The Value of Three-Dimensional Printing Spine Model in Severe Spine Deformity Correction Surgery.
Retrospective case-control study. We aimed to evaluate the value of 3-dimensional printing (3DP) spine model in the surgical treatment of severe spinal deformity since the prosperous development of 3DP technology. Severe scoliosis or hyper-kyphosis patients underwent posterior fixation and fusion surgery using the 3DP spine models were reviewed (3DP group). Spinal deformity surgeries operated by free-hand screw implantation during the same period were selected as the control group after propensity score matching (PSM). The correction rate, pedicle screw accuracy, and complications were analyzed. Class A and B screws were defined as accurate according to Gertzbein and Robbins criteria. 35 patients were enrolled in the 3DP group and 35 matched cases were included in the control group. The perioperative baseline data and deformity correction rate were similar between both groups (P > .05). However, the operation time and blood loss were significantly less in the 3DP group (296.14 ± 66.18 min vs. 329.43 ± 67.16 min, 711.43 ± 552.28 mL vs. 1322.29 ± 828.23 mL, P < .05). More three-column osteotomies (Grade 3-6) were performed in the 3DP group (30/35, 85.7% vs. 21/35, 60.0%. P = .016). The screw placement accuracy was significantly higher in the 3DP group (422/582, 72.51% vs. 397/575, 69.04%. P = .024). The screw misplacement related complication rate was significantly higher in the free-hand group (6/35 vs. 1/35, P = .046). The study provided solid evidence that 3DP spine models can enhance surgeons' confidence in performing higher grade osteotomies and improve the safety and efficiency in severe spine deformity correction surgery. 3D printing technology has a good prospect in spinal deformity surgery.


Individual fatty acids influence cultured human liver cells differently when compared to plant oils.
Most work to date using fatty acids and cultured cells has concentrated on single fatty acids, yet cells in vivo are exposed to fatty acid combinations in the form of triacylglycerols and phosphoglycerides. We have quantitated the influence of the individual fatty acids that make up three plant oils, as well as the plant oils themselves, on the growth of normal and transformed human liver cell lines. The effects of the individual fatty acids were then combined in proportion to their concentration in each plant oil and compared to the effects of the intact oils. The oils initially induced synergistic effects with low concentrations, but with high concentrations competitive effects were shown, when compared to the combined effects of the individual fatty acids.


Problematic mobile phone use and time management disposition in Chinese college students: the chain mediating role of sleep quality and cognitive flexibility.
With the widespread adoption of smartphones, there has been a notable increase in problematic mobile phone use (PMPU), particularly prevalent among college students. Research suggests that apart from being associated with various problematic behaviors, this excessive mobile phone use might also have an impact on individual personality traits, such as time management disposition (TMD), which plays a significant role in individual motivation and psychological well-being. While previous literature has identified a negative relationship between PMPU and TMD, few studies have delved into the underlying mediating mechanism. Thus, the main aim of this study was to examine the chain mediating effect of sleep quality and cognitive flexibility on the relationship between PMPU and TMD. A total of 921 Chinese college students completed the questionnaire. We collected basic information about the participants and assessed their PMPU, TMD, sleep quality, and cognitive flexibility using the Problematic Mobile Phone Use Scale-10, Adolescence Time Management Disposition Inventory, Pittsburgh Sleep Quality Index scale and Cognitive Flexibility Inventory. The results indicated a significant correlation among all the variables. Moreover, we noted that both sleep quality and cognitive flexibility fully mediated the association between PMPU and TMD. Additionally, a chain mediating effect involving sleep quality and flexibility in this relationship was also identified. We found that sleep quality and cognitive flexibility had a series of multiple mediating effects in the pathway from PMPU to TMD, and both significantly mediated TMD. These findings indicated that impaired cognitive function and sleep quality may contribute to time management difficulties resulting from PMPU, suggesting that problematic behaviors like PMPU can impact one's personality traits. Therefore, interventions should be enhanced to mitigate the adverse effects of PMPU.


Anaphylactoid reactions to tolmetin after interrupted dosage.
Seven patients had anaphylactoid reactions after ingesting tolmetin sodium. In each case the reaction followed readministration of the drug after an interrupted period of at least three days and within 90 minutes of taking a single 400-mg capsule. None of the patients had had prior anaphylaxis and none were judged atopic. Skin tests to an extract of the drug (0.02 mg) were uniformly negative, whereas a higher concentration produced a nonspecific irritant reaction. In vitro tests in one patient for IgE antibodies to tolmetin and for complement activation were negative.


The effect of maternal sleep deprivation on differentiation of mesenchymal stem cells in the presence of neonates brain cerebrospinal fluid of Wistar rats.
Cerebrospinal fluid (CSF) contains proliferation, differentiation and maturation signals that are essential factors for brain development. Due to presence of such factors this fluid has proliferative and differentiation potential. A previous study showed that maternal sleep deprivation (MSD) decrease the number of newborn neurons in development of hippocampus. Also, it impairs hippocampus-dependent spatial learning and memory in the young offspring rat. MSD can change CSF factors. In the present study, the effect of MSD-CSF on differentiation of mesenchymal stem cells to neural cells was examined, and expression of Nestin, Neun, and NeurD1 that are neurogenic markers was investigated. In this study, bone marrow mesenchymal stem cells were aspirated from the femur and tibia of young male Wistar rats. Then cell suspension was cultured in DMEM medium supplemented with 10 % FBS and 1 % antibiotics. Pregnant rats were subjected to sleep deprivation for 6 h by gentle handling during 4th, 7th, and 18th day of pregnancy. CSF was collected from cisterna magna of neonate rats. CSF was added to culture media with a 5 % ratio (v/v). Then cell viability was determined with MTT assay. Total cellular RNA was extracted, cDNA was synthesized and NeuN, Nestin, NeuroD1 and IL-6 genes were analyzed by Real-time PCR. Real time-PCR analysis showed that expression of Neun and NeurD1gene decreased compared with culture in normal CSF (N-CSF), and also showed that expression of Nestin did not decrease, inflammatory gene (IL-6) showed high expression compared to culture with N-CSF. Based on our results, MSD-CSF could inhibit neurogenesis process in mesenchymal stem cells and also, this result suggests that MSD can suppress neural differentiation and decrease of neurogenesis gene expression. Overall these findings suggest that insomnia and sleep loss may active inflammatory responses in the brain and change CSF-markers (Fig. 3, Ref. 34).


Association between breastfeeding, host genetic factors, and calicivirus gastroenteritis in a Nicaraguan birth cohort.
Norovirus and sapovirus are important causes of childhood acute gastroenteritis (AGE). Breastfeeding prevents AGE generally; however, it is unknown if breastfeeding prevents AGE caused specifically by norovirus and sapovirus. We investigated the association between breastfeeding and norovirus or sapovirus AGE episodes in a birth cohort. Weekly data on breastfeeding and AGE episodes were captured during the first year of life. Stools were collected from children with AGE and tested by RT-qPCR for norovirus and sapovirus. Time-dependent Cox models estimated associations between weekly breastfeeding and time to first norovirus or sapovirus AGE. From June 2017 to July 2018, 444 newborns were enrolled in the study. In the first year of life, 69 and 34 children experienced a norovirus and a sapovirus episode, respectively. Exclusive breastfeeding lasted a median of 2 weeks, and any breastfeeding lasted a median of 43 weeks. Breastfeeding in the last week did not prevent norovirus (HR: 1.09, 95% CI: 0.62, 1.92) or sapovirus (HR: 1.00, 95% CI: 0.82, 1.21) AGE in a given week, adjusting for household sanitation, consumption of high-risk foods, and mother's and child's histo-blood group phenotypes. Maternal secretor-positive phenotype was protective against norovirus AGE, whereas child's secretor-positive phenotype was a risk factor for norovirus AGE. Exclusive breastfeeding in this population was short-lived, and no conclusions could be drawn about its potential to prevent norovirus or sapovirus AGE. Non-exclusive breastfeeding did not prevent norovirus or sapovirus AGE in the first year of life. However, maternal secretor-positive phenotype was associated with a reduced hazard of norovirus AGE.


Cassava is not a goitrogen in mice.
To examine the effect of cassava on the thyroid function of mice, we fed fresh cassava root to mice and compared this diet with low iodine diet and Purina. Cassava provided a low iodine intake and increased urine thiocyanate excretion and serum thiocyanate levels. Mice on cassava lost weight. The thyroid glands of mice on cassava were not enlarged, even when normalized for body weight. The 4- and 24-hr thyroid uptakes of mice on cassava were similar to those of mice on low iodine diets. Protein-bound [125I]iodine at 24 hr was high in mice on either the cassava or low iodine diets. The thyroid iodide trap (T/M) was similar in mice on cassava and low iodine diets. When thiocyanate was added in vitro to the incubation medium, T/M was reduced in all groups of mice; under these conditions, thiocyanate caused a dose-related inhibition of T/M. The serum thyroxine (T4) and triiodothyronine (T3) concentrations of mice on cassava were reduced compared with mice on Purina diet. Thyroid T4 and T3 contents of mice on cassava were relatively low compared with mice on Purina diet. Hepatic T3 content and T4 5'-monodeiodination in liver homogenates were reduced in mice on cassava compared with other groups. The data show that cassava does not cause goiter in mice. The thiocyanate formed from ingestation of cassava is insufficient to inhibit thyroid iodide transport or organification of iodide. The cassava diet leads to rapid turnover of hormonal iodine because it is a low iodine diet. It also impairs 5'-monodeiodination of T4 which may be related to nutritional deficiency. These data in mice do not support the concept that cassava per se has goitrogenic action in man.


Mannich-Benzimidazole Derivatives as Antioxidant and Anticholinesterase Inhibitors: Synthesis, Biological Evaluations, and Molecular Docking Study.
A series of Mannich bases of benzimidazole derivatives having a phenolic group were designed to assess their anticholinesterase and antioxidant activities. The acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities were evaluated in vitro by using Ellman's method. According to the activity results, all of the compounds exhibited moderate to good AChE inhibitory activity (except for 2a), with IC50 values ranging from 0.93 to 10.85 μM, and generally displayed moderate BuChE inhibitory activity. Also, most of the compounds were selective against BuChE. Compound 4b was the most active molecule on the AChE enzyme and also selective. In addition, we investigated the antioxidant effects of the synthesized compounds against FeCl2 /ascorbic acid-induced oxidative stress in the rat brain in vitro, and the activity results showed that most of the compounds are effective as radical scavengers. Molecular docking studies and molecular dynamics simulations were also carried out.


Comparison of five commonly-available, lidocaine-containing topical anesthetics and their effect on serum levels of lidocaine and its metabolite monoethylglycinexylidide (MEGX).
Topical anesthetics are commonly applied for a variety of indications. Several lidocaine-containing topical anesthetics are available for purchase over the counter (OTC). Recently, the authors' group has shown that there is great interindividual discrepancy in the manner in which lidocaine is absorbed and metabolized for even a single OTC product. The authors compare five commonly-available lidocaine preparations and their levels of absorption when applied to the face. Three of the preparations are available OTC; two require prescriptions and were compounded in a pharmacy. Twenty-five subjects enrolled in this Institutional Review Board-approved study were randomly assigned to one of five groups. The five topical anesthetics were LMX-4 (4% lidocaine; Biopelle/Ferndale Laboratories, Ferndale, Michigan), Topicaine (4% lidocaine; Ebsa Laboratories, Jupiter, Florida), 2.5% lidocaine/2.5% prilocaine (generic EMLA preparation; High Tech Pharmaceuticals, Amityville, New York), LET (4% lidocaine, 1:2000 epinephrine, and 0.5% tetracaine), and BLT (20% benzoncaine, 6% lidocaine, and 4% tetracaine). After a patch test for adverse reactions, the topical anesthetic was applied to each patient's face and neck and covered with an occlusive dressing for 60 minutes. Blood was drawn at 90, 120, 150, 240, and 480 minutes to measure serum levels of lidocaine and monoethylglycinexylidide (MEGX). The average age of the 17 women and eight men included in the study was 26 years (range, 22-62 years), and the average weight was 70.9 kg (range, 46.4-96.4 kg). The OTC preparations had the highest serum lidocaine and MEGX levels. Topicaine had the greatest serum levels of individual lidocaine absorption (0.808 µg/mL), followed by generic EMLA (0.72 µg/mL), LMX-4 (0.44 µg/mL), BLT (0.17 µg/mL), and LET (0.13 µg/mL). On average, Topicaine had the highest serum lidocaine and MEGX levels: 0.438 µg/mL and 0.0678 µg/mL, respectively. There were significant interindividual differences between the serum levels of MEGX and lidocaine in all groups except LET (P<.0001). There were significant differences between the 4% lidocaine-containing preparations (P=.0439); the 2.5% preparation had a greater absorption than the 4% lidocaine-containing preparation and the 6% lidocaine preparation (P=.0016). There were three adverse reactions in patients who received OTC preparations, one of which resulted in postinflammatory hyperpigmentation. This study demonstrates that although topical anesthetics are considered safe, some individuals have unpredictably high absorption levels. This study also demonstrates that the concentration of lidocaine, the formulation of the drug, and the individual patient all have significant effects on serum levels of lidocaine. The authors recommend that even OTC topical anesthetics be used under the supervision of a healthcare professional to avoid adverse toxic effects and, in rare cases, death. 2.


Subarachnoid haemorrhage: older patients have low cerebral blood flow.
Daily estimations of hemispheral cerebral blood flow using the xenon-133 inhalation technique was made in 116 patients during the first three weeks after subarachnoid haemorrhage. The patients' cerebral perfusion on average remained less than the normal perfusion expected for their age (based on a single estimation of cerebral blood flow in 67 volunteers). On each separate day after subarachnoid haemorrhage cerebral blood flow was inversely related to the patient's age. Older patients seem especially at risk of developing cerebral ischaemia after subarachnoid haemorrhage. The clinical outcome was more often unfavourable in older patients--that is, in those who tended to have the lowest cerebral blood flow. Present results support the view that episodes of low cerebral blood flow lead to a poor outcome after subarachnoid haemorrhage. Because of the risk of inducing cerebral ischaemia great care should be exercised by physicians administering hypotensive drugs to older patients after subarachnoid haemorrhage.


Blood manipulation: current challenges from an anti-doping perspective.
The delivery of oxygen is the limiting factor during whole-body endurance exercise in well-trained individuals, so manipulating the amount of hemoglobin in the blood results in changes in endurance exercise capacity. Athletes began using novel erythropoiesis-stimulating agents well before they were approved for medical use. Older manipulation practices, such as autologous blood transfusions or the administration of first-generation recombinant human erythropoietins, are still widely abused due to challenges in their detection. More recent performance enhancement maneuvers include efforts to mask doping and to induce increased endogenous erythropoietin expression. Confessions by athletes have revealed an ongoing yet extremely sophisticated modus operandi when manipulating the blood. In this review, weaknesses in detection methods and sample collection procedures are scrutinized and strategies developed to circumvent the test system discussed.


Transporter associated with antigen processing preselection of peptides binding to the MHC: a bioinformatic evaluation.
TAP is responsible for the transit of peptides from the cytosol to the lumen of the endoplasmic reticulum. In an immunological context, this event is followed by the binding of peptides to MHC molecules before export to the cell surface and recognition by T cells. Because TAP transport precedes MHC binding, TAP preferences may make a significant contribution to epitope selection. To assess the impact of this preselection, we have developed a scoring function for TAP affinity prediction using the additive method, have used it to analyze and extend the TAP binding motif, and have evaluated how well this model acts as a preselection step in predicting MHC binding peptides. To distinguish between MHC alleles that are exclusively dependent on TAP and those exhibiting only a partial dependence on TAP, two sets of MHC binding peptides were examined: HLA-A*0201 was selected as a representative of partially TAP-dependent HLA alleles, and HLA-A*0301 represented fully TAP-dependent HLA alleles. TAP preselection has a greater impact on TAP-dependent alleles than on TAP-independent alleles. The reduction in the number of nonbinders varied from 10% (TAP-independent) to 33% (TAP-dependent), suggesting that TAP preselection is an important component in the successful in silico prediction of T cell epitopes.


Coherent broadband light effects in grating spectrometer studies of emission from alkali atoms at surfaces.
With ordinary grating spectrometers, strong bands that are due to broadband coherent light emission from samples containing various amounts of alkali atoms can be observed. The coherent light is proposed to be emitted by the alkali Rydberg states that are easily formed in these systems. The edges of the bands are observed at angles corresponding to low numbers of standing waves along the grating surface and perpendicular to it. This type of band is observed both with thermal sources and with broadband light sources created by pulsed laser light, and it is observed only with s-polarized (TE-mode) light. The band intensities are independent of the entrance slit width in the spectrometer, which shows that strong interference effects exist. The number of interference fringes observed on top of the most intense band is directly proportional to the width of the entrance slit. The time-resolved signal shows that large photon peaks from thermal sources are emitted in bursts within 2 mus, probably corresponding to the lifetime of the emitting Rydberg states and Rydberg clusters.


Imaging of cerebral blood flow in patients with severe traumatic brain injury in the neurointensive care.
Ischemia is a common and deleterious secondary injury following traumatic brain injury (TBI). A great challenge for the treatment of TBI patients in the neurointensive care unit (NICU) is to detect early signs of ischemia in order to prevent further advancement and deterioration of the brain tissue. Today, several imaging techniques are available to monitor cerebral blood flow (CBF) in the injured brain such as positron emission tomography (PET), single-photon emission computed tomography, xenon computed tomography (Xenon-CT), perfusion-weighted magnetic resonance imaging (MRI), and CT perfusion scan. An ideal imaging technique would enable continuous non-invasive measurement of blood flow and metabolism across the whole brain. Unfortunately, no current imaging method meets all these criteria. These techniques offer snapshots of the CBF. MRI may also provide some information about the metabolic state of the brain. PET provides images with high resolution and quantitative measurements of CBF and metabolism; however, it is a complex and costly method limited to few TBI centers. All of these methods except mobile Xenon-CT require transfer of TBI patients to the radiological department. Mobile Xenon-CT emerges as a feasible technique to monitor CBF in the NICU, with lower risk of adverse effects. Promising results have been demonstrated with Xenon-CT in predicting outcome in TBI patients. This review covers available imaging methods used to monitor CBF in patients with severe TBI.


Endobronchial ultrasound-guided transbronchial needle injection of cisplatin results in dynamic changes in the tumor immune microenvironment.
Direct intratumoral delivery of cisplatin via endobronchial ultrasound guided-transbronchial needle injections (EBUS-TBNI) is a novel approach for salvage treatment of advanced stage non-small cell lung cancer (NSCLC). The goal of this study was to evaluate changes in the tumor immune microenvironment during the course of EBUS-TBNI cisplatin therapy. Under an IRB approved protocol, patients with recurrence after radiation therapy who were not receiving other cytotoxic therapy, were prospectively enrolled, and underwent weekly treatments with EBUS-TBNI with additional biopsies obtained for research. Needle aspiration was performed prior to cisplatin delivery at each procedure. Samples were evaluated by flow cytometry for the presence of immune cell types. Three of the six patients responded to the therapy based on RECIST criteria. Compared to the pre-treatment baseline, intratumoral neutrophils increased in 5 of the 6 patients (p = 0.041), with an average increase of 27.1%, but was not associated with response. A lower pre-treatment CD8+/CD4+ ratio at baseline was associated with response (P = 0.01). Responders demonstrated a lower final proportion of PD-1+ CD8+ T cells compared to non-responders (8.6% vs. 62.3%, respectively, P<0.001. Lower doses of intratumoral cisplatin were associated with subsequent increases in CD8+ T cells within the tumor microenvironment (P = 0.008). EBUS-TBNI cisplatin resulted in significant alterations in the tumor immune microenvironment. Further studies are needed to determine if the changes seen here generalize to larger cohorts.


Non-invasive prenatal testing for trisomy 21: a cross-sectional survey of service users' views and likely uptake.
To assess the views and likely uptake of non-invasive prenatal testing (NIPT) for trisomy 21 among potential service users in the UK. Cross-sectional survey. Four antenatal clinics in England and two websites. A total of 1131 women and partners. Questionnaire conducted with women (and partners) recruited through antenatal clinics, a random sample of members of the website Mumsnet, and viewers of the website and Facebook page of the support group Antenatal Results and Choices (ARC). Factors impacting decision-making towards prenatal testing; views on NIPT, including service delivery and likely uptake; hypothetical scenarios, focused on current screening, invasive testing, and NIPT offered to women with a high-risk screening result. The vast majority (95.7%; 1071/1119; 95% CI 94.4-96.8%) thought NIPT was a positive development in prenatal care, with 88.2% (972/1103; 95% CI 86.1-90%) indicating that they would use the test, including respondents who would currently decline trisomy 21 screening (P < 0.001). Of the respondents who would have NIPT, 30.7% (299/973; 95% CI = 27.8-33.7%) said that they were 'likely' to terminate an affected pregnancy (including those who would currently decline screening or invasive testing), and 36.5% (355/973; 95% CI 33.5-39.6%) were 'not likely' to terminate an affected pregnancy. Respondents overwhelmingly indicated that safety for the baby was the most important attribute of NIPT (70.1%; 712/1015; 95% CI 67.2-73%). Respondents were overwhelmingly positive towards the introduction of NIPT. Uptake is likely to be high, and includes women who currently decline screening as well as those who will use the test for information only. Pre-test counselling to ensure that women understand the implications of the test result is essential.


Where to Stand with Stromal Cells and Chronic Synovitis in Rheumatoid Arthritis?
The synovium exercises its main function in joint homeostasis through the secretion of factors (such as lubricin and hyaluronic acid) that are critical for the joint lubrication and function. The main synovium cell components are fibroblast-like synoviocytes, mesenchymal stromal/stem cells and macrophage-like synovial cells. In the synovium, cells of mesenchymal origin modulate local inflammation and fibrosis, and interact with different fibroblast subtypes and with resident macrophages. In pathologic conditions, such as rheumatoid arthritis, fibroblast-like synoviocytes proliferate abnormally, recruit mesenchymal stem cells from subchondral bone marrow, and influence immune cell activity through epigenetic and metabolic adaptations. The resulting synovial hyperplasia leads to secondary cartilage destruction, joint swelling, and pain. In the present review, we summarize recent findings on the molecular signature and the roles of stromal cells during synovial pannus formation and rheumatoid arthritis progression.


Genetic structure and relationships within and between cultivated and wild sorghum (Sorghum bicolor (L.) Moench) in Kenya as revealed by microsatellite markers.
Understanding the extent and partitioning of diversity within and among crop landraces and their wild/weedy relatives constitutes the first step in conserving and unlocking their genetic potential. This study aimed to characterize the genetic structure and relationships within and between cultivated and wild sorghum at country scale in Kenya, and to elucidate some of the underlying evolutionary mechanisms. We analyzed at total of 439 individuals comprising 329 cultivated and 110 wild sorghums using 24 microsatellite markers. We observed a total of 295 alleles across all loci and individuals, with 257 different alleles being detected in the cultivated sorghum gene pool and 238 alleles in the wild sorghum gene pool. We found that the wild sorghum gene pool harbored significantly more genetic diversity than its domesticated counterpart, a reflection that domestication of sorghum was accompanied by a genetic bottleneck. Overall, our study found close genetic proximity between cultivated sorghum and its wild progenitor, with the extent of crop-wild divergence varying among cultivation regions. The observed genetic proximity may have arisen primarily due to historical and/or contemporary gene flow between the two congeners, with differences in farmers' practices explaining inter-regional gene flow differences. This suggests that deployment of transgenic sorghum in Kenya may lead to escape of transgenes into wild-weedy sorghum relatives. In both cultivated and wild sorghum, genetic diversity was found to be structured more along geographical level than agro-climatic level. This indicated that gene flow and genetic drift contributed to shaping the contemporary genetic structure in the two congeners. Spatial autocorrelation analysis revealed a strong spatial genetic structure in both cultivated and wild sorghums at the country scale, which could be explained by medium- to long-distance seed movement.


[Cytological structure of the lymph nodes after administration of foreign protein].
Changes in the cellular compositon of popliteal lymphatic nodes of albino rats were studied in normality and after injection of a foreign protein. Allergic inflammation was caused by injection of 0,25 ml of the horse serum. Observations were made on the 1, 3, 8, 19 and 30 days after the beginning of experiment. The cellular composition was counted in 30 visual fileds in the cortical plateu, follicles and myelinated cords. The average percentage of cellular elements has been determined. It has been shown that the reticular cells have less changes as compared with other cells in response to antigen stimulation. An inverse correlative relationship was noted between small and medium-sized lymphocytes. The greatest percentage of plasmic cells was noted in myelinated cords; in the follicles they are found as solitary units. In the primary immunization the cellular composition of follicles was little changed as compared with the cellular composition of the cortical and medullary substance. The cellular reaction was most pronounced in myelinated cords. Thus, the investigation of the cellular composition of different structure components of the rat's lymphatic nodes after injection of the foreign protein has revealed a different degree of their reaction.


Lack of correlation between start codon polymorphism of the vitamin D receptor gene and bone mineral density in premenopausal French women: the OFELY study.
Previous studies have demonstrated an association between bone mineral density (BMD) and a start codon polymorphism (SCP) of the vitamin D receptor (VDR) gene in pre- and postmenopausal Caucasian and Japanese women. The SCP can be determined by a restriction fragment length polymorphism defined by the FokI restriction endonuclease. VDR alleles containing the FokI site are denoted by f and alleles lacking the site by F. In this study, the association between BMD and the SCP was examined in a group of 174 premenopausal French women who previously had been studied for a relationship between BMD and the VDR BsmI polymorphism. The SCP genotypes of the French women were FF 40%, Ff 44%, and ff 16% and they were independent of the BsmI genotype. BMD was measured by dual-energy X-ray absorptiometry at the lumbar spine, proximal femur, forearm, and total body. In contrast to previous reports, there was no association of BMD with SCP genotype in this group of Caucasian women at any site. We also measured several biochemical indices of calcium homeostasis and bone turnover. We found no statistically significant associations between SCP genotype and calcium, parathyroid hormone, or vitamin D levels. There was a 33.5% higher level of the skeletal resorption marker N-telopeptides of type I collagen in the women with the ff genotype when compared with women with the FF genotype (p = 0.004). Other bone turnover markers failed to show an association with SCP genotype. In summary, the SCP genotype may not be associated with reduced BMD in all geographical or ethnic populations.


Risk factors for isolated talipes equinovarus in Western Australia, 1980-1994.
A case-control study based on routinely collected data for a West Australian (WA) cohort of births born 1980-94 inclusive was used to identify potential risk factors for the foot deformity isolated talipes equinovarus. Race and sex were identified as risk factors and risk modifiers of the condition in WA infants. In comparison with their female counterparts, Aboriginal males were at greater risk of the deformity than Caucasian males (odd ratio [OR] 7.15, 95% confidence interval [CI] 2.68, 21.12 vs. OR 1.63, 95% CI 1.24, 2.15 respectively). Sex-specific risk estimates of the deformity showed that Aboriginal males were at more than four times the risk of Caucasian males (OR 4.27, 95% CI 2.30, 8.25 respectively), but the risk amongst Aboriginal females was not dissimilar to the risk amongst Caucasian females (OR 1.12, 95% CI 0.49, 2.45). Intrauterine constraint of the fetus was not found to be an important contributing factor to the deformity after accounting for the effect of all risk factors modelled in multivariable analyses. Indicators of intrauterine constraint including prolonged gestation, high infant birthweight, young maternal age (< 20 years) and breech presentation were not associated with excess numbers of isolated talipes equinovarus cases. Limited numbers of cases associated with other indicators of intrauterine constraint, including the specific categories of short maternal stature and a twin pregnancy prevented definitive conclusions regarding their association with the deformity. Improved data quality for gestational age estimates and oligohydramnios status are required before determining the prenatal risk impact of season of conception and of reduced amniotic fluid levels, respectively, on the occurrence of the deformity.


Patient-controlled analgesia with oral methadone in cancer pain: preliminary report.
Methadone is a very useful drug in cancer pain because of its low cost, lack of active metabolites, high oral availability, and the rapid onset of its analgesic effect. It seems to be well tolerated in patients with difficult pain syndromes who are receiving high doses of opioids, and it may deter the development of tolerance, but a high individual variation in terminal elimination half-life can result in different rates and extents of drug accumulation. For this reason, oral patient-controlled analgesia with methadone was used in 24 advanced-disease patients with pain. A regimen of self-administered oral methadone at fixed doses and flexible patient-controlled dosage intervals to achieve adequate analgesia, while avoiding toxic effects of methadone accumulation, was used in 24 patients requiring opioid therapy. After a priming period of three days with fixed doses of 3-5 mg three times a day for naïve patients and 50% of the morphine equivalent of methadone in patients switched from morphine, patients and relatives were instructed to maintain the night-time dose and to administer a second dose when the pain recurred. When more than four doses of methadone a day were used, an increase of the dosage was prescribed. Continuous pain assessment and monitoring of symptoms were offered. The majority of patients achieved good pain relief until death, and three were switched to very low doses of subcutaneous morphine in their final days. The methadone escalation index was about 2% a day, with a mean dosage increase of 0.3 mg a day for an average of 60 days of treatment at doses ranging from 9 to 80 mg. The plasma concentration in 14 patients ranged from 0.013 to 0.273 mcg/ml with dosages of 20-80 mg during chronic treatment. A mean of 2.4 doses a day was reported, including the fixed night-time dose. The extent of side effects was considered acceptable. Patient-administered analgesia with oral methadone appears to be a simple, cheap and relatively safe technique for controlling cancer pain, permitting individualization by the patient him- or herself and avoiding the risk of accumulation. Continuous assessment is necessary.


Increased Levels of miR-15b-5p and miR-20b-5p in Pancreatic Ductal Adenocarcinoma with Hepatic Metastases.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and lethal forms of cancer. The symptoms appear in advanced stages, and diagnostic and prognostic tests for the early detection of PDAC and disease evolution are not available. The dysregulation of microRNAs (miRNAs) has been associated with cancer development and progression, and some miRNAs have been reported to promote specific metastasis. In this study we aimed to identify the miRNAs dysregulated in PDAC tumoral tissues and a subset of miRNAs associated with tumoral characteristics, mainly metastasis presence and site. For this, the expression of 84 miRNAs was evaluated by qPCR in 30 tumoral tissues and 16 samples of non-tumoral pancreatic tissues. The comparison revealed 32 dysregulated miRNAs (19 upregulated and 13 downregulated) in the PDAC group. Reactome pathway over-representation analysis revealed that these miRNAs are involved in several biological pathways, including "ESR-mediated signaling", "PIP3 activates AKT signaling", and "Regulation of PTEN", among others. Moreover, our study identified an upregulation of miR-15b-5p and miR-20b-5p in the tumoral tissues of patients with hepatic metastasis, outlining these miRNAs as potential markers for hepatic metastasis. No significant difference in miRNA expression was observed in relation to anatomic location, lymphovascular invasion, lung metastasis, and the presence of diabetes.


A population-based examination of trends and disparities in medication treatment for opioid use disorders among Medicaid enrollees.
Medication treatment (MT) with methadone and buprenorphine are effective treatments for opioid use disorders, but little information is available regarding the extent to which buprenorphine's approval resulted in more individuals receiving MT nor to what extent receipt of such treatment was equitable across communities. To examine changes in MT utilization and the association between MT utilization and county-level indicators of poverty, race/ethnicity, and urbanicity, we used Medicaid claims of non-dually eligible Medicaid enrollees aged 18-64 from 14 states for 2002-2009. We generated county-level aggregate counts of MT (methadone, buprenorphine, and any MT) by year (N = 7760 county-years). We estimated count data models to identify associations between MT and county characteristics, including levels of poverty and racial/ethnic concentration. The number of Medicaid enrollees receiving MT increased 62% from 2002 to 2009. The number of enrollees receiving methadone increased 20%, with the remaining increase resulting from buprenorphine. Urban county residents were significantly more likely to receive MT in both 2002 and 2009 than rural county residents. However, buprenorphine substantially increased MT in rural counties from 2002 to 2009. Receipt of MT increased at a much higher rate for residents of counties with lower poverty rates and lower concentrations of black and Hispanic individuals than for residents of counties without those characteristics. The increase in Medicaid enrollees receiving MT in the years following buprenorphine's approval is encouraging. However, it is concerning that MT trends varied so dramatically by characteristics of the county population and that increases in utilization were substantially lower in counties with populations that historically have been disadvantaged with respect to health care access and quality. Concerted efforts are needed to ensure that MT benefits are equitably distributed across society and reach disadvantaged individuals who may be at higher risk of experiencing opioid use disorders.


Cooperative atom-light interaction in a blockaded Rydberg ensemble.
By coupling a probe transition to a Rydberg state using electromagnetically induced transparency (EIT) we map the strong dipole-dipole interactions onto an optical field. We characterize the resulting cooperative optical nonlinearity as a function of probe strength and density. We demonstrate good quantitative agreement between the experiment and an N-atom cooperative model for N=3 atoms per blockade sphere and the n=60 Rydberg state. The measured linewidth of the EIT resonance places an upper limit on the dephasing rate of the blockade spheres of <110 kHz.


[Nonsurgical and surgical management of osteoporotic vertebral body fractures : Recommendations of the Spine Section of the German Society for Orthopaedics and Trauma (DGOU)].
Prospective clinical cohort study (data collection); expert opinion (recommendation development). Treatment options for nonsurgical and surgical management of osteoporotic vertebral body fractures differ widely. Based on the current literature, the knowledge of the experts, and their classification for osteoporotic fractures (OF classification), the Spine Section of the German Society for Orthopaedics and Trauma has now introduced general treatment recommendations. A total of 707 clinical cases from 16 hospitals were evaluated. An OF classification-based score was developed for guidance in the option of nonsurgical versus surgical management. For every classification type, differentiated treatment recommendations were deduced. Diagnostic prerequisites for reproducible treatment recommendations were defined: conventional X‑rays with consecutive follow-up images (standing position whenever possible), magnetic resonance imaging, and computed tomography scans. OF classification allows for upgrading of fracture severity during the course of radiographic follow-up. The actual classification type is decisive for the score. A score of less than 6 points advocates nonsurgical management; in cases with more than 6 points, surgical management is recommended. The primary goal of treatment is fast and painless mobilization. Because of the expected comorbidities in this age group, minimally invasive procedures are preferred. As a general rule, stability is more important than motion preservation. It is mandatory to restore the physiological loading capacity of the spine. If the patient was in a compensated unbalanced state at the time of fracture, reconstruction of the individual prefracture sagittal profile is sufficient. The instrumentation technique has to account for compromised bone quality. We recommend the use of cement augmentation or high purchase screws. The particular situations of injuries with neurological impairment, the necessity to fuse, multiple level fractures, consecutive and adjacent fractures and fractures in ankylosing spondylitis are addressed separately. The therapeutic recommendations presented here provide a reliable and reproducible basis to decide for the treatment choices available. However, intermediate clinical situations with a score of 6 points remain, allowing for both nonsurgical and surgical options. As a result, individualized treatment decisions may still be necessary. In the subsequent step, the recommendations presented will be further evaluated in a multicentre controlled clinical trial.


Analysis of burst activity of the buccal ganglion of Aplysia depilans.
1. Certain nonlinear properties of molluscan neurones suggest that network activity could be described in relation to nonlinear relaxation oscillators. 2. A specific example is fitted by the Van der Pol equation with an exponentially decaying damping coefficient, and changes in the associated energy and sensitivity are considered. 3. More precise details of the interaction pattern determined by analysis of a single cycle of burst activity, and it is shown that there are two states of activity, depending on the balance of presumed inhibitory components. 4. Previous results are discussed in relation to an overall decaying oscillation. 5. Changes in subcomponents of a cycle are described for decaying sequences and the constancy of synaptic interaction is demonstrated. The results are briefly discussed in relation to catastrophe theory and learning.


Cardiovascular imaging with nuclear magnetic resonance.
This article provides some examples of what the previous article outlined. The sections on Image Display and Myocardial Characterization contain numerous illustrations of gating, cross-sectional images, flow signals, and ischemic injury. The possibility of metabolic imaging with NMR and quantitating blood flow is also considered.


The CD27L and CTP1L endolysins targeting Clostridia contain a built-in trigger and release factor.
The bacteriophage ΦCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, and represents a promising alternative as a bactericide. To better understand the lysis mechanism, we have determined the crystal structure of an autoproteolytic fragment of the CD27L endolysin. The structure covers the C-terminal domain of the endolysin, and represents a novel fold that is identified in a number of lysins that target Clostridia bacteria. The structure indicates endolysin cleavage occurs at the stem of the linker connecting the catalytic domain with the C-terminal domain. We also solved the crystal structure of the C-terminal domain of a slow cleaving mutant of the CTP1L endolysin that targets C. tyrobutyricum. Two distinct dimerization modes are observed in the crystal structures for both endolysins, despite a sequence identity of only 22% between the domains. The dimers are validated to be present for the full length protein in solution by right angle light scattering, small angle X-ray scattering and cross-linking experiments using the cross-linking amino acid p-benzoyl-L-phenylalanine (pBpa). Mutagenesis on residues contributing to the dimer interfaces indicates that there is a link between the dimerization modes and the autocleavage mechanism. We show that for the CTP1L endolysin, there is a reduction in lysis efficiency that is proportional to the cleavage efficiency. We propose a model for endolysin triggering, where the extended dimer presents the inactive state, and a switch to the side-by-side dimer triggers the cleavage of the C-terminal domain. This leads to the release of the catalytic portion of the endolysin, enabling the efficient digestion of the bacterial cell wall.


Changing bodies, changing narratives and the consequences of tellability: a case study of becoming disabled through sport.
This article explores the life story of a young man who experienced a spinal cord injury (SCI) and became disabled though playing the sport of rugby union football. His experiences post SCI illuminate the ways in which movement from one form of embodiment to another connects him to a dominant cultural narrative regarding recovery from SCI that is both tellable and acceptable in terms of plot and structure to those around him. Over time, the obdurate facts of his impaired and disabled body lead him to reject this dominant narrative and move into a story line that is located on Norrick's (2005) upper-bounding side of tellability. This makes it transgressive, frightening, difficult to hear, and invokes the twin processes of deprivation of opportunity and infiltrated consciousness as described by Nelson (2001). These, and the effects of impairment, are seen to have direct consequences for the tellability of embodied experiences along with identity construction and narrative repair over time. Finally, some reflections are offered on how the conditions that negate the telling of his story might be challenged.


Bacterial biofilm and associated infections.
Microscopic entities, microorganisms that drastically affect human health need to be thoroughly investigated. A biofilm is an architectural colony of microorganisms, within a matrix of extracellular polymeric substance that they produce. Biofilm contains microbial cells adherent to one-another and to a static surface (living or non-living). Bacterial biofilms are usually pathogenic in nature and can cause nosocomial infections. The National Institutes of Health (NIH) revealed that among all microbial and chronic infections, 65% and 80%, respectively, are associated with biofilm formation. The process of biofilm formation consists of many steps, starting with attachment to a living or non-living surface that will lead to formation of micro-colony, giving rise to three-dimensional structures and ending up, after maturation, with detachment. During formation of biofilm several species of bacteria communicate with one another, employing quorum sensing. In general, bacterial biofilms show resistance against human immune system, as well as against antibiotics. Health related concerns speak loud due to the biofilm potential to cause diseases, utilizing both device-related and non-device-related infections. In summary, the understanding of bacterial biofilm is important to manage and/or to eradicate biofilm-related diseases. The current review is, therefore, an effort to encompass the current concepts in biofilm formation and its implications in human health and disease.


Topology of Euglena chloroplast protein precursors within endoplasmic reticulum to Golgi to chloroplast transport vesicles.
Euglena chloroplast protein precursors are transported as integral membrane proteins from the endoplasmic reticulum (ER) to the Golgi apparatus prior to chloroplast localization. All Euglena chloroplast protein precursors have functionally similar bipartite presequences composed of an N-terminal signal peptide domain and a stromal targeting domain containing a hydrophobic region approximately 60 amino acids from the predicted signal peptidase cleavage site. Asparagine-linked glycosylation reporters and presequence deletion constructs of the precursor to the Euglena light-harvesting chlorophyll a/b-binding protein of photosystem II (pLHCPII) were used to identify presequence regions translocated into the ER lumen and stop transfer membrane anchor domains. An asparagine-linked glycosylation site present at amino acid 148 of pLHCPII near the N terminus of mature LHCPII was not glycosylated in vitro by canine microsomes while an asparagine-linked glycosylation site inserted at amino acid 40 was. The asparagine at amino acid 148 was glycosylated upon deletion of amino acids 46-146, which contain the stromal targeting domain, indicating that the hydrophobic region within this domain functions as a stop transfer membrane anchor sequence. Protease protection assays indicated that for all constructs, mature LHCPII was not translocated across the microsomal membrane. Taken together with the structural similarity of all Euglena presequences, these results demonstrate that chloroplast precursors are anchored within ER and Golgi transport vesicles by the stromal targeting domain hydrophobic region oriented with the presequence N terminus formed by signal peptidase cleavage in the vesicle lumen and the mature protein in the cytoplasm.


Facilitation of gastric motility induced by portal infusion of hyper- and hypotonic solution in rats.
The effects of the portal infusion of hyper- and hypotonic solution on gastric motility in rats were investigated. The infusion of hypertonic saline into the portal vein (portal infusion) elicited a significant enhancement of gastric contractile activity. The portal infusion of water also produced this enhancement. However, the portal infusion of isotonic saline showed no significant enhancement; nor did the infusion of water and hypertonic saline into the jugular vein. Sectioning of the hepatic branch of the vagus nerve (hepatic vagus) eliminated the enhanced responses of the gastric motility. It is therefore concluded that hepatoportal osmoreceptive afferent signals affect the gastric motility by way of the hepatic vagus. These effects on osmolarity revealed that hypotonic stimulation is more effective than hypertonic stimulation for the enhancement of motility. Sectioning of the dorsal subdiaphragmatic vagus, which includes the dorsal gastric and celiac branch, did not eliminate these responses. Sectioning of the ventral gastric vagus, in contrast, did eliminate the responses. These results suggest that vagal preganglionic neurons in the left dorsal motor nucleus of the vagus play a role in enhancement of gastric motility observed in the present research.


What did domestication do to dogs? A new account of dogs' sensitivity to human actions.
Over the last two decades increasing evidence for an acute sensitivity to human gestures and attentional states in domestic dogs has led to a burgeoning of research into the social cognition of this highly familiar yet previously under-studied animal. Dogs (Canis lupus familiaris) have been shown to be more successful than their closest relative (and wild progenitor) the wolf, and than man's closest relative, the chimpanzee, on tests of sensitivity to human social cues, such as following points to a container holding hidden food. The "Domestication Hypothesis" asserts that during domestication dogs evolved an inherent sensitivity to human gestures that their non-domesticated counterparts do not share. According to this view, sensitivity to human cues is present in dogs at an early age and shows little evidence of acquisition during ontogeny. A closer look at the findings of research on canine domestication, socialization, and conditioning, brings the assumptions of this hypothesis into question. We propose the Two Stage Hypothesis, according to which the sensitivity of an individual animal to human actions depends on acceptance of humans as social companions, and conditioning to follow human limbs. This offers a more parsimonious explanation for the domestic dog's sensitivity to human gestures, without requiring the use of additional mechanisms. We outline how tests of this new hypothesis open directions for future study that offer promise of a deeper understanding of mankind's oldest companion.


[The restoration of the enzyme activity of chernozem soil after gamma-irradiation].
The Influence of gamma-radiation by dozes 1, 5, 10 and 20 kGy on enzyme activity of ordinary chemozem were studied. Dynamics of the restoration of the enzyme activity after the influence of gamma-radiation in model experiments in 3, 30, 90 and 180 days was investigated. The doze 1 kGy did no statistically significant influence on the investigated enzymes. Dehydrogenase is more radiosensitive enzyme than catalase. Values of the saccharase activity differed a significant variation so in most cases it has not been registered statistically significant changes. In 90-180 days of the incubation enzymes activity was restored up to control values. Dehydrogenase activity in 180 days in variants with dozes 10 and 20 kGy was restored up to a level of the control, over variants with dozes 1 and 5 kGy--is higher than the control over 78% and 23% accordingly. Saccharase activity in 180 days after the influence of gamma-radiation with a doze 20 kGy was on 61% lower than the control.


Impact of treatment strategies on cephalosporin and tetracycline resistance gene quantities in the bovine fecal metagenome.
The study objective was to determine the effects of two treatment regimens on quantities of ceftiofur and tetracycline resistance genes in feedlot cattle. The two regimens were ceftiofur crystalline-free acid (CCFA) administered to either one or all steers within a pen and subsequent feeding/not feeding of therapeutic doses of chlortetracycline. A 26-day randomized controlled field trial was conducted on 176 steers. Real-time PCR was used to quantify bla(CMY-2), bla(CTX-M), tet(A), tet(B), and 16S rRNA gene copies/gram of feces from community DNA. A significant increase in ceftiofur resistance and a decrease in tetracycline resistance elements were observed among the treatment groups in which all steers received CCFA treatment, expressed as gene copies/gram of feces. Subsequent chlortetracycline administration led to rapid expansion of both ceftiofur and tetracycline resistance gene copies/gram of feces. Our data suggest that chlortetracycline is contraindicated when attempting to avoid expansion of resistance to critically important third-generation cephalosporins.


The role of newer antiarrhythmic drugs in the management of patients with ventricular arrhythmias.
The purpose of this review is to discuss the role of mexiletine, tocainide, flecainide, and amiodarone in the treatment of patients with symptomatic ventricular ectopy or sustained ventricular tachycardia (VT) and fibrillation (VF). The pharmacologic properties, electrophysiologic effects, clinical efficacy, adverse reactions, and costs of these new agents are compared with those of procainamide, quinidine, and disopyramide. With the exception of more convenient dosing intervals, these new agents do not afford novel or advantageous pharmacologic properties. The incidences and severity of adverse reactions are similar to those reported with conventional antiarrhythmic drugs. Results of clinical studies demonstrate that these newer drugs are comparable to procainamide, quinidine, and disopyramide in suppressing ventricular ectopy. Accordingly, the choice of an antiarrhythmic agent for patients with symptomatic ventricular ectopy can be based on the side-effect profile. The propensity for developing certain adverse effects can be defined if patients are subgrouped by age and left ventricular function. The adverse effects of amiodarone preclude its use in patients manifesting only ventricular ectopy. Treatment of patients with sustained VT/VF is more complex. Mexiletine and tocainide are generally not effective. The efficacy of flecainide is similar to that of procainamide, quinidine, and disopyramide. The negative inotropic effects of flecainide and disopyramide preclude their use in patients with severe ventricular dysfunction. Amiodarone is the most effective of these drugs for preventing recurrences of sustained VT/VF. Its use should, however, be restricted to patients refractory or intolerant to other antiarrhythmic drugs.


Extradural S(-)-bupivacaine: comparison with racemic RS-bupivacaine.
Bupivacaine has a chiral centre and is currently available as a racemic mixture of its two enantiomers: R(+)-bupivacaine and S(-)-bupivacaine. Preclinical studies have demonstrated that there is enantiomer selectivity of action with the bulk of central nervous system and cardiovascular toxicity residing with the R(+) isomer. The aim of this study was to compare the clinical efficacy and safety of S(-)-bupivacaine with racemic RS-bupivacaine for extradural anaesthesia. We studied 88 patients undergoing elective lower limb surgery under lumbar extradural anaesthesia who received 15 ml of 0.5% or 0.75% S(-)-bupivacaine, or 0.5% RS-bupivacaine in a randomized, double-blind study. There was no difference in onset time, maximum spread of sensory block or intensity of motor block between the three groups. Duration of sensory block was significantly longer for 0.75% S(-)-bupivacaine. We conclude that S(-)-bupivacaine has similar local anaesthetic characteristics to RS-bupivacaine when used for extradural anaesthesia.


Exploiting QSAR models in lead optimization.
QSAR models can play a vital role in both the opening phase and the endgame of lead optimization. In the opening phase, there is often a large quantity of data from high-throughput screening (HTS), and potential leads need to be selected from several distinct chemotypes. In the endgame, the throughput of the final, critical ADMET and pharmacokinetic assays is often not sufficient to allow full experimental characterization of all the structures in the available time. A considerable amount of the current research toward new QSAR models is based on the modeling of the general ADMET phenomena, with the aim of constructing globally applicable models. The process to construct QSAR models is relatively straightforward; however, it is also simple to build misleading, or even incorrect, models. This review considers the key developments in the field of QSAR modeling: how QSAR models are constructed, how they can be validated, their reliability and their applicability. If applied carefully and appropriately, the QSAR technique has a valuable role to play during lead optimization.


Exploring patient and public involvement (PPI) and co-production approaches in mental health research: learning from the PARTNERS2 research programme.
Patient and Public Involvement (PPI) in research is a growing field of work, incorporating experiential knowledge within research processes. Co-production is a more recent PPI approach that emphasises the importance of power-sharing to promote inclusive research practices, valuing and respecting knowledge from different sources, and relationship building. Applying co-production principles in research trials can be difficult, and there are few detailed worked examples or toolkits. This paper explores the successes and challenges encountered by one research team. Our paper is written by a team of 21 people working on PARTNERS2, led by a smaller co-ordinating group. Using a co-operative style inquiry, the authors have reflected on and written about their experiences; analysis of the resulting 15 accounts provided examples of how PPI and co-production were delivered in practice. We reveal varied and complicated experiences as we developed our collaborative approach across the entire research programme. Four main themes emerge from reflective accounts which describe aspects of this process: (1) recognising the importance of 'emotional work'; (2) developing safe spaces to create and share knowledge; (3) some challenges of using our personal identities in research work; and (4) acknowledging power-sharing within the research hierarchy. We also found continual relationship building, how different forms of expertise were valued, and stigma were central to shaping what work was possible together. Other important practices were transparency, particularly over decision making, and clear communication. Our work provides one example of the 'messy' nature of collaborative research in practice. The learning we surface was contextual, generated within a large-scale research programme, but applicable to other studies. We found for success there needs to be an acknowledgement of the importance of emotional work, creating safe spaces to co-produce, transparency in decision making and reflection on the difficulties of using personal identities in research work including for service user researchers. These elements are more important than existing guidelines suggest. Implementation of actions to support emotional work, will require changes within individual teams as well as institutions. Introducing reflective practice in teams may be helpful in identifying further improvements to inclusive research practice.


Analysis of suspected trace human remains from an indoor concrete surface.
This paper describes the sequence of analyses used to determine the nature of a stain located on the floor of room in the former Athens Mental Health and Retardation Hospital in Athens, OH. The location of the stain was reported to be the position in which a decomposing body was discovered on January 11, 1979. The current stain is found to contain strong evidence for both natural decomposition products and deliberate adulteration. Microscopic analyses, solubility tests, FTIR, ICP-OES, pyrolysis-MS, and derivatization GC-MS were consistent in determining the removable parts of the stain to be composed mostly of calcium and sodium salts of free fatty acids, such as palmitic acid, consistent with previous descriptions of adipocere. The free fatty acids could have been formed via known bacterial degradation pathways or via saponification through the basic environment caused through contact with the concrete. To our knowledge, adipocere formation on an exposed indoor environment has not been described before. The stain and concrete also show signs of being chemically modified with an acidic reagent, such as Blu-Lite--a phosphoric acid-based cleaner that was a commonly used cleaner in the building from the time of discovery to the present day. The chemical etching appears to have been restricted to an area resembling the shape of a human body, which is consistent with deliberate adulteration of the appearance of the stain.


Effects of Strong Acidic Electrolyzed Water in Wound Healing via Inflammatory and Oxidative Stress Response.
Strong acidic electrolyzed water (StAEW) is known to inactivate microorganisms but is not fully explored in the medical field. This study is aimed at exploring StAEW as a potential wound care agent and its mechanism. StAEW (pH: 2.65, ORP: 1159 mV, ACC: 32.1 ppm) was sprayed three times a day to the cutaneous wounds of hairless mice for seven days. Wound morphological and histological features and immune-redox markers were compared with saline- (Sal-) and alcohol- (Alc-) treated groups. Results showed that the StAEW group showed a significantly higher wound healing percentage than the Sal group on days 2, 4, 5, and 6 and the Alc group on day 4. The StAEW group also showed earlier mediation on proinflammatory cytokines such as tumor necrosis factor-α, interleukin- (IL-) 6, IL-1β, and keratinocyte chemoattractant. In addition, basic fibroblast growth factor and platelet-derived growth factor were found to be significantly changed in favor of the fibroblast synthesis and angiogenesis. In line, the StAEW group showed a controlled amount of ROS and significantly decreased compared to the Alc group. The StAEW group also favored oxidative stress balance through antioxidant responses. Additionally, matrix metalloproteinases (MMP) 9 and MMP1 were also modulated for keratinocyte and cell migration. Taken together, this study has proven the wound healing effect of StAEW and its earlier mediation through oxidative and inflammatory responses.


Transforming growth factor beta can be a parameter of aggressiveness of pT1 colorectal cancer.
To evaluate the significance of transforming growth factor beta (TGF beta) expression, in correlation with histopathological parameters, at the front of invasion in T1 colorectal cancer (CRC) and presence of metastases. TGF beta immunohistochemical expression was studied in 34 specimens of colorectal adenocarcinomas (pT1). A three-step avidin-biotinylated immuno-peroxidase (ABCu-NCL) staining technique was performed on 4-mum paraffin-embedded tissue sections with a monoclonal antibody to TGF beta (Novocastra, NCL-TGFB, clone TGFB 17, dilution 1:40). Seventeen (50%) out of 34 lesions were positive for TGF beta expression. The TGF beta-positive rate in patients with vascular invasion was significantly higher than in those without vascular invasion (11/14 cases, P<0.01, P = 0.005). The TGF beta-positive rate was observed in 91.7% of patients with presence of tumor budding at the front of invasion (11/12 cases, P<0.01, P = 0.0003). A statistically significant correlation was found between the presence of lymph node metastases and positive expression of TGF beta (14/16 cases, P<0.01, P = 0.0001). We also observed that the TGF beta-positive rates in groups with distant and non-distant metastases were 92.8% and 20% respectively, and a significant correlation between TGF beta expression and distant metastasis was shown (P<0.01, P = 0.00003). The evaluation of TGF beta expression of protein in association with histological parameters can be used as a parameter of the aggressiveness of pT1 CRC.


Usefulness of current sgRNA design guidelines and in vitro cleavage assays for plant CRISPR/Cas genome editing: a case targeting the polyphenol oxidase gene family in eggplant (Solanum melongena L.).
The advent of genome editing platforms such as the CRISPR/Cas9 system ushers an unprecedented speed in the development of new crop varieties that can withstand the agricultural challenges of the 21st century. The CRISPR/Cas9 system depends on the specificity of engineered single guide RNAs (sgRNAs). However, sgRNA design in plants can be challenging due to the multitude of design tools to choose from, many of which use guidelines that are based on animal experiments yet allow the use of plant genomes. Upon choosing sgRNAs, it is also unclear whether an in vitro assay is needed to validate the targeting efficiency of a particular sgRNA before in vivo delivery of the CRISPR/Cas9 system. Here, we demonstrate the in vitro and in vivo activity of four different sgRNAs that we selected based on their ability to target multiple members of the eggplant polyphenol oxidase gene family. Some sgRNAs that have high in vitro cleavage activity did not produce edits in vivo, suggesting that an in vitro assay may not be a reliable basis to predict sgRNAs with highly efficient in vivo cleavage activity. Further analysis of our sgRNAs using other design algorithms suggest that plant-validated criteria such as the presence of necessary secondary structures and appropriate base-pairing may be the reason for the discrepancy between our observed in vitro and in vivo cleavage efficiencies. However, recent reports and our data suggests that there is no guaranteed way to ensure the in vivo cleavage of chosen sgRNAs.


[A new method of DNA and RNA transfer onto nitrocellulose filters during blot hybridization].
A quick (1-2 hour) method of DNA and RNA transfer onto nitrocellulose filters for subsequent blot-hybridization was elaborated. The main features of the method proposed are, firstly, almost complete exclusion of the mechanical impact on the gel and, secondly, addition to the transfer medium (20 X SSC) of a chaotropic agent, 0.5 M NaClO4. The latter results in a slight dissolution of the gel matrix and, on the other hand, somewhat increases the binding of the nucleic acid to the nitrocellulose. The method shortens significantly the time of DNA or RNA transfer at equal, or even higher, quality of hybridization.


Value of contrast-enhanced magnetic resonance cholangiography in patients undergoing laparoscopic cholecystectomy.
Laparoscopic cholecystectomy (LC) is the preferred treatment for symptomatic gallstone disease. Biliary injury during LC is still a serious problem. Knowledge of anatomic detail is important for not encountering the injury. Magnetic resonance cholangiography (MRC) is a noninvasive method for imaging the biliary ducts. However, MRC has many drawbacks such as not showing anatomic structures in detail and respiratory motion. In this study, contrast-enhanced MRC was used to show cystic ducts that are not seen on MRC. Reasons for patient referral for MRC and contrast-enhanced MRC included suspicion of cholecystolithiasis, adenomyomatosis, and gallbladder polyp. Our results show that routine MRC revealed cystic ducts in 38 patients (77.5%) and contrast-enhanced MRC in 46 patients (93.8%). Intraoperative cholangiography (IOC) was taken as gold standard for all patients. We found that contrast-enhanced MRC can provide a useful supplement to MRC in patients with cystic ducts not seen on MRC. To our knowledge, this is the first study of visualization of a cystic duct in patients undergoing LC depicted by both MRC and contrast-enhanced MRC.


Factors influencing the deaggregation of human and rabbit platelets.
The mechanisms involved in platelet deaggregation are unclear. Washed platelets from rabbits or humans aggregated by ADP can be deaggregated by EDTA or PGI2 if the release reaction has not occurred; during deaggregation 125I-fibrinogen dissociates from the platelets. Human platelets suspended in a medium without calcium undergo the release reaction during ADP-induced aggregation; EDTA, PGE1 or PGI2 do not deaggregate these platelets although EDTA displaces much of the 125I-fibrinogen that associates with them during aggregation. Rabbit platelets aggregated by low concentrations of release-inducing stimuli (sodium arachidonate, collagen or thrombin) can be deaggregated by EDTA, PGI2 or PGE1 and 125I-fibrinogen dissociates from them; with high concentrations of collagen or thrombin, deaggregation and dissociation of 125I-fibrinogen is slower. Human platelets that have undergone the release reaction in response to thrombin, collagen or a combination of sodium arachidonate and ADP are not readily deaggregated by EDTA or PGE1. Since aggregation and fibrinogen binding involving the glycoprotein IIb/IIIa complex are readily reversed by EDTA, and since Ca2+ is required for thrombospondin binding to activated platelets, there may be a third type of platelet-platelet adherence that is not disrupted by EDTA; this type of binding plays a greater role with human than with rabbit platelets.


The interplay of self-acceptance, social comparison and attributional style in adolescent mental health: cross-sectional study.
Adolescence is a pivotal stage vulnerable to mental health problems such as anxiety and depression. Although self-acceptance and social comparison are known to affect adolescent mental health, their interactive and moderating roles are not fully understood. To explore the role of self-acceptance, social comparison and attributional style in predicting these mental health outcomes among adolescents in clinical settings. A cross-sectional study was conducted on a sample of 242 adolescents. Participants completed measures assessing self-acceptance, social comparison, attributional style and mental health outcomes (depression and anxiety). Mediation models and multi-group analysis were used to examine the relationships among these variables. Our findings demonstrated a significant relationship between self-acceptance, social comparison, depression and anxiety (rs = 0.32-0.88). Specifically, lower self-acceptance and higher social comparison were associated with higher levels of depression and anxiety. Additionally, individuals with external attributional tendencies reported higher depression (Cohen's d = 0.61) and anxiety (d = 0.58) compared with those with internal tendencies. Mediation modelling showed that social comparison is a mediator between self-acceptance and depression (effect size -0.04, 95% CI -0.08 to -0.01) and anxiety (effect size -0.06, 95% CI -0.10 to -0.02). Crucially, multi-group analysis showed that the impact of social comparison on mental health outcomes varied significantly based on attributional style. These findings underscore the importance of considering self-acceptance, social comparison and attributional style in understanding and addressing mental health challenges during adolescence. This could inform the development of targeted interventions to promote mental health and well-being among adolescents. However, further research is needed to confirm these findings in diverse populations and to explore the underlying mechanisms in greater detail.


[MR spectroscopy in dementia].
With an increasingly aging population we are faced with the problem of an increasing number of dementia patients. In addition to clinical, neuropsychological and laboratory procedures, MRI plays an important role in the early diagnosis of dementia. In addition to various morphological changes functional changes can also help in the diagnosis and differential diagnosis of dementia. Overall the diagnosis of dementia can be improved by using parameters from MR spectroscopy. This article focuses on MR spectroscopic changes in the physiological aging process as well as on changes in mild cognitive impairment a precursor of Alzheimer's dementia, in Alzheimer's dementia, frontotemporal dementia, vascular dementia and Lewy body dementia.


Gastric Cancer Treatments and Survival Trends in the United States.
Gastric cancer is the third most common cause of cancer deaths worldwide. Despite evidence-based recommendation for treatment, the current treatment patterns for all stages of gastric cancer remain largely unexplored. This study investigates trends in the treatments and survival of gastric cancer. The National Cancer Database was used to identify gastric adenocarcinoma patients from 2004-2016. Chi-square tests were used to examine subgroup differences between disease stages: Stage I, II/III and IV. Multivariate analyses identified factors associated with the receipt of guideline concordant care. The Kaplan-Meier method was used to assess three-year overall survival. The final cohort included 108,150 patients: 23,584 Stage I, 40,216 Stage II/III, and 44,350 Stage IV. Stage specific guideline concordant care was received in only 73% of patients with Stage I disease and 51% of patients with Stage II/III disease. Patients who received guideline consistent care had significantly improved survival compared to those who did not. Overall, we found only moderate improvement in guideline adherence and three-year overall survival during the 13-year study time period. This study showed underutilization of stage specific guideline concordant care for stage I and II/III disease.


Atropine-like actions of muscarine isomers.
Molecular pharmacology of muscarine isomers has been studied in the rat intestine and frog heart. The significance of the peculiar finding of atropinelike action of some of the isomers is discussed.


[Clinical guideline for management of patients with low risk differentiated thyroid carcinoma].
Incidence of thyroid cancer is increasing in Spain and worldwide. Overall thyroid cancer survival is very high, and stratification systems to reliably identify patients with worse prognosis have been developed. However, marked differences exist between the different specialists in clinical management of low-risk patients with thyroid carcinoma. Almost half of all papillary thyroid carcinomas are microcarcinomas, and 90% are tumors < 2 cm that have a particularly good prognosis. However, they are usually treated more aggressively than needed, despite the lack of adequate scientific support. Surgery remains the gold standard treatment for these tumors. However, lobectomy may be adequate in most patients, without the need for total thyroidectomy. Similarly, prophylactic lymph node dissection of the central compartment is not required in most cases. This more conservative approach prevents postoperative complications such as hypoparathyroidism or recurrent laryngeal nerve injury. Postoperative radioiodine remnant ablation and strict suppression of serum thyrotropin, although effective for the more aggressive forms of thyroid cancer, have not been shown to be beneficial for the treatment of low risk patients, and may impair their quality of life. This guideline provides recommendations from the task force on thyroid cancer of the Spanish Society of Endocrinology and Nutrition for adequate management of patients with low-risk thyroid cancer.


Subsequent Injuries and Early Recurrent Diagnoses in elite Rugby Union Players.
An eight-season (2005/06-2012/13) prospective cohort design was used to record time-loss injuries in 15 English Premiership teams. Data pertaining to a total of 1 556 players and 9 597 injuries (8 180 subsequent) were included in the analysis. Injuries subsequent to an index injury were classified as (1) New: different site; (2) Local: same site (and different type); or (3) Recurrent: same site and type. The severity of subsequent injuries (days missed) was compared with their related index injury. The proportions of early (<2 months), late (2-12 months) and delayed (>12 months) subsequent injuries were compared across injury classifications and diagnosis groupings. The majority of subsequent injuries (70%) were classified as new injuries, with 14% local and 16% recurrent. A large proportion of recurrent subsequent injuries (42%) occurred within two months of return-to-play. Subsequent injuries were not more severe than their corresponding index injury (effect sizes <0.20). Specific local and recurrent subsequent injury diagnoses with the highest risk of occurring within two months of return-to-play were: 'neck muscle strain', 'ankle joint capsule sprain', and 'cervical nerve root' injuries. These findings may be used to drive targeted secondary prevention efforts, such as reconsideration of return-to-play protocols for neck muscle strain injuries.


Adherence to an injury prevention program in male amateur football players is affected by players' age, experience and perceptions.
Adherence to injury prevention programmes in football remains low, which is thought to drastically reduce the effects of injury prevention programmes. Reasons why (medical) staff and players implement injury prevention programmes, have been investigated, but player's characteristics and perceptions about these programmes might influence their adherence. Therefore, this study investigated the relationships between player's characteristics and adherence and between player's perceptions and adherence following an implemented injury prevention programme. Data from 98 of 221 football players from the intervention group of a cluster randomised controlled trial concerning hamstring injury prevention were analysed. Adherence was better among older and more experienced football players, and players considered the programme more useful, less intense, more functional and less time-consuming. Previous hamstring injuries, educational level, the programme's difficulty and intention to continue the exercises were not significantly associated with adherence. These player's characteristics and perceptions should be considered when implementing injury prevention programmes.


Study of the role of steroid receptors in human breast cancer.
Cytoplasmic estrogen receptors (ER) were studied in 200 female patients with primary breast cancer. Both ER and progesterone receptors (PgR) were studied in 38 patients. Of the 200 tumors, 55% were estrogen receptor-positive, of the 38 tumors, 34.2% were progesterone receptor-positive. The level of steroid receptors was compared with some parameters characterizing the host and the tumor. It was found that the level of ER correlated with the age factors and the menstrual status. The level of ER was higher in women with adrenal or involutive pathogenetic forms of breast cancer and in women with great body weight. There was not found any relation between the level of estrogen receptors and the stage of menstrual cycle as well as between the level of estrogen receptors and the content of sex chromatin in the tumors.


[Langerhans cell histiocytosis].
We describe the case of male diagnosed with histiocytosis of Langerhans cells. Histiocytosis X is an interstitial disease with a real incidence and an unknown prevalence that can be suspected due to epidemiological and radiological data. The diagnosis can be realized using BAL and/or anatomo-pathological study of a biopsy. The principal treatment is to give up the tobacco habit. Evolution can be favourable.


Compact setup for the production of (87)Rb |F = 2, m = + 2〉 Bose-Einstein condensates in a hybrid trap.
We present a compact experimental apparatus for Bose-Einstein condensation of (87)Rb in the |F = 2, mF = + 2〉 state. A pre-cooled atomic beam of (87)Rb is obtained by using an unbalanced magneto-optical trap, allowing controlled transfer of trapped atoms from the first vacuum chamber to the science chamber. Here, atoms are transferred to a hybrid trap, as produced by overlapping a magnetic quadrupole trap with a far-detuned optical trap with crossed beam configuration, where forced radiofrequency evaporation is realized. The final evaporation leading to Bose-Einstein condensation is then performed by exponentially lowering the optical trap depth. Control and stabilization systems of the optical trap beams are discussed in detail. The setup reliably produces a pure condensate in the |F = 2, mF = + 2〉 state in 50 s, which includes 33 s loading of the science magneto-optical trap and 17 s forced evaporation.


PrimerBank: a PCR primer database for quantitative gene expression analysis, 2012 update.
Optimization of primer sequences for polymerase chain reaction (PCR) and quantitative PCR (qPCR) and reaction conditions remains an experimental challenge. We have developed a resource, PrimerBank, which contains primers that can be used for PCR and qPCR under stringent and allele-invariant amplification conditions. A distinguishing feature of PrimerBank is the experimental validation of primer pairs covering most known mouse genes. Here, we describe a major update of PrimerBank that includes the design of new primers covering 17,076 and 18,086 genes for the human and mouse species, respectively. As a result of this update, PrimerBank contains 497,156 primers (an increase of 62% from the previous version) that cover 36,928 human and mouse genes, corresponding to around 94% of all known protein-coding gene sequences. An updated algorithm based on our previous approach was used to design new primers using current genomic information available from the National Center for Biotechnology Information (NCBI). PrimerBank primers work under uniform PCR conditions, and can be used for high-throughput or genome-wide qPCR. Because of their broader linear dynamic range and greater sensitivity, qPCR approaches are used to reanalyze changes in expression suggested by exploratory technologies such as microarrays and RNA-Seq. The primers and all experimental validation data can be freely accessed from the PrimerBank website, http://pga.mgh.harvard.edu/primerbank/.


SHOOT GROWTH1 maintains Arabidopsis epigenomes by regulating IBM1.
Maintaining correct DNA and histone methylation patterns is essential for the development of all eukaryotes. In Arabidopsis, we identified SHOOT GROWTH1 (SG1), a novel protein involved in the control of gene methylation. SG1 contains both a Bromo-Adjacent Homology (BAH) domain found in several chromatin regulators and an RNA-Recognition Motif (RRM). The sg1 mutations are associated with drastic pleiotropic phenotypes. The mutants degenerate after few generations and are similar to mutants of the histone demethylase INCREASE IN BONSAI METHYLATION1 (IBM1). A methylome analysis of sg1 mutants revealed a large number of gene bodies hypermethylated in the cytosine CHG context, associated with an increase in di-methylation of lysine 9 on histone H3 tail (H3K9me2), an epigenetic mark normally found in silenced transposons. The sg1 phenotype is suppressed by mutations in genes encoding the DNA methyltransferase CHROMOMETHYLASE3 (CMT3) or the histone methyltransferase KRYPTONITE (KYP), indicating that SG1 functions antagonistically to CMT3 or KYP. We further show that the IBM1 transcript is not correctly processed in sg1, and that the functional IBM1 transcript complements sg1. Altogether, our results suggest a function for SG1 in the maintenance of genome integrity by regulating IBM1.


Online Bayesian Phylodynamic Inference in BEAST with Application to Epidemic Reconstruction.
Reconstructing pathogen dynamics from genetic data as they become available during an outbreak or epidemic represents an important statistical scenario in which observations arrive sequentially in time and one is interested in performing inference in an "online" fashion. Widely used Bayesian phylogenetic inference packages are not set up for this purpose, generally requiring one to recompute trees and evolutionary model parameters de novo when new data arrive. To accommodate increasing data flow in a Bayesian phylogenetic framework, we introduce a methodology to efficiently update the posterior distribution with newly available genetic data. Our procedure is implemented in the BEAST 1.10 software package, and relies on a distance-based measure to insert new taxa into the current estimate of the phylogeny and imputes plausible values for new model parameters to accommodate growing dimensionality. This augmentation creates informed starting values and re-uses optimally tuned transition kernels for posterior exploration of growing data sets, reducing the time necessary to converge to target posterior distributions. We apply our framework to data from the recent West African Ebola virus epidemic and demonstrate a considerable reduction in time required to obtain posterior estimates at different time points of the outbreak. Beyond epidemic monitoring, this framework easily finds other applications within the phylogenetics community, where changes in the data-in terms of alignment changes, sequence addition or removal-present common scenarios that can benefit from online inference.


Effect of repeated anterior chamber perfusion on intraocular pressure and total outflow facility in the cynomolgus monkey.
Total outflow facility was determined by two-level constant pressure perfusion of the anterior chamber in surgically virgin, aniridic, and ciliary muscle disinserted cynomolgus monkey eyes 6 to 13 times at 1- to 2-month intervals over periods of 8 to 24 months. Facility decreased approximately 15 to 20% between consecutive thirds of the perfusion history, independent of eye type. The facility decreases were too large to be explained by decreased uveoscleral facility or pseudofacility, and were not mediated by the iris, ciliary muscle, or gonioscopically or ultrastructurally apparent chamber angle alterations. They most probably reflected functional alterations in the trabecular meshwork/inner wall of Schlemm's canal. Intraocular pressure as measured by applanation tonometry did not increase progressively, most probably due to a decreased rate of aqueous humor formation.


Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy.
Whilst there is a large body of evidence looking at the design of cationic liposomes as transfection agents, correlates of formulation to function remain elusive. In this research, we investigate if lipid packaging can give further insights into transfection efficacy. DNA lipoplexes composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method. Each of the formulations was prepared by hydration in dH2O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. In addition, Langmuir monolayer studies were performed to investigate any possible correlation between lipid packaging and liposome attributes. Using PBS, rather than dH2O, to prepare the lipoplexes increased the size of vesicles in most of formulations and resulted in variation in transfection efficacies. However, one combination of lipids (DSPE:DOTAP) could not form liposomes in PBS, whilst the DSPE:DSTAP combination could not form liposomes in either aqueous media. Monolayer studies demonstrated saturated lipid combinations offered dramatically closer molecular packing compared to the other combinations which could suggest why this lipid combination could not form vesicles. Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent.


Brain CRMP forms heterotetramers similar to liver dihydropyrimidinase.
The cytoplasmic collapsin response mediator protein CRMP62 is involved in the signaling cascade initiated by collapsin-1, which collapses neuronal growth cones. To investigate the mechanism of CRMP action, we screened mouse and human fetal cDNA libraries by the yeast two-hybrid method with CRMP as bait. Clones encoding CRMP1 and CRMP4 were isolated, suggesting that the CRMPs form multimers. This finding was confirmed by expressing various rat CRMP cDNAs in the yeast two-hybrid system. Rat CRMP isoforms show differential association with one another. Heterooligomerization is preferred in both two-hybrid and in vitro binding assays. Purified bovine brain CRMP migrates as a tetramer during size exclusion chromatography. Examination of binding with truncated forms of CRMPs indicates that the avid association of CRMPs requires nearly intact proteins. Through the analysis of CRMP chimeras, CRMP amino acids 8-134 and 281-435 are found to be essential for CRMP oligomerization. The tetrameric structure of CRMP resembles that of liver dihydropyrimidinase (DHPase), a protein that shares sequence similarity with the CRMPs. Although purified brain CRMP does not hydrolyze several DHPase substrates, it is likely that a related activity accounts for CRMP participation in neuronal growth cone signaling.


Response of lactating cows to pelleted and unpelleted soybean meal after partial protein depletion.
Twenty-four first-lactation cows, averaging 82 d postpartum, were protein depleted for 20 d by consuming a 9.4% crude protein diet. The cows were then assigned randomly and equally to a 2 X 3 factorial arrangement of dietary treatments in which either pelleted or unpelleted soybean meal containing 1% sodium bentonite was supplemented to corn or corn silage-based diets to yield 12.2, 15.4, or 18.1% crude protein, dry basis. Between 9 and 15 d of the depletion period, milk, milk protein production, and dry matter consumption decreased 20 to 25%. Blood hematocrit increased. On refeeding three amounts of protein, dry matter intake was greatest during week 4 of repletion. Milk production rapidly increased to a plateau averaging approximately 88% of preexperimental production with a somewhat greater response to the higher soybean rations. Neither intake, milk, milk protein production, plasma urea, serum albumin, or blood hematocrit provided any evidence that pelleted or unpelleted soybean meal differed nutritionally.


Noninvasive investigations of the normal ocular circulation in humans.
To compare the reproducibility of laser interferometric measurements of fundus pulsation, pneumatonometric measurement of pulse amplitude (PA) and pulsatile ocular blood flow (POBF), and Doppler ultrasonic measurements of blood flow velocity in the ophthalmic artery (OA) and the posterior ciliary arteries (PCAs) and to investigate the association of the results obtained with these methods and to characterize ocular hemodynamics during Valsalva maneuver and isometric handgrip. All studies were performed in healthy subjects. Fundus pulsation (n = 48), POBF (n = 24), and blood flow velocities (n = 24) were measured at baseline and during the Valsalva maneuver and isometric handgrip. Intraclass correlation coefficients were calculated for test/retest variability, for short-term variability, and for interobserver variability. Intraclass correlation coefficients were between 0.95 and 0.98 for fundus pulsation measurements, between 0.54 and 0.76 for pneumatonometric measurements, between 0.44 and 0.88 for Doppler sonographic measurements in the OA and between 0.32 and 0.60 in the PCAs. There was a high degree of association between pneumatonometric parameters and fundus pulsation amplitude. Valsalva maneuver significantly reduced fundus pulsations, PA, and POBF, whereas isometric handgripping did not change these parameters. The reproducibility of fundus pulsation measurements was excellent. Reproducibility of pneumatonometric parameters and ultrasonographic measurements in the OA was satisfactory. Reproducibility of ultrasonographic measurements in the PCAs was low. Data obtained during the autonomic stimuli experiments argue against the sole use of systems to measure pulsatile blood flow, if no additional data on flow pulsatility are available. At least in cases when changes in blood pressure are likely to occur the ratio of pulsatile to nonpulsatile blood flow might not be constant and POBF might not be an adequate measure of total ocular blood flow.


Reasons of drop-out from rehabilitation in patients of Turkish and Moroccan origin with chronic low back pain in The Netherlands: a qualitative study.
To explore which factors led to drop-out in patients of Turkish and Moroccan origin with chronic non-specific low back pain who participated in a rehabilitation programme. Patients of Turkish or Moroccan origin with chronic non-specific low back pain (n = 23) from 2 rehabilitation centres and 4 rehabilitation departments of general hospitals in the Netherlands. In-depth semi-structured interviews were conducted with patients of Turkish and Moroccan origin (n = 23), rehabilitation physicians (n = 8) and rehabilitation therapists (n = 2). Interviews were transcribed or summarized and subsequently coded and analysed according to themes. Most patients dropped out due to expectations of a specific medical diagnosis and pain relief as the main aims of rehabilitation treatment. Other reasons for drop-out detected in the interviews were: lack of acknowledgement of the patient's complaints, lack of trust in the rehabilitation physician, contradicting views to those of the physician from the patients' country of origin with regard to the cause and treatment of pain, and communication problems. The major reason for drop-out was patients having different expectations, from those of their health providers, of the aim of treatment, as a result of a different view of the origin and treatment of low back pain.


Intricacies and inter-relationships between HIV disclosure and HAART: a qualitative study.
This study aimed to understand whether and how highly active antiretroviral treatment (HAART) affects views and patterns of disclosure and how disclosure interacts with treatment decisions. One hundred and fifty-two HIV-positive adults (52 MSM, 56 women and 44 IDU men) from four US cities participated in two to three-hour, semi-structured interviews in 1998-99. Results indicate that HAART interacts with and shapes HIV disclosure issues in several ways. Medications may 'out' people living with HIV. Thus, in different settings (e.g. work, prisons, drug rehabs and public situations), some try to hide medications or modify dosing schedules, which can contribute to non-adherence, and affect sexual behaviours. Disclosure of HIV and/or HAART may also result in antagonism from others who hold negative attitudes and beliefs about HAART, potentially impeding adherence. Observable side effects of medications can also 'out' individuals. Conversely, medications may improve appearance, delaying or impeding disclosure. Some wait until they are on HAART and look 'well' before disclosing; some who look healthy as a result of medication deny being HIV-positive. Alternatively, HIV disclosure can lead to support that facilitates initiation of, and adherence to, treatment. HIV disclosure and adherence can shape one another in critical ways. Yet these interactions have been under-studied and need to be further examined. Interventions and studies concerning each of these domains have generally been separate, but need to be integrated, and the importance of relationships between these two areas needs to be recognized.