honoring Ricardo Miledi - outstanding neuroscientist of XX-XXI centuries.


Cannabidiol in sport: Ergogenic or else?
In the sports domain, cannabis is prohibited by the World Anti-Doping Agency (WADA) across all sports in competition since 2004. The few studies on physical exercise and cannabis focused on the main compound i.e. Δ9-tetrahydrocannabinol. Cannabidiol (CBD) is another well-known phytocannabinoid present in dried or heated preparations of cannabis. Unlike Δ9-tetrahydrocannabinol, CBD is non-intoxicating but exhibits pharmacological properties that are interesting for medical use. The worldwide regulatory status of CBD is complex and this compound is still a controlled substance in many countries. Interestingly, however, the World Anti-Doping Agency removed CBD from the list of prohibited substances - in or out of competition - since 2018. This recent decision by the WADA leaves the door open for CBD use by athletes. In the present opinion article we wish to expose the different CBD properties discovered in preclinical studies that could be further tested in the sport domain to ascertain its utility. Preclinical studies suggest that CBD could be useful to athletes due to its anti-inflammatory, analgesic, anxiolytic, neuroprotective properties and its influence on the sleep-wake cycle. Unfortunately, almost no clinical data are available on CBD in the context of exercise, which makes its use in this context still premature.


Three pairs of cysteine residues mediate both redox and zn2+ modulation of the nmda receptor.
NMDA receptor activity is modulated by various compounds, including sulfhydryl redox agents and Zn(2+). In addition to a slow and persistent component of redox modulation common to all NMDA receptors, NR1/NR2A receptors uniquely have a rapid and reversible component that has been variously attributed to redox or Zn(2+) effects. Here we show that this rapid modulatory effect can be described by two time constants with relatively fast ( approximately 6 sec) and intermediate (60 sec) half lives, and it is likely to be attributable to both redox agents and Zn(2+). Using site-directed mutagenesis, we identified three pairs of cysteine residues that underlie the various kinetic components of redox modulation of NMDA-evoked currents in Xenopus oocytes expressing NR1/NR2A receptors: (1) Cys 87 and Cys 320 in NR2A underlie the fast component, (2) Cys 79 and Cys 308 in NR1 underlie the intermediate component, and (3) Cys 744 and Cys 798 in NR1 underlie the persistent component. Mutation of these redox-sensitive cysteine residues also affects high-affinity, voltage-independent Zn(2+) inhibition that is specific to NR1/NR2A receptors. Exposure to methanethiosulfonate agents that modify cysteine residues did not block the Zn(2+) inhibition. Thus, these cysteine residues do not appear to coordinate Zn(2+) directly. Instead, the redox status of these cysteine residues may modulate the sensitivity of the receptor to Zn(2+).


Design of retrodiffraction gratings for polarization-insensitive and polarization-sensitive characteristics by using the Taguchi method.
We present the design of retrodiffraction gratings that utilize total internal reflection (TIR) in a lamellar configuration to achieve high performance for both TE and TM polarized light and polarization-sensitive performance for gratings behaving as polarizer filters; the design was based on rigorous coupled wave analysis (RCWA) and the Taguchi method. The components can thus be fabricated from a single dielectric material and do not have to be coated with a metallic or dielectric film layer to enhance the reflectance. The effects of the structural and optical parameters of lamellar gratings were investigated, and the TIR gratings in a lamellar configuration were structurally and optically optimized in terms of the signal-to-noise ratio (S/N) and a statistical analysis of variance (ANOVA) of the refractive index, grating period, filling factor, and grating depth as control factors and the estimated efficiency by RCWA as a noise factor. For more accurate robustness, a two-step optimization process was used for each purpose. For TIR gratings designed to perform similarly for TE and TM incident polarization, the -1st-order efficiencies were estimated to be up to 92.0% and 88.5% for TE and TM polarization, respectively. Also, for the TIR gratings designed to achieve polarization-sensitive performance when behaving as a polarizer filters, the -1st-order diffraction efficiencies for TE and TM polarization were estimated to be up to 95.5% and 2.7%, respectively. From these analysis results, it was confirmed that the Taguchi method shows feasibility for an optimization approach to a technique for designing optical devices.


A Systematic Review of Hospital-to-School Reintegration Interventions for Children and Youth with Acquired Brain Injury.
We reviewed the literature on interventions that aimed to improve hospital-to-school reintegration for children and youth with acquired brain injury (ABI). ABI is the leading cause of disability among children and youth. A successful hospital-to-school reintegration process is essential to the rehabilitative process. However, little is known about the effective components of of such interventions. Our research team conducted a systematic review, completing comprehensive searches of seven databases and selected reference lists for relevant articles published in a peer-reviewed journal between 1989 and June 2014. We selected articles for inclusion that report on studies involving: a clinical population with ABI; sample had an average age of 20 years or younger; an intentional structured intervention affecting hospital-to-school transitions or related components; an experimental design; and a statistically evaluated health outcome. Two independent reviewers applied our inclusion criteria, extracted data, and rated study quality. A meta-analysis was not feasible due to the heterogeneity of the studies reported. Of the 6933 articles identified in our initial search, 17 articles (reporting on 350 preadolescents and adolescents, aged 4-19, (average age 11.5 years, SD: 2.21) met our inclusion criteria. They reported on interventions varying in number of sessions (one to 119) and session length (20 minutes to 4 hours). The majority of interventions involved multiple one-to-one sessions conducted by a trained clinician or educator, homework activities, and parental involvement. The interventions were delivered through different settings and media, including hospitals, schools, and online. Although outcomes varied (with effect sizes ranging from small to large), 14 of the articles reported at least one significant improvement in cognitive, social, psychological, or behavioral functioning or knowledge of ABI. Cognitive, behavioral, and problem-solving interventions have the potential to improve school reintegration for youth with ABI. However, more comprehensive interventions are needed to help link rehabilitation clinicians, educators, adolescents, and families.


Bcl-2 inhibits apoptosis induced by mitochondrial uncoupling but does not prevent mitochondrial transmembrane depolarization.
Bcl-2 overexpression protects cells from apoptosis induced by many cytotoxic agents. In this study, we investigated the effects of uncoupling mitochondrial electron transport in both HL60 wild-type and Bcl-2-overexpressing cells using the protonophore carbonyl cyanide m-chlorophenylhydrazone. We found that uncoupling mitochondrial electron transport induced apoptosis in wild-type, but not in Bcl-2-overexpressing cells. To investigate the mechanism of action of Bcl-2-mediated inhibition of cyanide m-chlorophenylhydrazone-induced apoptosis, we measured the mitochondrial transmembrane potential (DeltaPsi(m)) after uncoupling mitochondrial electron transport and found that both HL-60 wild-type and Bcl-2-overexpressing cells similarly depolarize following cyanide m-chlorophenylhydrazone exposure. Western blot analysis demonstrated that Bcl-2 overexpression did not completely block cytochrome c release from mitochondria after uncoupling mitochondrial electron transport. Since Bcl-2 may act as an antioxidant, we studied the effect of altering the cellular redox state prior to uncoupling mitochondrial electron transport in Bcl-2-overexpressing cells. Depletion of mitochondrial (but not cytosolic) glutathione induced apoptosis in Bcl-2-overexpressing cells and negated the protective effect of Bcl-2. Furthermore, following glutathione depletion, Bcl-2-overexpressing cells were sensitized to undergo cyanide m-chlorophenylhydrazone-induced apoptosis. These data suggest that the action of Bcl-2 is dependent, in part, on the cellular and mitochondrial redox state.


A systematic survey of the quality of research reporting in general orthopaedic journals.
The poor reporting and use of statistical methods in orthopaedic papers has been widely discussed by both clinicians and statisticians. A detailed review of research published in general orthopaedic journals was undertaken to assess the quality of experimental design, statistical analysis and reporting. A representative sample of 100 papers was assessed for compliance to CONSORT and STROBE guidelines and the quality of the statistical reporting was assessed using a validated questionnaire. Overall compliance with CONSORT and STROBE guidelines in our study was 59% and 58% respectively, with very few papers fulfilling all criteria. In 37% of papers patient numbers were inadequately reported; 20% of papers introduced new statistical methods in the 'results' section not previously reported in the 'methods' section, and 23% of papers reported no measurement of error with the main outcome measure. Taken together, these issues indicate a general lack of statistical rigour and are consistent with similar reviews undertaken in a number of other scientific and clinical research disciplines. It is imperative that the orthopaedic research community strives to improve the quality of reporting; a failure to do so could seriously limit the development of future research.


Irregular S-cone mosaics in felid retinas. Spatial interaction with axonless horizontal cells, revealed by cross correlation.
In most mammals short-wavelength-sensitive (S) cones are arranged in irregular patterns with widely variable intercell distances. Consequently, mosaics of connected interneurons either may show some type of correlation to photoreceptor placement or may establish an independent lattice with compensatory dendritic organization. Since axonless horizontal cells (A-HC's) are supposed to direct all dendrites to overlying cones, we studied their spatial interaction with chromatic cone subclasses. In the cheetah, the bobcat, and the leopard, anti-S-opsin antibodies have consistently colabeled the A-HC's in addition to the S cones. We investigated the interaction between the two cell mosaics, using autocorrelation and cross-correlation procedures, including a Voronoi-based density probe. Comparisons with simulations of random mosaics show significantly lower densities of S cones above the cell bodies and primary dendrites of A-HC's. The pattern results in different long-wavelength-sensitive-L- and S-cone ratios in the central versus the peripheral zones of A-HC dendritic fields. The existence of a related pattern at the synaptic level and its potential significance for color processing may be investigated in further studies.


PLK1 (polo like kinase 1) inhibits MTOR complex 1 and promotes autophagy.
Mechanistic target of rapamycin complex 1 (MTORC1) and polo like kinase 1 (PLK1) are major drivers of cancer cell growth and proliferation, and inhibitors of both protein kinases are currently being investigated in clinical studies. To date, MTORC1's and PLK1's functions are mostly studied separately, and reports on their mutual crosstalk are scarce. Here, we identify PLK1 as a physical MTORC1 interactor in human cancer cells. PLK1 inhibition enhances MTORC1 activity under nutrient sufficiency and in starved cells, and PLK1 directly phosphorylates the MTORC1 component RPTOR/RAPTOR in vitro. PLK1 and MTORC1 reside together at lysosomes, the subcellular site where MTORC1 is active. Consistent with an inhibitory role of PLK1 toward MTORC1, PLK1 overexpression inhibits lysosomal association of the PLK1-MTORC1 complex, whereas PLK1 inhibition promotes lysosomal localization of MTOR. PLK1-MTORC1 binding is enhanced by amino acid starvation, a condition known to increase autophagy. MTORC1 inhibition is an important step in autophagy activation. Consistently, PLK1 inhibition mitigates autophagy in cancer cells both under nutrient starvation and sufficiency, and a role of PLK1 in autophagy is also observed in the invertebrate model organism Caenorhabditis elegans. In summary, PLK1 inhibits MTORC1 and thereby positively contributes to autophagy. Since autophagy is increasingly recognized to contribute to tumor cell survival and growth, we propose that cautious monitoring of MTORC1 and autophagy readouts in clinical trials with PLK1 inhibitors is needed to develop strategies for optimized (combinatorial) cancer therapies targeting MTORC1, PLK1, and autophagy.


Phosphazene vs.diazaphospholene PN-bond cleavage in spirocyclic cyclodiphosphazenes.
Thermolysis of 2-azido-1,3,2-diazaphospholenes offers access to novel and rare spirocyclic cyclodiphosphazenes. The spectroscopic data and X-ray structure of one representative of the 2-azido-1,3,2-diazaphospholenes reveals an ionic bonding situation explaining sufficiently its rather high thermal stability. The cyclodiphosphazenes were characterised by NMR, mass spectrometry, and X-ray diffractometry. The results of ESI-FT-ICR studies demonstrate the potential of these compounds to undergo reductive elimination at a phosphazene unit via [1,4]-cycloreversion of a lambda5-diazaphospholene ring, as well as symmetrical cleavage of the P2N2-unit. The unexpected inclusion of benzene in the crystal of one of the cyclodiphosphazenes was interpreted in terms of molecular recognition. Chemical reaction studies comprise the proof of double N-protonation at a phosphazene ring, and hydrolytic degradation via selective cleavage of a phosphazene P-N bond.


Large-scale ab initio simulations of binary transition metal clusters for storage media materials.
In the quest for ultra-high-density magnetic recording, new materials in the nanometre range have attracted much interest over the last decade involving intense studies of L1(0) phases of contemporary or future storage media materials like FePt or CoPt nanoparticles. Based on large-scale density functional theory calculations, we provide a systematic overview of the structural and magnetic properties of various morphologies of FePt and CoPt nanoclusters with diameters up to 3 nm. In this size range, the ordered multiply twinned morphologies are energetically favoured over the nanoclusters with the desired layer type L1(0) and high magnetocrystalline anisotropy. Other nanoparticles of interest, like FePd, also show a preference for multiply twinned structures or exhibit, as in the case of MnPt nanoclusters, a strong tendency for antiferromagnetic ordering instead of ferromagnetic order. The compositional trends of the various nanoparticles can be traced back to differences in the partial electronic density of states of the 3d element.


Connected, Respected and Contributing to Their World: The Case of Sexual Minority and Non-Minority Young People in Ireland.
Outcome 5 of the Irish Better Outcomes, Brighter Futures national youth policy framework ("Connected, respected, and contributing to their world") offers a suitable way to study psychosocial determinants of adolescent health. The present study (1) provides nationally representative data on how 15- to 17-year-olds score on these indicators; (2) compares sexual minority (same- and both-gender attracted youth) with their non-minority peers. We analyzed data from 3354 young people (aged 15.78 ± 0.78 years) participating in the Health Behaviour in School-aged Children (HBSC) study in Ireland. Age and social class were associated with the indicators only to a small extent, but girls were more likely than boys to report discrimination based on gender and age. Frequency of positive answers ranged from 67% (feeling comfortable with friends) to 12% (being involved in volunteer work). Sexual minority youth were more likely to feel discriminated based on sexual orientation, age, and gender. Both-gender attracted youth were less likely than the other groups to report positive outcomes. Same-gender attracted youth were twice as likely as non-minority youth to volunteer. The results indicate the importance of a comprehensive approach to psycho-social factors in youth health, and the need for inclusivity of sexual minority (especially bisexual) youth.


Sex differences in patients with acute ischemic stroke in Tuzla region, Bosnia and Herzegovina.
Although many aspects of stroke are similar at both sexes, however, there are some differences and characteristics as well. The aim of this study was to analyze sex differences in patients with acute ischemic stroke (IS) regarding to risk factors, subtypes, stroke severity and outcome. From January 1st 2001 to December 31st 2005 at the Department of Neurology Tuzla 2833 patients were admitted with acute ischemic stroke (IS). We were analyzed risk factors, subtypes, stroke severity (Scandinavian Stroke Scale), and thirty-day outcome. There were 1484 (52.3%) female, and they were older than male (67.8 +/- 10.6 vs. 65.7 +/- 10.5, p<0.0001). Hypertension (78% vs. 67%, p<0.0001), heart diseases (50% vs. 45%, p=0.009), atrial fibrillation (22% vs. 14%, p<0.0001) and diabetes mellitus (33% vs. 21%, p<0.0001) were frequently in female, while smoking (45% vs. 14%) and alcohol overuse (18% vs. 0,6%) in male (p<0.0001). Atherothrombotic type of ischemic stroke was frequently in male (37.4% vs. 31.6%, p=0.0013) and cardioembolic in female (21.7% vs. 15.5%, p<0.0001). At admission female had lower SS (SS 31.0 +/- 15 vs. 34.0 +/- 15, p<0.0001). Thirty-day mortality was significantly higher in female (23.3% vs. 18.4%, p=0.0015), and favourable outcome within one month (Rankin Scale <or= 2) had 58% male and 51% female (p=0.001). The frequency of ischemic stroke is higher in female who are older than male. There are some sex differences according to the distribution of risk factors and subtypes of ischemic stroke. Stroke severity at admission, thirty-day mortality, and disability are higher in female.


Rationale and design of the POLEM trial: avelumab plus fluoropyrimidine-based chemotherapy as adjuvant treatment for stage III mismatch repair deficient or POLE exonuclease domain mutant colon cancer: a phase III randomised study.
10%-15% of early-stage colon cancers harbour either deficient mismatch repair (dMMR), microsatellite instability high (MSI-H) or POLE exonuclease domain mutations, and are characterised by high tumour mutational burden and increased lymphocytic infiltrate. Metastatic dMMR colon cancers are highly sensitive to immune checkpoint inhibition, and recent data show POLE-mutant tumours are similarly responsive. We are conducting a phase III randomised trial to determine if the addition of the anti-PD-L1 antibody avelumab following adjuvant chemotherapy improves disease-free survival (DFS) in patients with stage III dMMR/MSI-H or POLE mutant colon cancer and is a cost-effective approach for the UK National Health Service (NHS). We are recruiting patients with completely resected, stage III colon cancer confirmed to have dMMR/MSI-H, locally or POLE exonuclease domain mutation on central testing. Eligible patients are randomised in a 1:1 ratio to standard fluoropyrimidine-based chemotherapy (capecitabine, oxaliplatin for 12 weeks or capecitabine for 24 weeks) or chemotherapy, followed by avelumab (10 mg/kg, 2 weekly for 24 weeks). Stratification is by chemotherapy received and MMR/MSI-H status. The primary endpoint is DFS. Secondary endpoints include overall survival, toxicity, quality of life and health resource use. The 3-year DFS rate in the control arm is expected to be ~75%. Avelumab is expected to improve the 3-year DFS rate by 12% (ie, 87%). Target accrual is 402 patients, which provides 80% power to detect an HR of 0.48 for DFS at a two-sided alpha of 0.05. This national, multicentre phase III trial is sponsored by the Royal Marsden NHS Foundation Trust and it is anticipated that approximately 40 centres in the UK will participate. This study opened to recruitment in August 2018. NCT03827044.


What were the advantages of microendoscopic discectomy for lumbar disc herniation comparing with open discectomy: a meta-analysis?
The purpose of this study was to compare the safety and efficacy of micro-endoscopic discectomy (MED) and open discectomy (OD) for lubmar disc herniation (LDH). Randomised controlled trials (RCTs) comparing MED with OD for LDH were searched comprehensively in PubMed, EMBASE, the Cochrane Library. Relevant studies retrieved, data extracted and the quality of included studies were independently performed by two authors. RevMan software (Version 5.2.0) was used to analyse and synthesis relevant data of the included studies. Nine RCTs involving 774 patients were obtained and reported the relevant outcome measures. Compared with OD group, there were significant difference in the general operation indicators including operation time, blood loss, site of incision, hospital stay and time of return to work, biochemical indexes including C-reactive protein (CRP) and interleukin-6 (IL-6) in MED group. Meanwhile, there were no difference in effective rate, complication including total complications, dural leaks occurred and recurrence of the disc herniation, compared MED group with OD group. MED had slighter trauma, milder blood loss and shorter healing time than OD. The results demonstrated MED has great efficacy and safety comparable to OD. So we think that MED can be used routinely for LDH patients, especially the patients of old and intolerable major surgery. Meanwhile, it is necessary for surgeon to master indication and contraindication of MED and improve the operative technique.


Generation of an artificial ring chromosome in Arabidopsis by Cre/LoxP-mediated recombination.
A eukaryotic chromosome consists of a centromere, two telomeres and a number of replication origins, and 'artificial chromosomes' may be created in yeast and mammals when these three elements are artificially joined and introduced into cells. Plant artificial chromosomes (PACs) have been suggested as new vectors for the development of new crops and as tools for basic research on chromosomes. However, indisputable PAC formation has not yet been confirmed. Here, we present a method for generating PACs in the model plant Arabidopsis thaliana using the Cre/LoxP and Activator/Dissociation element systems. The successfully generated PAC, designated AtARC1 (A. thaliana artificial ring chromosome 1), originated from a centromeric edge of the long arm of chromosome 2, but its size (2.85 Mb) is much smaller than that of the original chromosome (26.3 Mb). Although AtARC1 contains only a short centromere domain consisting of 180 bp repeats approximately 250 kb in length, compared with the 3 Mb domain on the original chromosome 2, centromere-specific histone H3 (HTR12) was detected on the centromeric region. This result supported the observed stability of the PAC during mitosis in the absence of selection, and transmission of the PAC to the next generation through meiosis. Because AtARC1 contains a unique LoxP site driven by the CaMV 35S promoter, it is possible to introduce a selectable marker and desired transgenes into AtARC1 at the LoxP site using Cre recombinase. Therefore, AtARC1 meets the criteria for a PAC and is a promising vector.


Finishing the finished human chromosome 22 sequence.
Although the human genome sequence was declared complete in 2004, the sequence was interrupted by 341 gaps of which 308 lay in an estimated approximately 28 Mb of euchromatin. While these gaps constitute only approximately 1% of the sequence, knowledge of the full complement of human genes and regulatory elements is incomplete without their sequences. We have used a combination of conventional chromosome walking (aided by the availability of end sequences) in fosmid and bacterial artificial chromosome (BAC) libraries, whole chromosome shotgun sequencing, comparative genome analysis and long PCR to finish 8 of the 11 gaps in the initial chromosome 22 sequence. In addition, we have patched four regions of the initial sequence where the original clones were found to be deleted, or contained a deletion allele of a known gene, with a further 126 kb of new sequence. Over 1.018 Mb of new sequence has been generated to extend into and close the gaps, and we have annotated 16 new or extended gene structures and one pseudogene. Thus, we have made significant progress to completing the sequence of the euchromatic regions of human chromosome 22 using a combination of detailed approaches. Our experience suggests that substantial work remains to close the outstanding gaps in the human genome sequence.


Enzymatic synthesis and properties of glycoconjugates with legionaminic acid as a replacement for neuraminic acid.
In addition to sialic acid, bacteria produce several other nonulosonic acids, including legionaminic acid (Leg). This has exactly the same stereochemistry as sialic acid, with the added features of 9-deoxy and 7-amino groups. In order to explore the biological effects of replacing sialic acid residues (Neu5Ac) in glycoconjugates with Leg in its diacetylated form, diacetyllegionaminic acid (Leg5Ac7Ac), we tested CMP-Leg5Ac7Ac as a donor substrate with a selection of bacterial and mammalian sialyltransferases. The CMP-Leg5Ac7Ac was synthesized in vitro by means of cloned enzymes from the bacillosamine portion of the Campylobacter jejuni N-glycan pathway and from the Leg pathway of Legionella pneumophila. Using fluorescent derivatives of lactose, Galβ1,4GlcNAcβ and T-antigen (Galβ1,3GalNAcα) as acceptors, we tested eight different sialyltransferases and found that the Pasteurella multocida PM0188h and porcine ST3Gal1 sialyltransferases were significantly active with CMP-Leg5Ac7Ac, showing ∼60% activity when compared with CMP-Neu5Ac. The Photobacterium α2,6 sialyltransferase was weakly active, with ∼6% relative activity. The Leg5Ac7Ac-α-2,3-lactose product was then tested as a substrate with six sialidases of viral, bacterial and mammalian origin. All showed much lower activities than with the corresponding sialic acid substrate, with the influenza virus N1 being the most active and human NEU2 being the least active. These results show the feasibility of producing glycoconjugates with Leg5Ac7Ac residues as the terminal sugars, which should display novel biological properties.


American Society of Hematology living guidelines on the use of anticoagulation for thromboprophylaxis in patients with COVID-19: July 2021 update on postdischarge thromboprophylaxis.
COVID-19-related acute illness is associated with an increased risk of venous thromboembolism (VTE). These evidence-based guidelines of the American Society of Hematology (ASH) are intended to support patients, clinicians, and other health care professionals in decisions about the use of anticoagulation for thromboprophylaxis in patients with COVID-19 who do not have confirmed or suspected VTE. ASH formed a multidisciplinary guideline panel, including 3 patient representatives, and applied strategies to minimize potential bias from conflicts of interest. The McMaster University GRADE Centre supported the guideline development process, including performing systematic evidence reviews (up to March 2021). The panel prioritized clinical questions and outcomes according to their importance for clinicians and patients. The panel used the grading of recommendations assessment, development, and evaluation (GRADE) approach to assess evidence and make recommendations, which were subject to public comment. The panel agreed on 1 additional recommendation. The panel issued a conditional recommendation against the use of outpatient anticoagulant prophylaxis in patients with COVID-19 who are discharged from the hospital and who do not have suspected or confirmed VTE or another indication for anticoagulation. This recommendation was based on very low certainty in the evidence, underscoring the need for high-quality randomized controlled trials assessing the role of postdischarge thromboprophylaxis. Other key research priorities include better evidence on assessing risk of thrombosis and bleeding outcomes in patients with COVID-19 after hospital discharge.


A comprehensive model for right-left heart interaction under the influence of pericardium and baroreflex.
A phenomenological model of the cardiopulmonary circulation is developed with a focus on the interaction between the right heart and the left heart. The model predicts the hemodynamic consequences of changing circulatory parameters in terms of a broad spectrum of pressure and flow waveforms. Hemodynamics are characterized by use of an electrical analog incorporating mechanisms for transseptal pressure coupling, pericardial volume coupling, intrathoracic pressure, and baroreflex control of heart rate. Computer simulations are accomplished by numerically integrating 28 differential equations that contain nonlinear and time-varying coefficients. Validity of the model is supported by its accurate fit to clinical pressure and Doppler echocardiographic recordings. The model characterizes the hemodynamic waveforms for mitral stenosis, mitral regurgitation, left heart failure, right heart failure, cardiac tamponade, pulsus paradoxus, and the Valsalva maneuver. The wave shapes of pulmonary capillary wedge pressure under the above conditions are also accurately represented. Sensitivity analysis reveals that simulated hemodynamics are insensitive to most individual model parameters with the exception of afterload resistance, preload capacitances, intrathoracic pressure, contractility, and pericardial fluid volume. Baseline hemodynamics are minimally affected by transseptal coupling (up to 2%) and significantly affected by pericardial coupling (up to 20%). The model should be useful for quantitative studies of cardiopulmonary dynamics related to the right-left heart interaction under normal and disease conditions.


An Overview on Principles for Energy Efficient Robot Locomotion.
Despite enhancements in the development of robotic systems, the energy economy of today's robots lags far behind that of biological systems. This is in particular critical for untethered legged robot locomotion. To elucidate the current stage of energy efficiency in legged robotic systems, this paper provides an overview on recent advancements in development of such platforms. The covered different perspectives include actuation, leg structure, control and locomotion principles. We review various robotic actuators exploiting compliance in series and in parallel with the drive-train to permit energy recycling during locomotion. We discuss the importance of limb segmentation under efficiency aspects and with respect to design, dynamics analysis and control of legged robots. This paper also reviews a number of control approaches allowing for energy efficient locomotion of robots by exploiting the natural dynamics of the system, and by utilizing optimal control approaches targeting locomotion expenditure. To this end, a set of locomotion principles elaborating on models for energetics, dynamics, and of the systems is studied.


Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture.
The equine granulocytic ehrlichiosis agent, Ehrlichia equi, is closely related or identical to the human granulocytic ehrlichiosis (HGE) agent. Both are suspected of being transmitted by ticks. We have successfully isolated E. equi in a cell line, IDE8, derived from a putative vector, the tick Ixodes scapularis. Peripheral blood leukocytes from an experimentally infected horse were inoculated onto IDE8 monolayers. Cultures were incubated in a candle jar at 34 degrees C in tick cell culture medium with NaHCO3 and an organic buffer [3-(N-morpholino)-propanesulfonic acid] (MOPS). Within 2 weeks, infected cells were detected in Giemsa-stained culture samples, and the organisms subsequently spread to uninfected cells in the cultures. E. equi was passaged serially by transferring a portion of an infected culture to new cell layers every 2 to 3 weeks. The identity of the organisms was confirmed by PCR using oligonucleotide primers specific for E. equi and the HGE agent and by immunocytology. Homologous equine antibodies and human anti-HGE convalescent serum recognized E. equi grown in tick cell culture. Electron microscopy revealed electron-lucent and -dense ehrlichia-like forms developing within host cell endosomes. E. equi passaged twice in tick cell culture retained infectivity and pathogenicity for the equine host, as demonstrated by intravenous inoculation of a suspension of infected tick cells and subsequent reisolation from peripheral blood, in fulfillment of Koch's postulates. The horse developed severe clinical signs, i.e., fever, inappetence, thrombocytopenia, icterus, and limb edema, typical of granulocytic equine ehrlichiosis, within 1 week.


Adverse events during testosterone replacement therapy in 95 young hypogonadal thalassemic men.
Hormonal treatment of hypogonadism in thalassaemia major (TM) is a complex issue due to the co-existence of other contributing factors such as severity of iron overload, associated chronic liver disease and other endocrine complications. Data about adverse events (AEs) of testosterone replacement therapy (TRT) in hypogonadal males with TM is scarce.We report the adverse events registered during TRT in 95 young patients with TM. These AEs included gynecomastia, documented in 41/95 (43.1%) TM patients of mild to moderate severity (90%). Persistent pain in the injection site and local reactions to testosterone (T) skin patch occurred in a third of patients. Priapism was reported in 2 patients on treatment with intramuscular T enanthate. In both patients, substitution with T gel was successful, and no recurrence during the following 24 months was observed. Clinicians should exercise caution when considering TRT for hypogonadal men with TM. (www.actabiomedica.it).


Melioidosis in the Lao People's Democratic Republic.
Melioidosis is clearly highly endemic in Laos, although the disease has only been diagnosed regularly in humans (1359 cases) since 1999, and only a single animal case has been microbiologically confirmed. Burkholderia pseudomallei is extensively and abundantly present in soil and surface water in central and southern Laos, but the true distribution of the disease across the country remains to be determined. Surveillance is almost non-existent and diagnostic microbiology services are not yet well established, whilst awareness of melioidosis is low amongst policy-makers, healthcare providers, and the public. It is hoped that this situation will improve over the next decade as the country rapidly develops, especially as this is likely to be accompanied by a further increase in the prevalence of diabetes, meaning that more people in this predominantly agricultural population will be at risk of contracting melioidosis.


Incidence and clinical predictors of psoriatic arthritis in patients with psoriasis: a population-based study.
To determine the incidence and disease-specific predictors of clinically recognized psoriatic arthritis (PsA) in patients with psoriasis. We identified an incidence cohort of psoriasis subjects age >/=18 years diagnosed between January 1, 1970 and December 31, 1999 in a population-based setting. Psoriasis diagnoses were validated by confirmatory diagnosis in the medical record. Incident and clinically recognized PsA subjects were classified according to the Classification of Psoriatic Arthritis (CASPAR) criteria. Cox proportional hazards models were used to identify predictors of PsA within the psoriasis cohort. The psoriasis incidence cohort comprised 1,633 subjects. Of these, 40 were diagnosed with PsA concurrently with psoriasis and were excluded from analysis. The remaining 1,593 psoriasis subjects had a mean age of 43 years and 50% were men. Over 20,936 person-years of followup, 57 subjects were clinically recognized with new-onset PsA, with a cumulative incidence of 1.7% (95% confidence interval [95% CI] 1.0-2.3%), 3.1% (95% CI 2.2-4.1%), and 5.1% (95% CI 3.7-6.6%) at 5, 10, and 20 years following psoriasis incidence, respectively. Psoriasis features associated with higher risk of PsA were scalp lesions (hazard ratio [HR] 3.89, 95% CI 2.18-6.94), nail dystrophy (HR 2.93, 95% CI 1.68-5.12), and intergluteal/perianal lesions (HR 2.35, 95% CI 1.32-4.19). Calendar year was not associated with risk of PsA (P = 0.15), indicating that the likelihood of PsA in psoriasis subjects did not change over time. In this population-based study, <10% of patients with psoriasis developed clinically recognized PsA during a 30-year period. Psoriasis features associated with a higher likelihood of PsA were nail dystrophy, scalp lesions, and intergluteal/perianal psoriasis.


Further evidence for genetic heterogeneity in the fragile X syndrome.
The X-linked fragile X [fra(X)] syndrome, associated with a fragile site at Xq27.3, is the most common Mendelian inherited form of mental deficiency. Approximately 1 in 1060 males and 1 in 677 females carry the fra(X) chromosome. However, diagnosis of carrier status can be difficult since about 20% of males and 44% of females are nonpenetrant for mental impairment and/or expression of fra(X). We analyzed DNA from 327 individuals in 23 families segregating fra(X) for linkage to three flanking polymorphic probes: 52A, F9, and ST14. This allowed probable nonpenetrant, transmitting males and carrier females to be identified. A combined linkage analysis was conducted using these families and published probe information on F9 in 27 other families, 52A in six families, and ST14 in five families. The two-point recombination fraction for 52A-F9 was 0.13 (90% confidence interval, 0.10-0.16), for F9-fra(X) was 0.21 (0.17-0.24), and for fra(X)-ST14 was 0.12 (0.07-0.17). Tight linkage between F9 and fra(X) was observed in some families; in others loose linkage was seen suggesting genetic linkage heterogeneity. Risk analysis of carrier status using flanking DNA probes showed that probable nonpenetrant transmitting males were included in families showing both tight and loose linkage. Thus, in contrast to our previous conclusions, it appears that the presence or absence of nonpenetrant, transmitting males in a family is not an indicator of heterogeneity. To determine if heterogeneity was present, we employed the admixture test. Evidence for linkage heterogeneity between F9 and fra(X) was found, significant at P less than 0.0005. Nonsignificant heterogeneity was seen for 52A-F9 linkage. No heterogeneity was found for fra(X)-ST14. The frequency of fra(X) expression was significantly lower in families with tight F9-fra(X) linkage than in families with loose linkage. Cognition appeared to relate to linkage type: affected males in tight linkage families had higher IQs than those in loose linkage families. These findings of genetic heterogeneity can account in part for the high prevalence and apparent high new mutation rate of fra(X). They will affect genetic counseling using RFLPs. An understanding of the basis for genetic heterogeneity in fra(X) will help to clarify the nature of the unusual pattern of inheritance seen in this syndrome.


Genome-Wide Identification of BAHD Acyltransferases and In vivo Characterization of HQT-like Enzymes Involved in Caffeoylquinic Acid Synthesis in Globe Artichoke.
Globe artichoke (Cynara cardunculus L. var. scolymus) is a rich source of compounds promoting human health (phytonutrients), among them caffeoylquinic acids (CQAs), mainly represented by chlorogenic acid (CGA), and dicaffeoylquinic acids (diCQAs). The enzymes involved in their biosynthesis belong to the large family of BAHD acyltransferases. Following a survey of the globe artichoke genome, we identified 69 BAHD proteins carrying the catalytic site (HXXXD). Their phylogenetic analysis together with another 43 proteins, from 21 species, representative of the BAHD family, highlighted their grouping in seven major clades. Nine globe artichoke acyltransferases clustered in a sub-group of Clade V, with 3 belonging to hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase (HQT) and 2 to hydroxycinnamoyl-CoA:shikimate/quinate hydroxycinnamoyl transferase (HCT) like proteins. We focused our attention on the former, HQT1, HQT2, and HQT3, as they are known to play a key role in CGA biosynthesis. The expression of genes coding for the three HQTs and correlation of expression with the CQA content is reported for different globe artichoke tissues. For the first time in the globe artichoke, we developed and applied the virus-induced gene silencing approach with the goal of assessing in vivo the effect of HQT1 silencing, which resulted in a marked reduction of both CGA and diCQAs. On the other hand, when the role of the three HQTs was assessed in leaves of Nicotiana benthamiana through their transient overexpression, significant increases in mono- and diCQAs content were observed. Using transient GFP fusion proteins expressed in N. benthamiana leaves we also established the sub-cellular localization of these three enzymes.


SLN, NLC, LDC: state of the art in drug and active delivery.
Drug delivery system focuses on the regulation of the in vivo dynamics, in order to improve the effectiveness and safety of the incorporated drugs by use of novel drug formulation technologies. Lipids such as fatty acids, triglycerides, vegetable oils and their derivatives, used for developing multiparticulate dosage forms, may be available in solid, semi-solid or liquid state. Solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs) and lipid drug conjugate (LDCs) nanoparticles are novel lipid drug delivery systems. They were devised to address some of the challenges of conventional drug delivery systems ranging from low drug encapsulation efficiency to low bioavailability of Biopharmaceutical Classification Systems (BCS) class II and class IV drugs. SLNs are based on melt-emulsified lipids, which are solid at room temperature and consist of physiologically well tolerated ingredients often generally recognised as safe. NLCs are colloidal carriers characterized by a solid lipid core consisting of a mixture of solid and liquid lipids, and having a mean particle size in the nanometer range. LDC are nanoparticles contain drugs linked to lipid particles. This minireview highlights these three different but related technologies in lipid drug delivery. The objectives of their introduction, current applications, major challenges and some patented formulations are highlighted.


A community-based cluster randomised controlled trial in rural Bangladesh to evaluate the impact of the use of iron-folic acid supplements early in pregnancy on the risk of neonatal mortality: the Shonjibon trial.
Iron-deficiency is the most common nutritional deficiency globally. Due to the high iron requirements for pregnancy, it is highly prevalent and severe in pregnant women. There is strong evidence that maternal iron deficiency anaemia increases the risk of adverse perinatal outcomes. However, most of the evidence is from observational epidemiological studies except for a very few randomised controlled trials. IFA supplements have also been found to reduce the preterm delivery rate and neonatal mortality attributable to prematurity and birth asphyxia. These results combined indicate that IFA supplements in populations of iron-deficient pregnant women could lead to a decrease in the number of neonatal deaths mediated by reduced rates of preterm delivery. In this paper, we describe the protocol of a community-based cluster randomised controlled trial that aims to evaluate the impact of maternal antenatal IFA supplements on perinatal outcomes. The effect of the early use of iron-folic acid supplements on neonatal mortality will be examined using a community based, cluster randomised controlled trial in five districts with 30,000 live births. In intervention clusters trained BRAC village volunteers will identify pregnant women & provide iron-folic acid supplements. Groundwater iron levels will be measured in all study households using a validated test kit. The analysis will follow the intention to treat principle. We will compare neonatal mortality rates & their 95% confidence intervals adjusted for clustering between treatment groups in each groundwater iron-level group. Cox proportional hazards mixed models will be used for mortality outcomes & will include groundwater iron level as an interaction term in the mortality model. This paper aims to describe the study protocol of a community based randomised controlled trial evaluating the impact of the use of iron-folic acid supplements early in pregnancy on the risk of neonatal mortality. This study is critical because it will determine if antenatal IFA supplements commenced in the first trimester of pregnancy, rather than later, will significantly reduce neonatal deaths in the first month of life, and if this approach is cost-effective. This trial has been registered with the Australian New Zealand Clinical Trials Registry (ANZCTR) on 31 May 2012. The registration ID is ACTRN12612000588897 .


Online mass spectrometric analysis of proteins/peptides following electrolytic cleavage of disulfide bonds.
The disulfide bond bridge is an important post-translational modification for proteins. This study presents a structural analysis of biologically active peptides and proteins containing disulfide bonds using electrochemistry (EC) online combined with desorption electrospray ionization mass spectrometry (DESI-MS), in which the sample undergoes electrolytic disulfide cleavage in an electrochemical flow cell followed by MS detection. Using this EC/DESI-MS method, the disulfide-containing peptides can be quickly identified from enzymatic digestion mixtures, simply based on the abrupt decrease in their relative ion abundances after electrolysis. Peptide mass mapping and tandem MS analysis of the ions of the resulting free peptide chains can possibly establish the disulfide linkage pattern and sequence the precursor peptides. In this regard, the method provides much more chemical information than previous analogous electrochemical analyses. In addition, derivatization of thiols by selective selenamide reagents is useful for easy recognition of reduced peptide ions and the number of their free thiols. Furthermore, electrolytic reduction of proteins (e.g., α-lactalbumin) leads to increased charges on the detected protein ions, revealing the role of disulfide bonds on maintaining protein conformation. This electrochemical mass spectrometric method is fast (completed in few minutes) and does not need chemical reductants, potentially having valuable applications in proteomics research.