The effects of post-ganglionic axotomy on selective synaptic connexions in the superior cervical ganglion of the guinea-pig.
Stimulation of preganglionic axons arising from different levels of the thoracic spinal cord causes different effects on end-organs supplied by the superior cervical ganglion (Langley, 1892; Nja & Purves, 1977a; Lichtman, Purves & Yip, 1979). For example, stimulation of the first thoracic ventral root (T1) causes pupillary dilatation and widening of the palpebral fissure; stimulation of T4, on the other hand, has little effect on the eye, even though axons arising from this level innervate about as many superior cervical ganglion cells as those from T1. Thus ganglion cell innervation is selective. (1) Three months after crushing the major post-ganglionic branches of the superior cervical ganglion this differential effectiveness is lost: T1 and T4 stimulation have approximately equal effects on the end-organs of the eye. (2) In normal animals, the cellular counterpart of selective end-organ effects is the innervation of each ganglion cell by a contiguous subset of the spinal segments that innervate the ganglion as a whole. One of these segments is usually dominant, the strength of innervation from adjacent segments falling off as a function of distance from the dominant one (Nja & Purves, 1977a). Intracellular recordings from ganglion cells 3 months after post-ganglionic axotomy showed that this selective pattern is re-established. (3) Since the innervation of ganglion cells appears normal, the abnormal end-organ responses after post-ganglionic axotomy suggest that ganglion cell axons are not limited to their original targets during peripheral re-innervation. This suggestion is supported by the finding that ganglion cells sending axons to different peripheral destinations via the second and third cervical spinal nerves were no longer distinguishable on the basis of their segmented inputs 3 months after post-ganglionic axotomy. (4) Similar results were obtained when the preganglionic cervical trunk was cut at the same time as the post-ganglionic axons were crushed; the pattern of end-organ responses was abnormal, whereas individual ganglion cells were re-innervated according to the rules of contiguity and segmental dominance. (5) These results indicate that ganglion cells do not undergo a compensatory change in the segmental innervation they receive when their axons regenerate to targets different from, or in addition to those they originally innervated, even when an entirely new set of ganglionic connexions is formed. This suggests that ganglion cells, or some aspect of their immediate environment, possess a permanent label that determines the segmental innervation they receive.


The breadth, but not the magnitude, of circulating memory B cell responses to P. falciparum increases with age/exposure in an area of low transmission.
Malaria caused by Plasmodium falciparum remains a major cause of death in sub-Saharan Africa. Immunity against symptoms of malaria requires repeated exposure, suggesting either that the parasite is poorly immunogenic or that the development of effective immune responses to malaria may be impaired. We carried out two age-stratified cross-sectional surveys of anti-malarial humoral immune responses in a Gambian village where P. falciparum malaria transmission is low and sporadic. Circulating antibodies and memory B cells (MBC) to four malarial antigens were measured using ELISA and cultured B cell ELISpot. The proportion of individuals with malaria-specific MBC and antibodies, and the average number of antigens recognised by each individual, increased with age but the magnitude of these responses did not. Malaria-specific antibody levels did not correlate with either the prevalence or median number of MBC, indicating that these two assays are measuring different aspects of the humoral immune response. Among those with immunological evidence of malaria exposure (defined as a positive response to at least one malarial antigen either by ELISA or ELISPOT), the median number of malaria-specific MBC was similar to median numbers of diphtheria-specific MBC, suggesting that the circulating memory cell pool for malaria antigens is of similar size to that for other antigens.


Psychiatric problems in fibromyalgia: clinical and neurobiological links between mood disorders and fibromyalgia.
To review the literature addressing the relationship between mood disorders and fibromyalgia/chronic pain and our current understanding of overlapping pathophysiological processes and pain and depression circuitry. We selectively reviewed articles on the co-occurrence of mood disorders and fibromyalgia/chronic pain published between 1990 and July 2012 in PubMed. Bibliographies and cross references were considered and included when appropriate. Forty-nine out of 138 publications were retained for review. The vast majority of the studies found an association between depression and fibromyalgia. There is evidence that depression is often accompanied by symptoms of opposite polarity characterised by heights of mood, thinking and behaviour that have a considerable impact on pharmacological treatment. Recent developments support the view that the high rates of fibromyalgia and mood disorder comorbidity is generated by largely overlapping pathophysiological processes in the brain, that provide a neurobiological basis for the bidirectional, mutually exacerbating and disabling relationship between pain and depression. The finding of comparable pathophysiological characteristics of pain and depression provides a framework for understanding the relationship between the two conditions and sheds some light on neurobiological and therapeutic aspects.


Efficient Ex vivo stimulation of Mycobacterium tuberculosis-specific T cells by genetically detoxified Bordetella pertussis adenylate cyclase antigen toxoids.
Mycobacterium tuberculosis is a significant threat to global health. Mycobacterium bovis BCG vaccine provides only partial protection, and the skin test reagent used to aid diagnosis of both active and latent tuberculosis, purified protein derivative (PPD), lacks specificity and sensitivity. The use of genetically detoxified Bordetella pertussis adenylate cyclase toxin (CyaA) as a delivery system for two immunodominant proteins of M. tuberculosis that are of greater specificity than PPD, early-secreted antigenic target 6-kDa protein (ESAT-6) and culture filtrate protein 10 (CFP-10), was therefore investigated. CyaA toxoids incorporating these antigens were able to restimulate T cells from more than 91% tuberculosis patients and healthy sensitized donors. Delivery of antigen by CyaA decreased by 10-fold the amount of ESAT-6 and CFP-10 required to restimulate T cells, and in low responders, the overall frequency of gamma interferon-producing cells detected by enzyme-linked immunospot assay was increased (P < 0.01 for both antigens). Delivery of ESAT-6 and CFP-10 by CyaA enabled the detection of both CD4(+) and CD8(+) T cells: these responses could be blocked by inhibition of major histocompatibility complex class II or class I, respectively. Covalent linkage of antigen to the CyaA vector was required for enhancement to occur, as a mixture of mock CyaA toxoid plus recombinant ESAT-6 did not lead to enhancement. In a simplified whole-blood model to detect tuberculosis infection, the frequency of positive responses to CFP-10 was increased by CyaA delivery, a potentially important attribute that could facilitate the identification of latent infection.


Mitochondrial generation of reactive oxygen species after brain ischemia in the rat.
Brain mitochondria have a substantial capacity to generate reactive oxygen species after ischemia when the components of the respiratory chain are reduced and molecular oxygen is present. We tested the hypothesis that brain mitochondria in vivo produce reactive oxygen species after ischemia/reperfusion (IR) in rats at a rate sufficient to escape endogenous antioxidant defenses. Ischemia-dependent production of hydroxyl radical in the hippocampus of the anesthetized rat was monitored with the use of intracerebral microdialysis. Transient global ischemia was produced by bilateral carotid artery occlusion and hemorrhagic hypotension to a mean arterial pressure of 35 mm Hg for 15 minutes followed by reperfusion for 60 minutes. Salicylic acid was infused into the hippocampus during the experiments, and changes in the recovery of its hydroxylated product, 2,3-dihydroxybenzoic acid (2,3-DHBA), were used to assess the effects of inhibitors of mitochondrial complex I on formation of hydroxyl radical during IR. Hydroxylation data from control groups of animals were compared with data from animals undergoing IR during treatment with either a mitochondrial complex I inhibitor alone or the inhibitor plus succinic acid. Transient ischemia led to a fivefold increase in the recovery of 2,3-DHBA by microdialysis after 1 hour relative to control animals (P < .05). Inhibition of mitochondrial complex I prevented 2,3-DHBA formation after IR; this effect could be reversed by infusion of succinic acid by microdialysis during IR. The data indicate that reactive oxygen species generated by mitochondrial electron transport escape cellular antioxidant defenses and promote highly damaging hydroxyl radical activity after transient brain ischemia in the rat.


Myeloid translocation gene 16 is required for maintenance of haematopoietic stem cell quiescence.
The t(8;21) and t(16;21) that are associated with acute myeloid leukaemia disrupt two closely related genes termed Myeloid Translocation Genes 8 (MTG8) and 16 (MTG16), respectively. Many of the transcription factors that recruit Mtg16 regulate haematopoietic stem and progenitor cell functions and are required to maintain stem cell self-renewal potential. Accordingly, we found that Mtg16-null bone marrow (BM) failed in BM transplant assays. Moreover, when removed from the animal, Mtg16-deficient stem cells continued to show defects in stem cell self-renewal assays, suggesting a requirement for Mtg16 in this process. Gene expression analysis indicated that Mtg16 was required to suppress the expression of several key cell-cycle regulators including E2F2, and chromatin immunoprecipitation assays detected Mtg16 near an E2A binding site within the first intron of E2F2. BrdU incorporation assays indicated that in the absence of Mtg16 more long-term stem cells were in the S phase, even after competitive BM transplantation where normal stem and progenitor cells are present, suggesting that Mtg16 plays a role in the maintenance of stem cell quiescence.


Disclosing individual CDKN2A research results to melanoma survivors: interest, impact, and demands on researchers.
Whether to return individual research results from cancer genetics studies is widely debated, but little is known about how participants respond to results disclosure or about its time and cost burdens on investigators. We recontacted participants at one site of a multicenter genetic epidemiologic study regarding their CDKN2A gene test results and implications for melanoma risk. Interested participants were disclosed their results by telephone and followed for 3 months. Among 39 patients approached, 27 were successfully contacted, and 19 (70% uptake) sought results, including three with mutations. Prior to disclosure, participants endorsed numerous benefits of receiving results (mean=7.7 of 9 posed), including gaining information relevant to their children's disease risk. Mean psychological well-being scores did not change from baseline, and no decreases to melanoma prevention behaviors were noted. Fifty-nine percent of participants reported that disclosure made participation in future research more likely. Preparation for disclosure required 40 minutes and $611 per recontact attempt. An additional 78 minutes and $68 was needed to disclose results. Cancer epidemiology research participants who received their individual genetic research results showed no evidence of psychological harm or false reassurance from disclosure and expressed strong trust in the accuracy of results. Burdens to our investigators were high, but protocols may differ in their demands and disclosure may increase participants' willingness to enroll in future studies. Providing individual study results to cancer genetics research participants poses potential challenges for investigators, but many participants desire and respond positively to this information.


Delineation of the timing of second-line therapy post-autologous stem cell transplant in patients with AL amyloidosis.
Among patients with immunoglobulin light chain (AL) amyloidosis, there is little consensus on when reinstitution of chemotherapy should occur. We conducted a retrospective study to evaluate the patterns of relapse or progression (R/P) and the timing of reinitiating therapy among 235 patients initially treated with autologous stem cell transplant (ASCT) at Mayo Clinic. The median time from ASCT to second-line therapy was 24.3 months. At the time of restarting therapy, median difference of free light chain (dFLC) was 9.9 mg/dL (42% of diagnosis value), 32% had a dFLC <5 mg/dL, and 63% met criteria for organ R/P. The indications for retreatment were (1) clinical suspicion of R/P, 10%; 92) hematologic R/P only, 23%; (3) organ R/P only, 32%; (4) both hematologic and organ R/P, 31%; and (5) suboptimal response to ASCT and second-line therapy as consolidation, 4%. Patients with organ progression at the time of second-line therapy had inferior survival. Although a dFLC of >5 mg/dL at the time of reinstituting therapy was associated with risk, patients relapsing from very good partial response (VGPR) or better had a longer time to develop organ progression after hematologic R/P (24.2 vs 3.2 months, P = .007). These data suggest that the best candidates for clinical trials testing novel plasma cell-directed chemotherapy beyond first line may be those patients who are either relapsing from VGPR or better (dFLC at diagnosis was >5 mg/dL) or having inadequate response to prior therapy. This strategy should allow for hematologic response assessment while avoiding the risk of deleterious organ progression. Implementation of more stringent progression criteria may also be warranted.


An in vitro setup to test the relevance and the accuracy of low-order vocal folds models.
An experimental setup and human vocal folds replica able to produce self-sustained oscillations are presented. The aim of the setup is to assess the relevance and the accuracy of theoretical vocal folds models. The applied reduced mechanical models are a variation of the classical two-mass model, and a simplification inspired on the delayed mass model for which the coupling between the masses is expressed as a fixed time delay. The airflow is described as a laminar flow with flow separation. The influence of a downstream resonator is taken into account. The oscillation pressure threshold and fundamental frequency are predicted by applying a stability analysis to the mechanical models. The measured frequency response of the mechanical replica together with the initial (rest) area allows us to determine the model parameters (spring stiffness, damping, geometry, masses). Validation of theoretical model predictions to experimental data shows the relevance of low-order models in gaining a qualitative understanding of phonation. However, quantitative discrepancies remain large due to an inaccurate estimation of the model parameters and the crudeness in either flow or mechanical model description. As an illustration it is shown that significant improvements can be made by accounting for viscous flow effects.


Correlates of depressive symptoms among North Korean refugees adapting to South Korean society: the moderating role of perceived discrimination.
Although the prevalence of depressive disorders among North Korean (NK) refugees living in South Korea has been reported to be twice the rate of their South Korean counterparts, little is known about the correlates of depressive symptoms among this population. Despite their escape from a politically and economically repressive setting, NK refugees continue to face multidimensional hardships during their adaptation process in South Korea, which can adversely affect their mental health. However, to our knowledge, no empirical research exists to date on depressive symptoms in the context of adaptation or perceived discrimination among NK refugees. To fill this gap, this study used a sample of 261 NK refugees in South Korea from the 2010 National Survey on Family Violence to examine associations between sociocultural adaptation, perceived discrimination, and depressive symptoms, as well as the moderation effect of discrimination on adaptation to depressive symptoms. We found that poor sociocultural adaptation and perception of discrimination were associated with increased levels of depressive symptoms. Perception of discrimination attenuated the association between better adaptation and fewer depressive symptoms, when compared to no perception of discrimination. These findings highlight the need to improve NK refugees' adaptation and integration as well as their psychological well-being in a culturally sensitive and comprehensive manner. They also underscore the importance of educating South Koreans to become accepting hosts who value diversity, yet in a homogeneous society.


Dog model for cerebrovascular studies of the proximal-to-distal distribution of sequentially injected emboli.
This study was undertaken to determine whether microemboli injected in a predetermined sequence would maintain that sequence once they came to rest in brain microvessels. If so, the injection of different-colored microspheres at different times could be used to bracket-in-time emboli that are known to be released into the circulation during cardiopulmonary bypass. We injected different-colored microspheres into the arterial circulation of anesthetized dogs before and after the injection of fat emboli and before and after cardiopulmonary bypass. Coronal slices of the dog brains were embedded in celloidin, sectioned at 100 microns, and stained for alkaline phosphatase. The afferent cerebrovasculature stained dark brown against a light background, and the proximal/distal orientation of many of the arterioles could be determined by following their course within the thick sections. When different types of emboli were found in a single arteriole, they appeared in the order injected or the order of occurrence in the bypass protocol in 99.3% of the 867 such arterioles counted. Therefore, the microemboli maintained their ordered sequence with only a very small degree of mixing. Once they came to rest, there was not sufficient collateral blood flow in the brain microvessels to move them into disordered positions. This dog model should facilitate studies of the time of release of microemboli within narrower windows of time during cardiopulmonary bypass.


CARE+ user study: usability and attitudes towards a tablet pc computer counseling tool for HIV+ men and women.
CARE+ is a tablet PC-based computer counseling tool designed to support medication adherence and secondary HIV prevention for people living with HIV. Thirty HIV+ men and women participated in our user study to assess usability and attitudes towards CARE+. We observed them using CARE+ for the first time and conducted a semi-structured interview afterwards. Our findings suggest computer counseling may reduce social bias and encourage participants to answer questions honestly. Participants felt that discussing sensitive subjects with a computer instead of a person reduced feelings of embarrassment and being judged, and promoted privacy. Results also confirm that potential users think computers can provide helpful counseling, and that many also want human counseling interaction. Our study also revealed that tablet PC-based applications are usable by our population of mixed experience computer users. Computer counseling holds great potential for providing assessment and health promotion to individuals with chronic conditions such as HIV.


Nephroprotective Effect of Essential Oils from Ginger (Zingiber officinale) and Turmeric (Curcuma longa) Rhizomes against Cadmium-induced Nephrotoxicity in Rats.
Several studies have shown that cadmium (Cd) induces nephrotoxicity and many plant foods phytochemicals have been found useful but their possible mechanism of action still remains unexplored. Hence, this study aimed to investigate the nephroprotective effect of essential oils from Nigeria ginger and turmeric rhizomes in cadmium-treated rats by examining their effect on renal function biomarkers (creatinine, urea and BUN), inflammatory cytokines (IL-6, IL-10 and TNF-Alpha) and renal adenosine deaminase (ADA) activity. The result revealed that essential oils from ginger and turmeric rhizomes exert anti-inflammatory effect by preventing alterations of renal function markers and cytokines (IL-6, IL-10 and TNF-Alpha) levels in Cd-treated rats. In addition, the essential oils inhibited renal ADA activity in Cdtreated rats. In conclusion, inhibition of ADA activity and modulation of inflammatory cytokines could be suggested as the possible mechanism of action by which essential oils from ginger and turmeric rhizomes exert their nephroprotective activities.


The determinants of trust and credibility in environmental risk communication: an empirical study.
This study examines a key component of environmental risk communication; trust and credibility. The study was conducted in two parts. In the first part, six hypotheses regarding the perceptions and determinants of trust and credibility were tested against survey data. The hypotheses were supported by the data. The most important hypothesis was that perceptions of trust and credibility are dependent on three factors: perceptions of knowledge and expertise; perceptions of openness and honesty; and perceptions of concern and care. In the second part, models were constructed with perceptions of trust and credibility as the dependent variable. The goal was to examine the data for findings with direct policy implications. One such finding was that defying a negative stereotype is key to improving perceptions of trust and credibility.


Comorbidity of Alcohol and Gambling Problems in Emerging Adults: A Bifactor Model Conceptualization.
Addictive disorders, such as pathological gambling and alcohol use disorders, frequently co-occur at greater than chance levels. Substantive questions stem from this comorbidity regarding the extent to which shared variance between gambling and alcohol use reflects a psychological core of addictive tendencies, and whether this differs as a function of gender. The aims of this study were to differentiate both common and unique variance in alcohol and gambling problems in a bifactor model, examine measurement invariance of this model by gender, and identify substantive correlates of the final bifactor model. Undergraduates (N = 4475) from a large northwestern university completed an online screening questionnaire which included demographics, quantity of money lost and won when gambling, the South Oaks Gambling Screen, the AUDIT, gambling motives, drinking motives, personality, and the Brief Symptom Inventory. Results suggest that the bifactor model fit the data well in the full sample. Although the data suggest configural invariance across gender, factor loadings could not be constrained to be equal between men and women. As such, general and specific factors were examined separately by gender with a more intensive subsample of females and males (n = 264). Correlations with motivational tendencies, personality traits, and mental health symptoms indicated support for the validity of the bifactor model, as well as gender-specific patterns of association. Results suggest informative distinctions between shared and unique attributes related to problematic drinking and gambling.


Sexting and the Definition Issue.
Sexting among youths has become a necessary topic of interest in research because of the negative consequences that this activity could create, especially when content is shared with others. Indeed, this loss of control could lead to humiliation, (cyber)bullying, or harassment. The development of new technologies, press coverage, and increase of prevalence rates could also explain the growth of interest in sexting. However, its definition is still a gray area. This review examines the different definitions of sexting used in the literature and its correlates. Several elements of the definition of sexting were assessed: actions (sending, receiving, and forwarding); media types (text, images, and videos); sexual characteristics; and transmission modes. Nine databases were searched for studies on sexting among youths up to 18 years of age. Eighteen studies published between 2012 and 2015 were included. Prevalence rates of sexting ranged between .9% and 60% partly depending on the definition. Most studies assessed sending, but when sending and receiving were measured, prevalence rates were higher for receiving. Some articles found associations with age, gender, race, sexual behavior, romantic relationships, risky behaviors, online activity, psychological difficulties, and social pressure. Finding a consensus regarding the definition is essential to assess accurately the activity and adapt prevention. Adolescents' interpretations of the activity are important as sexting could be used as a sexual behavior between two consenting persons. Prevention strategies should focus on sexting that goes wrong when it is forwarded to a third party and when it occurs in a context of pressure or harassment.


Cyclic AMP potentiates glucose-induced insulin release from mouse pancreatic islets without increasing cytosolic free Ca2+.
The effects of various stimulants of insulin release on cytosolic free Ca2+, [Ca2+]i, in dispersed and cultured pancreatic beta-cells from ob/ob-mice were studied using the indicator quin-2, which in itself has only slight effects on the glucose-induced insulin release and the metabolism of the sugar. The resting [Ca2+]i was 158 +/- 7 nM. After increasing glucose to 20 mM there was a lag-period of 1-2 min before [Ca2+]i gradually rose, reaching a new plateau 60% higher after 5-6 min. Increasing intracellular cyclic AMP by adding forskolin did not further increase [Ca2+]i; on the contrary there was a slight temporary reduction despite a doubling of insulin secretion. The maintenance of the beta-cell function was evident from a marked increase of cytosolic [Ca2+]i after depolarization evoked by high extracellular K+. Also dibutyryl cyclic AMP and theophylline lacked the ability to raise [Ca2+]i beyond that obtained by glucose. The results suggest that cyclic AMP potentiates glucose-induced insulin release by sensitizing the secretory machinery to changes of [Ca2+]i rather than by increasing the cytosolic concentration of the ion.


Using genome-wide associations to identify metabolic pathways involved in maize aflatoxin accumulation resistance.
Aflatoxin is a potent carcinogen that can contaminate grain infected with the fungus Aspergillus flavus. However, resistance to aflatoxin accumulation in maize is a complex trait with low heritability. Here, two complementary analyses were performed to better understand the mechanisms involved. The first coupled results of a genome-wide association study (GWAS) that accounted for linkage disequilibrium among single nucleotide polymorphisms (SNPs) with gene-set enrichment for a pathway-based approach. The rationale was that the cumulative effects of genes in a pathway would give insight into genetic differences that distinguish resistant from susceptible lines of maize. The second involved finding non-pathway genes close to the most significant SNP-trait associations with the greatest effect on reducing aflatoxin in multiple environments. Unlike conventional GWAS, the latter analysis emphasized multiple aspects of SNP-trait associations rather than just significance and was performed because of the high genotype x environment variability exhibited by this trait. The most significant metabolic pathway identified was jasmonic acid (JA) biosynthesis. Specifically, there was at least one allelic variant for each step in the JA biosynthesis pathway that conferred an incremental decrease to the level of aflatoxin observed among the inbred lines in the GWAS panel. Several non-pathway genes were also consistently associated with lowered aflatoxin levels. Those with predicted functions related to defense were: leucine-rich repeat protein kinase, expansin B3, reversion-to-ethylene sensitivity1, adaptor protein complex2, and a multidrug and toxic compound extrusion protein. Our genetic analysis provided strong evidence for several genes that were associated with aflatoxin resistance. Inbred lines that exhibited lower levels of aflatoxin accumulation tended to share similar haplotypes for genes specifically in the pathway of JA biosynthesis, along with several non-pathway genes with putative defense-related functions. Knowledge gained from these two complementary analyses has improved our understanding of population differences in aflatoxin resistance.


Combined vaccination with major histocompatibility class I and interleukin 2 gene-transduced melanoma cells synergizes the cure of postsurgical established lung metastases.
We have analyzed and compared in detail the malignant phenotypes of, the immune mechanisms induced by, and the immunotherapeutic potentials of B16-F10.9 melanoma cells manipulated by gene transfer to express syngeneic H-2Kb molecules or to secrete the cytokines interleukin 2 (IL-2) or IL-6. Local tumor growth in the footpad of transduced cells is mainly retarded by expression of H-2Kb and IL-2 genes and less by expression of IL-6. Mice given injections intrafootpad of tumorigenic doses of transduced clones manifested significantly reduced postsurgical spontaneous metastasis. After i.v. inoculation, mice given injections of F10.9-Kb expressors did not develop experimental lung metastases; mice given injections of F10.9-IL-6 secretors developed reduced metastatic loads; whereas mice given injections of F10.9-IL-2 secretors developed high loads of lung metastases. On the basis of injections into nude mice, in vivo depletions of CD4+, CD8+, and NK1.1+ cells, and in vitro CTL and natural killer (NK) assays, we show that all F10.9-modified cells induce CD8+ tumor-specific CTL activity and that F10.9-IL-2 secretors also induce nonspecific NK/lymphokine-activated killer cell activity. Vaccinations with F10.9-modified cells were capable of significantly reducing metastatic spread from small established F10.9 footpad tumors. However, in mice carrying preestablished lung metastases, a highly therapeutic effect was achieved only when H-2Kb expressors and IL-2 secretors were combined in vaccination, whereas individual vaccines or other combinations had marginal effects. This higher efficiency of the combined vaccine is due to the combined effect of efficient CTL induction and NK/lymphokine-activated killer cell activity as concluded from depletion of CD8+ and NK1.1 cells during immunotherapy. Thus, the cure of established metastasis can be achieved by the synergistic effects of vaccination with class I and IL-2-transduced tumor cells.


DNA minor groove binding-directed poisoning of human DNA topoisomerase I by terbenzimidazoles.
We have employed a broad range of spectroscopic, calorimetric, DNA cleavage, and DNA winding/unwinding measurements to characterize the DNA binding and topoisomerase I (TOP1) poisoning properties of three terbenzimidazole analogues, 5-phenylterbenzimidazole (5PTB), terbenzimidazole (TB), and 5-(naphthyl[2,3-d]imidazo-2-yl)bibenzimidazole (5NIBB), which differ with respect to the substitutions at their C5 and/or C6 positions. Our results reveal the following significant features. (i) The overall extent to which the three terbenzimidazole analogues poison human TOP1 follows the hierarchy 5PTB > TB >> 5NIBB. (ii) The impact of the three terbenzimidazole analogues on the superhelical state of plasmid DNA depends on the [total ligand] to [base pair] ratio (rbp), having no effect on DNA superhelicity at rbp ratios < or = 0.1, while weakly unwinding DNA at rbp ratios > 0.1. This weak DNA unwinding activity exhibited by the three terbenzimidazoles does not appear to be correlated with the abilities of these compounds to poison TOP1. (iii) Upon complexation with both poly(dA).poly(dT) and salmon testes DNA, the three terbenzimidazole analogues exhibit flow linear dichroism properties characteristic of a minor groove-directed mode of binding to these host DNA duplexes. (iv) The apparent minor groove binding affinities of the three terbenzimidazole analogues for the d(GA4T4C)2 duplex follow a qualitatively similar hierarchy to that noted above for ligand-induced poisoning of human TOP1-namely, 5PTB > TB > 5NIBB. In the aggregate, our results suggest that DNA minor groove binding, but not DNA unwinding, is important in the poisoning of TOP1 by terbenzimidazoles.


Childhood weight gain and thyroid autoimmunity at age 60-64 years: the 1946 British birth cohort study.
Complex bidirectional relationships have been described between body weight, thyroid function, and risk of thyroid disorders, including thyroid autoimmunity. We used a life-course approach to examine the potential association of childhood or adult body weight with the risk of thyroid autoimmunity and other thyroid disorders at age 60-64 years in a large population-based birth cohort study. In the UK Medical Research Council 1946 British Birth Cohort study, at age 60-64 years, 1277 women and 1185 men (78% of the target sample) responded to a postal questionnaire, which included questions on thyroid disease and thyroid medication. Circulating antithyroid peroxidase antibodies, free T4, and TSH concentrations were measured in 1057 women and 997 men at a subsequent clinic visit. Birth weight was recorded, and height and weight were measured at ages 2, 4, 6, 7, 11, 15 years and also repeatedly in adulthood. At age 60-64 years, 10.9% of women (139 of 1277) and 2.3% of men (27 of 1185) reported they were taking T4, and 11.5% of women (122 of 1057) and 3.3% of men (33 of 997) had positive anti-TPO antibodies (>100 IU/mL), consistent with thyroid autoimmunity. Among women, both T4 use and positive anti-TPO antibodies at age 60-64 years were positively associated with childhood body weight, childhood overweight, and adult body mass index. Childhood weight gain between 0 and 14 years of age was positively associated with later T4 use (odds ratio 1.21, 95% confidence interval 1.03-1.42) and positive anti-TPO antibodies (1.21, 1.00-1.47). Women who were overweight or obese at age 14 years (127 of 972) had a higher risk of later positive anti-TPO antibodies (2.05, 1.12-3.76). In men and women without any thyroid disorders, serum free T4 concentrations were inversely associated with concurrent body mass index (P = .002). Childhood weight gain and childhood overweight conferred an increased susceptibility to later hypothyroidism and thyroid autoimmunity, particularly in women.


An improved kinematic model of the spine for three-dimensional motion analysis in the Vicon system.
The mechanism of creation and pathomechanics of lateral spinal deformation is still not fully explained. Modern medical imaging techniques give scientists possibility to understand some aspects, but vast majority of those techniques is based on static trials. A motion capture system belongs to techniques which enable visualization of a spine during dynamic trials; however, due to lack of appropriate computational model, it is unsuitable for scoliosis imaging. A few years ago our group has proposed a kinematic model of the spine to be used with Vicon Motion Capture System, which was based on Bézier curves. That model allowed for much more precise investigation of spinal kinematics during dynamic trials as compared with other computational models. However, it did not allowed to restrict only selected movements for particular segments of the spine (e.g. axial rotation for lumbar spine). The aim of the current work is to improve the proposed model in order to be able to restrict selected movements according to the knowledge concerning spinal anatomy and spinal range of motion. The new kinematic model of the spine was written in BodyBuilder for Biomechanics Language. For the purpose of visualization also an accurate graphical representation of each vertebra (polygon mesh) was computed and adapted to be compatible with the kinematic model. Using a new version of the model it is possible to perform precise analysis of movement of all vertebrae during such dynamic activities as e.g. gait and forward or lateral bending, as well as to present the results not only on the charts, but also as a 3D animation of movements of a realistically looking spine. The paper describes the new kinematic model and the process of creating graphical representation of the vertebrae. Also sample results obtained using that model are presented.


Connexin43 contributes to electrotonic conduction across scar tissue in the intact heart.
Studies have demonstrated non-myocytes, including fibroblasts, can electrically couple to myocytes in culture. However, evidence demonstrating current can passively spread across scar tissue in the intact heart remains elusive. We hypothesize electrotonic conduction occurs across non-myocyte gaps in the heart and is partly mediated by Connexin43 (Cx43). We investigated whether non-myocytes in ventricular scar tissue are electrically connected to surrounding myocardial tissue in wild type and fibroblast-specific protein-1 driven conditional Cx43 knock-out mice (Cx43fsp1KO). Electrical coupling between the scar and uninjured myocardium was demonstrated by injecting current into the myocardium and recording depolarization in the scar through optical mapping. Coupling was significantly reduced in Cx43fsp1KO hearts. Voltage signals were recorded using microelectrodes from control scars but no signals were obtained from Cx43fsp1KO hearts. Recordings showed significantly decreased amplitude, depolarized resting membrane potential, increased duration and reduced upstroke velocity compared to surrounding myocytes, suggesting that the non-excitable cells in the scar closely follow myocyte action potentials. These results were further validated by mathematical simulations. Optical mapping demonstrated that current delivered within the scar could induce activation of the surrounding myocardium. These data demonstrate non-myocytes in the scar are electrically coupled to myocytes, and coupling depends on Cx43 expression.


History of upper gastrointestinal cancers in relatives: a community-based estimate.
The present study aimed to evaluate the prevalence of positive family history of these cancers in a large population-based sample of Tehran province, capital of Iran. Upper gastrointestinal (UGI) cancers (gastric and esophagus cancer) constitute a major health problem worldwide. A family history of cancer can increase the risk for developing cancer and recognized as one of the most important risk factors in predicting personal cancer risk. This study designed as a cross-sectional survey in general population (2006-2007) of Tehran province. Totally 7,300 persons (age > = 20 years) sampled by random sampling on the basis of the list of postal, of whom 6,700 persons agreed to participate (response rate 92%). Respondents were asked if any first-degree (FDR) or second-degree (SDR) relatives had gastric or esophageal cancer. Totally, 6,453 respondents (48% male) entered to the study. The mean age of responders with positive FH was significantly higher than those with negative FH (P < 0.05). In total, 341 respondents (5.3%) reporting a history of UGI cancers in their relatives, 134(2.1%) in FDRs, and 207(3.2%) in SDRs. Our findings showed that the reported prevalence of FH of UGI cancers was relatively low and varied by specific respondent characteristics such as age and sex. However, the estimates of prevalence presented here are likely to be conservative compared with actual prevalence because of self-reported data gathering.


Can images of pain enhance patient-clinician rapport in pain consultations?
A variety of treatment outcomes in chronic pain are influenced by patient-clinician rapport. Patients often report finding it difficult to explain their pain, and this potential obstacle to mutual understanding may impede patient-clinician rapport. Previous research has argued that the communication of both patients and clinicians is facilitated by the use of pain-related images in pain assessments. This study investigated whether introducing pain-related images into pain assessments would strengthen various components of patient-clinician rapport, including relative levels of affiliation and dominance, and interpersonal coordination between patient and clinician behaviour. Videos of 35 pain assessments in which pain images were present or absent were used to code behavioural displays of patient and clinician rapport at fixed intervals across the course of the assessment. Mixed modelling was used to examine patterns of patient and clinician affiliation and dominance with consultation type (Image vs Control) as a moderator. When pain images were present, clinicians showed more affiliation behaviour over the course of the consultation and there was greater correspondence between the affiliation behaviour of patient and clinician. However, relative levels of patient and clinician dominance were unaffected by the presence of pain images in consultations. Additional analyses revealed that clinicians responded directly to patients' use of pain images with displays of affiliation. Based on the results of this study, we recommend further investigation into the utility and feasibility of incorporating pain images into pain assessments to enhance patient-clinician communication.


Current trends in wireless mesh sensor networks: a review of competing approaches.
Finding a complete mesh-based solution for low-rate wireless personal area networks (LR-WPANs) is still an open issue. To cope with this concern, different competing approaches have emerged in the Wireless Mesh Sensor Networks (WMSNs) field in the last few years. They are usually supported by the IEEE 802.15.4 standard, the most commonly adopted LR-WPAN recommendation for point-to-point topologies. In this work, we review the most relevant and up-to-date WMSN solutions that extend the IEEE 802.15.4 standard to multi-hop mesh networks. To conduct this review, we start by identifying the most significant WMSN requirements (i.e., interoperability, robustness, scalability, mobility or energy-efficiency) that reveal the benefits and shortcomings of each proposal. Then, we re-examine thoroughly the group of proposals following different design guidelines which are usually considered by end-users and developers. Among all of the approaches reviewed, we highlight the IEEE 802.15.5 standard, a recent recommendation that, in its LR-WPAN version, fully satisfies the greatest number of WMSN requirements. As a result, IEEE 802.15.5 can be an appropriate solution for a wide-range of applications, unlike the majority of the remaining solutions reviewed, which are usually designed to solve particular problems, for instance in the home, building and industrial sectors. In this sense, a description of IEEE 802.15.5 is also included, paying special attention to its efficient energy-saving mechanisms. Finally, possible improvements of this recommendation are pointed out in order to offer hints for future research.


Identification of the stereospecific hexose transporter from starved and fed chicken embryo fibroblasts.
When deprived of D-glucose for 24 hr, chicken embryo fibroblasts exhibit a marked increase in hexose transport activity compared with that of control cells. Scatchard analysis of [3H]cytochalasin B binding to starved cell plasma membranes (46 pmol/mg) indicated a six-fold increase compared with fed cell plasma membranes (7.5 pmol/mg). Irradiation of starved cell plasma membranes with high-intensity UV light in the presence of 0.5 microM [3H]cytochalasin B resulted in covalent labeling of polypeptides of Mr 52,000 and 46,000. In fed cell plasma membranes irradiated under the same conditions, both polypeptides were labeled but at greatly decreased levels. In fact, labeling of the Mr 52,000 polypeptide was barely detectable. The amount of D-glucose-sensitive [3H]cytochalasin B covalent insertion into these membrane components was increased 11 +/- 2 (n = 4)-fold in starved versus fed cell plasma membranes. Photoaffinity labeling of both polypeptides in starved cell plasma membranes was inhibited by D-glucose, 3-O-methylglucose, 2-deoxyglucose, cytochalasin B, and cytochalasin A but not by D-sorbitol, L-glucose, or cytochalasin E. Half-maximal inhibition of labeling of the Mr 52,000 polypeptide occurred at 8 mM D-glucose whereas, for the Mr 46,000 polypeptide, half-maximal inhibition occurred at 40 mM D-glucose. It is concluded that (i) two hexose transport proteins, one of Mr 46,000 and one of Mr 52,000, have been identified in chicken embryo fibroblasts and (ii) the increased affinity labeling of these transporter components after cell starvation may reflect increased numbers of transporters in the plasma membrane.


Intramolecular Coupling of Terminal Alkynes by Atom Manipulation.
Glaser-like coupling of terminal alkynes by thermal activation is extensively used in on-surface chemistry. Here we demonstrate an intramolecular version of this reaction performed by atom manipulation. We used voltage pulses from the tip to trigger a Glaser-like coupling between terminal alkyne carbons within a custom-synthesized precursor molecule adsorbed on bilayer NaCl on Cu(111). Different conformations of the precursor molecule and the product were characterized by molecular structure elucidation with atomic force microscopy and orbital density mapping with scanning tunneling microscopy, accompanied by density functional theory calculations. We revealed partially dehydrogenated intermediates, providing insight into the reaction pathway.


[Local governance in the decentralized health care system in Brazil].
To analyze the changes in local health care governance resulting from the decentralization process associated with the Unified Health System (SUS) in Brazil between 1996 and 2006. A questionnaire was answered in 1996 and again in 2006 by all city officials involved in health care management in Brazil. Information was collected on the innovative characteristics of administrative practices in terms of three dimensions: social, management, and care. The present article analyzes the results relating to the social dimension (relationship between municipal officials and the various community actors) according to four attributes: preparing the budget (degree of influence of various actors), establishing priorities, accountability, and flow of information to the community. The influence of municipal secretaries of health and health councils on budget preparation has increased, with a decrease of local politician influence. In prioritizing health issues, local politicians and spontaneous demands have also become less influential, with strengthening of the influence of technical opinions and proposals by health councils and conferences. Public disclosure of results has become institutionalized as a result of the diversification of stakeholders (especially municipal secretaries and health councils) and of the methods available for disclosure, even though balance sheets are still the most common type of information disclosed (which imply technical knowledge for interpretation of results). Finally, the information conveyed to the community still mainly refers to health actions and campaigns and functioning of health services, even though a larger amount of innovative information is being communicated. This was observed in all regions and in cities of all sizes, with a more progressive trend in the South of Brazil. The relationship between government and society has changed toward a more democratic standard of local governance, despite the maintenance of centralized government decision-making practices. The process of decentralization still faces important obstacles to the establishment of a more participative model, with enhanced social control, accountability and interaction between government and society.


Interfascial block at the serratus muscle plane versus conventional analgesia in breast surgery: a randomized controlled trial.
In the context of opioid-sparing perioperative management, there is still little evidence from randomized controlled trials regarding the effectiveness of interfascial thoracic blocks. This study hypothesizes that receiving a serratus plane block reduces opioid requirements, pain scores, and rescue medication needs. This double-blind, randomized controlled study was conducted on 60 adult females undergoing oncologic breast surgery. After general anesthesia, patients were randomly allocated to either conventional analgesia (control group, n=30) or single-injection serratus block with L-bupivacaine 0.25% 30mL (study group, n=30). First 24-hour total morphine consumption (primary outcome), pain scores at 1, 3, 6, 12, and 24 hours, time-to-first opioid rescue analgesia, and adverse effects were recorded. Median 24 hours' opioid dose was greater in the control group (median difference 9 mg (95% CI 4 to 14.5 mg); p<0.001). Proportional odds model showed that the study group has a lower probability of receiving opioid drugs (OR=0.26 (95% CI 0.10 to 0.68); p<0.001), while mastectomies have a higher probability of receiving them (OR=4.11 (95% CI 1.25 to 13.58); p=0.002). Pain scores in the study group were significantly lower throughout the follow-up period (p<0.001). Control group subjects needed earlier morphine rescue and had a higher risk of rescue dose requirement (p=0.002). Interfascial serratus plane block reduces opioid requirements and is associated with better pain scores and lower and later rescue analgesia needs in the first 24 hours, compared with conventional intravenous analgesia, in breast surgery. NCT02905149.


Analysis of immune regulatory genes in familial and sporadic Graves' disease.
Graves' disease (GD) is seen in apparently sporadic and familial forms. At least two immune regulatory genes are associated with GD, human leukocyte antigen (HLA) and cytotoxic T lymphocyte antigen-4 (CTLA-4). The aim of our study was to examine the contributions of HLA and CTLA-4 to the familial clustering of GD by analyzing them for association with familial and sporadic GD. We analyzed 160 Caucasian GD patients (69 familial and 91 sporadic), and 150 matched controls. Analysis of all GD patients demonstrated significant associations between GD and HLA-DR3 [P = 9.0 x 10(-7); relative risk (RR) = 3.8] and two CTLA-4 single nucleotide polymorphisms (SNPs), A/G(49) SNP (P = 0.03; RR = 1.5), and CT60 SNP (P = 0.03; RR = 1.4). Moreover, there was evidence for joint susceptibility to risk between HLA-DR3 and CTLA-4, giving a combined RR of 5.9. Subset analysis demonstrated no significant difference between the frequencies of HLA-DR3 and the susceptibility alleles of CTLA-4 A/G(49) and CT60 SNPs in the familial and sporadic GD subsets (P > 0.05). These results suggested that HLA-DR3 and CTLA-4 conferred a general increased risk for GD in both the sporadic and familial forms, and that the risk conferred by them was additive. However, HLA-DR3 and CTLA-4 did not have a stronger effect in the familial GD patients, suggesting that additional genes must contribute to the aggregation of GD within families.


Further Characterization of Expression of Auxin-Induced Genes in Tobacco (Nicotiana tabacum) Cell-Suspension Cultures.
We have described the modulation of four auxin-regulated genes during the growth cycle of suspension-cultured tobacco (Nicotiana tabacum [L.] var White Burley) cells. The genes were transiently expressed 2 to 8 h after transfer of stationary phase cells to fresh medium, during the transition from the quiescent phase of cells leaving the mitotic cycle to the synthesis phase of the cell cycle. After this transient induction, the cells showed a decreased sensitivity to auxin. Although the expression pattern suggests that induction of these genes might be important for cell division, over-production of antisense mRNA for one of these genes (pCNT103) did not influence cell division in transgenic tobacco cells. Furthermore, stimuli such as salicylic acid were capable of inducing gene expression but were unable to restore cell division. Although these data do not conclusively exclude a role for these genes in cell division, their significance in this process is discussed in view of their homology with other auxin-induced genes and in view of the specificity of hormone-induced early responses.


Stochastic organization of output codes in multiclass learning problems.
The best-known decomposition schemes of multiclass learning problems are one per class coding (OPC) and error-correcting output coding (ECOC). Both methods perform a prior decomposition, that is, before training of the classifier takes place. The impact of output codes on the inferred decision rules can be experienced only after learning. Therefore, we present a novel algorithm for the code design of multiclass learning problems. This algorithm applies a maximum-likelihood objective function in conjunction with the expectation-maximization (EM) algorithm. Minimizing the augmented objective function yields the optimal decomposition of the multiclass learning problem in two-class problems. Experimental results show the potential gain of the optimized output codes over OPC or ECOC methods.


Stability of Chimpanzee Adenovirus Vectored Vaccines (ChAdOx1 and ChAdOx2) in Liquid and Lyophilised Formulations.
Adenovirus vectored vaccines have entered global use during the COVID-19 pandemic, and are in development for multiple other human and veterinary applications. An attraction of the technology is the suitability of the vaccines for storage at 2-8 °C for months. Widely used COVID-19 vaccine ChAdOx1 nCoV-19 (University of Oxford/AstraZeneca) is based on a species E simian adenovirus. Species E simian serotypes have been used in a wide range of other development programs, but the stability of such vectors has not been extensively described in the peer-reviewed literature. Here, we explore the stability of two candidate vaccines based on two species E serotypes: a Rift Valley fever vaccine based upon the ChAdOx1 vector (Y25 serotype) used in ChAdOx1 nCoV-19, and a rabies vaccine based upon a ChAdOx2 vector (AdC68 serotype). We describe each vector's stability in liquid and lyophilised formulations using in vitro and in vivo potency measurements. Our data support the suitability of liquid formulations of these vectors for storage at 2-8 °C for up to 1 year, and potentially for nonrefrigerated storage for a brief period during last-leg distribution (perhaps 1-3 days at 20 °C-the precise definition of acceptable last-leg storage conditions would require further product-specific data). Depending upon the level of inprocess potency loss that is economically acceptable, and the level of instorage loss that is compatible with maintenance of acceptable end-of-storage potency, a previously reported lyophilised formulation may enable longer term storage at 20 °C or storage for a number of days at 30 °C.


MRI Atlas of the Human Deep Brain.
Mastering detailed anatomy of the human deep brain in clinical neurosciences is challenging. Although numerous pioneering works have gathered a large dataset of structural and topographic information, it is still difficult to transfer this knowledge into practice, even with advanced magnetic resonance imaging techniques. Thus, classical histological atlases continue to be used to identify structures for stereotactic targeting in functional neurosurgery. Physicians mainly use these atlases as a template co-registered with the patient's brain. However, it is possible to directly identify stereotactic targets on MRI scans, enabling personalized targeting. In order to help clinicians directly identify deep brain structures relevant to present and future medical applications, we built a volumetric MRI atlas of the deep brain (MDBA) on a large scale (infra millimetric). Twelve hypothalamic, 39 subthalamic, 36 telencephalic, and 32 thalamic structures were identified, contoured, and labeled. Nineteen coronal, 18 axial, and 15 sagittal MRI plates were created. Although primarily designed for direct labeling, the anatomic space was also subdivided in twelfths of AC-PC distance, leading to proportional scaling in the coronal, axial, and sagittal planes. This extensive work is now available to clinicians and neuroscientists, offering another representation of the human deep brain ([https://hal.archives-ouvertes.fr/] [hal-02116633]). The atlas may also be used by computer scientists who are interested in deciphering the topography of this complex region.


Patient-Derived Xenografts Can Be Reliably Generated from Patient Clinical Biopsy Specimens.
Patient-derived xenografts (PDX) are clinically relevant human cancer models that can be used to guide individualized medicine. We aimed to generate PDX models from clinically obtained biopsy specimens (surgical or image-guided) hypothesizing that low volume biopsy specimens could provide sufficient viable tissue to successfully engraft PDX models from patients with unresectable or metastatic disease. We maintain a prospective high volume gastrointestinal malignancy PDX program. With informed consent and institutional approval, biopsy specimens (surgical or image-guided) were obtained from patients with unresectable or metastatic tumors: pancreatic adenocarcinoma (PDAC), cholangiocarcinoma, gastric and gallbladder carcinoma. Biopsies were implanted into immunodeficient mice. Tumor growth was monitored, viable tumor was passed into subsequent generations, and histopathology was confirmed. In this study, biopsy specimens from 29 patients were used for PDX engraftment. Successful PDX engraftment was variable with highest engraftment rates in gastric and gallbladder carcinoma specimens (100%) compared to engraftment rates of 33% and 29% in PDAC and cholangiocarcinoma respectively. PDX models created from metastasis biopsies compared to unresectable primary tumor tissue demonstrated higher engraftment rates (69% versus 15.4%, p = 0.001). PDX models demonstrated higher engraftment rates when biopsies were obtained during surgical operations (n = 15) compared to image-guided (n = 14) (73% versus 14%, p = 0.003). Patient age, pretreatment status, or ischemic time was not different between biopsy methods. PDX models can be successfully created from clinical biopsy specimens in patients with metastatic or unresectable GI cancers. The use of clinical biopsy specimens for PDX engraftment can expand the repertoire of stage-specific PDX models for downstream basic/translational research.


Can psychological characteristics, football experience, and player status predict state anxiety before important matches in Danish elite-level female football players?
Elite football can make players feel nervous, and personality characteristics, as well as experience, affect how well pressure is handled before important games. Studying the psychological characteristics of female football players can provide information on how well psychological pressure is handled and generate knowledge on how to support players in order to improve performance. Based on a sample of 128 female elite football players from 8 top-level teams, the present study investigates whether psychological characteristics and football experience/player stus in elite female football players can predict state anxiety before important matches. Our results outline that high age and national team experience negatively predicted most of the trait anxiety subscales. In line with previous research, no psychological differences were found between goalkeepers, defenders, midfielders, and strikers while starting players revealed to have significantly lower trait anxiety. When measuring before important matches, we found that somatic state anxiety was negatively associated with senior national team experience and positively associated with worry trait anxiety and fear of failure. Cognitive state anxiety was negatively associated with hope for success and positively associated with somatic and worry trait anxiety. Self-confidence was positively associated with youth national team experience and negatively associated with worry trait anxiety. It can be concluded that psychological characteristics and national team experience are both important for optimal state anxiety before important matches in elite-level women's football. Implications for practice and future research are discussed.


A horse chestnut extract, which induces contraction forces in fibroblasts, is a potent anti-aging ingredient.
Contraction forces generated by non-muscle cells, such as fibroblasts, play important roles in determining cell morphology, vasoconstriction, and/or wound healing. We have searched among various plant extracts for ingredients that generate cell contraction forces using fibroblast-populated collagen gels. Using that model, we found that an extract of horse chestnuts (Aesculus hippocastanum) is able to generate such contraction forces in fibroblasts. The involvement of stress fiber formation in that response is suggested by the inhibition of such force generation by cytochalasin D and rhodamine phalloidin stain. Clinical testing of the extract was carried out using 40 healthy female volunteers. A gel formulation that included 3% of the extract was applied topically to the skin around the eye three times daily for nine weeks. The efficacy of the extract to diminish wrinkles was evaluated by visual scoring based on photo scales. After six weeks, significant decreases in the wrinkle scores at the corners of the eye or in the lower eyelid skin were observed compared with controls. After nine weeks, similar results were obtained. Taken together, our results suggest that an extract of horse chestnuts can generate contraction forces in fibroblasts and is a potent anti-aging ingredient.


Carbon tips for all-carbon single-molecule electronics.
We present here an exhaustive ab initio study of the use of carbon-based tips as electrodes in single-molecule junctions. Motivated by recent experiments, we show that carbon tips can be combined with other carbon nanostructures, such as graphene, to form all-carbon molecular junctions with molecules like benzene or C60. Our results show that the use of carbon tips can lead to relatively conductive molecular junctions. However, contrary to junctions formed with standard metals, the conductance traces recorded during the formation of the all-carbon single-molecule junctions do not exhibit clear conductance plateaus, which can be attributed to the inability of the hydrogenated carbon tips to form chemical bonds with the organic molecules. Additionally, we explore here the use of carbon tips for scanning tunneling microscopy and show that they are well suited for obtaining sample images with atomic resolution.


On Suggestibility and Placebo: A Follow-Up Study.
Identifying what makes some people respond well to placebos remains a major challenge. Here, we attempt to replicate an earlier study in which we found a relationship between hypnotic suggestibility and subjective ratings of relaxation following the ingestion of a placebo sedative (Sheiner, Lifshitz, & Raz, 2016). To assess the reliability of this effect, we tested 34 participants using a similar design. Participants ingested a placebo capsule in one of two conditions: (1) relaxation, wherein we described the capsule as a herbal sedative, or (2) control, wherein we described the capsule as inert. To index placebo response, we collected measures of blood pressure and heart rate, as well as self-report ratings of relaxation and drowsiness. Despite using a similar experimental design as in our earlier study, we were unable to replicate the correlation between hypnotic suggestibility and placebo response. Furthermore, whereas in our former experiment we observed a change in subjective ratings of relaxation but no change in physiological measures, here we found that heart rate dropped in the relaxation condition while subjective ratings remained unchanged. Even within a consistent context of relaxation, therefore, our present results indicate that placebos may induce effects that are fickle, tenuous, and unreliable. Although we had low statistical power, our findings tentatively accord with the notion that placebo response likely involves a complex, multifaceted interaction between traits, expectancies, and contexts.


Comparative sensitivity and specificity of ELISA and IHA for serodiagnosis of strongyloidiasis with larval antigens.
Sera from 68 patients with parasitologically proven strongyloidiasis were tested by the ELISA and IHA tests using larval antigens prepared from Strongyloides stercoralis and Strongyloides ratti. The ELISA using the S. stercoralis antigen detected the greatest number of sero-reactors (83.8%), whereas the IHA using the S. ratti antigen detected the fewest (55.9%). In addition, the S. stercoralis antigen had higher geometric mean titers than the S. ratti antigen in both the ELISA and the IHA tests. Sera from 37 presumed normal individuals also were tested by IHA and ELISA and nonspecific reactions were seen only with the IHA test. When sera from patients with parasitic infections other than strongyloidiasis were tested, the only consistent cross-reactions were with those sera from patients who had occult filariasis and acute schistosomiasis.


Diet composition, adherence to calorie restriction, and cardiometabolic disease risk modification.
Calorie restriction (CR) is a promising approach for attenuating the risk of age-related disease. However, the role of diet composition on adherence to CR and the effects of CR on cardiometabolic markers of healthspan remains unknown. We used the Geometric Framework for Nutrition approach to examine the association between macronutrient composition and CR adherence during the 2-year CALERIE trial. Adult participants without obesity were randomized to a 25% CR intervention or an ad libitum intake control. Correlations of cardiometabolic risk factors with macronutrient composition and standard dietary pattern indices [Alternate Mediterranean Diet Index (aMED), Dietary Inflammatory Index (DII), and Healthy Eating Index (HEI)] were also evaluated by Spearman's correlation at each time point. The mean age was 38.1 ± 7.2 years at baseline and the mean BMI was 25.1 ± 1.7. The study population was 70% female. The CR group, but not the control, consumed a higher percentage reported energy intake from protein and carbohydrate and lower fat at 12 months compared to baseline; comparable results were observed at 24 months. Protein in the background of higher carbohydrate intake was associated with greater adherence at 24 months. There was no correlation between macronutrient composition and cardiometabolic risk factors in the CR group. However, statistically significant correlations were observed for the DII and HEI. These findings suggest that individual self-selected macronutrients have an interactive but not independent role in CR adherence. Additional research is required to examine the impact of varying macronutrient compositions on adherence to CR and resultant modification to cardiometabolic risk factors.


Misoprostol dosage in the prevention of nonsteroidal anti-inflammatory drug-induced gastric and duodenal ulcers: a comparison of three regimens.
To compare the effectiveness and tolerability of three misoprostol dosing regimens for the prevention of gastric and duodenal ulcers associated with long-term nonsteroidal anti-inflammatory drug (NSAID) therapy. A multicenter, 12-week, randomized, double-blind, placebo-controlled, parallel, four-limb study. Eligibility criteria included upper gastrointestinal symptoms during NSAID therapy and no endoscopic evidence of gastric or duodenal ulcers. A total of 1623 patients was enrolled; 1197 of these met major accession and regimen-compliance criteria and completed the trial. These 1197 patients composed the evaluable group. Patients were randomly assigned to one of four regimens: placebo four times daily; 200 micrograms of misoprostol twice daily and placebo twice daily; 200 micrograms of misoprostol three times daily and placebo once daily; and 200 micrograms of misoprostol four times daily. Upper gastrointestinal endoscopic examinations for ulcers were done after 4, 8, and 12 weeks of therapy. Tolerability and safety of the regimens were assessed by adverse-event monitoring. In the placebo group, the incidence of gastric ulcers was 15.7% and the incidence of duodenal ulcers was 7.5%. The incidence of gastric ulcers was significantly lower in the groups receiving misoprostol twice daily (8.1%; difference, 7.6% [95% CI, 2.7% to 12.5%]; P = 0.002), three times daily (3.9%; difference, 11.8% [CI, 7.4% to 16.3%]; P < 0.001), and four times daily (4%; difference, 11.7% [CI, 6.7% to 16.8%]; P < 0.001) compared with placebo. The gastric ulcer rate was significantly higher in patients receiving misoprostol twice daily compared with those receiving misoprostol three times daily (difference, 4.2% [95% CI, 0.7% to 7.7%]; P = 0.02). A significant (P = 0.02) misoprostol dose-response effect was noted in the prevention of gastric ulcers. The incidence of duodenal ulcers was significantly lower in the groups receiving misoprostol twice daily (2.6%; difference, 4.9% [CI, 1.5% to 8.2%]; P = 0.004), three times daily (3.3%; difference, 4.2% [CI, 0.6% to 7.7%]; P = 0.019), and four times daily (1.4%; difference, 6.1% [CI, 2.6% to 9.6%]; P = 0.007) compared with placebo. No significant difference was detected between patients receiving misoprostol twice daily and those receiving misoprostol three times daily, and no dose-response effect was noted with duodenal ulcers. The incidence of withdrawals for adverse events was significantly lower in the groups receiving misoprostol twice daily (12%) and three times daily (12%) than in the group receiving it four times daily (20%). The incidence of gastrointestinal adverse events was significantly higher in the group receiving misoprostol four times daily (74%) than in the placebo group (62%). Misoprostol, 200 micrograms twice or three times daily, offers substantial protection against gastric and duodenal ulcers in patients receiving long-term NSAID therapy. These dosages were better tolerated than the currently approved regimen of 200 micrograms four times daily.


The voices of older women in a disadvantaged community: issues of health and social capital.
The voices of older women are rarely heard in debates about the health of disadvantaged groups. Despite growing interest in health in old age, the health experiences of older women as gendered social beings have yet to be fully explored. Their potential to contribute positively to family and community health is seldom acknowledged. The aim of this article is to present findings from a qualitative British Health Development Agency funded project on the relationship between social capital, health and gender, focusing on the health and social networks of older women in a socially disadvantaged community in the north of England. Seventy-seven community members were interviewed, of these 19 were older women aged 55-78 years. Their accounts of ill health in the context of ageing were analysed to explore the intricate ways in which social capital was created, maintained and linked to health. Findings suggest that social constructions of motherhood and caring underpinned responsibility for their own and others' health. Their experiences of dealing with health matters, together with frequent health talk, gave the women confidence as lay health experts, enabling them to contest medical advice. Drawing on personal experiences of trust and reciprocity, they recognised the importance of social networking in alleviating the problems of loneliness and isolation. At stressful times in their lives they were able to draw on existing support networks and, in spite of occasional personal conflicts, some benefited from the empowering and health-enhancing role of formal and informal participation in community life. These findings indicate that older women can operate autonomously in health matters and can substantially influence the development of healthy communities, although this can sometimes be at a personal cost.


Shape engineering vs organic modification of inorganic nanoparticles as a tool for enhancing cellular internalization.
In nanomedicine, physicochemical properties of the nanocarrier affect the nanoparticle's pharmacokinetics and biodistribution, which are also decisive for the passive targeting and nonspecific cellular uptake of nanoparticles. Size and surface charge are, consequently, two main determining factors in nanomedicine applications. Another important parameter which has received much less attention is the morphology (shape) of the nanocarrier. In order to investigate the morphology effect on the extent of cellular internalization, two similarly sized but differently shaped rod-like and spherical mesoporous silica nanoparticles were synthesized, characterized and functionalized to yield different surface charges. The uptake in two different cancer cell lines was investigated as a function of particle shape, coating (organic modification), surface charge and dose. According to the presented results, particle morphology is a decisive property regardless of both the different surface charges and doses tested, whereby rod-like particles internalized more efficiently in both cell lines. At lower doses whereby the shape-induced advantage is less dominant, charge-induced effects can, however, be used to fine-tune the cellular uptake as a prospective 'secondary' uptake regulator for tight dose control in nanoparticle-based drug formulations.


Nickel in plants: I. Uptake kinetics using intact soybean seedlings.
The absorption of Ni(2+) by 21-day-old soybean plants (Glycine max cv. Williams) was investigated with respect to its concentration dependence, transport kinetics, and interactions with various nutrient cations. Nickel absorption, measured as a function of concentration (0.02 to 100 mum), demonstrated the presence of multiple absorption isotherms. Each of the three isotherms conforms to Michaelis-Menten kinetics; kinetic constants are reported for uptake by the intact plant and for transfer from root to shoot tissues. The absorption of Ni(2+) by the intact plant and its transfer from root to shoot were inhibited by the presence of Cu(2+), Zn(2+), Fe(2+), and Co(2+). Competition kinetic studies showed Cu(2+) and Zn(2+) to inhibit Ni(2+) absorption competitively, suggesting that Ni(2+), Cu(2+), and Zn(2+) are absorbed using the same carrier site. Calculated K(m) and K(i) constants for Ni(2+) in the presence and absence of Cu(2+) were 6.1 and 9.2 mum, respectively, whereas K(m) and K(i) constants were calculated to be 6.7 and 24.4 mum, respectively, for Ni(2+) in the presence and absence of Zn(2+). The mechanism of inhibition of Ni(2+) in the presence of Fe(2+) and Co(2+) was not resolved by classical kinetic relationships.


Alkaline-mediated differential interaction (AMDI): a simple automatable single-nucleotide polymorphism assay.
The key requirements for high-throughput single-nucleotide polymorphism (SNP) typing of DNA samples in large-scale disease case-control studies are automatability, simplicity, and robustness, coupled with minimal cost. In this paper we describe a fluorescence technique for the detection of SNPs that have been amplified by using the amplification refractory mutation system (ARMS)-PCR procedure. Its performance was evaluated using 32 sequence-specific primer mixes to assign the HLA-DRB alleles to 80 lymphoblastoid cell line DNAs chosen from our database for their diversity. All had been typed previously by alternative methods, either direct sequencing or gel electrophoresis. We believe the detection system that we call AMDI (alkaline-mediated differential interaction) satisfies the above criteria and is suitable for general high-throughput SNP typing.


Effect of cobalt content on the electrochemical properties and structural stability of NCA type cathode materials.
At present, the most common type of cathode materials, NCA (Li1-xNi0.80Co0.15Al0.05O2, x = 0 to 1), have a very high concentration of cobalt. Since cobalt is toxic and expensive, the existing design of cathode materials is neither cost-effective nor environmentally benign. We have performed density functional theory (DFT) calculations to investigate electrochemical, electronic, and structural properties of four types of NCA cathode materials with the simultaneous decrease in Co content along with the increase in Ni content. Our results show that even if the cobalt concentration is significantly decreased from 16.70% (NCA_I) to 4.20% (NCA_IV), variation in intercalation potential and specific capacity is not significant. For example, in the case of 50% Li concentration, the voltage drop is only ∼17% while the change in specific capacity is negligible. Moreover, we have also explored the influence of sodium doping in the intercalation site on the electrochemical, electronic, and structural properties. By considering two extreme cases of NCAs (i.e., with highest and lowest Co content: NCA_I and NCA_IV, respectively), we have demonstrated the importance of Na doping from the structural and electronic point of view. Our results provide insight into the design of environmentally benign, low-cost cathode materials with reduced cobalt concentration.


Position Statement: A Pragmatic Approach for Medical Cannabis and Patients with Rheumatic Diseases.
Pain is one reason some rheumatology patients may consider use of medical cannabis, a product increasingly perceived as a safe and neglected natural treatment option for many conditions. Legalization of recreational cannabis in Canada will promote access to cannabis. Physicians must therefore provide patients with the best evidence-based information regarding the medicinal effects and harm of cannabis. The Canadian Rheumatology Association (CRA) mandated the development of a position statement for medical cannabis and the rheumatology patient. The current literature regarding the effects of medical cannabis for rheumatology patients was assessed, and a pragmatic position statement to facilitate patient care was developed by the Therapeutics Committee of the CRA and approved by the CRA board. There are no clinical trials of medical cannabis in rheumatology patients. Evidence is insufficient about the benefit of pharmaceutical cannabinoids in fibromyalgia, osteoarthritis, rheumatoid arthritis, and back pain, but there is evidence of a high risk of harm. Extrapolating from other conditions, medical cannabis may provide some symptom relief for some patients. Short-term risks of psychomotor effects can be anticipated, but longterm risks have not been determined and are of concern. Despite lack of evidence for use of medical cannabis in rheumatology patients, we acknowledge the need to provide empathetic and pragmatic guidance for patient care. This position statement aims to facilitate the dialogue between patients and healthcare professionals in a mutually respectful manner to ensure harm reduction for patients and society.


The major histocompatibility complex class Ib molecule HLA-E at the interface between innate and adaptive immunity.
The non-classical major histocompatibility complex (MHC) class I molecule human leucocyte antigen (HLA)-E is the least polymorphic of all the MHC class I molecules and acts as a ligand for receptors of both the innate and the adaptive immune systems. The recognition of self-peptides complexed to HLA-E by the CD94-NKG2A receptor expressed by natural killer (NK) cells represents a crucial checkpoint for immune surveillance by NK cells. However, HLA-E can also be recognised by the T-cell receptor expressed by alphabeta CD8 T cells and therefore can play a role in the adaptive immune response to invading pathogens. The recent resolution of HLA-E in complex with both innate and adaptive ligands has provided insight into the dual role of this molecule in immunity.


Cycles and synchrony in the Collared Lemming (Dicrostonyx groenlandicus) in Arctic North America.
Lemming populations are generally characterised by their cyclic nature, yet empirical data to support this are lacking for most species, largely because of the time and expense necessary to collect long-term population data. In this study we use the relative frequency of yearly willow scarring by lemmings as an index of lemming abundance, allowing us to plot population changes over a 34-year period. Scars were collected from 18 sites in Arctic North America separated by 2-1,647 km to investigate local synchrony among separate populations. Over the period studied, populations at all 18 sites showed large fluctuations but there was no regular periodicity to the patterns of population change. Over all possible combinations of pairs of sites, only sites that were geographically connected and close (<6 km) showed significant synchrony in fluctuations. The populations studied may not even be cyclic, at least for the time period 1960 to 1994, and although fluctuating, randomisation tests could not reject the null hypothesis of random fluctuations. These data have implications for the testing of hypotheses regarding lemming cycles and highlight the need for long-term trapping data to characterise the lemming cycle.


Control of HBV replication by antiviral microRNAs transferred by lentiviral vectors for potential cell and gene therapy approaches.
Because molecular mechanisms regulating host cell and virus interactions are not fully understood, we further defined roles of antiviral microRNAs (miRNAs) in HBV replication. We studied small interfering RNA sequences inserted into the miR-30 backbone in cell systems. Antiviral sequences were cloned into lentiviral vectors upstream of a green fluorescent protein reporter. Transduced cells included HepG2 or HepG2 2.2.15 cell lines and hTERT-FH-B fetal human liver cells. HBV replication was analysed by several assays. In 2.2.15 cells treated with constructs primarily targeting HBV polymerase and surface antigen or HBV polymerase and X open reading frames, HBV core protein, HBV DNA and HBV RNA expression decreased. This antiviral effect was more pronounced when the two constructs were expressed together. Similarly, antiviral constructs decreased HBV replication in HepG2 cells transduced with adenoviral vector to express HBV. Although antiviral sequences were expressed in hTERT-FH-B cells, these cells were non-permissive for HBV, possibly owing to expression of miRNAs reported to inhibit HBV replication, whereas these miRNAs were absent in HepG2 cells. Expression of antiviral miRNAs did not affect cell viability or proliferation and no deleterious changes were observed in expression of native cellular miRNAs. Moreover, expression of antiviral miRNA did not affect engraftment and survival of transplanted cells in mice. Identification of effective antiviral miRNAs and transfer of suitable constructs by lentiviral vectors will be helpful for pathophysiological studies of host cell-virus interactions. Simultaneously, this will advance potential mechanisms for cell/gene therapy in those afflicted with chronic hepatitis and refractory liver disease.


Update on forebrain evolution: From neurogenesis to thermogenesis.
Comparative developmental studies provide growing understanding of vertebrate forebrain evolution. This short review directs the spotlight to some newly emerging aspects, including the evolutionary origin of the proliferative region known as the subventricular zone (SVZ) and of intermediate progenitor cells (IPCs) that populate the SVZ, neural circuits that originated within homologous regions across all amniotes, and the role of thermogenesis in the acquisition of an increased brain size. These data were presented at the 8th European Conference on Comparative Neurobiology.


Physical aspects of sensory transduction on seeing, hearing and smelling.
What is the general principle of sensory transduction? Sensory transduction is defined as energy transformation from the external world to the internal world. The energy of the external world, such as thermal energy (heat), electro-magnetic energy (light), mechanical energy (sound) and the energy from molecules (chemicals), is converted into electrochemical events in the animal nervous system. The following five classes of special sense receptors are utilized for energy conversion: vision (photo); audition (sound); taste and smell (chemo); and tactile (mechano). There are also other special sense receptors, including thermo and noxious receptors. The focus of this study is on photoreceptors, sound-receptors and odorant-receptors because the transduction mechanisms of these receptors are explained biochemically and understood by a common physical principle; these biochemical models are well known in neuroscience. The following notable problems are inherent in these biochemical models: the cGMP ionophore model of the vertebrate photoreceptor cannot explain the fast photo-response (∼msec); the tip links connection model of stereocilia in the basilar membrane for opening the K(+) channel on the tip of a hair has difficulty explaining the high frequency vibration of hair cells without a damping of the oscillation, and the odorant shape-specific receptor model for olfactory transduction has difficulty in discriminating the minute differences among similar fragrant smells of essential oils with different molecular shapes. These difficulties might arise from a lack of the physical sense when the transduction models were proposed. This article will reconsider these problems and propose rational models for visual, olfactory and auditory transduction.


Changes in the Racial Composition of Phytophthora sojae in Australia Between 1979 and 1996.
Surveys of commercial soybean fields, disease nurseries, and trial plots of soybean were conducted throughout eastern Australia between 1979 and 1996, and 694 isolates of Phytophthora sojae were collected and classified into races. Fourteen races, 1, 2, 4, 10, 15, and 25, and eight new races, 46 to 53, were identified, but only races 1, 4, 15, 25, 46, and 53 were found in commercial fields. Races 1 and 15 were the only races found in commercial fields in the soybean-growing areas of Australia up until 1989, with race 1 being the dominant race. Race 4 was found in central New South Wales in 1989 on cultivars with the Rps1a gene, and it is now the dominant race in central and southern New South Wales. Races 46 and 53 have only been found once, in southern New South Wales, and race 25 was identified in the same region in 1994 on a cultivar with the Rps1k gene. Only races 1 and 15 have been found in the northern soybean-growing regions, with the latter dominating, which coincides with the widespread use of cultivars with the Rps2 gene. Changes in the race structure of the P. sojae population from commercial fields in Australia follow the deployment of specific resistance genes.


Functional identification of an aggression locus in the mouse hypothalamus.
Electrical stimulation of certain hypothalamic regions in cats and rodents can elicit attack behaviour, but the exact location of relevant cells within these regions, their requirement for naturally occurring aggression and their relationship to mating circuits have not been clear. Genetic methods for neural circuit manipulation in mice provide a potentially powerful approach to this problem, but brain-stimulation-evoked aggression has never been demonstrated in this species. Here we show that optogenetic, but not electrical, stimulation of neurons in the ventromedial hypothalamus, ventrolateral subdivision (VMHvl) causes male mice to attack both females and inanimate objects, as well as males. Pharmacogenetic silencing of VMHvl reversibly inhibits inter-male aggression. Immediate early gene analysis and single unit recordings from VMHvl during social interactions reveal overlapping but distinct neuronal subpopulations involved in fighting and mating. Neurons activated during attack are inhibited during mating, suggesting a potential neural substrate for competition between these opponent social behaviours.


An examination of willingness to participate and willingness to pay for a universal school food program in the Canadian context.
To examine parents'/caregivers' willingness to participate and willingness to pay (WTP) for a cost-shared school food program (SFP) and its associated factors. A quantitative survey design was used where WTP for a hypothetical SFP was elicited using a double-bounded dichotomous choice elicitation method. We used a double hurdle (logistic and truncated regression) model to examine WTP and positively or negatively associated factors. Saskatoon Public School Division elementary schools situated in high-, mid- or low-median-income neighbourhoods. Parents or caregivers of children attending grades 1 to grade 8 in the Saskatoon Public School Division elementary schools. 94 % respondents were willing to participate in a SFP while less than two-thirds of participants were willing to pay for such a program. Over 90 % respondents from all the socio-economic groups were willing to participate. Multiple household income earners, higher household income, higher number of children, household food security status and higher academic attainment of parents'/caregivers predicted greater willingness to pay. Mean willingness to pay was $4·68 (CAN), and households reporting moderate or severe food insecurity were likely to be willing to pay significantly less for a SFP. A cost-shared program might be financially sustainable in Canada if community characteristics such as household food insecurity status, economic participation of women and average household size are kept in mind while determining the price of the program.


Regulation of carbonic anhydrase expression by zinc, cobalt, and carbon dioxide in the marine diatom Thalassiosira weissflogii.
TWCA1 is the major Zn-requiring isoform of carbonic anhydrase (CA) in the marine diatom Thalassiosira weissflogii. We have examined the roles that trace metals and CO(2) play in the regulation of TWCA1 expression over ranges of concentrations that bracket those encountered in the marine environment. Both steady-state levels of TWCA1 and the kinetics of induction were measured by western analysis. TWCA1 levels correlated well with cellular CA activity levels. TWCA1 was induced at a low CO(2) concentration but the level of induction, as determined by western analysis, was dependent on the availability of Zn. Co effectively substituted for Zn in regulating TWCA1 expression and promoting TWCA1 activity. Upon shift from low to high CO(2), the concentration of TWCA1 decreased. The expression of TWCA1 is diel cycle regulated, and cellular TWCA1 decreased during the dark phase. These results provide the basis for studying the expression of CA in field populations and, taken together with previous radiolabeling studies, provide strong evidence of in vivo metal substitution of Co for Zn in a CA. Our data also support the conclusion that TWCA1 plays a central role in carbon acquisition in T. weissflogii.


A randomized controlled trial comparing intrathecal sustained-release cytarabine (DepoCyt) to intrathecal methotrexate in patients with neoplastic meningitis from solid tumors.
Standard treatment for neoplastic meningitis requires frequent intrathecal (IT) injections of chemotherapy and is only modestly effective. DepoCyt is a sustained-release formulation of cytarabine that maintains cytotoxic concentrations of the drug in the cerebrospinal fluid (CSF) for more than 14 days after a single 50-mg injection. We conducted a randomized, controlled trial of DepoCyt versus methotrexate in patients with solid tumor neoplastic meningitis. Sixty-one patients with histologically proven cancer and positive CSF cytologies were randomized to receive IT DepoCyt (31 patients) or IT methotrexate (30 patients). Patients received up to six 50-mg doses of DepoCyt or up to sixteen 10-mg doses of methotrexate over 3 months. Treatment arms were well balanced with respect to demographic and disease-related characteristics. Responses occurred in 26% of DepoCyt-treated and 20% of methotrexate-treated patients (P = 0.76). Median survival was 105 days in the DepoCyt arm and 78 days in the methotrexate arm (log-rank P = 0.15). The DepoCyt group experienced a greater median time to neurological progression (58 versus 30 days; log-rank P = 0.007) and longer neoplastic meningitis-specific survival (log-rank P = 0.074; median meningitis-specific survival, 343 versus 98 days). Factors predictive of longer progression-free survival included absence of visible central nervous system disease on neuroimaging studies (P<0.001), longer pretreatment duration of CSF disease (P<0.001), history of intraparenchymal tumor (P<0.001), and treatment with DepoCyt (P = 0.002). The frequency and grade of adverse events were comparable between treatment arms. In patients with solid tumor neoplastic meningitis, DepoCyt produced a response rate comparable to that of methotrexate and significantly increased the time to neurological progression while offering the benefit of a less demanding dose schedule.


Lipoamide influences substrate selection in post-ischaemic perfused rat hearts.
We investigated whether lipoamide and diacetyl-lipoamide are able to change the substrate selection in post-ischaemic myocardium. This can be important, because shifting heart metabolism from fatty acid to carbohydrate oxidation can decrease ischaemic injury. Studying the metabolism of [1,2-13C]diacetyl-lipoamide in situ in perfused rat heart by 13C n.m.r., we found intense 13C labelling in glutamate and aspartate, showing that acetyl groups from diacetyl-lipoamide are effectively transferred to CoA and metabolized in heart tissue. From analysis of glutamate C-3 and C-4 isotopomers, we determined the [1,2-13C]acetate/[3-13C]lactate utilization ratio in normoxic and post-ischaemic hearts, where under our experimental conditions the acetate/lactate utilization ratios were 1.2 +/- 0.2 and 2.4 +/- 0.3 in normoxic and post-ischaemic hearts respectively. When 0.25 mM lipoamide was added to the perfusate the acetate/lactate utilization ratio decreased to 1.4 +/- 0.1, which is almost equal to that found for normoxic hearts, showing that lipoamide increased the lactate utilization. In accordance with these data, we found that lipoamide activated pyruvate dehydrogenase by 50% in post-ischaemic myocardium. Competition between [3-13C]lactate and unlabelled octanoate was also studied in post-ischaemic hearts, and we found that lipoamide increased lactate utilization by 100% and increased the rate of the tricarboxylic acid cycle by 64%. Under the same experimental conditions, lipoamide significantly promoted the recovery of post-ischaemic unpaced hearts, showing the positive effect of increased lactate oxidation in post-ischaemic myocardium.


Minimal mechanical load and tissue culture conditions preserve native cell phenotype and morphology in tendon-a novel ex vivo mouse explant model.
Appropriate mechanical load is essential for tendon homeostasis and optimal tissue function. Due to technical challenges in achieving physiological mechanical loads in experimental tendon model systems, the research community still lacks well-characterized models of tissue homeostasis and physiological relevance. Toward this urgent goal, we present and characterize a novel ex vivo murine tail tendon explant model. Mouse tail tendon fascicles were extracted and cultured for 6 days in a load-deprived environment or in a custom-designed bioreactor applying low magnitude mechanical load (intermittent cycles to 1% strain, at 1 Hz) in serum-free tissue culture. Cells remained viable, as did collagen structure and mechanical properties in all tested conditions. Cell morphology in mechanically loaded tendon explants approximated native tendon, whereas load-deprived tendons lost their native cell morphology. These losses were reflected in altered gene expression, with mechanical loading tending to maintain tendon specific and matrix remodeling genes phenotypic of native tissue. We conclude from this study that ex vivo load deprivation of murine tendon in minimal culture medium results in a degenerative-like phenotype. We further conclude that onset of tissue degeneration can be suppressed by low-magnitude mechanical loading. Thus a minimal explant culture model featuring serum-free medium with low mechanical loads seems to provide a useful foundation for further investigations. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1383-1390, 2018.