Reductions in Cerebral Blood Flow Can Be Provoked by Sitting in Severe Myalgic Encephalomyelitis/Chronic Fatigue Syndrome Patients.
In a large study with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) patients, we showed that 86% had symptoms of orthostatic intolerance in daily life and that 90% had an abnormal reduction in cerebral blood flow (CBF) during a standard tilt test. A standard head-up tilt test might not be tolerated by the most severely affected bed-ridden ME/CFS patients. Sitting upright is a milder orthostatic stress. The present study examined whether a sitting test, measuring cerebral blood flow by extracranial Doppler, would be sufficient to provoke abnormal reductions in cerebral blood flow in severe ME/CFS patients. 100 severe ME/CFS patients were studied, (88 females) and were compared with 15 healthy controls (HC) (13 females). CBF was measured first while seated for at least one hour, followed by a CBF measurement in the supine position. Fibromyalgia was present in 37 patients. Demographic data as well as supine heart rate and blood pressures were not different between ME/CFS patients and HC. Heart rate and blood pressure did not change significantly between supine and sitting both in patients and HC. Supine CBF was not different between patients and HC. In contrast, absolute CBF during sitting was lower in patients compared to HC: 474 (96) mL/min in patients and 627 (89) mL/min in HC; p < 0.0001. As a result, percent CBF reduction while seated was -24.5 (9.4)% in severe ME/CFS patients and -0.4 (1.2)% in HC (p < 0.0001). In the ten patients who had no orthostatic intolerance complaints in daily life, the CBF reduction was -2.7 (2.1)%, which was not significantly different from HC (p = 0.58). The remaining 90 patients with orthostatic intolerance complaints had a -26.9 (6.2)% CBF reduction. No difference in CBF parameters was found in patients with and without fibromyalgia. Patients with a previous diagnosis of postural orthostatic tachycardia syndrome (POTS) had a significantly larger CBF reduction compared with those without POTS: 28.8 (7.2)% vs. 22.3 (9.7)% (p = 0.0008). A sitting test in severe ME/CFS patients was sufficient to provoke a clinically and statistically significant mean CBF decline of 24.5%. Patients with a previous diagnosis of POTS had a larger CBF reduction while seated, compared to patients without POTS. The magnitude of these CBF reductions is similar to the results in less severely affected ME/CFS patients during head-up tilt, suggesting that a sitting test is adequate for the diagnosis of orthostatic intolerance in severely affected patients.


Thromboprophylaxis in medical-surgical critically ill patients.
Although critically ill patients are at high risk of venous thromboembolism and bleeding, and thromboprophlyaxis is of proven effectivity in other settings, there remain relatively few data to assist clinicians in providing evidence-based care for medical-surgical patients in the intensive care unit. Deep vein thrombosis occurs in 5-10% of critically ill patients even if they receive unfractionated heparin for prophylaxis. Both heparin and low molecular weight heparin can be safely administered to the majority of critically ill patients and the low molecular weight heparin dalteparin does not appear to bioaccumulate even when administered to patients with severe renal dysfunction. Further research is currently underway to better define how these conditions can be optimally treated. Despite the high morbidity and mortality because of critical illness, the risk of venous thromboembolism in these patients, and adverse outcomes due to venous thromboembolism, much more methodologically rigorous data are required in the form of large, well designed randomized trials before firm recommendations about prophylaxis can be provided to this highly vulnerable population.


Community-based, point-of-care hepatitis C testing: perspectives and preferences of people who inject drugs.
A barrier to hepatitis C treatment for people who inject drugs (PWID) is needing to attend multiple appointments for diagnosis. Point-of-care hepatitis C tests provide results within 20 to 105 minutes and can be offered opportunistically in nonclinical settings such as needle syringe programmes. In this nested qualitative study, we explored the acceptability of point-of-care testing for PWID. PWID attending participating needle syringe programmes were screened using the OraQuick HCV antibody mouth swab (result in 20 minutes); those with a reactive result then underwent venepuncture for a point-of-care RNA test: the Xpert HCV Viral Load (result in 105 minutes). Convenience sampling was used to select participants for a semi-structured interview. A hybrid thematic analysis was performed, guided by Sekhon's "Theoretical Framework of Acceptability." Nineteen participants were interviewed. Three core themes emerged: "people and place," "method of specimen collection," and "rapidity of result return." It was highly acceptable to be offered testing at the needle syringeprogrammes by nurses and community health workers, who were described as competent and nonjudgemental. Most participants reported that even if a finger-stick point-of-care RNA test were an option in the future, they would prefer venepuncture, as the sample could be used for pre-treatment workup and bundled testing. Waiting 20 minutes to receive the antibody test result was acceptable, whereas the 105 minutes required for the RNA result was unacceptable. Offering point-of-care hepatitis C testing at needle syringe programmes is acceptable to PWID, however tests that avoid venepuncture are not necessarily the most attractive to PWID.


Increased expression of platelet-derived growth factor type B receptors in the skin of patients with systemic sclerosis.
The expression of B-type receptors for platelet-derived growth factor (PDGF) was investigated in skin biopsy samples from patients with systemic sclerosis (SSc), by immunohistochemical staining using monoclonal antibodies specific for the receptor. Whereas skin from healthy individuals lacked expression of PDGF-B receptors, receptor expression was seen in sclerodermatous skin lesions from 13 of 14 patients. Increased receptor expression was observed in dermal vessels, as well as on many stromal fibroblast-like cells close to these vessels. PDGF-B receptor expression was most pronounced within and around dermal vessels in which perivascular infiltrates of Leu-4-positive T lymphocytes and HLA-DR-positive, RFD7-positive activated macrophages were present. Both perivascular inflammatory cell infiltrates and PDGF-B receptor expression were generally also seen in macroscopically normal areas of the skin of the SSc patients, indicating that the observed phenotypic alterations may precede the macroscopically observable features of scleroderma in the skin. The observed induction of PDGF-B receptors, together with indirect indications of increased synthesis and release of PDGF, would be compatible with altered PDGF-mediated control of connective tissue cell growth as part of the molecular basis for development of the skin lesions in SSc.


Adhesion of human and animal Escherichia coli strains in association with their virulence-associated genes and phylogenetic origins.
Intestinal colonization is influenced by the ability of the bacterium to inhabit a niche, which is based on the expression of colonization factors. Escherichia coli carries a broad range of virulence-associated genes (VAGs) which contribute to intestinal (inVAGs) and extraintestinal (exVAGs) infection. Moreover, initial evidence indicates that inVAGs and exVAGs support intestinal colonization. We developed new screening tools to genotypically and phenotypically characterize E. coli isolates originating in humans, domestic pigs, and 17 wild mammal and avian species. We analyzed 317 isolates for the occurrence of 44 VAGs using a novel multiplex PCR microbead assay (MPMA) and for adhesion to four epithelial cell lines using a new adhesion assay. We correlated data for the definition of new adhesion genes. inVAGs were identified only sporadically, particularly in roe deer (Capreolus capreolus) and the European hedgehog ( Erinaceus europaeus). The prevalence of exVAGs depended on isolation from a specific host. Human uropathogenic E. coli isolates carried exVAGs with the highest prevalence, followed by badger (Meles meles) and roe deer isolates. Adhesion was found to be very diverse. Adhesion was specific to cells, host, and tissue, though it was also unspecific. Occurrence of the following VAGs was associated with a higher rate of adhesion to one or more cell lines: afa-dra, daaD, tsh, vat, ibeA, fyuA, mat, sfa-foc, malX, pic, irp2, and papC. In summary, we established new screening methods which enabled us to characterize large numbers of E. coli isolates. We defined reservoirs for potential pathogenic E. coli. We also identified a very broad range of colonization strategies and defined potential new adhesion genes.


Bilateral Deep Brain Stimulation of the Subthalamic Nucleus under Sedation with Propofol and Fentanyl.
Awakening during deep brain stimulation (DBS) surgery may be stressful to patients. The aim of the current study was to evaluate the effect on MER signals and their applicability to subthalmic nucleus (STN) DBS surgery for patients with Parkinson's disease (PD) under sedation with propofol and fentanyl. Sixteen consecutive patients with PD underwent STN-DBS surgery with propofol and fentanyl. Their MER signals were achieved during the surgery. To identify the microelectrodes positions, the preoperative MRI and postoperative CT were used. Clinical profiles were also collected at the baseline and at 6 months after surgery. All the signals were slightly attenuated and contained only bursting patterns, compared with our previous report. All electrodes were mostly located in the middle one third part of the STN on both sides of the brain in the fused images. Six months later, the patients were improved significantly in the medication-off state and they met with less dyskinesia and less off-duration. Our study revealed that the sedation with propofol and fentanyl was applicable to STN-DBS surgery. There were no significant problems in precise positioning of bilateral electrodes. The surgery also improved significantly clinical outcomes in 6-month follow-up.


[Reference values for lead levels in blood for the urban population].
The lead reference values for blood used in Brazil come from studies conducted in other countries, where socioeconomic, clinical, nutritional and occupational conditions are significantly different. In order to guarantee an accurate biomonitoring of the population which is occupationally exposed to lead, a major health concern of the studied community, reference values for individuals who are not occupationally exposed and who live in the southern region of the city were established. The sample was composed of 206 subjects of at least 15 years of age. Various strategies were employed to assure good-quality sampling. Subjects who presented values outside clinical or laboratory norms were excluded, as well as those whose specific activities might interfere with the results. Lead reference values for blood were found to be from 2.40 to 16.6 micrograms.dL-1, obtained by the interval x +/- 2s (where x is the mean and s is the standard deviation form observed values) and the median was 7.9 micrograms.dL-1.


Atherosclerotic occlusive disease of the lower extremity: prospective evaluation with two-dimensional time-of-flight MR angiography.
A prospective, blinded comparison of two-dimensional time-of-flight (TOF) magnetic resonance (MR) angiography and conventional arteriography was performed in 25 patients who underwent routine arteriography for symptomatic atherosclerotic occlusive disease of the lower extremity. MR angiography was performed from the distal abdominal aorta through the popliteal trifurcation. The native arterial tree was divided into nine segments; each segment was assessed for patency (defined as stenosis < 50% of arterial diameter), moderate stenosis (50%-69%), severe stenosis (70%-99%), or occlusion (100%). In all 206 segments examined, the sensitivity of MR angiography in diagnosis of occlusion was 100%; the specificity, 98%. All long occlusions were correctly classified. In specific categories of occlusive disease, the sensitivity and specificity were as follows: in all segments with 70% or greater stenosis, including occlusions, 90% and 97%; in all segments with 50% or greater stenosis, including occlusions, 92% and 88%; in femoropopliteal segments with 50% or greater stenosis, 92% and 93%; and in iliac segments, 93% and 83%. Long occlusions were distinguished from short occlusions and stenoses; hence, MR angiography enabled identification of patients with lesions suitable for angioplasty.


Structure and evolution of human apolipoprotein genes: identification of regulatory elements of the human apolipoprotein E gene.
The structures of the major human apolipoprotein genes have been determined. The genes for apoE, apoC-I, apoC-II, apoC-III, apoA-I, apoA-II and apoA-IV have similar structures, consisting of four exons and three introns, which suggests that they evolved from a common ancestral gene. The third and fourth exons of the ancestral gene appear to have evolved from the duplication of a 66-nucleotide repeat unit that encodes a 22-residue alpha-helical peptide element of amphipathic character. The apoA-I, apoC-III and apoA-IV genes are linked closely within a 20-kilobase (kb) span of chromosome 11. The apoE and apoC-I genes, together with an apoC-I' pseudogene, are linked closely within a 25-kb span of chromosome 19. To characterize potential functional relationships among the apolipoprotein genes, initial studies have been done to identify the molecular elements involved in the regulation of the human apoE gene. Fragments of the 5'-flanking portion of this gene were inserted into appropriate plasmid vectors, which contained the bacterial chloramphenicol acetyl transferase gene, and were examined for promoter activity and potential enhancer activity after transfection into cultured mammalian cells. Deletion mapping of the promoter region has identified multiple functional elements, including an enhancer, two G-C boxes (Sp 1 transcription factor binding sites) and an upstream control element. In addition, there is an enhancer located in the first intron. Interactions among these various control elements are likely to determine the ways in which the expression of the apoE gene is regulated.


A meta-ethnography investigating relational influences on mental health and cancer-related health care interventions for racially minoritised people in the UK.
Despite calls to increase the 'cultural competence' of health care providers, racially minoritised people continue to experience a range of problems when it comes to health care, including discrimination. While relevant qualitative meta-syntheses have suggested better ways forward for health care for racialised minorities, many have lacked conceptual depth, and none have specifically investigated the relational dimensions involved in care. We set out to investigate the social and cultural influences on health care interventions, focusing on psychological approaches and/or cancer care to inform the trial of a new psychological therapy for those living with or beyond cancer. A meta-ethnography approach was used to examine the relevant qualitative studies, following Noblit and Hare, and guided by patient involvement throughout. Papers were analysed between September 2018 and February 2023, with some interruptions caused by the Covid pandemic. The following databases were searched: Ovid MEDLINE, EBSCO CINAHL, Ovid Embase, EBSCO PsycINFO, Proquest Sociology Collection (including Applied Social Sciences Index & Abstracts (ASSIA), Sociological Abstracts and Sociology Database), EBSCO SocINDEX, Ovid AMED, and Web of Science. The systematic review protocol was registered with the International Prospective Register of Systematic Reviews (PROSPERO) (ID: CRD42018107695), and reporting follows the eMERGe Reporting Guidance for meta-ethnographies (France et al. 2019). Twenty-nine journal papers were included in the final review. Themes (third-order constructs) developed in the paper include the centrality of the patient-practitioner relationship; how participants give meaning to their illness in connection to others; how families (rather than individuals) may make health decisions; how links with a higher power and spiritual/religious others can play a role in coping; and the ways in which a hierarchy of help-seeking develops, frequently with the first port of call being the resources of oneself. Participants in studies had a need to avoid being 'othered' in their care, valuing practitioners that connected with them, and who were able to recognise them as whole and complex (sometimes described in relational languages like 'love'). Complex family-based health decision-making and/or the importance of relations with non-human interactants (e.g. God, spiritual beings) were frequently uncovered, not to mention the profoundly emergent nature of stigma, whereby families could be relatively safe havens for containing and dealing with health challenges. A conceptual framework of 'animated via (frequently hidden) affective relationality' emerged in the final synthesis, bringing all themes together, and drawing attention to the emergent nature of the salient issues facing minoritised patients in health care interactions. Our analysis is important because it sheds light on the hitherto buried relational forces animating and producing the specific issues facing racially minoritised patients, which study participants thought were largely overlooked, but to which professionals can readily relate (given the universal nature of human relations). Thus, training around the affective relationality of consultations could be a fruitful avenue to explore to improve care of diverse patients.


Selenium and coenzyme Q10 interrelationship in cardiovascular diseases--A clinician's point of view.
A short review is given of the potential role of selenium deficiency and selenium intervention trials in atherosclerotic heart disease. Selenium is an essential constituent of several proteins, including the glutathione peroxidases and selenoprotein P. The selenium intake in Europe is generally in the lower margin of recommendations from authorities. Segments of populations in Europe may thus have a deficient intake that may be presented by a deficient anti-oxidative capacity in various illnesses, in particular atherosclerotic disease, and this may influence the prognosis of the disease. Ischemic heart disease and heart failure are two conditions where increased oxidative stress has been convincingly demonstrated. Some of the intervention studies of anti-oxidative substances that have focused on selenium are discussed in this review. The interrelationship between selenium and coenzyme Q10, another anti-oxidant, is presented, pointing to a theoretical advantage in using both substances in an intervention if there are deficiencies within the population. Clinical results from an intervention study using both selenium and coenzyme Q10 in an elderly population are discussed, where reduction in cardiovascular mortality, a better cardiac function according to echocardiography, and finally a lower concentration of the biomarker NT-proBNP as a sign of lower myocardial wall tension could be seen in those on active treatment, compared to placebo.


Electronic health information exchange in underserved settings: examining initiatives in small physician practices & community health centers.
Health information exchange (HIE) is an important tool for improving efficiency and quality and is required for providers to meet Meaningful Use certification from the United States Centers for Medicare and Medicaid Services. However widespread adoption and use of HIE has been difficult to achieve, especially in settings such as smaller-sized physician practices and federally qualified health centers (FQHCs). We assess electronic data exchange activities and identify barriers and benefits to HIE participation in two underserved settings. We conducted key-informant interviews with stakeholders at physician practices and health centers. Interviews were recorded, transcribed, and then coded in two waves: first using an open-coding approach and second using selective coding to identify themes that emerged across interviews, including barriers and facilitators to HIE adoption and use. We interviewed 24 providers, administrators and office staff from 16 locations in two states. They identified barriers to HIE use at three levels-regional (e.g., lack of area-level exchanges; partner organizations), inter-organizational (e.g., strong relationships with exchange partners; achieving a critical mass of users), and intra-organizational (e.g., type of electronic medical record used; integration into organization's workflow). A major perceived benefit of HIE use was the improved care-coordination clinicians could provide to patients as a direct result of the HIE information. Utilization and perceived benefit of the exchange systems differed based on several practice- and clinic-level factors. The adoption and use of HIE in underserved settings appears to be impeded by regional, inter-organizational, and intra-organizational factors and facilitated by perceived benefits largely at the intra-organizational level. Stakeholders should consider factors both internal and external to their organization, focusing efforts in changing modifiable factors and tailoring HIE efforts based on all three categories of factors. Collective action between organizations may be needed to address inter-organizational and regional barriers. In the interest of facilitating HIE adoption and use, the impact of interventions at various levels on improving the use of electronic health data exchange should be tested.


Safety and efficacy of endovascular ultrasound renal denervation in resistant hypertension: 12-month results from the ACHIEVE study.
First-generation radiofrequency renal denervation devices may have had limited efficacy due to incomplete circumferential ablation and dependence on individual operator technique. This study evaluated a next-generation catheter-based technology using ultrasound designed to maximize nerve coverage using circumferential ultrasound energy. This was a prospective, multicenter, nonrandomized, postmarket study evaluating the safety and efficacy of an endovascular ultrasound renal denervation system (Paradise) in patients who met the European Society of Hypertension/European Society of Cardiology definition of resistant hypertension. Major exclusion criteria included renal artery stenosis and renal insufficiency. Patients were followed for 12 months. Safety and efficacy endpoints included procedural safety and renal artery patency, as well as changes in systolic office and 24-h ambulatory bllod pressure (BP) measurement. A total of 96 patients from eight European sites were included. Mean age was 63.9 years and 41% of patients were female. At baseline, mean 24-h ambulatory BP was 156.2/88.4 ± 15.4/12.7 mmHg, and mean office BP was 176.2/95.0 ± 20.6/16.0 mmHg. Patients were on average on 5.1 ± 2.5 antihypertensive agents. At 12 months, the average 24-h ambulatory BP change was -7.5/-3.8 ± 18.3/10.6 mmHg (P = 0.0007; P = 0.0024) with an average office BP change of -15.0/-7.0 ± 27.0/12.3 mmHg (P < 0.0001 for both). At 12 months, there was a single patient death unrelated to the device or procedure. This single-arm study represents the largest cohort of patients treated with endovascular ultrasound renal denervation to date. Within this trial, the therapy appeared safe and resulted in sustained reductions in both office BP and 24-h ambulatory BP through 12 months.


Worldwide Organization of Neurocritical Care: Results from the PRINCE Study Part 1.
Neurocritical care focuses on the care of critically ill patients with an acute neurologic disorder and has grown significantly in the past few years. However, there is a lack of data that describe the scope of practice of neurointensivists and epidemiological data on the types of patients and treatments used in neurocritical care units worldwide. To address these issues, we designed a multicenter, international, point-prevalence, cross-sectional, prospective, observational, non-interventional study in the setting of neurocritical care (PRINCE Study). In this manuscript, we analyzed data from the initial phase of the study that included registration, hospital, and intensive care unit (ICU) organizations. We present here descriptive statistics to summarize data from the registration case report form. We performed the Kruskal-Wallis test followed by the Dunn procedure to test for differences in practices among world regions. We analyzed information submitted by 257 participating sites from 47 countries. The majority of those sites, 119 (46.3%), were in North America, 44 (17.2%) in Europe, 34 (13.3%) in Asia, 9 (3.5%) in the Middle East, 34 (13.3%) in Latin America, and 14 (5.5%) in Oceania. Most ICUs are from academic institutions (73.4%) located in large urban centers (44% > 1 million inhabitants). We found significant differences in hospital and ICU organization, resource allocation, and use of patient management protocols. The highest nursing/patient ratio was in Oceania (100% 1:1). Dedicated Advanced Practiced Providers are mostly present in North America (73.7%) and are uncommon in Oceania (7.7%) and the Middle East (0%). The presence of dedicated respiratory therapist is common in North America (85%), Middle East (85%), and Latin America (84%) but less common in Europe (26%) and Oceania (7.7%). The presence of dedicated pharmacist is highest in North America (89%) and Oceania (85%) and least common in Latin America (38%). The majority of respondents reported having a dedicated neuro-ICU (67% overall; highest in North America: 82%; and lowest in Oceania: 14%). The PRINCE Study results suggest that there is significant variability in the delivery of neurocritical care. The study also shows it is feasible to undertake international collaborations to gather global data about the practice of neurocritical care.


Association of cognitive impairment and elderly mortality: differences between two cohorts ascertained 6-years apart in China.
Cognitive impairment is a major contributor to mortality among the elderly. However, the relationship between cognitive impairment evaluated by educational levels and mortality and the trend between cognitive impairment and mortality with time are unclear. We aim to evaluate the differences in associations of cognitive impairment, taking the stratification by educational levels into account, with all-cause mortality and further explore the relationship of cognitive impairment with mortality in different age and sex groups in two cohorts ascertained 6 years apart in China. A total of 13,906 and 13,873 Chinese elderly aged 65 years and older were included in the 2002-2008 and 2008-2014 cohorts from the Chinese Longitudinal Healthy Longevity Survey (CLHLS). Mortality data was ascertained from interviews with family members or relatives of participants. Cognitive function, evaluated by the Mini-Mental State Examination (MMSE), were defined by different cut-offs taking educational background into account. Cox models were used to explore the relationship of cognitive impairment with mortality. For the 2002-2008 and 2008-2014 cohorts, 55,277 and 53,267 person-years were followed up, and the mean (SD) age were 86.5 (11.6) and 87.2 (11.3) years, respectively. Compared to normal cognition, cognitive impairment was independently associated with higher mortality risk after controlling for potential confounders, with hazard ratios (HRs) of 1.32 (95% confidence interval [CI], 1.25-1.39) in 2002-2008 cohort and 1.26 (95% CI, 1.19-1.32) in 2008-2014 cohort, stratified by educational levels. The trend of cognitive impairment with all-cause mortality risk decreased from 2002 to 2008 to 2008-2014 cohort, while no significant interaction of cognitive impairment with cohort for all-cause mortality was observed. The associations of cognitive impairment and mortality were decreased with age in the two cohorts. Cognitive impairment evaluated by different cut-offs were associated with increased risk of mortality, especially among those aged 65-79 years in the two cohorts; this advocates that periodic screening for cognitive impairment among the elderly is warranted.


Molecular composition and distribution of gap junctions in the sensory epithelium of the human cochlea-a super-resolution structured illumination microscopy (SR-SIM) study.
Mutations in the GJB2 gene, which encodes the Connexin26 (Cx26) protein, are the most common cause of childhood hearing loss in American and European populations. The cochlea contains a gap junction (GJ) network in the sensory epithelium and two connective tissue networks in the lateral wall and spiral limbus. The syncytia contain the GJ proteins beta 2 (GJB2/Cx26) and beta 6 (GJB6/Cx30). Our knowledge of their expression in humans is insufficient due to the limited availability of tissue. Here, we sought to establish the molecular arrangement of GJs in the epithelial network of the human cochlea using surgically obtained samples. We analyzed Cx26 and Cx30 expression in GJ networks in well-preserved adult human auditory sensory epithelium using confocal, electron, and super-resolution structured illumination microscopy (SR-SIM). Cx30 plaques (<5 μm) dominated, while Cx26 plaques were subtle and appeared as 'mini-junctions' (2-300 nm). 3-D volume rendering of Z-stacks and orthogonal projections from single optical sections suggested that the GJs are homomeric/homotypic and consist of assemblies of identical GJs composed of either Cx26 or Cx30. Occasionally, the two protein types were co-expressed, suggesting functional cooperation. Establishing the molecular composition and distribution of the GJ networks in the human cochlea may increase our understanding of the pathophysiology of Cx-related hearing loss. This information may also assist in developing future strategies to treat genetic hearing loss.


Selection of tool diameter by New Caledonian crows Corvus moneduloides.
One important element of complex and flexible tool use, particularly where tool manufacture is involved, is the ability to select or manufacture appropriate tools anticipating the needs of any given task-an ability that has been rarely tested in non-primates. We examine aspects of this ability in New Caledonian crows-a species known to be extraordinary tool users and manufacturers. In a 2002 study, Chappell and Kacelnik showed that these crows were able to select a tool of the appropriate length for a task among a set of different lengths, and in 2002, Weir, Chappell and Kacelnik showed that New Caledonian crows were able to shape unfamiliar materials to create a usable tool for a specific task. Here we examine their handling of tool diameter. In experiment 1, we show that when facing three loose sticks that were usable as tools, they preferred the thinnest one. When the three sticks were presented so that one was loose and the other two in a bundle, they only disassembled the bundle when their preferred tool was tied. In experiment 2, we show that they manufacture, and modify during use, a tool of a suitable diameter from a tree branch, according to the diameter of the hole through which the tool will have to be inserted. These results add to the developing picture of New Caledonian crows as sophisticated tool users and manufacturers, having an advanced level of folk physics.


Release behaviour of propranolol HCl from hydrophilic matrix tablets containing psyllium powder in combination with hydrophilic polymers.
The objective of this study was to investigate the release behaviour of propranolol hydrochloride from psyllium matrices in the presence hydrophilic polymers. The dissolution test was carried out at pH 1.2 and pH 6.8. Binary mixtures of psyllium and hydroxypropyl methylcellulose (HPMC) used showed that an increase in the percentage of HPMC in the binary mixtures caused a significant decrease in the release rate of propranolol. Psyllium-alginate matrices produced lower drug release as compared to when the alginate was the matrix former alone. When sodium carboxy methyl cellulose (NaCMC) was incorporated into the psyllium, the results showed that matrices containing the ratio of psyllium-NaCMC in the 1:1 ratio are able to slow down the drug release significantly as compared to matrices made from only psyllium or NaCMC as retardant agent suggesting that there could be a synergistic effect between psyllium and NaCMC. The double-layered tablets showed that the psyllium and HPMC in the outer shell of an inner formulation of psyllium alone had the greatest effect of protecting the inner core and thus producing the lowest drug release (DE = 38%, MDT = 93 min). A significant decrease in the value of n in Q = kt(n) from 0.70 to 0.51 as the psyllium content was increased from 50 to 150 mg suggests that the presence of psyllium in HPMC matrices affected the release mechanism. Psyllium powder had the ability in the combination with other hydrophilic polymers to produce controlled release profiles. Care and consideration should as such be taken when formulating hydrophilic matrices in different combinations.


Years of potential life lost and productivity costs due to premature mortality from six priority diseases in Tanzania, 2006-2015.
Mortality statistics are traditionally used to quantify the burden of disease and to determine the relative importance of the various causes of death. Some of the most frequently used indices to quantify the burden of disease are the years of potential life lost (YPLL) and years of potential productive life lost (YPPLL). These two measures reflect the mortality trends in younger age groups and they provide a more accurate picture of premature mortality. This study was carried out to determine YPLL, YPPLL and cost of productivity lost (CPL) due to premature mortality caused by selected causes of deaths in Tanzania. Malaria, respiratory diseases, HIV/AIDS, tuberculosis, cancers and injuries were selected for this analysis. The number of deaths by sex and age groups were obtained from hospital death registers and ICD-10 reporting forms in 39 public hospitals in Tanzania, covering a period of 2006-2015. The life expectancy method and human capital approach were used to estimate the YPLL, YPPLL and CPL due to premature mortality. During 2006-2015, malaria, HIV/AIDS, tuberculosis, respiratory diseases, HIV+tuberculosis, cancer and injury were responsible for a total of 96,834 hospital deaths, of which 46.4% (n = 57,508) were among individuals in the productive age groups (15-64 years). The reported deaths contributed to 2,850,928 YPLL (female = 1,326,724; male = 1,524,205) with an average of 29 years per death. The average YPLL among females (32) was higher than among males (28). Malaria (YPLL = 38 per death) accounted for over one-third (35%) of the total YPLL. There was a significant increase in YPLL due to the selected underlying causes of death over the 10-year period. Deaths from the selected causes resulted into 1,207,499 YPPLL (average = 21 per death). Overall, HIV/AIDS contributed to the highest YPPLL (323,704), followed by malaria (243,490) and injuries (196,505). While there was a general decrease in YPPLL due to malaria, there was an increase of YPPLL due to HIV/AIDS, respiratory diseases, cancer and injuries during the 10-year period. The total CPL due to the six diseases was US$ 148,430,009 for 10 years. The overall CPL was higher among males than females by 29.1%. Over half (58%) of the losses were due to deaths among males. HIV/AIDS accounted for the largest (29.2%) CPL followed by malaria (17.8%) and respiratory diseases (14.6%). The CPL increased from US$11.4 million in 2006 to US$17.9 million in 2016. The YPLL, YPPLL and CPL due to premature death associated with the six diseases in Tanzania are substantially high. While malaria accounted for highest YPLL, HIV/AIDS accounted for highest YPPLL and CPL. The overall CPL was higher among males than among females. Setting resource allocation priorities to malaria, HIV/AIDS and respiratory diseases that are responsible for the majority of premature deaths could potentially reduce the costs of productivity loss in Tanzania.


E-cadherin binding prevents beta-catenin nuclear localization and beta-catenin/LEF-1-mediated transactivation.
Beta-catenin is a multifunctional protein found in three cell compartments: the plasma membrane, the cytoplasm and the nucleus. The cell has developed elaborate ways of regulating the level and localization of beta-catenin to assure its specific function in each compartment. One aspect of this regulation is inherent in the structural organization of beta-catenin itself; most of its protein-interacting motifs overlap so that interaction with one partner can block binding of another at the same time. Using recombinant proteins, we found that E-cadherin and lymphocyte-enhancer factor-1 (LEF-1) form mutually exclusive complexes with beta-catenin; the association of beta-catenin with LEF-1 was competed out by the E-cadherin cytoplasmic domain. Similarly, LEF-1 and adenomatous polyposis coli (APC) formed separate, mutually exclusive complexes with beta-catenin. In Wnt-1-transfected C57MG cells, free beta-catenin accumulated and was able to associate with LEF-1. The absence of E-cadherin in E-cadherin-/- embryonic stem (ES) cells also led to an accumulation of free beta-catenin and its association with LEF-1, thereby mimicking Wnt signaling. beta-catenin/LEF-1-mediated transactivation in these cells was antagonized by transient expression of wild-type E-cadherin, but not of E-cadherin lacking the beta-catenin binding site. The potent ability of E-cadherin to recruit beta-catenin to the cell membrane and prevent its nuclear localization and transactivation was also demonstrated using SW480 colon carcinoma cells.


[New aspects of recurrent pseudocroup].
Three consecutive studies were concerned with clinical, pathophysiologic, and immunologic aspects of recurrent croup. The first investigation demonstrated that recurrent croup is a distinct disease entity which shares many clinical features with asthma and should be distinguished from viral laryngotracheobronchitis. In the second study a high incidence of airway hyperreactivity was found in children with recurrent croup. This specific type of hyperreactivity suggested an involvement of both the lower and the upper respiratory tract. The third study demonstrated a tendency towards low serum IgA levels in children with recurrent croup; hence immunoregulatory defects may be conducive to the development of this disorder.


The relation between multilocus population genetics and social evolution theory.
Evolution at multiple gene positions is complicated. Direct selection on one gene disturbs the evolutionary dynamics of associated genes. Recent years have seen the development of a multilocus methodology for modeling evolution at arbitrary numbers of gene positions with arbitrary dominance and epistatic relations, mode of inheritance, genetic linkage, and recombination. We show that the approach is conceptually analogous to social evolutionary methodology, which focuses on selection acting on associated individuals. In doing so, we (1) make explicit the links between the multilocus methodology and the foundations of social evolution theory, namely, Price's theorem and Hamilton's rule; (2) relate the multilocus approach to levels-of-selection and neighbor-modulated-fitness approaches in social evolution; (3) highlight the equivalence between genetical hitchhiking and kin selection; (4) demonstrate that the multilocus methodology allows for social evolutionary analyses involving coevolution of multiple traits and genetical associations between nonrelatives, including individuals of different species; (5) show that this methodology helps solve problems of dynamic sufficiency in social evolution theory; (6) form links between invasion criteria in multilocus systems and Hamilton's rule of kin selection; (7) illustrate the generality and exactness of Hamilton's rule, which has previously been described as an approximate, heuristic result.


Exercise as a Countermeasure to Declining Central Nervous System Function in Multiple Sclerosis.
The decline of central nervous system (CNS) function is a hallmark characteristic of multiple sclerosis (MS) that can manifest as cognitive impairment. We believe that exercise represents a potential behavioral approach for counteracting the declines in CNS structure and associated function among persons with MS (ie, exercise as a countermeasure of CNS decline). This theory is important because disease-modifying drugs represent a first-line approach for modifying the immune system and its effects on the CNS, but these drugs do not generally demonstrate robust improvements in cognitive performance. To the best of our knowledge, this article presents the first argument positioning exercise as a countermeasure for CNS decline in MS. The reviewed research indicates a proliferating body of evidence describing physical fitness, physical activity, and exercise effects on cognitive performance and neuroimaging outcomes (ie, CNS functioning) in MS, with the consistent and strong association between cognitive performance and neuroimaging outcomes in this population as a backdrop. We further present a framework and future research directions for better positioning exercise as a possible neuroprotective behavior against declining CNS function in MS.


Previous ethanol experience enhances synaptic plasticity of NMDA receptors in the ventral tegmental area.
Alcohol addiction (alcoholism) is one of the most prevalent substance abuse disorders worldwide. Addiction is thought to arise, in part, from a maladaptive learning process in which enduring memories of drug experiences are formed. However, alcohol (ethanol) generally interferes with synaptic plasticity mechanisms in the CNS and thus impairs various types of learning and memory. Therefore, it is unclear how powerful memories associated with alcohol experience are formed during the development of alcoholism. Here, using brain slice electrophysiology in mice, we show that repeated in vivo ethanol exposure (2 g/kg, i.p., three times daily for 7 d) causes increased susceptibility to the induction of long-term potentiation (LTP) of NMDA receptor (NMDAR)-mediated transmission in mesolimbic dopamine neurons, a form of synaptic plasticity that may drive the learning of stimuli associated with rewards, including drugs of abuse. Enhancement of NMDAR plasticity results from an increase in the potency of inositol 1,4,5-trisphosphate (IP(3)) in producing facilitation of action potential-evoked Ca(2+) signals, which is critical for LTP induction. This increase in IP(3) effect, which lasts for a week but not a month after ethanol withdrawal, occurs through a protein kinase A (PKA)-dependent mechanism. Corticotropin-releasing factor, a stress-related neuropeptide implicated in alcoholism and other addictions, further amplifies the PKA-mediated increase in IP(3) effect in ethanol-treated mice. Finally, we found that ethanol-treated mice display enhanced place conditioning induced by the psychostimulant cocaine. These data suggest that repeated ethanol experience may promote the formation of drug-associated memories by enhancing synaptic plasticity of NMDARs in dopamine neurons.


OXI1 and DAD Regulate Light-Induced Cell Death Antagonistically through Jasmonate and Salicylate Levels.
Singlet oxygen produced from triplet excited chlorophylls in photosynthesis is a signal molecule that can induce programmed cell death (PCD) through the action of the OXIDATIVE STRESS INDUCIBLE 1 (OXI1) kinase. Here, we identify two negative regulators of light-induced PCD that modulate OXI1 expression: DAD1 and DAD2, homologs of the human antiapoptotic protein DEFENDER AGAINST CELL DEATH. Overexpressing OXI1 in Arabidopsis (Arabidopsis thaliana) increased plant sensitivity to high light and induced early senescence of mature leaves. Both phenomena rely on a marked accumulation of jasmonate and salicylate. DAD1 or DAD2 overexpression decreased OXI1 expression, jasmonate levels, and sensitivity to photooxidative stress. Knock-out mutants of DAD1 or DAD2 exhibited the opposite responses. Exogenous applications of jasmonate upregulated salicylate biosynthesis genes and caused leaf damage in wild-type plants but not in the salicylate biosynthesis mutant Salicylic acid induction-deficient2, indicating that salicylate plays a crucial role in PCD downstream of jasmonate. Treating plants with salicylate upregulated the DAD genes and downregulated OXI1 We conclude that OXI1 and DAD are antagonistic regulators of cell death through modulating jasmonate and salicylate levels. High light-induced PCD thus results from a tight control of the relative activities of these regulating proteins, with DAD exerting a negative feedback control on OXI1 expression.


Characterization of a rediscovered haplosporidian parasite from cultured Penaeus vannamei.
Mortalities of Penaeus vannamei, cultured in ponds in Belize, Central America, began during the last part of the grow-out cycle during the cold weather months from September 2004 through February 2005. Tissue squashes of infected hepatopancreata and histological examination of infected shrimp revealed that the mortalities might have been caused by an endoparasite. To confirm the diagnosis, DNA was extracted from ethanol preserved hepatopancreata and the small-subunit rRNA gene was sequenced. The 1838 bp sequence was novel and phylogenetic analysis placed the P. vannamei parasite within the phylum Haplosporidia as a sister taxon to a clade that includes Bonamia and Minchinia species. In situ hybridization was performed using anti-sense DNA probes that were designed to hybridize specifically with the parasite's nucleic acid. This organism presents similar characteristics to those of a haplosporidian that infected cultured P. vannamei imported from Nicaragua into Cuba, as described by Dyková et al. (1988; Fish Dis 11:15-22).


Uptake of screening for breast cancer in patients with mental health problems.
Mental illness is associated with physical illness and mortality from a variety of causes including cancer. There is little information on screening attendance among the mentally ill population. An audit was conducted of a breast screening service in inner London to determine uptake rates in women with mental illness. Cross sectional data linkage study of the local screening register and patients of the local psychiatric units. Screening uptake rates in all patients, those with a history of multiple detention in hospital, and those with psychosis were compared with the local reference population. Women in three inner London boroughs. Screening records for 933 psychiatric patients and 44 195 women without mental health problems aged 50 to 64 years. Overall, psychiatric patients were as likely as the reference group to attend breast screening. Patients with a history of multiple detention were significantly less likely to attend (OR = 0.40, 0.29 to 0.55; p<0.001), as were patients with a diagnosis of psychosis (OR = 0.33, 0.18 to 0.61; p<0.01). Increasing age, a history of detention in hospital, and social deprivation remained independent predictors for non-attendance. Women with severe mental health problems may be less likely to attend national screening programmes such as breast screening, and action should be taken to overcome the barriers to attendance.


Allicin ameliorates obesity comorbid depressive-like behaviors: involvement of the oxidative stress, mitochondrial function, autophagy, insulin resistance and NOX/Nrf2 imbalance in mice.
The increased prevalence of obesity has been a major medical and public health problem in the past decades. In obese status, insulin resistance and sustained oxidative stress damage might give rise to behavioral deficits. The anti-obesity and anti-oxidant effects of allicin have been previously reported in peripheral tissues. In the present study, the functions and mechanisms of allicin involved in the prevention of high-fat diet (HFD)-induced depressive-like behaviors were investigated to better understand the pharmacological activities of allicin. Obese mice (five weeks of age) were treated with allicin (50, 100, and 200 mg/kg) by gavage for 15 weeks and behavioral test (sucrose preference, open field, and tail suspension) were performed. Furthermore, markers of oxidative stress, mitochondrial function, autophagy, and insulin resistance were measured in the hippocampal tissue. Finally, the levels of NADPH oxidase (NOX2, NOX4) and the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway were evaluated in the hippocampus. The body weight, metabolic disorders, and depressive-like behaviors in obese mice were ameliorated by allicin. The depressive-like behaviors presented in the obese mice were accompanied by remarkably excessive reactive oxygen species (ROS) production and oxidative stress, damaged mitochondrial function, imbalanced autophagy, and enhanced insulin resistance in the hippocampus. We found that allicin improved the above undesirable effects in the obese mice. Furthermore, allicin significantly decreased NOX2 and NOX4 levels and activated the Nrf2 pathway. Allicin attenuated depressive-like behaviors triggered by long-term HFD consumption by inhibiting ROS production and oxidative stress, improving mitochondrial function, regulating autophagy, and reducing insulin resistance in the hippocampus via optimization of NOX/Nrf2 imbalance.


Simultaneous Acute Anterior ST-Elevation Myocardial Infarction and Acute Ischemic Stroke of Left Middle Cerebral Artery: A Case Report.
BACKGROUND Acute ST-elevation myocardial infarction and acute ischemic stroke are both life-threatening conditions with high risk for morbidity and mortality without timely intervention. This simultaneous event has been reported to be as rare as 0.009%. The treatments of both conditions individually have been well documented in the literature and guidelines, but when presenting concomitantly, it poses a unique therapeutic challenge. Immediate treatment of one condition can delay management or even have deleterious effects on the other condition. CASE REPORT We present the case of a 41-year-old female with simultaneous acute anterior ST-elevation myocardial infarction and acute left middle cerebral artery ischemic stroke. Due to a low National Institute of Health score she was not a candidate for endovascular treatment and received alteplase per acute ischemic stroke protocol with delayed percutaneous coronary intervention. She was eventually discharged to a long-term acute care facility for continued rehabilitation. CONCLUSIONS The co-existence of ST elevation myocardial infarction and acute ischemic stroke is rare, but when these 2 common conditions present simultaneously, it provides a unique therapeutic challenge. Although infrequent, this challenging scenario deserves more recognition and a discussion among the medical community.


Bioartificial sinus node constructed via in vivo gene transfer of an engineered pacemaker HCN Channel reduces the dependence on electronic pacemaker in a sick-sinus syndrome model.
The normal cardiac rhythm originates in the sinoatrial (SA) node that anatomically resides in the right atrium. Malfunction of the SA node leads to various forms of arrhythmias that necessitate the implantation of electronic pacemakers. We hypothesized that overexpression of an engineered HCN construct via somatic gene transfer offers a flexible approach for fine-tuning cardiac pacing in vivo. Using various electrophysiological and mapping techniques, we examined the effects of in situ focal expression of HCN1-DeltaDeltaDelta, the S3-S4 linker of which has been shortened to favor channel opening, on impulse generation and conduction. Single left ventricular cardiomyocytes isolated from guinea pig hearts preinjected with the recombinant adenovirus Ad-CMV-GFP-IRES-HCN1-DeltaDeltaDelta in vivo uniquely exhibited automaticity with a normal firing rate (237+/-12 bpm). High-resolution ex vivo optical mapping of Ad-CGI-HCN1-DeltaDeltaDelta-injected Langendorff-perfused hearts revealed the generation of spontaneous action potentials from the transduced region in the left ventricle. To evaluate the efficacy of our approach for reliable atrial pacing, we generated a porcine model of sick-sinus syndrome by guided radiofrequency ablation of the native SA node, followed by implantation of a dual-chamber electronic pacemaker to prevent bradycardia-induced hemodynamic collapse. Interestingly, focal transduction of Ad-CGI-HCN1-DeltaDeltaDelta in the left atrium of animals with sick-sinus syndrome reproducibly induced a stable, catecholamine-responsive in vivo "bioartificial node" that exhibited a physiological heart rate and was capable of reliably pacing the myocardium, substantially reducing electronic pacing. The results of the present study provide important functional and mechanistic insights into cardiac automaticity and have further refined an HCN gene-based therapy for correcting defects in cardiac impulse generation.


Quantitative genetic variation of odor-guided behavior in a natural population of Drosophila melanogaster.
Quantitative genetic variation in behavioral response to the odorant, benzaldehyde, was assessed among a sample of 43 X and 35 third chromosomes extracted from a natural population and substituted into a common inbred background. Significant genetic variation among chromosome lines was detected. Heritability estimates for olfactory response, however, were low, as is typical for traits under natural selection. Furthermore, the loci affecting naturally occurring variation in olfactory response to benzaldehyde were not the same in males and females, since the genetic correlation between the sexes was low and not significantly different from zero for the chromosome 3 lines. Competitive fitness, viability and fertility of the chromosome 3 lines were estimated using the balancer equilibrium technique. Genetic correlations between fitness and odor-guided behavior were not significantly different from zero, suggesting the number of loci causing variation in olfactory response is small relative to the number of loci causing variation in fitness. Since different genes affect variation in olfactory response in males and females, genetic variation for olfactory response could be maintained by genotype x sex environment interaction. This unusual genetic architecture implies that divergent evolutionary trajectories for olfactory behavior may occur in males and females.


Isolation from Klebsiella and characterization of two rcs genes that activate colanic acid capsular biosynthesis in Escherichia coli.
Two genes, designated rcsA (regulation of capsule synthesis) and rcsB, that had been cloned from the chromosome of Klebsiella aerogenes (K. pneumoniae) capsular serotype K21 were capable of activating expression of colanic acid capsular polysaccharide in Escherichia coli K12. The Klebsiella rcsA gene encoded a polypeptide of 23 kDa that was required for the induction of a mucoid phenotype at less than or equal to 30 degrees C but not at greater than or equal to 37 C. The Klebsiella rcsB locus encoded no apparent polypeptides and was not capable by itself of causing the overproduction of colanic acid. However, when present in the same cell with rcsA, either in cis or in trans, rcsB caused expression of mucoidy in E. coli at all growth temperatures. These findings are best explained if the Klebsiella rcsA gene product acts as a positive regulator of colanic acid biosynthesis in E. coli and that activity of this protein is in turn subject to regulation by Lon protease. The Klebsiella rcsB locus may exert its effect by preferentially binding a negative regulator of capsular biosynthesis, possibly Lon itself. DNA sequences homologous to the Klebsiella K21b rcsA and rcsB genes were found in the genomes of all other capsular serotypes of klebsiellae examined, including K2, K12, K36 and K43. However, there was no homology between such genes and the chromosome of E. coli. The ability of these rcs genes to induce a mucoid phenotype explains the apparent conjugative transfer from klebsiellae to E. coli of the ability to produce K21 or other Klebsiella capsular polysaccharides that are structurally and antigenically related to colanic acid.


A comparison of methods and materials for the analysis of leaded wipes.
The purposes of this study are: (1) to determine whether proficiency analytical test (PAT) materials from the American Industrial Hygiene Association can be used to provide quality data for portable X-ray fluorescence analysis (XRF) of lead in dust wipe surface samples; (2) to provide data to determine whether the on-site analysis of field dust wipe samples by XRF and the laboratory method of inductively coupled plasma emission analysis (ICP) are comparable; and (3) to determine if differences exist between different wipe materials. Several wipes meet the ASTM E1792 performance requirements of lead background level less than 5 microg per wipe, be only one layer thick, yield recovery rates of 80- 120% from spiked samples, remain damp throughout the sampling procedure, and do not contain aloe. The wipes used in this study were Pace Wipes, which are used for the PAT materials, and, for the field samples, Palintest Wipes, which were supplied by the instrument manufacturer, and Ghost Wipes, which are popular because they digest in hot, concentrated acid, so that chemical analysis is simplified. Twenty PAT wipe samples were obtained from four different proficiency test rounds. Surface wipe samples were taken at three different locations representing different industry types. All samples were analyzed using a portable XRF spectrometer and by ICP. Strong linear relationships were found for the analysis of wipe samples by ICP and by portable XRF. For the PAT samples, the results from the ICP and XRF analysis were not statistically equivalent, which indicates a bias in the ICP analysis. The bias was not excessive, since all ICP analyses fell within the acceptable range for the proficiency samples. The good correlation between the proficiency sample reference values and the XRF determinations is not surprising considering similar proficiency samples were used to calibrate the instrument response. Users of this portable XRF analyzer could enroll in the proficiency test program as part of their quality assurance program. For field samples, the relationship was strongest for Palintest wipes, and the values found for all three industries could be combined. However, the results from the ICP and XRF analysis were not statistically equivalent using the correction factor in the calculation algorithm as supplied with the instrument, and a new coefficient was derived. The mean relative error for the XRF analysis versus the ICP analysis was greater than 25%, such that the method falls within the realm of screening procedures. For Ghost Wipe samples, the precision was different for different industries, and the results could not be pooled. Differences between the two wipe materials may be related to the number of folds required for analysis.


Strategies for Coping with Chronic Lower Back Pain in Patients with Long Physiotherapy Wait Time.
BACKGROUND Treatment efficacy for the increasing prevalence of back pain is a great challenge for both health care providers and individuals coping with this problem. This study aimed to evaluate pain coping strategies used by primary care patients with chronic lower back pain (CLBP) as a supplementation of medical diagnosis before a physiotherapy programme. MATERIAL AND METHODS A total of 88 people were divided into 3 age groups: young adults (21-40 years old), middle-aged adults (41-60 years old), and the elderly (over 60 years old). Data was gathered from rehabilitation centers and primary medical care facilities. A cross-sectional design was used. The Coping Strategies Questionnaire (CSQ) was completed before the physiotherapy course. RESULTS Patients complained of CLBP for 11.32±6.81 years on average. The most common strategies to cope with back pain included declaring that the pain is manageable, praying and hoping, as well as increased behavioral activity. Statistically significant differences in coping strategies were found between age groups. The elderly patients were more likely to "declare coping with pain" in comparison to the younger age groups (p<0.01). People over 60 years of age were more likely to declare active coping with pain, while young people reported catastrophizing. CONCLUSIONS Patients in different age groups had various difficulties in pain coping. Most of them required support in self-management of pain in addition to physiotherapy. The basic assessment of pain coping strategies should be consistently taken into account and included in rehabilitation protocols in chronic pain treatment.


Molecular Chaperone HSP70 and Key Regulators of Apoptosis - A Review.
Identified as a molecular chaperone constitutively being synthesized due to enhanced elevated temperature change, this heat shock protein HSP70 has shown to be intimately involved in many protein biogenesis, facilitating the synthesis and folding of proteins and trafficking of nascent peptides during cell growth. HSP70 also plays a vital role in protein assembly, regulation and interaction with a wide variety of proteins. Stress-induced cell death is under the control of the Bcl-2 family of apoptotic regulators and display either pro-apoptotic or anti-apoptotic activities. Subjected to stress conditions such as heat shock, cells have been reported to express elevated expressions of HSP70. Moreover, this molecular chaperon has shown to act at multiple levels to suppress stressed-induced apoptotic signals of some Bcl-2 members by repairing, re-synthesizing damaged proteins, and stabilizing unfolded proteins. Therefore, HSP70 synthesis can act as an essential recovery mode for cellular survival and adaptation during lethal conditions.


Genetic studies of the sulfonylurea receptor gene locus in NIDDM and in morbid obesity among French Caucasians.
The sulfonylurea receptor (SUR) is a key component in glucose-stimulated insulin secretion. Obesity and NIDDM are frequently associated and share some metabolic abnormalities, suggesting that they might also share some susceptibility genes. Thus, the SUR encoding gene is a plausible candidate for a primary pancreatic beta-cell defect and thus for hyperglycemia and weight gain. Through association and linkage studies, we have investigated the potential role of the SUR gene in families with NIDDM and in two independent sets of morbidly obese families. The exon 22 T-allele at codon 761 was more common in patients with NIDDM (7.7%) and morbid obesity (7.8%) than in control subjects (1.8%, P = 0.030 and P = 0.023, respectively). This variant was associated with morbid obesity (odds ratio 3.71, P = 0.017) and NIDDM (odds ratio 2.20, P = 0.04; association dependent on BMI). Although the frequencies for intron 24 variant were similar in all groups, morbidly obese patients homozygous for the c-allele had a more deleterious form of obesity. Sib-pair linkage studies with NIDDM in French Caucasian families gave no evidence for linkage to the SUR locus. However, in one set of the obese families, we found an indication for linkage with a SUR-linked microsatellite marker (D11S419, P = 0.0032). We conclude that in Caucasians, the SUR locus may contribute to the genetic susceptibility to NIDDM and obesity.


Full length Vpu from HIV-1: combining molecular dynamics simulations with NMR spectroscopy.
Based on structures made available by solution NMR, molecular models of the protein Vpu from HIV-1 were built and refined by 6 ns MD simulations in a fully hydrated lipid bilayer. Vpu is an 81 amino acid type I integral membrane protein encoded by the human immunodeficiency virus type-1 (HIV-1) and closely related simian immunodeficiency viruses (SIVs). Its role is to amplify viral release. Upon phosphorylation, the cytoplasmic domain adopts a more compact shape with helices 2 and 3 becoming almost parallel to each other. A loss of helicity for several residues belonging to the helices adjacent to both ends of the loop region containing serines 53 and 57 is observed. A fourth helix, present in one of the NMR-based structures of the cytoplasmic domain and located near the C-terminus, is lost upon phosphorylation.


Changes in Mobility and Muscle Function of Children with Cerebral Palsy after Gait Training: A Pilot Study.
The goal of this pilot study was to characterize the effects of gait training on the capacity of muscles to produce body accelerations and relate these changes to mobility improvements seen in children with cerebral palsy (CP). Five children (14 years ± 3 y; GMFCS I-II) with spastic diplegic CP participated in a 6-week gait training program. Changes in 10-m fast-as-possible walking speed and 6-minute walking endurance were used to assess changes in mobility. In addition, musculoskeletal modeling was used to determine the potential of lower-limb muscles to accelerate the body's center of mass vertically and forward during stance. The mobility changes after the training were mixed, with some children demonstrating vast improvements, while others appeared to be minimal. However, the musculoskeletal results revealed unique responses for each child. The most common changes occurred in the capacity for the hip and knee extensors to produce body support and the hip flexors to produce body propulsion. These results cannot yet be generalized to the broad population of children with CP, but demonstrate that therapy protocols may be enhanced by modeling analyses. The pilot study results provide motivation for gait training emphasizing upright leg posture, mediolateral balance, and ankle push-off.


Annexin A2-STAT3-Oncostatin M receptor axis drives phenotypic and mesenchymal changes in glioblastoma.
Glioblastoma (GBM) is characterized by extensive tumor cell invasion, angiogenesis, and proliferation. We previously established subclones of GBM cells with distinct invasive phenotypes and identified annexin A2 (ANXA2) as an activator of angiogenesis and perivascular invasion. Here, we further explored the role of ANXA2 in regulating phenotypic transition in GBM. We identified oncostatin M receptor (OSMR) as a key ANXA2 target gene in GBM utilizing microarray analysis and hierarchical clustering analysis of the Ivy Glioblastoma Atlas Project and The Cancer Genome Atlas datasets. Overexpression of ANXA2 in GBM cells increased the expression of OSMR and phosphorylated signal transducer and activator of transcription 3 (STAT3) and enhanced cell invasion, angiogenesis, proliferation, and mesenchymal transition. Silencing of OSMR reversed the ANXA2-induced phenotype, and STAT3 knockdown reduced OSMR protein expression. Exposure of GBM cells to hypoxic conditions activated the ANXA2-STAT3-OSMR signaling axis. Mice bearing ANXA2-overexpressing GBM exhibited shorter survival times compared with control tumor-bearing mice, whereas OSMR knockdown increased the survival time and diminished ANXA2-mediated tumor invasion, angiogenesis, and growth. Further, we uncovered a significant relationship between ANXA2 and OSMR expression in clinical GBM specimens, and demonstrated their correlation with tumor histopathology and patient prognosis. Our results indicate that the ANXA2-STAT3-OSMR axis regulates malignant phenotypic changes and mesenchymal transition in GBM, suggesting that this axis is a promising therapeutic target to treat GBM aggressiveness.


The role of neutrophyl lymphocyte count ratio as an inflammatory marker in systemic lupus erythematosus.
to evaluate neutrophyl lymphocyte count ratio (NLCR) as an inflammatory marker in systemic lupus erythematosus (SLE). this was a cross sectional study involving 21 SLE patients with mild and moderate disease activity according to Mex SLEDAI score and 30 normal subjects. White blood cells counts were examined with Sysmex XE-5000. The absolute neutrophyl count and absolute lymphocyte count were tabulated and compared between SLE and normal subjects using unpaired t-test. The comparison of NLCR between SLE and normal subject was calculated using Mann-Whitney test. This study was conducted in the Department of Internal Medicine Hasan Sadikin Hospital from November 2011 until January 2012. there was no significant difference in absolute neutrophyl count between SLE and normal subjects (4158+1517 vs 4031+1218 /mm3, p=0.74). The absolute lymphocyte count was significantly lower in SLE than normal subjects (1721+600 vs 2397+587/mm3, p=0.000). NLCR was significantly higher in SLE than normal subjects (2.52 (1.01-10.92) vs 1.65 (0.77-4.59), p=0.007). There was no significant difference in NLCR between SLE with mild and moderate activity (2.59 (1.01-10.92) vs 2.01 (1.38-3.98), p= 0.412). Based on ROC curve, with AUC 0.727, cut off NLCR value >1.93 had 70% sensitivity and 67% specificity in differentiating between SLE and normal subjects. NLCR could be used as simple inflammatory parameter in SLE.


Health Impacts of Climate Change as Contained in Economic Models Estimating the Social Cost of Carbon Dioxide.
The health impacts of climate change are substantial and represent a primary motivating factor to mitigate climate change. However, the health impacts in economic models that estimate the social cost of carbon dioxide (SC-CO2) have generally been made in isolation from health experts and have never been rigorously evaluated. Version 3.10 of the Framework for Uncertainty, Negotiation and Distribution (FUND) model was used to estimate the health-based portion of current SC-CO2 estimates across low-, middle-, and high-income regions. In addition to the base model, three additional experiments assessed the sensitivity of these estimates to changes in the socio-economic assumptions in the model. Economic impacts from adverse health outcomes represent ∼8.7% of current SC-CO2 estimates. The majority of these health impacts (74%) were attributable to diarrhea mortality (from both low- and high-income regions) followed by diarrhea morbidity (12%) and malaria mortality (11%); no other health impact makes a meaningful contribution to SC-CO2 estimates in current economic models. The results of the socio-economic experiments show that the health-based portion of SC-CO2 estimates are highly sensitive to assumptions regarding income elasticity of health effects, income growth, and use of equity weights. Improving the health-based portion of SC-CO2 estimates could have substantial impacts on magnitude of the SC-CO2. Incorporating additional health impacts not previously included in estimates of SC-CO2 will be a critical component of model updates. This effort will be most successful through coordination between economists and health researchers and should focus on updating the form and function of concentration-response functions.


Selection on Accessible Chromatin Regions in Capsella grandiflora.
Accurate estimates of genome-wide rates and fitness effects of new mutations are essential for an improved understanding of molecular evolutionary processes. Although eukaryotic genomes generally contain a large noncoding fraction, functional noncoding regions and fitness effects of mutations in such regions are still incompletely characterized. A promising approach to characterize functional noncoding regions relies on identifying accessible chromatin regions (ACRs) tightly associated with regulatory DNA. Here, we applied this approach to identify and estimate selection on ACRs in Capsella grandiflora, a crucifer species ideal for population genomic quantification of selection due to its favorable population demography. We describe a population-wide ACR distribution based on ATAC-seq data for leaf samples of 16 individuals from a natural population. We use population genomic methods to estimate fitness effects and proportions of positively selected fixations (α) in ACRs and find that intergenic ACRs harbor a considerable fraction of weakly deleterious new mutations, as well as a significantly higher proportion of strongly deleterious mutations than comparable inaccessible intergenic regions. ACRs are enriched for expression quantitative trait loci (eQTL) and depleted of transposable element insertions, as expected if intergenic ACRs are under selection because they harbor regulatory regions. By integrating empirical identification of intergenic ACRs with analyses of eQTL and population genomic analyses of selection, we demonstrate that intergenic regulatory regions are an important source of nearly neutral mutations. These results improve our understanding of selection on noncoding regions and the role of nearly neutral mutations for evolutionary processes in outcrossing Brassicaceae species.


Molecular analysis of anaplastic oligodendroglial tumors in a prospective randomized study: A report from EORTC study 26951.
Recent studies have shown that the clinical outcome of anaplastic oligodendroglial tumors is variable, but also that the histological diagnosis is subject to interobserver variation. We investigated whether the assessment of 1p/19q codeletion, polysomy of chromosome 7, epidermal growth factor receptor (EGFR) gene amplification (EGFR(amp)), and loss of chromosome 10 or 10q offers additional prognostic information to the histological diagnosis and would allow molecular subtyping. For this study, we used the clinical data and tumor samples of the patients included in multicenter prospective phase III European Organisation for Research and Treatment of Cancer (EORTC) study 26951 on the effects of adjuvant procarbazine, chloroethyl cyclohexylnitrosourea (lomustine), and vincristine chemotherapy in anaplastic oligodendroglial tumors. Fluorescence in situ hybridization was used to assess copy number aberrations of chromosome 1p, 19q, 7, 10, and 10q and EGFR. Three different analyses were performed: on all included patients based on local pathology diagnosis, on the patients with confirmed anaplastic oligodendroglial tumors on central pathology review, and on this latter group but after excluding anaplastic oligoastrocytoma (AOA) with necrosis. As a reference set for glioblastoma multiforme (GBM), patients from the prospective randomized phase III study on GBM (EORTC 26981) were used as a benchmark. In 257 of 368 patients, central pathology review confirmed the presence of an anaplastic oligodendroglial tumor. Tumors with combined 1p and 19q loss (1p(loss)19q(loss)) were histopathologically diagnosed as anaplastic oligodendroglioma, were more frequently located in the frontal lobe, and had a better outcome. Anaplastic oligodendroglial tumors with EGFR(amp) were more frequently AOA, were more often localized outside the frontal lobe, and had a survival similar to that for GBM. Survival of patients with AOA harboring necrosis was in a similar range as for GBM, while patients with AOA with only endothelial proliferation had better overall survival. In univariate analyses, all molecular factors except loss of 10q were of prognostic significance, but on multivariate analysis a histopathological diagnosis of AOA, necrosis, and 1p(loss)19q(loss) remained independent prognostic factors. AOA tumors with necrosis are to be considered WHO grade IV tumors (GBM). Of all molecular markers analyzed in this study, especially loss of 1p/19q carried prognostic significance, while the others contributed little prognostic value to classical histology.


Avian leukosis virus subgroup J-associated myelocytoma in a hobby chicken.
The avian leukosis viruses (ALVs) are a major group of retroviruses associated with neoplastic diseases in poultry. The ALV-J strain was identified as a cause of myelocytomas in broiler breeder and broiler chickens in the UK in the 1980s; however, following eradication of the virus,commercial broilers have remained free of infection since the early 2000s. A pet chicken was submitted to Animal and Plant Health Agency (APHA) in 2013 with a history of croaking respirations, abnormality of the left eye and apparent paralysis. Postmortem examination of the bird showed widespread tumour-like infiltration of many organs, including the pectoral muscles, internal organs, sternum and ribs. Histopathological examination of the affected tissues revealed myelocytoma formation typical of the lesions associated with ALV-J, and the virus was confirmed by PCR testing and sequencing. Virus was not detected in blood samples in the other five chickens remaining in the flock. The source of infection was not established. This was the first time ALV-J had been seen in the UK since its eradicationand the case highlights the importance of continued surveillance of backyard and hobby chickens to detect potential new and re-emerging disease threats, such as ALV-J, which may be of significance to the wider poultry population.


Analysis of mutations from SCID and Omenn syndrome patients reveals the central role of the Rag2 PHD domain in regulating V(D)J recombination.
Rag2 plays an essential role in the generation of antigen receptors. Mutations that impair Rag2 function can lead to severe combined immunodeficiency (SCID), a condition characterized by complete absence of T and B cells, or Omenn syndrome (OS), a form of SCID characterized by the virtual absence of B cells and the presence of oligoclonal autoreactive T cells. Here, we present a comparative study of a panel of mutations that were identified in the noncanonical plant homeodomain (PHD) of Rag2 in patients with SCID or OS. We show that PHD mutant mouse Rag2 proteins that correspond to those found in these patients greatly impaired endogenous recombination of Ig gene segments in a Rag2-deficient pro-B cell line and that this correlated with decreased protein stability, impaired nuclear localization, and/or loss of the interaction between Rag2 and core histones. Our results demonstrate that point mutations in the PHD of Rag2 compromise the functionality of the entire protein, thus explaining why the phenotype of cells expressing PHD point mutants differs from those expressing core Rag2 protein that lacks the entire C-terminal region and is therefore devoid of the regulation imposed by the PHD. Together, our findings reveal the various deleterious effects of PHD Rag2 mutations and demonstrate the crucial role of this domain in regulating antigen receptor gene assembly. We believe these results reveal new mechanisms of immunodeficiency in SCID and OS.


Two-dimensional correlation analysis of nuclear magnetic resonance metabonomics data.
Two-dimensional (2D) correlation analysis has been used in this study to identify changes in complex nuclear magnetic resonance (NMR) metabonomics spectra of rat urine samples obtained during a study in which vasculitis (vascular injury), an important safety element in preclinical trials, was induced. Two types of correlation analysis were performed, along the variables and along the samples, and both 2D covariance and correlation coefficient maps were calculated. The binned and ''raw'' NMR spectra were analyzed (0.04 and 0.001 ppm resolution, respectively). Good correlation was found among the major peaks of the binned spectra, and two groups of samples were identified using sample-sample 2D correlation maps. Much more complex correlation features were obtained from the ''raw'' spectra, in which the specific, butterfly-like patterns were obtained in the covariance map but with only a few significant correlation coefficients in the corresponding 2D correlation maps. In terms of classification, the same group of the last nine spectra that indicated the end of the process and clustered in the 2D sample-sample covariance map of the binned data was also found in the 2D sample-sample covariance map of the raw NMR spectra but, again, not in the 2D correlation coefficient map. A discussion is given on the details of the application of the correlation analysis with regard to spectral data resolution, alignment, the effect of actual intensities of the NMR signal, and reference to various results from 2D correlation analysis of vibrational spectra.


Is Non-Contrast CT Adequate for the Evaluation of Hepatic Metastasis in Patients Who Cannot Receive Iodinated Contrast Media?
To evaluate the appropriateness of follow-up with only non-enhanced CT (NECT) in patients with gastrointestinal cancer. This retrospective study included 323 patients with colorectal and gastric cancer who underwent two consecutive CT examinations (CT1 and CT2), including non-contrast and portal venous phase CT images, with an interval of 1 year. Patients were divided into 2 groups: Group A included patients with no hepatic metastasis on CT1 and with or without newly developed metastasis on CT2 to evaluate the diagnostic performance of NECT for detecting newly developed hepatic metastasis; Group B included patients with known hepatic metastasis both on CT1 and CT2 to evaluate the accuracy of NECT for the assessment of hepatic metastasis based on RECIST criteria (version 1.1). Contrast-enhanced CT (CECT) images were considered as reference standards. Group A included 172 patients (M:F = 107:65; mean age, 62.6 years). Among them, 57 patients had 95 metastases (mean size, 2.2 ± 1.3 cm). Per patient and per lesion sensitivity for diagnosing newly developed hepatic metastasis was 56.1-66.7% and 52.6-56.8%, respectively. In terms of small metastases (<1.5 cm), per lesion sensitivity was significantly decreased to 28.1-34.4% (P < 0.05). Metastasis size measurements were significantly smaller on NECT (P < 0.001) compared with reference standards. In Group B, the accuracy of response evaluation based on RECIST criteria was 65.6-72.2%. NECT showed inadequate diagnostic performances in both detecting newly developed hepatic metastasis and evaluating the response of hepatic metastasis based on RECIST criteria.


Measuring equity in disability and healthcare utilization in Afghanistan.
This paper analyses equity in health and healthcare utilization in Afghanistan based on a representative national household survey. Equitable access is a cornerstone of the Afghan health policy. We measured socioeconomic-related equity in access to public health care, using disability--because people with disabilities are poorer and more likely to use health care--and a concentration index (CI) and its decomposition. The socioeconomic-related equity in healthcare utilization was measured using a probit model and compared with an OLS model providing the horizontal inequity index (HI). We found a low rate of healthcare facilities utilization (25%). Disabled persons are using more healthcare facilities and have higher medical expenses. Disability is more frequently associated with older age, unemployed heads of household and lower education. The Cl of disability is 0.0221 indicating a pro-rich distribution of health. This pro-rich effect is higher in small households (CI decreases with size of the household, -0.0048) and safe (0.0059) areas. The CI of healthcare utilization is -0.0159 indicating a slightly pro-poor distribution of healthcare utilization but, overall, there is no difference in healthcare utilization by wealth status. Our study does not show major socioeconomic related inequity in disability and healthcare utilization in Afghanistan. This is due to the extreme and pervasive poverty found in Afghanistan. The absence of inequity in health access is explained by the uniform poverty of the population and the difficulty of accessing BPHS facilities (a basic package of health services), despite alarming health indicators.


Common pathological processes in Alzheimer disease and type 2 diabetes: a review.
Alzheimer disease (AD) and type 2 diabetes mellitus (T2DM) are conditions that affect a large number of people in the industrialized countries. Both conditions are on the increase, and finding novel treatments to cure or prevent them are a major aim in research. Somewhat surprisingly, AD and T2DM share several molecular processes that underlie the respective degenerative developments. This review describes and discusses several of these shared biochemical and physiological pathways. Disturbances in insulin signalling appears to be the main common impairment that affects cell growth and differentiation, cellular repair mechanisms, energy metabolism, and glucose utilization. Insulin not only regulates blood sugar levels but also acts as a growth factor on all cells including neurons in the CNS. Impairment of insulin signalling therefore not only affects blood glucose levels but also causes numerous degenerative processes. Other growth factor signalling systems such as insulin growth factors (IGFs) and transforming growth factors (TGFs) also are affected in both conditions. Also, the misfolding of proteins plays an important role in both diseases, as does the aggregation of amyloid peptides and of hyperphosphorylated proteins. Furthermore, more general physiological processes such as angiopathic and cytotoxic developments, the induction of apoptosis, or of non-apoptotic cell death via production of free radicals greatly influence the progression of AD and T2DM. The increase of detailed knowledge of these common physiological processes open up the opportunities for treatments that can prevent or reduce the onset of AD as well as T2DM.


Development of a polydimethylsiloxane-thymol/nitroprusside composite based sensor involving thymol derivatization for ammonium monitoring in water samples.
This report describes a polydimethylsiloxane (PDMS)-thymol/nitroprusside delivery composite sensor for direct monitoring of ammonium in environmental water samples. The sensor is based on a PDMS support that contains the Berthelot's reaction reagents. To prepare the PDMS-thymol/nitroprusside composite discs, thymol and nitroprusside have been encapsulated in the PDMS matrix, forming a reagent release support which significantly simplifies the analytical measurements, since it avoids the need to prepare derivatizing reagents and sample handling is reduced to the sampling step. When, the PDMS-thymol/nitroprusside composite was introduced in water samples spontaneous release of the chromophore and catalyst was produced, and the derivatization reaction took place to form the indothymol blue. Thus, qualitative analysis of NH4(+) could be carried out by visual inspection, but also, it can be quantified by measuring the absorbance at 690 nm. These portable devices provided good sensitivity (LOD<0.4 mg L(-1)) and reproducibility (RSD <10%) for the rapid detection of ammonium. The PDMS-NH4(+) sensor has been successfully applied to determine ammonium in water samples and in the aqueous extracts of particulate matter PM10 samples. Moreover, the reliability of the method for qualitative analysis has been demonstrated. Finally, the advantages of the PDMS-NH4(+) sensor have been examined by comparing some analytical and complementary characteristics with the properties of well-established ammonium determination methods.


Dissecting the diverse actions of pro- and mature neurotrophins.
The neurotrophins mediate diverse actions in the developing peripheral and central nervous systems. They are initially synthesized as precursor forms, or proneurotrophins, that are cleaved to release C-terminal mature forms that bind to Trk receptor tyrosine kinases to enhance synaptic plasticity and neuronal survival. Recent studies suggest that proneurotrophins are not inactive precursors, but signaling proteins that can activate the p75 receptor to mediate diverse responses. Proneurotrophins can activate a heteromeric receptor complex of p75 and sortilin to initiate cell death, or bind to p75 in hippocampal neurons to enhance long term depression. Thus, neurotrophin actions are regulated by the form of the neurotrophin (pro- or mature) secreted by cells, by extracellular proteolytic cleavage of proneurotrophins to generate mature forms, and by the expression of neurotrophin receptors Trk, or p75 and sortilin, that are selectively activated by mature or proneurotrophins, respectively. Here, recent studies are reviewed that reveal that pro- and mature neurotrophins have distinct and sometimes opposing actions in regulating cell death and survival in development and in pathophysiologic states, in regulating neurotrophin secretion, and in modulating synaptic plasticity.


Factors That Increase Risk of Celiac Disease Autoimmunity After a Gastrointestinal Infection in Early Life.
Little is known about the pathogenic mechanisms of gluten immunogenicity in patients with celiac disease. We studied temporal associations between infections and the development of celiac disease autoimmunity, and examined effects of HLA alleles, rotavirus vaccination status, and infant feeding. We monitored 6327 children in the United States and Europe carrying HLA risk genotypes for celiac disease from 1 to 4 years of age for presence of tissue transglutaminase autoantibodies (the definition of celiac disease autoimmunity), until March 31, 2015. Parental reports of gastrointestinal and respiratory infections were collected every third month from birth. We analyzed time-varying relationships among reported infections, rotavirus vaccination status, time to first introduction of gluten, breastfeeding, and risk of celiac disease autoimmunity using proportional hazard models. We identified 13,881 gastrointestinal infectious episodes (GIE) and 79,816 respiratory infectious episodes. During the follow-up period, 732 of 6327 (11.6%) children developed celiac disease autoimmunity. A GIE increased the risk of celiac disease autoimmunity within the following 3 months by 33% (hazard ratio [HR], 1.33; 95% confidence interval [CI], 1.11-1.59). This risk increased 2-fold among children born in winter and introduced to gluten before age 6 months (HR, 2.08; 95% CI, 1.46-2.98), and increased 10-fold among children without HLA-DQ2 alleles and breastfed for fewer than 4 months (HR, 9.76; 95% CI, 3.87-24.8). Risk of celiac disease autoimmunity was reduced in children vaccinated against rotavirus and introduced to gluten before age 6 months (HR, 0.57; 95% CI, 0.36-0.88). Gastrointestinal infections increase the risk of celiac disease autoimmunity in children with genetic susceptibility to this autoimmune disorder. The risk is modified by HLA genotype, infant gluten consumption, breastfeeding, and rotavirus vaccination, indicating complex interactions among infections, genetic factors, and diet in the etiology of celiac disease in early childhood.


Relationship of the recurrent laryngeal nerve to the superior parathyroid gland during thyroidectomy.
Design: The relationship of the recurrent laryngeal nerve to the superior parathyroid gland during consecutive thyroidectomies was prospectively evaluated. When one structure was noted, careful dissection was performed to locate the other structure, to preserve their natural anatomical relationship. Patients: In total, 103 consecutive thyroid lobectomies were performed on 73 patients. The distance from the superior parathyroid gland to the recurrent laryngeal nerve was recorded. Results: In 88 cases (88.9 per cent), the superior parathyroid gland was identified within 5 mm of the recurrent laryngeal nerve. In 62 cases (62.6 per cent), the gland was within 1 mm of the recurrent laryngeal nerve. The height of the thyroid lobe was positively associated with the distance between the two structures (p = 0.001), as was the incidence of cancer (p = 0.033). The incidence of recurrent laryngeal nerve paresis was less than 4 per cent. Conclusion: In most cases, the recurrent laryngeal nerve was found in close proximity to the superior parathyroid gland. In a thyroid gland with a large height, or in a cancerous lobe, this relationship is less reliable.


Cellular immune responses of human cadaver donor bone marrow cells and their susceptibility to commonly used immunosuppressive drugs in transplantation.
The cascade of immunological effects brought about by donor bone marrow cell (DBMC) infusions in human organ transplantation, especially in the context of continuous pharmacologic immunosuppression, is not fully understood. Yet, in inbred rodents and even primates, administration of specific bone marrow cells has caused a state of acquired immunologic tolerance. In vitro mixed lymphocyte culture (MLC) and cell-mediated lympholysis (CML) culture systems were used to compare the responding and regulatory properties of DBMC and individual bone marrow cell subsets versus spleen cells in the presence or absence of pharmacologic immunosuppression. In the absence of immunosuppressive drugs, the DBMC proliferated in MLC and in response to phytohemagglutinin, but to a lower magnitude than donor spleen cells. In CML assays, DBMC failed to function as cytotoxic cells. Removal of both CD3+ and CD34+ cells together (not just singly) had to occur for complete abrogation of the proliferative response of DBMC evoked in the presence of allogeneic stimulating cells. Testing several experimental variables using flow cytometric analysis led to the conclusion that when purified DBMC CD34+ cells were placed in coculture with irradiated allogeneic peripheral blood mononuclear cells, such CD34+ cells give rise both to CD3- TCRalphabeta+ as well as to dimly staining CD3+ TCRalphabeta+ cells. Low pharmacologic concentrations of tacrolimus/cyclosporine (CsA) and mycophenolic acid (MPA) singly or in combination had no effect on the spontaneous proliferation of DBMC and had significantly less inhibitory activity on MLC responses of DBMC and its purified CD3+ or CD34+ subpopulations, compared with the responses of spleen cells. Moreover, the previously described regulatory effects of DBMC on the MLC responses of peripheral blood or splenic responding cells were not inhibited by these immunosuppressive drugs. Taken together, these results support the notion that in vitro DBMC subpopulations, which proliferate as responding cells in co-culture with x-irradiated allogeneic cells and which cause regulatory effects when added as a third component to MLC reactions, seem to be culture-generated lymphoid cell lineage(s) progeny of CD34+ cells. This possibly includes unique CD3+ "primitive" (dimly staining) T cells, which are not as inhibited in their function by tacrolimus/CsA and MPA, as are postthymic (splenic) T cells.


High glucose and low specific cell growth but not mild hypothermia improve specific r-protein productivity in chemostat culture of CHO cells.
In the biopharmaceutical sector, Chinese hamster ovary (CHO) cells have become the host of choice to produce recombinant proteins (r-proteins) due to their capacity for correct protein folding, assembly, and posttranslational modification. However, the production of therapeutic r-proteins in CHO cells is expensive and presents insufficient production yields for certain proteins. Effective culture strategies to increase productivity (qp) include a high glucose concentration in the medium and mild hypothermia (28-34 °C), but these changes lead to a reduced specific growth rate. To study the individual and combined impacts of glucose concentration, specific growth rate and mild hypothermia on culture performance and cell metabolism, we analyzed chemostat cultures of recombinant human tissue plasminogen activator (rh-tPA)-producing CHO cell lines fed with three glucose concentrations in feeding media (20, 30 and 40 mM), at two dilution rates (0.01 and 0.018 1/h) and two temperatures (33 and 37 °C). The results indicated significant changes in cell growth, cell cycle distribution, metabolism, and rh-tPA productivity in response to the varying environmental culture conditions. High glucose feed led to constrained cell growth, increased specific rh-tPA productivity and a higher number of cells in the G2/M phase. Low specific growth rate and temperature (33 °C) reduced glucose consumption and lactate production rates. Our findings indicated that a reduced specific growth rate coupled with high feed glucose significantly improves r-protein productivity in CHO cells. We also observed that low temperature significantly reduced qp, but not cell growth when dilution rate was manipulated, regardless of the glucose concentration or dilution rate. In contrast, we determined that feed glucose concentration and consumption rate were the dominant aspects of the growth and productivity in CHO cells by using multivariate analysis.


Histochemical demonstration of neural carbonic anhydrase activity within the mandibular molar and incisor tooth pulps of the rat.
Carbonic anhydrase (CA) activity was enzyme histochemically assessed in: (1) neuronal cell bodies in the rat trigeminal ganglion that were retrogradely labeled with FITC-WGA from the mandibular molar and incisor tooth pulps; (2) cell bodies retrogradely labeled with HRP-WGA from the mandibular molar tooth pulp; and (3) nerve fibers within the decalcified mandibular molar and incisor tooth pulps. The cell size spectrum of the FITC-WGA-labeled neurons was similar for both molar and incisor tooth pulps with about 90% being medium or large (greater than or equal to 300 microns2 in cross-sectional area). About 30% of the FITC-WGA-labeled cell bodies exhibited CA activity for both tooth pulps. The HRP-WGA-labeled cells also showed CA activity. In the molar root pulp, finely myelinated nerve fibers exhibited CA activity. Histochemical reaction products were distributed in the axoplasm and the cytoplasmic pocket of Schwann cell inside the compact myelin. In the incisor pulp, CA-reactive unmyelinated axons coexisted with non-reactive axons within the same Schwann units. The Schwann cell cytoplasm directly surrounding reactive axons also exhibited histochemical stainability. In the light of known distribution of CA in the peripheral nervous system, the present data suggest that the rat molar and incisor tooth pulps receive substantial innervation of large myelinated primary afferent fibers. The finely myelinated and unmyelinated axons, that have been repeatedly shown to predominate the intrapulpal nerve fibers, are considered to represent intrapulpal, terminal and preterminal specialization of otherwise large myelinated nerve fibers.