Targeting the Insulin-Like Growth Factor (IGF) System Is Not as Simple as Just Targeting the Type 1 IGF Receptor.
Increased signaling of the insulin-like growth factor (IGF) system via alterations in expression levels of its components has been demonstrated in various tumor types. Numerous experimental studies have supported the involvement of the IGF system signaling axis in tumor initiation and progression. These studies, combined with data that link alterations in the levels of circulating IGFs with cancer risk and prognosis, have focused on the IGF-1 receptor (IGF-1R) as a therapeutic target for patients with cancer. As a consequence, most therapeutic strategies have been designed to specifically inhibit IGF-1R but have for the most part ignored the insulin receptor (IR), based on concerns that targeting IR would lead to unacceptable toxicity both because of its role in physiologic metabolism and because we frequently try to oversimplify biologic complexity whenever we are urged to find practical, friendly solutions for clinical practice. Although this is an understandable and necessary starting point in the complex and long-lasting processes that leads to translational biology, the crude reality of the results obtained from phase I and II studies suggest a need for researchers to be humble and go back to the drawing board. Cancer research has substantially neglected the role of IR, and it remains unclear whether and to what extent avoiding the inhibition of IR has compromised the efficacy of anti-IGF-1R therapy. Clarifying its role might also help us take advantage of older drugs that could offer new perspectives in cancer care.


Preparing synthetic biology for the world.
Synthetic Biology promises low-cost, exponentially scalable products and global health solutions in the form of self-replicating organisms, or "living devices." As these promises are realized, proof-of-concept systems will gradually migrate from tightly regulated laboratory or industrial environments into private spaces as, for instance, probiotic health products, food, and even do-it-yourself bioengineered systems. What additional steps, if any, should be taken before releasing engineered self-replicating organisms into a broader user space? In this review, we explain how studies of genetically modified organisms lay groundwork for the future landscape of biosafety. Early in the design process, biological engineers are anticipating potential hazards and developing innovative tools to mitigate risk. Here, we survey lessons learned, ongoing efforts to engineer intrinsic biocontainment, and how different stakeholders in synthetic biology can act to accomplish best practices for biosafety.


Toxoplasma gondii infection enhances testicular steroidogenesis in rats.
The protozoan parasite Toxoplasma gondii enhances the sexual attractiveness of infected male rats and attenuates the innate fear of cat odour in infected individuals. These behavioural changes plausibly lead to greater transmission of parasites through sexual and trophic routes, respectively. Testosterone, a testicular steroid, is known to reduce fear and enhance sexual attractiveness in males. Here, we show that Toxoplasma gondii infection enhances expression of genes involved in facilitating synthesis of testosterone, resulting in greater testicular testosterone production in male rats. In several species, testosterone mediates trade-offs between sexually selected traits and life history decisions. Augmentation of testosterone synthesis by Toxoplasma gondii suggests that parasites may manipulate these trade-offs in rats.


Effects of dimethylsulfoxide and pentoxifylline in the vitality of cutaneous flaps in rats.
to verify the influence of dimethylsulfoxide and pentoxifylline on the vitality of cutaneous flaps in rats and the tissue repair process. were studied 30 Wistar rats, submitting them to a 2cm wide by 8cm long dorsal cutaneous flap, of caudal base. We distributed the animals in three groups: Control Group (n=10) with application gauze moistened with 0.9% Saline in the flap bed for 30 seconds; Dimethylsulfoxide group (n=10), with administration of 1ml of 5% dimethylsulfoxide divided into five injections of 0.2ml in the transition of the flap segments; Pentoxifylline group (n=10), with administration of pentoxifylline 20mg/kg, diluted to 1ml and divided into five injections of 0.2ml in the transition of the flap segments. Drugs were administered intraoperatively, in a single dose and subcutaneously. We observed the skin flaps for changes in color and texture. On the 10th postoperative day, we checked the dimensions of viable and necrotic tissues, followed by excision of the specimen for histological analysis. the measurements of length of the viable and necrotic tissues between groups showed no differences. Histological analysis showed that the Dimethylsulfoxide group presented neovascularization, inflammatory infiltrate with leukocytes and more structured conjunctival stroma. The Pentoxifylline group showed neovascularization and inflammatory infiltrate, with moderate to intense granulation. The control group evolved with a higher rate of necrosis in the distal segment. dimethylsulfoxide and pentoxifylline influenced the vitality of the flap and the tissue repair process. However, they did not prevent necrosis macroscopically.


Structure of Staphylococcus aureus cytidine monophosphate kinase in complex with cytidine 5'-monophosphate.
The crystal structure of Staphylococcus aureus cytidine monophosphate kinase (CMK) in complex with cytidine 5'-monophosphate (CMP) has been determined at 2.3 angstroms resolution. The active site reveals novel features when compared with two orthologues of known structure. Compared with the Streptococcus pneumoniae CMK solution structure of the enzyme alone, S. aureus CMK adopts a more closed conformation, with the NMP-binding domain rotating by approximately 16 degrees towards the central pocket of the molecule, thereby assembling the active site. Comparing Escherichia coli and S. aureus CMK-CMP complex structures reveals differences within the active site, including a previously unreported indirect interaction of CMP with Asp33, the replacement of a serine residue involved in the binding of CDP by Ala12 in S. aureus CMK and an additional sulfate ion in the E. coli CMK active site. The detailed understanding of the stereochemistry of CMP binding to CMK will assist in the design of novel inhibitors of the enzyme. Inhibitors are required to treat the widespread hospital infection methicillin-resistant S. aureus (MRSA), currently a major public health concern.


Cerebral collateral therapeutics in acute ischemic stroke: A randomized preclinical trial of four modulation strategies.
Cerebral collaterals are dynamically recruited after arterial occlusion and highly affect tissue outcome in acute ischemic stroke. We investigated the efficacy and safety of four pathophysiologically distinct strategies for acute modulation of collateral flow (collateral therapeutics) in the rat stroke model of transient middle cerebral artery (MCA) occlusion. A composed randomization design was used to assign rats (n = 118) to receive phenylephrine (induced hypertension), polygeline (intravascular volume load), acetazolamide (cerebral arteriolar vasodilation), head down tilt (HDT) 15° (cerebral blood flow diversion), or no treatment, starting 30 min after MCA occlusion. Compared to untreated animals, treatment with collateral therapeutics was associated with lower infarct volumes (62% relative mean difference; 51.57 mm3 absolute mean difference; p < 0.001) and higher chance of good functional outcome (OR 4.58, p < 0.001). Collateral therapeutics acutely increased cerebral perfusion in the medial (+40.8%; p < 0.001) and lateral (+19.2%; p = 0.016) MCA territory compared to pretreatment during MCA occlusion. Safety indicators were treatment-related mortality and cardiorespiratory effects. The highest efficacy and safety profile was observed for HDT. Our findings suggest that acute modulation of cerebral collaterals is feasible and provides a tissue-saving effect in the hyperacute phase of ischemic stroke prior to recanalization therapy.


The form of folate affects the mechanisms of methotrexate resistance in Enterococcus hirae [corrected].
Several mechanisms have been described to explain the resistance of cells to methotrexate (MTX); however, the basis for the heterogeneity of mechanisms has been obscure. It was hypothesized that the type of MTX resistance in a single species can be influenced by the form of extracellular folate supplied during the development of resistance. Two strains of MTX-resistant Enterococcus hirae [corrected] were developed by transferring the bacteria to media containing increasing concentrations of MTX in the presence of constant concentrations of either 5-formyl-5,6,7,8-tetrahydropteroylglutamic acid (5-HCO-H4PteGlu) or pteroylglutamic acid (PteGlu). These resistant strains were designated E. hirae/MTX/5-HCO-H4PteGlu and E. hirae/MTX/PteGlu, respectively [corrected]. The mechanisms of MTX resistance included: (1) increased folic acid reductase (FAR) activity in both resistant strains but increased dihydrofolate reductase (DHFR) activity only in E. hirae/MTX/PteGlu [corrected]; (2) decreased synthesis and intracellular retention of MTX containing two glutamyl residues; (3) decreased uptake of MTX accompanied by decreased uptake of folates; and (4) reduction of folate-binding capacity. Among these, the form of folate present in the media during the development of resistance affected DHFR and FAR activities and the transport of folates. These findings, together with data from other laboratories, suggest that it may be important to use a reduced form of folate, a more physiological form than oxidized PteGlu, in the media during the development of resistance for the study of the mechanisms of MTX resistance in cultured cells.


Generalized chronotaxic systems: time-dependent oscillatory dynamics stable under continuous perturbation.
Chronotaxic systems represent deterministic nonautonomous oscillatory systems which are capable of resisting continuous external perturbations while having a complex time-dependent dynamics. Until their recent introduction in Phys. Rev. Lett. 111, 024101 (2013) chronotaxic systems had often been treated as stochastic, inappropriately, and the deterministic component had been ignored. While the previous work addressed the case of the decoupled amplitude and phase dynamics, in this paper we develop a generalized theory of chronotaxic systems where such decoupling is not required. The theory presented is based on the concept of a time-dependent point attractor or a driven steady state and on the contraction theory of dynamical systems. This simplifies the analysis of chronotaxic systems and makes possible the identification of chronotaxic systems with time-varying parameters. All types of chronotaxic dynamics are classified and their properties are discussed using the nonautonomous Poincaré oscillator as an example. We demonstrate that these types differ in their transient dynamics towards a driven steady state and according to their response to external perturbations. Various possible realizations of chronotaxic systems are discussed, including systems with temporal chronotaxicity and interacting chronotaxic systems.


Multiphasic temporal dynamics in responses of midbrain dopamine neurons to appetitive and aversive stimuli.
The transient response of dopamine neurons has been described as reward prediction error (RPE), with activation or suppression by events that are better or worse than expected, respectively. However, at least a minority of neurons are activated by aversive or high-intensity stimuli, casting doubt on the generality of RPE in describing the dopamine signal. To overcome limitations of previous studies, we studied neuronal responses to a wider variety of high-intensity and aversive stimuli, and we quantified and controlled aversiveness through a choice task in which macaques sacrificed juice to avoid aversive stimuli. Whereas most previous work has portrayed the RPE as a single impulse or "phase," here we demonstrate its multiphasic temporal dynamics. Aversive or high-intensity stimuli evoked a triphasic sequence of activation-suppression-activation extending over a period of 40-700 ms. The initial activation at short latencies (40-120 ms) reflected sensory intensity. The influence of motivational value became dominant between 150 and 250 ms, with activation in the case of appetitive stimuli, and suppression in the case of aversive and neutral stimuli. The previously unreported late activation appeared to be a modest "rebound" after strong suppression. Similarly, strong activation by reward was often followed by suppression. We suggest that these "rebounds" may result from overcompensation by homeostatic mechanisms in some cells. Our results are consistent with a realistic RPE, which evolves over time through a dynamic balance of excitation and inhibition.


Panning for gold in mould: can we increase the odds for fungal genome mining?
Fungi are a rich source of bioactive small molecules. However, the large number of biosynthetic gene clusters (BGCs) encoding these molecules in their genomes suggests their biosynthetic potential is far greater than we previously appreciated. The mining of fungal genomes therefore holds great promise for the discovery of new chemical entities for pharmaceutical and agricultural applications. As more and more fungal genomes become available, the accompanying number of BGCs is quickly becoming unmanageable. Along with improving molecular genetic tools to accelerate the translation of BGCs to small molecules, we must devise strategies to prioritise BGCs most likely to encode the biosynthesis of novel small molecules and molecules with new or improved bioactivities or functions. In this perspective, we discuss existing and emerging strategies for prioritisation of BGCs to increase the odds of fruitful genome mining in fungi.


Toxicology and Carcinogenesis Studies of C.I. Pigment Red 23 (CAS No. 6471-49-4) in F344 Rats and B6C3F1 Mice (Feed Studies).
C.I. Pigment Red 23 is a bluish red commercial dye used as a coloring agent in paints, inks, rubber, plastics, lacquers, and paper. Toxicology and carcinogenicity studies were conducted by feeding groups of rats and mice diets containing C.I. Pigment Red 23 (greater than 96% pure) for 17 days, 13 weeks, and 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and in Chinese hamster ovary cells. 17-Day Studies: Groups of five rats and five mice of each sex were fed diets containing 0, 6,000, 12,500, 25,000, 50,000, or 100,000 ppm C.I. Pigment Red 23 for 15 to 17 days. All rats and all female mice lived until the end of the studies. Two male mice in the 12,500 ppm dose group died accidentally. No other deaths occurred among male mice. Final mean body weights of rats and mice receiving C.I. Pigment Red 23 were within 10% of those of the controls. Feed consumption by exposed animals was similar to that of the controls. Hematocrit value, hemoglobin concentration, and erythrocyte count were decreased in the 50,000 and 100,000 ppm groups of rats. A corresponding decrease was not seen in mice. Absolute and relative organ weights of exposed animals were generally similar to those of the controls. No chemical-related gross lesions were seen in rats or mice. 13-Week Studies: Groups of 10 rats and 10 mice of each sex were fed diets containing 0, 3,000, 6,000, 12,500, 25,000, or 50,000 ppm C.I. Pigment Red 23 for 13 weeks. All rats and mice lived until the end of the studies. Final mean body weights of rats and mice receiving C.I. Pigment Red 23 were within 10% of those of the controls. Feed consumption by exposed animals was similar to that of the controls. In 50,000 ppm male rats, hematocrit and hemoglobin concentrations and erythrocyte counts were significantly less than those of the controls. In female rats receiving 3,000, 6,000 and 50,000 ppm C.I. Pigment Red 23, lymphocyte counts were significantly higher than the control values. Leukocyte counts in 3,000 ppm females were also significantly increased. Female mice in the 6,000 ppm dose group had significantly lower hematocrit and hemoglobin concentrations than did untreated females. Hematology parameters in exposed males were similar to those of untreated males. There were no biologically significant differences in organ weights among dosed and control rats. Absolute and relative liver weights of male mice receiving 12,500 ppm C.I. Pigment Red 23 were significantly increased compared to those of the controls. Absolute and relative thymus weights for all but 12,500 ppm female mice were significantly lower than those of the controls. No chemical-related gross or histopathologic lesions occurred in rats or mice. 2-Year Studies: Survival, Body Weights, Feed Consumption, and Clinical Findings Because levels of C.I. Pigment Red 23 as high as 50,000 or 100,000 ppm in the feed did not adversely affect survival and mean body weights in the 17-day and 13-week studies, nor cause any chemical- related lesions, doses of 0, 10,000, 25,000, or 50,000 ppm were selected for the 2-year studies. Doses higher than 50,000 ppm (5%) are not used in 2-year studies because they may lead to excessive dilution of nutrients in feed which in turn could produce nutritional deficiencies. Survival rates of mid- and high-dose male and of high-dose female rats were significantly greater than those of the controls, due primarily to a chemical related decreased incidence of mononuclear cell leukemia in these groups (survival in male rats: control, 22/50, low-dose, 29/50, mid-dose, 36/50, high-dose, 35/51; female rats: 29/50, 34/50, 33/50, 40/50). Survival of mice was not affected by the administration of C.I. Pigment Red 23, although survival of low-dose male mice was significantly lower than that of controls (male mice: 29/51, 17/53, 27/52, 30/51; female mice: 35/50, 34/49, 36/50, 35/49). The decreased survival in the low- dose males was associated with evidence of body trauma and secondary septicemia caused by fighting. From approximately week 20 of the study, the group mean body weight, the group mean body weights of exposed female rats were consistently lower than those of controls; at week 101, mean body weights of mid- dose (25,000 ppm) and high-dose (50,000 ppm) females were 6% and 8% less, respectively. The final mean body weights of exposed male rats and male and female mice were similar to those of controls. Feed consumption values for exposed male and female rats and mice were similar to those of the controls and there were no clinical signs associated with the administration of C.I. Pigment Red 23. Pathology Findings: Renal tubule adenomas occurred in two high- dose male rats. Renal tubule carcinomas occurred in one high-dose male and one mid-dose male rat. No renal tubule neoplasms were seen in the controls. Renal tubule neoplasms are uncommon and have occurred in 8/499 (1.6%) untreated historical controls with a range of 0% to 6%. The residual halves of kidneys from control and high-dose males were step sectioned and examined; renal tubule adenomas were observed in a control male and in two additional high- dose males. Because of the low numbers of renal neoplasms, it is uncertain if they were related to chemical administration. The incidence of renal tubule hyperplasia (3/50, 6/48, 5/50, 8/50) and the mean severity of nephropathy were also slightly increased in high-dose male rats. The incidence of mononuclear cell leukemia occurred with a significant negative trend in exposed male and female rats. No chemical-related increases in the incidence of neoplasms were observed in mice of either sex. There was a chemical-related increase in the incidence of hyperplasia (male mice: 0/49, 1/48, 1/50, 7/48; female mice: 6/49, 14/49, 43/50, 47/49) and hyperkeratosis of the forestomach epithelium attributed to chemical administration. Genetic Toxicology: C.I. Pigment Red 23 was mutagenic in Salmonella typhimurium strains TA100, TA1537, and TA98 with and without exogenous metabolic activation (S9), but it was not mutagenic in strain TA1535. C.I. Pigment Red 23 induced sister chromatid exchanges in Chinese hamster ovary cells in the absence of S9, but not with S9 activation. The pigment was negative for the induction of chromosomal aberrations in Chinese hamster ovary cells both in the presence and absence of S9. Conclusions: Under the conditions of these 2-year feed studies, there was equivocal evidence of carcinogenic activity of C.I. Pigment Red 23 in male F344 rats as evidenced by a marginally increased incidence of renal tubule cell neoplasms. There was no evidence of carcinogenic activity of C.I. Pigment Red 23 in female F344 rats fed diets containing 10,000, 25,000, or 50,000 ppm. Mononuclear cell leukemia occurred with a decreased incidence in male and female rats receiving C.I. Pigment Red 23. There was no evidence of carcinogenic activity of C.I. Pigment Red 23 in male and female B6C3F1 mice fed diets containing 10,000, 25,000 or 50,000 ppm. The severity of kidney nephropathy was increased in exposed male rats. In mice, C.I. Pigment Red 23 caused an increase in hyperkeratosis and epithelial hyperplasia of the fore- stomach. Synonyms: 2-Naphthalenecarboxamide; 3-hydroxy-4-((2-methoxy-5- nitrophenyl)azo)-N-(3- nitrophenyl); 3-hydroxy-4-((2-methoxy-5- nitrophenyl)azo)-3 -2-naphthanilide; Alkali Resistant Red Dark; Calcotone Red 3B; Carnation Red Toner B; CI 12355; Congo Red R- 138; Fenalac Red FKB Extra; Malta Red X2284; Naphthol Red B; Naphthol Red T Toner 35- 6001; Naphthol Red Deep 10459; Pigment Red BH; Rubescence Red MT-21; Sanyo Fast Red 10B; Sapona Red Lake RL-6280; Sengale Light Rubin RG; Textile Red WD-263


Lipid functions in skin: Differential effects of n-3 polyunsaturated fatty acids on cutaneous ceramides, in a human skin organ culture model.
Ceramides are important for skin health, with a multitude of species found in both dermis and epidermis. The epidermis contains linoleic acid-Ester-linked Omega-hydroxylated ceramides of 6-Hydroxy-sphingosine, Sphingosine and Phytosphingosine bases (CER[EOH], CER[EOS] and CER[EOP], respectively), that are crucial for the formation of the epidermal barrier, conferring protection from environmental factors and preventing trans-epidermal water loss. Furthermore, a large number of ceramides, derivatives of the same sphingoid bases and various fatty acids, are produced by dermal and epidermal cells and perform signalling roles in cell functions ranging from differentiation to apoptosis. Supplementation with the n-3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have shown promise as therapeutic agents in a number of inflammatory skin conditions, altering the lipid profile of the skin and production of bioactive lipids such as the eicosanoids, docosanoids and endocannabinoids. In this study we wished to investigate whether EPA and DHA could also affect the ceramide profile in epidermis and dermis, and, in this way, contribute to formation of a robust lipid barrier and ceramide-mediated regulation of skin functions. Ex vivo skin explants were cultured for 6days, and supplemented with EPA or DHA (50μM). Liquid chromatography coupled to tandem mass spectrometry with electrospray ionisation was used to assess the prevalence of 321 individual ceramide species, and a number of sphingoid bases, phosphorylated sphingoid bases, and phosphorylated ceramides, within the dermis and epidermis. EPA augmented dermal production of members of the ceramide families containing Non-hydroxy fatty acids and Sphingosine or Dihydrosphingosine bases (CER[NS] and CER[NDS], respectively), while epidermal CER[EOH], CER[EOS] and CER[EOP] ceramides were not affected. DHA did not significantly affect ceramide production. Ceramide-1-phosphate levels in the epidermis, but not the dermis, increased in response to EPA, but not DHA. This ex vivo study shows that dietary supplementation with EPA has the potential to alter the ceramide profile of the skin, and this may contribute to its anti-inflammatory profile. This has implications for formation of the epidermal lipid barrier, and signalling pathways within the skin mediated by ceramides and other sphingolipid species. This article is part of a Special Issue entitled: Membrane Lipid Therapy: Drugs Targeting Biomembranes edited by Pablo V. Escribá.


The role of self-association in Fin1 function on the mitotic spindle.
Stabilization of spindle microtubules during anaphase is essential for proper chromosome segregation. Fin1 is a budding yeast protein that localizes to the poles and microtubules of the spindle during anaphase and contributes to spindle stability. The N-terminal half of Fin1 is phosphorylated at multiple sites by the cyclin-dependent kinase Clb5-Cdk1, and dephosphorylation in anaphase triggers its localization to the spindle. The C-terminal half of Fin1 contains coiled-coil motifs that are required for its self-association. Here we investigated the functional importance of the two regions of Fin1. Fin1 mutants lacking the C-terminal coiled-coil domains localized to spindle pole bodies but not along spindle microtubules. These mutants failed to self-associate and displayed reduced binding to microtubules in vitro but were functional in vivo and stabilized anaphase spindles when dephosphorylated. Deletion of the Fin1 C terminus suppressed the lethal phenotypes of the phospho-mutant Fin15A. Our findings suggest that the N-terminal region of Fin1 is sufficient for its regulated function as a spindle-stabilizing factor and that this function involves association with the spindle pole body. The ability of the C-terminal region to promote Fin1 self-association and microtubule binding may underlie the lethal effects of the deregulated Fin15A mutant.


Testosterone Threshold for Increased Cardiovascular Risk in Middle-Aged and Elderly Men: A Locally Weighted Regression Analysis.
Although testosterone deficiency has a well-known association with increased risk of cardiovascular disease (CVD), the threshold remains to be determined. To investigate whether there is a discriminatory testosterone level below which the CVD risk increases. The study included 876 men 45 to 74 years old who underwent a general health checkup. The Framingham Risk Score was used to estimate the 10-year CVD risk; a high-sensitivity C-reactive protein (hsCRP) level of at least 1 mg/L was considered an indicator of increased CVD risk. Aging symptoms and sexual function were evaluated with the Aging Males' Symptom Scale. Locally weighted regression was performed to determine the testosterone threshold for Framingham CVD risk and increased hsCRP. The mean age was 56.6 ± 7.0 years. The mean total testosterone level was 394.3 ± 115.7 ng/dL. The mean 10-year Framingham CVD risk was 16.6 ± 10.7%, and 169 (19.3%) had increased hsCRP. The locally weighted regression showed that total testosterone levels of 440 and 480 ng/dL were associated with increased Framingham CVD risk and an increased probability of increased hsCRP, respectively. Men with sexual dysfunction (poor sexual performance, decreased morning erection, and loss of libido) had significantly greater CVD risk. Their risk appeared to increase at a relatively higher testosterone level, and it reached a plateau at a testosterone level of 300 to 350 ng/dL. In contrast, the risk in those with no or less sexual dysfunction remained low at a higher testosterone level, and a threshold level of 425 to 475 ng/dL was associated with increased CVD risk. A similar pattern and threshold were identified in the analyses of the relation between testosterone and hsCRP. These data showed that a testosterone threshold of 440 ng/dL was associated with increased Framingham 10-year CVD risk in middle-aged and elderly men. Poor sexual performance, decreased morning erection, and loss of libido had an impact on the testosterone threshold for CVD risk. The threshold level was higher in men with sexual dysfunction. Further study is required to evaluate the validity of these testosterone thresholds for CVD risk.


The crucial role of thiamine in the development of syntax and lexical retrieval: a study of infantile thiamine deficiency.
This study explored the effect of thiamine deficiency during early infancy on the development of syntax and lexical retrieval. We tested syntactic comprehension and production, lexical retrieval abilities and conceptual abilities of 59 children aged 5-7 years who had been fed during their first year of life with a thiamine-deficient milk substitute. We compared them to 35 age-matched control children who were fed with other milk sources. Experiment 1 tested the comprehension of relative clauses using a sentence-picture-matching task. Experiment 2 tested the production of relative clauses using a preference elicitation task. Experiment 3 tested the repetition of various syntactic structures with various types of syntactic movement and embedding. Experiment 4 tested picture naming and Experiment 5 tested lexical substitutions in a sentence repetition task. Experiments 6 and 7 tested the children's conceptual abilities using a picture association task and a picture absurdity description task. The results indicated a very high rate of syntactic and lexical retrieval deficits in the group of children who were exposed to thiamine deficiency in early infancy: 57 of the 59 thiamine-deficient children examined had language impairment, compared with three of the 35 controls (9%). Importantly, unlike the impairment this group sustained in their language abilities, the conceptual abilities of most of the children were intact (only six children, 10%, were conceptually impaired). These findings indicate that thiamine deficiency in infancy causes severe and long-lasting language disorders and that nutrition may be one of the causes for language impairment.


Tuneable ultra high specific surface area Mg/Al-CO3 layered double hydroxides.
We report the synthesis of tuneable ultra high specific surface area Aqueous Miscible Organic solvent-Layered Double Hydroxides (AMO-LDHs). We have investigated the effects of different solvent dispersion volumes, dispersion times and the number of re-dispersion cycles specific surface area of AMO-LDHs. In particular, the effects of acetone dispersion on two different morphology AMO-LDHs (Mg3Al-CO3 AMO-LDH flowers and Mg3Al-CO3 AMO-LDH plates) was investigated. It was found that the amount of acetone used in the dispersion step process can significantly affect the specific surface area of Mg3Al-CO3 AMO-LDH flowers while the dispersion time in acetone is critical factor to obtain high specific surface area Mg3Al-CO3 AMO-LDH plates. Optimisation of the acetone washing steps enables Mg3Al-CO3 AMO-LDH to have high specific surface area up to 365 m(2) g(-1) for LDH flowers and 263 m(2) g(-1) for LDH plates. In addition, spray drying was found to be an effective and practical drying method to increase the specific surface area by a factor of 1.75. Our findings now form the basis of an effective general strategy to obtain ultrahigh specific surface area LDHs.


Direct estimation of the mitochondrial DNA mutation rate in Drosophila melanogaster.
Mitochondrial DNA (mtDNA) variants are widely used in evolutionary genetics as markers for population history and to estimate divergence times among taxa. Inferences of species history are generally based on phylogenetic comparisons, which assume that molecular evolution is clock-like. Between-species comparisons have also been used to estimate the mutation rate, using sites that are thought to evolve neutrally. We directly estimated the mtDNA mutation rate by scanning the mitochondrial genome of Drosophila melanogaster lines that had undergone approximately 200 generations of spontaneous mutation accumulation (MA). We detected a total of 28 point mutations and eight insertion-deletion (indel) mutations, yielding an estimate for the single-nucleotide mutation rate of 6.2 x 10(-8) per site per fly generation. Most mutations were heteroplasmic within a line, and their frequency distribution suggests that the effective number of mitochondrial genomes transmitted per female per generation is about 30. We observed repeated occurrences of some indel mutations, suggesting that indel mutational hotspots are common. Among the point mutations, there is a large excess of G-->A mutations on the major strand (the sense strand for the majority of mitochondrial genes). These mutations tend to occur at nonsynonymous sites of protein-coding genes, and they are expected to be deleterious, so do not become fixed between species. The overall mtDNA mutation rate per base pair per fly generation in Drosophila is estimated to be about 10x higher than the nuclear mutation rate, but the mitochondrial major strand G-->A mutation rate is about 70x higher than the nuclear rate. Silent sites are substantially more strongly biased towards A and T than nonsynonymous sites, consistent with the extreme mutation bias towards A+T. Strand-asymmetric mutation bias, coupled with selection to maintain specific nonsynonymous bases, therefore provides an explanation for the extreme base composition of the mitochondrial genome of Drosophila.


Microbes follow Humboldt: temperature drives plant and soil microbial diversity patterns from the Amazon to the Andes.
More than 200 years ago, Alexander von Humboldt reported that tropical plant species richness decreased with increasing elevation and decreasing temperature. Surprisingly, coordinated patterns in plant, bacterial, and fungal diversity on tropical mountains have not yet been observed, despite the central role of soil microorganisms in terrestrial biogeochemistry and ecology. We studied an Andean transect traversing 3.5 km in elevation to test whether the species diversity and composition of tropical forest plants, soil bacteria, and fungi follow similar biogeographical patterns with shared environmental drivers. We found coordinated changes with elevation in all three groups: species richness declined as elevation increased, and the compositional dissimilarity among communities increased with increased separation in elevation, although changes in plant diversity were larger than in bacteria and fungi. Temperature was the dominant driver of these diversity gradients, with weak influences of edaphic properties, including soil pH. The gradients in microbial diversity were strongly correlated with the activities of enzymes involved in organic matter cycling, and were accompanied by a transition in microbial traits towards slower-growing, oligotrophic taxa at higher elevations. We provide the first evidence of coordinated temperature-driven patterns in the diversity and distribution of three major biotic groups in tropical ecosystems: soil bacteria, fungi, and plants. These findings suggest that interrelated and fundamental patterns of plant and microbial communities with shared environmental drivers occur across landscape scales. These patterns are revealed where soil pH is relatively constant, and have implications for tropical forest communities under future climate change.


Dual targeting of the cyclin/Rb/E2F and mitochondrial pathways in mantle cell lymphoma with the translation inhibitor silvestrol.
During cell-cycle progression, D-cyclins activate cyclin-dependent kinases (CDKs) 4/6 to inactivate Rb, permitting E2F1-mediated S-phase gene transcription. This critical pathway is typically deregulated in cancer, and novel inhibitory strategies would be effective in a variety of tumors. The protein synthesis inhibitor silvestrol has potent activity in B-cell leukemias via the mitochondrial pathway of apoptosis, and also reduces cyclin D1 expression in breast cancer and lymphoma cell lines. We hypothesized that this dual activity of silvestrol would make it especially effective in malignancies driven by aberrant cyclin D1 expression. Mantle cell lymphoma (MCL), characterized by elevated cyclin D1, was used as a model to test this approach. The cyclin D/Rb/E2F1 pathway was investigated in vitro using MCL cell lines and primary tumor cells. Silvestrol was also evaluated in vivo using an aggressive model of MCL. Silvestrol showed low nanomolar potency both in MCL cell lines and primary MCL tumor cells. D-cyclins were depleted with just 10 nmol/L silvestrol at 16 hours, with subsequent reductions of phosphorylated Rb, E2F1 protein, and E2F1 target transcription. As showed in other leukemias, silvestrol caused Mcl-1 depletion followed by mitochondrial depolarization and caspase-dependent apoptosis, effects not related to inhibition of CDK4/6. Silvestrol significantly (P < 0.0001) prolonged survival in a MCL xenograft model without detectable toxicity. These data indicate that silvestrol effectively targets the cyclin/CDK/Rb pathway, and additionally induces cytotoxicity via intrinsic apoptosis. This dual activity may be an effective therapeutic strategy in MCL and other malignancies.


Dormancy as a survival strategy of the fish pathogen Streptococcus parauberis in the marine environment.
The fate of Streptococcus parauberis in seawater and sediment microcosms at different temperatures (6 and 22 degrees C) was investigated by comparing the survival dynamics of 2 strains of this bacterial species, isolated respectively from diseased turbot and cattle. The turbot and the bovine isolate showed similar survival kinetics, remaining culturable for approximately 1 mo in water and 6 mo in sediment. A slight influence of temperature on the stability of the cells was observed, in that the number of culturable cells was about 1 log10 unit higher at 6 than at 22 degrees C. During the starvation period, the metabolic activity of the cells, after suffering a strong reduction during the first 12 d, stabilized at levels ranging from 20 to 40% of the initial values. However, in all the microcosms, the acridine orange (AO) and 4',6-diamidino-2-phenilindole (DAPI) counts remained at about 10(5) cells ml(-1) throughout the experimental period, even when cells became undetectable by standard plate count methods. The addition of fresh medium to microcosms containing nonculturable cells induced the return to culturability of S. parauberis strains. On the basis of these results, it seems that S. parauberis has the ability to enter into a viable but nonculturable (VBNC) state. Dormant cells of the turbot isolate maintained their infectivity and pathogenic potential for fish.


Hepatocellular ATP-binding cassette protein expression enhances ATP release and autocrine regulation of cell volume.
In a model liver cell line, recovery from swelling is mediated by a sensitive autocrine pathway involving conductive release of ATP, P2 receptor stimulation, and opening of membrane Cl- channels (Wang, Y., Roman, R. M., Lidofsky, S. D., and Fitz, J. G. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 12020-12025). However, the mechanisms coupling changes in cell volume to ATP release are not known. Based on evidence that certain ATP-binding cassette (ABC) proteins may function as ATP channels or channel regulators, we evaluated the potential role of ABC proteins by comparing ATP release and volume regulation in rat HTC and HTC-R hepatoma cells, the latter of which overexpress Mdr proteins. In both cell types, Cl- current activation (ICl-swell) and volume recovery following swelling were dependent on conductive ATP efflux. The rate of volume recovery was approximately 6-fold faster in HTC-R cells compared with HTC cells. This effect is likely due to enhanced ABC protein-dependent ATP release since (i) ICl-swell and cell volume recovery were eliminated by inhibition of P-glycoprotein transport (20 microM verapamil and 15 microM cyclosporin A); (ii) swelling-induced Cl- current density was similar in both cell types (approximately -50 pA/pF; not significant); and (iii) ATP conductance measured by whole-cell techniques was increased approximately 3-fold in HTC-R cells compared with HTC cells. Moreover, HTC-R cells exhibited enhanced survival during hypotonic stress. By modulating ATP release, hepatic ABC proteins may play a key role in the cellular pathways coupling changes in cell volume to ion permeability and secretion.


Test and treat: the early experiences in a clinic serving women at high risk of HIV infection in Kampala.
At the end of 2013, the Government of Uganda issued guidance recommending provision of Anti-Retroviral Treatment (ART) to HIV-positive people in key populations, including female sex workers, regardless of CD4 cell count. We describe the implementation of this new guidance in a clinic serving women at high risk of HIV infection in Kampala. Between July and December 2015, we conducted repeat in-depth interviews with 15 women attending the clinic after the change in guidelines, to explore their perceptions regarding prompt ART initiation. The sample included some women who were HIV-negative and women who had both started and deferred ART. We conducted a data-led thematic analysis of the material from the interviews. A total of 257 of 445 eligible women had started ART; others were undecided or had not returned to the clinic after receiving the new information. Participants recounted varying experiences with the provision of prompt treatment. At an individual level, a history of treatment for opportunistic infections and other illnesses, coupled with relatively poor health, encouraged some to initiate ART promptly. However, knowledge of friends/relatives already on ART who had experienced side effects caused others to delay starting, fearing the same experience for themselves. A number of women questioned why they should start treatment when they were not sick. Situational factors such as work and residence (with many sharing single rooms) caused discomfort among newly diagnosed women who feared disclosure and stigma that would result from taking ART when they were not ill. Alcohol consumption and irregular working hours affected perceptions of future adherence, making prompt ART harder to embrace for some. Our findings show the challenges that influence the delay of treatment initiation, and/or the decision to defer receiving information on ART, with implications for the success of the test and treat programmes and guidelines.


Linear summation of excitatory inputs by CA1 pyramidal neurons.
A fundamental problem in neurobiology is understanding the arithmetic that dendrites use to integrate inputs. The impact of dendritic morphology and active conductances on input summation is still unknown. To study this, we use glutamate iontophoresis and synaptic stimulation to position pairs of excitatory inputs throughout the apical, oblique, and basal dendrites of CA1 pyramidal neurons in rat hippocampal slices. Under a variety of stimulation regimes, we find a linear summation of most input combinations that is implemented by a surprising balance of boosting and shunting mechanisms. Active conductances in dendrites paradoxically serve to make summation linear. This "active linearity" can reconcile predictions from cable theory with the observed linear summation in vivo and suggests that a simple arithmetic is used by apparently complex dendritic trees.


Imaging diagnosis--congenital venous aneurysm of the left external jugular vein.
Primary congenital jugular vein aneurysms are rare with only one prior report in a canine patient. Herein, we describe an 18-week-old Flat Coated Retriever dog in which ultrasound was used to diagnose a primary congenital external jugular vein aneurysm with concurrent dilatation of the internal jugular vein. The aneurysm was successfully removed and the dog remains disease free 24 months postoperatively. Histopathologic analysis was used to validate the ultrasound-based diagnosis.


Microbial activity in the marine deep biosphere: progress and prospects.
The vast marine deep biosphere consists of microbial habitats within sediment, pore waters, upper basaltic crust and the fluids that circulate throughout it. A wide range of temperature, pressure, pH, and electron donor and acceptor conditions exists-all of which can combine to affect carbon and nutrient cycling and result in gradients on spatial scales ranging from millimeters to kilometers. Diverse and mostly uncharacterized microorganisms live in these habitats, and potentially play a role in mediating global scale biogeochemical processes. Quantifying the rates at which microbial activity in the subsurface occurs is a challenging endeavor, yet developing an understanding of these rates is essential to determine the impact of subsurface life on Earth's global biogeochemical cycles, and for understanding how microorganisms in these "extreme" environments survive (or even thrive). Here, we synthesize recent advances and discoveries pertaining to microbial activity in the marine deep subsurface, and we highlight topics about which there is still little understanding and suggest potential paths forward to address them. This publication is the result of a workshop held in August 2012 by the NSF-funded Center for Dark Energy Biosphere Investigations (C-DEBI) "theme team" on microbial activity (www.darkenergybiosphere.org).


Histologic Evaluation of Artificial Floors Under MTA and Nano-Filled Resin-Modified Glass Ionomer Used to Repair Furcation Perforations in Dogs.
This study aimed to compare the tissue reaction of two repair materials for furcation perforations, nano-filled resin modified glass ionomer (Nano-FRMGI) and mineral trioxide aggregate (MTA), used with or without an artificial floor. A total of 96 teeth in 6 dogs were used for this study. After access cavities, root canals were prepared and obturated with gutta percha using cold lateral condensation technique. Perforations were then created on the floors of the pulp chambers. The perforations divided into four groups n=24/group that were sealed with MTA alone, MTA with calcium sulphate artificial floor (CSAF), FRMGI alone and Nano-FRMGI with CSAF. All access cavities were filled with composite resin. Two dogs were sacrificed at 1, 3, and 6 month. The experimental tooth along with the surrounding alveolar bone were cut in block sections and histologically evaluated for tissue response. Data were analyzed by Chi-square (P≤0.05). MTA and MTA with CSAF showed more bone and cementum apposition when compared to Nano-FRMGI at 6-month interval. MTA and MTA with CSAF showed less bone resorption, epithelium proliferation and inflammation compared to Nano-FRMGI at 6-month interval. MTA with CSAF or MTA-alone show better outcomes in the repair of pulp chamber floor perforation.


Bile salt-dependent lipase interacts with platelet CXCR4 and modulates thrombus formation in mice and humans.
Bile salt-dependent lipase (BSDL) is an enzyme involved in the duodenal hydrolysis and absorption of cholesteryl esters. Although some BSDL is transported to blood, the role of circulating BSDL is unknown. Here, we demonstrate that BSDL is stored in platelets and released upon platelet activation. Because BSDL contains a region that is structurally homologous to the V3 loop of HIV-1, which binds to CXC chemokine receptor 4 (CXCR4), we hypothesized that BSDL might bind to CXCR4 present on platelets. In human platelets in vitro, both BSDL and a peptide corresponding to its V3-like loop induced calcium mobilization and enhanced thrombin-mediated platelet aggregation, spreading, and activated alpha(IIb)beta(3) levels. These effects were abolished by CXCR4 inhibition. BSDL also increased the production of prostacyclin by human endothelial cells. In a mouse thrombosis model, BSDL accumulated at sites of vessel wall injury. When CXCR4 was antagonized, the accumulation of BSDL was inhibited and thrombus size was reduced. In BSDL(-/-) mice, calcium mobilization in platelets and thrombus formation were attenuated and tail bleeding times were increased in comparison with those of wild-type mice. We conclude that BSDL plays a role in optimal platelet activation and thrombus formation by interacting with CXCR4 on platelets.


The effect of the Internet on the patient-doctor relationship from the patient's perspective: a survey from primary care.
Internet use by patients as a source of information on health and disease is expanding rapidly with obvious effects on the doctor-patient relationship. Many of these effects remain undocumented or are poorly understood. To assess the use of the internet for health information by patients in primary care in Israel and their perception of the effects of internet use on their relationship with their doctor. A cross-sectional survey was conducted among a convenience sample of patients visiting 10 primary care clinics in central Israel using a questionnaire developed for this survey. The survey examined attitudes to using the internet for health-related information and attitudes to sharing this information with doctors. Associations between demographic variables, internet use and patient satisfaction with the doctor's response were tested using the chi-square statistic and t-tests. Completed questionnaires were received from 138 patients; the response rate was 69%. Patients in the study sample had a high rate of internet access (87%), with many using the internet as a source of health information (41%) although most patients using the internet never share this information with their doctor (81%). Among those who share information with the doctor, most felt that this has a positive effect on the relationship (870/%). Few patients reported being referred to websites by the doctor (28%). Internet use is prevalent in this population, though physicians may be unaware of this. Future study could examine the effects of doctors who ask patients actively about their internet use and inform them of relevant health information sources online.


What makes Xanthomonas albilineans unique amongst xanthomonads?
Xanthomonas albilineans causes leaf scald, a lethal disease of sugarcane. Compared to other species of Xanthomonas, X. albilineans exhibits distinctive pathogenic mechanisms, ecology and taxonomy. Its genome, which has experienced significant erosion, has unique genomic features. It lacks two loci required for pathogenicity in other plant pathogenic species of Xanthomonas: the xanthan gum biosynthesis and the Hrp-T3SS (hypersensitive response and pathogenicity-type three secretion system) gene clusters. Instead, X. albilineans harbors in its genome an SPI-1 (Salmonella pathogenicity island-1) T3SS gene cluster usually found in animal pathogens. X. albilineans produces a potent DNA gyrase inhibitor called albicidin, which blocks chloroplast differentiation, resulting in the characteristic white foliar stripe symptoms. The antibacterial activity of albicidin also confers on X. albilineans a competitive advantage against rival bacteria during sugarcane colonization. Recent chemical studies have uncovered the unique structure of albicidin and allowed us to partially elucidate its fascinating biosynthesis apparatus, which involves an enigmatic hybrid PKS/NRPS (polyketide synthase/non-ribosomal peptide synthetase) machinery.


Rituximab in the Treatment of Interstitial Lung Disease Associated with Antisynthetase Syndrome: A Multicenter Retrospective Case Review.
To assess clinical outcomes including imaging findings on computed tomography (CT), pulmonary function testing (PFT), and glucocorticoid (GC) use in patients with the antisynthetase syndrome (AS) and interstitial lung disease (ILD) treated with rituximab (RTX). We retrospectively identified all patients at 2 institutions with AS-ILD who were treated with RTX. Baseline demographics, PFT, and chest CT were assessed before and after RTX. Two radiologists independently evaluated CT using a standardized scoring system. Twenty-five subjects at the Brigham and Women's Hospital (n = 13) and University of Pittsburgh Medical Center (n = 12) were included. Antisynthetase antibodies were identified in all patients (16 Jo1, 6 PL-12, 3 PL-7). In 21 cases (84%), the principal indication for RTX use was recurrent or progressive ILD, owing to failure of other agents. Comparing pre- and post-RTX pulmonary variables at 12 months, CT score and forced vital capacity were stable or improved in 88% and 79% of subjects, respectively. Total lung capacity (%) increased from 56 ± 13 to 64 ± 13 and GC dose decreased from 18 ± 9 to 12 ± 12 mg/day. Although DLCO (%) declined slightly at 1 year, it increased from 42 ± 17 to 70 ± 20 at 3 years. The most common imaging patterns on CT were nonspecific interstitial pneumonia (NSIP; n = 13) and usual interstitial pneumonia/fibrotic NSIP (n = 5), of which 5 had concurrent elements of cryptogenic organizing pneumonia. Stability or improvement in pulmonary function or severity of ILD on CT was seen in most patients. Use of RTX was well tolerated in the majority of patients. RTX may play a therapeutic role in patients with AS-ILD, and further clinical investigation is warranted.


Rapid targeted gene disruption in Bacillus anthracis.
Anthrax is a zoonotic disease recognized to affect herbivores since Biblical times and has the widest range of susceptible host species of any known pathogen. The ease with which the bacterium can be weaponized and its recent deliberate use as an agent of terror, have highlighted the importance of gaining a deeper understanding and effective countermeasures for this important pathogen. High quality sequence data has opened the possibility of systematic dissection of how genes distributed on both the bacterial chromosome and associated plasmids have made it such a successful pathogen. However, low transformation efficiency and relatively few genetic tools for chromosomal manipulation have hampered full interrogation of its genome. Group II introns have been developed into an efficient tool for site-specific gene inactivation in several organisms. We have adapted group II intron targeting technology for application in Bacillus anthracis and generated vectors that permit gene inactivation through group II intron insertion. The vectors developed permit screening for the desired insertion through PCR or direct selection of intron insertions using a selection scheme that activates a kanamycin resistance marker upon successful intron insertion. The design and vector construction described here provides a useful tool for high throughput experimental interrogation of the Bacillus anthracis genome and will benefit efforts to develop improved vaccines and therapeutics.


Influence of different treatment techniques and clinical factors over the intrafraction variation on lung stereotactic body radiotherapy.
In the present study we compared three different Stereotactic body radiation therapy (SBRT) treatment delivery techniques in terms of treatment time (TT) and their relation with intrafraction variation (IFV). Besides that, we analyzed if different clinical factors could have an influence on IFV. Finally, we appreciated the soundness of our margins. Forty-five patients undergoing SBRT for stage I lung cancer or lung metastases up to 5 cm were included in the study. All underwent 4DCT scan to create an internal target volume (ITV) and a 5 mm margin was added to establish the planning target volume (PTV). Cone-beam CTs (CBCTs) were acquired before and after each treatment to quantify the IFV. Three different treatment delivery techniques were employed: fixed fields (FF), dynamically collimated arcs (AA) or a combination of both (FA). We studied if TT was different among these modalities of SBRT and whether TT and IFV were correlated. Clinical data related to patients and tumors were recorded as potential influential factors over the IFV. A total of 52 lesions and 147 fractions were analyzed. Mean IFV for x-, y- and z-axis were 1 ± 1.16 mm, 1.29 ± 1.38 mm and 1.17 ± 1.08 mm, respectively. Displacements were encompassed by the 5 mm margin in 96.1 % of fractions. TT was significantly longer in FF therapy (24.76 ± 5.4 min), when compared with AA (15.30 ± 3.68 min) or FA (17.79 ± 3.52 min) (p < 0.001). Unexpectedly, IFV did not change significantly between them (p = 0.471). Age (p = 0.003) and left vs. right location (p = 0.005) were related to 3D shift ≥2 mm. In the multivariate analysis only age showed a significant impact on the IFV (OR = 1.07, p = 0.007). The choice of AA, FF or FA does not impact on IFV although FF treatment takes significantly longer treatment time. Our immobilization device offers enough accuracy and the 5 mm margin may be considered acceptable as it accounts for more than 95 % of tumor shifts. Age is the only clinical factor that influenced IFV significantly in our analysis.


Challenges of disseminating clinical practice guidelines in a weak health system: the case of HIV and infant feeding recommendations in Tanzania.
Clinical guidelines aim to improve patient outcomes by providing recommendations on appropriate healthcare for specific clinical conditions. Scientific evidence produced over time leads to change in clinical guidelines, and a serious challenge may emerge in the process of communicating the changes to healthcare practitioners and getting new practices adopted. There is very little information on the major barriers to implementing clinical guidelines in low-income settings. Looking at how continual updates to clinical guidelines within a particular health intervention are communicated may shed light on the processes at work. The aim of this paper is to explore how the content of a series of diverging infant feeding guidelines have been communicated to managers in the Prevention of Mother to Child Transmission of HIV Programme (PMTCT) with the aim of generating knowledge about both barriers and facilitating factors in the dissemination of new and updated knowledge in clinical guidelines in the context of weak healthcare systems. A total of 22 in-depth interviews and two focus group discussions were conducted in 2011. All informants were linked to the PMTCT programme in Tanzania. The informants included managers at regional and district levels and health workers at health facility level. The informants demonstrated partial and incomplete knowledge about the recommendations. There was lack of scientific reasoning behind various infant feeding recommendations. The greatest challenges to the successful communication of the infant feeding guidelines were related to slowness of communication, inaccessible jargon-ridden English language in the manuals, lack of summaries, lack of supportive supervision to make the guidelines comprehensible, and the absence of a reading culture. The study encountered substantial gaps in knowledge about the diverse HIV and infant feeding policies. These gaps were partly related to the challenges of communicating the clinical guidelines. There is a need for caution in assuming that important changes in guidelines for clinical practice can easily be translated to and implemented in local programme settings, not least in the context of weak healthcare systems.


Impact of Toceranib/Piroxicam/Cyclophosphamide Maintenance Therapy on Outcome of Dogs with Appendicular Osteosarcoma following Amputation and Carboplatin Chemotherapy: A Multi-Institutional Study.
We hypothesized that the addition of toceranib to metronomic cyclophosphamide/piroxicam therapy would significantly improve disease-free interval (DFI) and overall survival (OS) in dogs with appendicular osteosarcoma (OSA) following amputation and carboplatin chemotherapy. This was a randomized, prospective clinical trial in which dogs with OSA free of gross metastatic disease (n = 126) received carboplatin chemotherapy (4 doses) following amputation. On study entry, dogs were randomized to receive piroxicam/cyclophosphamide with or without toceranib (n = 63 each) after completing chemotherapy. Patient demographics were not significantly different between both groups. During or immediately following carboplatin chemotherapy, 32 dogs (n = 13 toceranib; n = 19 control) developed metastatic disease, and 13 dogs left the study due to other medical conditions or owner preference. Following carboplatin chemotherapy, 81 dogs (n = 46 toceranib; n = 35 control) received the metronomic treatment; 35 dogs (n = 20 toceranib; n = 15 control) developed metastatic disease during the maintenance therapy, and 26 dogs left the study due to other medical conditions or owner preference. Nine toceranib-treated and 11 control dogs completed the study without evidence of metastatic disease 1-year following amputation. Toceranib-treated dogs experienced more episodes of diarrhea, neutropenia and weight loss than control dogs, although these toxicities were low-grade and typically resolved with supportive care. More toceranib-treated dogs (n = 8) were removed from the study for therapy-associated adverse events compared to control dogs (n = 1). The median DFI for control and toceranib treated dogs was 215 and 233 days, respectively (p = 0.274); the median OS for control and toceranib treated dogs was 242 and 318 days, respectively (p = 0.08). The one year survival rate for control dogs was 35% compared to 38% for dogs receiving toceranib. The addition of toceranib to metronomic piroxicam/cyclophosphamide therapy following amputation and carboplatin chemotherapy did not improve median DFI, OS or the 1-year survival rate in dogs with OSA.


Global eradication rates for Helicobacter pylori infection: systematic review and meta-analysis of sequential therapy.
To do a systematic review and meta-analysis of studies comparing sequential therapy for eradication of Helicobacter pylori with pre-existing and new therapies, thus providing a glimpse of eradication success worldwide. Systematic review and meta-analysis. Medline, Embase, and Cochrane Central Register of Controlled Trials up to May 2013; abstract books of major European, American, and Asian gastroenterological meetings. Randomised controlled trials in previously untreated adults, in which sequential therapy was compared with a pre-existing or new therapy. 46 randomised controlled trials were reviewed and analysed. 5666 patients were randomised to sequential therapy and 7866 to other (established and new) treatments. The overall eradication rate of sequential therapy was 84.3% (95% confidence interval 82.1% to 86.4%). Sequential therapy was superior to seven day triple therapy (relative risk 1.21, 95% confidence interval 1.17 to 1.25; I(2)=29.3%; number needed to treat 6, 95% confidence interval 5% to 7%), marginally superior to 10 day triple therapy (1.11, 1.04 to 1.19; I(2)= 67.2%; NNT 10, 7 to 15), but not superior to 14 day triple therapy (1.00, 0.94 to 1.06; I(2)=54.3%), bismuth based therapy (1.01, 0.95 to 1.06; I(2)=21.1%), and non-bismuth based therapy (0.99, 0.94 to 1.05; I(2)=52.3%). Data on eradication according to pre-treatment antimicrobial susceptibility testing were available in eight studies, and sequential therapy was able to eradicate 72.8% (61.6% to 82.8%) of the strains resistant to clarithromycin. Eradication rates with pre-existing and new therapies for H pylori are suboptimal. Regional monitoring of resistance rates should help to guide treatment, and new agents for treatment need to be developed.


Erectogenic and neurotrophic effects of icariin, a purified extract of horny goat weed (Epimedium spp.) in vitro and in vivo.
Epimedium species (aka horny goat weed) have been utilized for the treatment of erectile dysfunction in Traditional Chinese Medicine for many years. Icariin (ICA) is the active moiety of Epimedium species. To evaluate the penile hemodynamic and tissue effects of ICA in cavernous nerve injured rats. We also studied the in vitro effects of ICA on cultured pelvic ganglia. Rats were subjected to cavernous nerve injury and subsequently treated for 4 weeks with daily gavage feedings of a placebo solution of normal saline and Dimethyl sulfoxide (DMSO) vs. ICA dissolved in DMSO at doses of 1, 5, and 10 mg/kg. A separate group underwent a single dose of ICA 10 mg/kg 2 hours prior to functional testing. Functional testing with cavernous nerve stimulation and real-time assessment of intracavernous pressure (ICP) was performed at 4 weeks. After functional testing, penile tissue was procured for immunohistochemistry and molecular studies. In separate experiments, pelvic ganglia were excised from healthy rats and cultured in the presence of ICA, sildenafil, or placebo culture media. Ratio of ICP and area under the curve (AUC) to mean arterial pressure (MAP) during cavernous nerve stimulation of subject rodents. We also assayed tissue expression of neuronal nitric oxide synthase (nNOS), eNOS: endothelial nitric oxide synthase (eNOS), calponin, and apoptosis via immunohistochemistry and Western blot. Serum testosterone and luteinizing hormone (LH) were assayed using enzyme-linked immunosorbant assay (ELISA). Differential length of neurite outgrowth was assessed in cultured pelvic ganglia. Rats treated with low-dose ICA demonstrated significantly higher ICP/MAP and AUC/MAP ratios compared with control and single-dose ICA animals. Immunohistochemistry and Western blot were revealing of significantly greater positivity for nNOS and calponin in penile tissues of all rats treated with ICA. ICA led to significantly greater neurite length in cultured specimens of pelvic ganglia. ICA may have neurotrophic effects in addition to known phosphodiesterase type 5 inhibiting effects.


Prevalence of Centrocestus formosanus Metacercariae in Ornamental Fish from Chiang Mai, Thailand, with Molecular Approach Using ITS2.
The prevalence of Centrocestus formosanus metacercariae was investigated in ornamental fish purchased from a pet shop in Chiang Mai, Thailand, including Carassius auratus (goldfish), Cyprinus carpio (Koi), Poecilia latipinna (Sailfin Molly), Danio rerio (Zebrafish), and Puntigrus tetrazona (Tiger barb). The parasite species was identified by the morphology of worms as well as by a molecular approach using ITS2. The results showed that 50 (33.3%) of 150 fish examined were infected with the metacercariae. The highest prevalence was found in C. auratus (83.3%), and the highest intensity was noted in C. carpio (70.8 metacercariae/fish). The most important morphological character was the presence of 32-34 circumoral spines on the oral sucker. The phylogenetic studies using the rRNA ITS2 region revealed that all the specimens of C. formosanus in this study were grouped together with C. formosanus in GenBank database. This is the first report on ornamental fish, C. carpio, P. latipinna, D. rerio, and P. tetrazona, taking the role of second intermediate hosts of C. formosanus in Thailand. Prevention and control of metacercarial infection in ornamental fish is urgently needed.


Time of Day of Vaccination Affects SARS-CoV-2 Antibody Responses in an Observational Study of Health Care Workers.
The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a global crisis with unprecedented challenges for public health. Vaccinations against SARS-CoV-2 have slowed the incidence of new infections and reduced disease severity. As the time of day of vaccination has been reported to influence host immune responses to multiple pathogens, we quantified the influence of SARS-CoV-2 vaccination time, vaccine type, participant age, sex, and days post-vaccination on anti-Spike antibody responses in health care workers. The magnitude of the anti-Spike antibody response is associated with the time of day of vaccination, vaccine type, participant age, sex, and days post-vaccination. These results may be relevant for optimising SARS-CoV-2 vaccine efficacy.


Meditation increases the depth of information processing and improves the allocation of attention in space.
During meditation, practitioners are required to center their attention on a specific object for extended periods of time. When their thoughts get diverted, they learn to quickly disengage from the distracter. We hypothesized that learning to respond to the dual demand of engaging attention on specific objects and disengaging quickly from distracters enhances the efficiency by which meditation practitioners can allocate attention. We tested this hypothesis in a global-to-local task while measuring electroencephalographic activity from a group of eight highly trained Buddhist monks and nuns and a group of eight age and education matched controls with no previous meditation experience. Specifically, we investigated the effect of attentional training on the global precedence effect, i.e., faster detection of targets on a global than on a local level. We expected to find a reduced global precedence effect in meditation practitioners but not in controls, reflecting that meditators can more quickly disengage their attention from the dominant global level. Analysis of reaction times confirmed this prediction. To investigate the underlying changes in brain activity and their time course, we analyzed event-related potentials. Meditators showed an enhanced ability to select the respective target level, as reflected by enhanced processing of target level information. In contrast with control group, which showed a local target selection effect only in the P1 and a global target selection effect in the P3 component, meditators showed effects of local information processing in the P1, N2, and P3 and of global processing for the N1, N2, and P3. Thus, meditators seem to display enhanced depth of processing. In addition, meditation altered the uptake of information such that meditators selected target level information earlier in the processing sequence than controls. In a longitudinal experiment, we could replicate the behavioral effects, suggesting that meditation modulates attention already after a 4-day meditation retreat. Together, these results suggest that practicing meditation enhances the speed with which attention can be allocated and relocated, thus increasing the depth of information processing and reducing response latency.


Comparison of CT and 18F-FDG pet for detecting peritoneal metastasis on the preoperative evaluation for gastric carcinoma.
The aim of our study was to compare the accuracy of CT and (18)FFDG PET for detecting peritoneal metastasis in patients with gastric carcinoma. One-hundred-twelve patients who underwent a histologic confirmative exam or treatment (laparotomy, n = 107; diagnostic laparoscopy, n = 4; peritoneal washing cytology, n = 1) were retrospectively enrolled. All the patients underwent CT and (18)F-FDG PET scanning for their preoperative evaluation. The sensitivities, specificities and accuracies of CT and (18)FFDG PET imaging for the detection of peritoneal metastasis were calculated and then compared using Fisher's exact probability test (p < 0.05), on the basis of the original preoperative reports. In addition, two board-certified radiologists and two board-certified nuclear medicine physicians independently reviewed the CT and PET scans, respectively. A receiver-operating characteristic curve analysis was performed to compare the diagnostic performance of CT and (18)F-FDG PET imaging for detecting peritoneal metastasis. Based on the original preoperative reports, CT and (18)F-FDG PET showed sensitivities of 76.5% and 35.3% (p = 0.037), specificities of 91.6% and 98.9% (p = 0.035), respectively, and equal accuracies of 89.3% (p = 1.0). The receptor operating characteristics curve analysis showed a significantly higher diagnostic performance for CT (Az = 0.878) than for PET (Az = 0.686) (p = 0.004). The interobserver agreement for detecting peritoneal metastasis was good (kappa value = 0.684) for CT and moderate (kappa value = 0.460) for PET. For the detection of peritoneal metastasis, CT was more sensitive and showed a higher diagnostic performance than PET, although CT had a relatively lower specificity than did PET.


Reliability and feasibility of a near patient test for C-reactive protein in primary care.
The applications of new diagnostic technologies such as near patient tests are relevant to the further development and potential of primary care. Through their use, doctors in the community may increase the accuracy of their diagnoses and improve their ability to monitor disease. A reliable indicator of disease activity in various clinical conditions is C-reactive protein (CRP) and a near patient test for this is now available, although there is little information on its use outside hospitals. A study was set up to evaluate the feasibility of using a novel near patient test for CRP in primary care to validate the results against the laboratory "gold standard' for CRP (Beckman Array) and to compare results with the usual inflammation test used in general practice. Prospective recording of CRP as a near patient test on an "intention to investigate' basis, with validation of results against the Beckman Array system for CRP and hospital laboratory erythrocyte sedimentation rate results, in six general medical practices in Birmingham. Main outcome measures were change in local laboratory usage, characteristics of patients chosen for testing, use of quality control, and comparison of readings with results from the same sample sent to an independent laboratory. Tests of CRP levels were rarely requested before the study was undertaken. During the 3-month study period, 181 near patient tests were carried out, 146 (81%) to establish a diagnosis and the remainder for disease monitoring. Out of the tests, 67% were performed by general practitioners, mostly during the consultation itself. Using a cut-off level of 10 mg I-1, the near patient test and the Beckman Array gave results which agreed in 84% of cases. The sensitivity and specificity of the near patient test results were 97 and 79%, respectively. The predictive value of a positive result was 59% and that of a negative result was 99%. Cohen's Kappa was 62% and the overall mean bias for results in the range of the test was 6.11 mg I-1 (SE = 3.07 mg I-1). Each test took 6 min on average to perform, including all preparations, blood letting, performing the test and averaging the time for quality control estimations. The cost per test averaged pounds 1.72, rising to pounds 4.17 including labour, capital costs, quality controls and consumables (general practitioner performing the assay at average frequency found in this study). Measurement of CRP is rarely used in primary care and awareness of its value could be raised. This near patient test proved feasible for use by general practitioners and practice nurses. Its reliability compared with a laboratory result was satisfactory overall, and excellent with adequate operator technique.


The WorldWide Antimalarial Resistance Network Clinical Trials Publication Library: A Live, Open-Access Database of Plasmodium Treatment Efficacy Trials.
Parasite resistance to antimalarial drugs poses a serious threat to malaria control. The WorldWide Antimalarial Resistance Network (WWARN) aims to provide a collaborative platform to support the global malaria research effort. Here, we describe the "WWARN clinical trials publication library," an open-access, up-to-date resource to streamline the synthesis of antimalarial safety and efficacy data. A series of iteratively refined database searches were conducted to identify prospective clinical trials assessing antimalarial drug efficacy with at least 28 days of follow-up. Of approximately 45,000 articles screened, 1,221 trials published between 1946 and 2018 were identified, representing 2,339 treatment arms and 323,819 patients. In trials from endemic locations, 75.7% (787/1,040) recruited patients with Plasmodium falciparum, 17.0% (177/1,040) Plasmodium vivax, 6.9% (72/1,040) both, and 0.4% (4/1,040) other Plasmodium species; 57.2% (585/1,022) of trials included under-fives and 5.3% (55/1,036) included pregnant women. In Africa, there has been a marked increase in both P. falciparum and P. vivax studies over the last two decades. The WHO-recommended artemisinin-based combination therapies alone or with a gametocidal drug were assessed in 39.5% (705/1,783) of P. falciparum treatment arms and 10.5% (45/429) of P. vivax arms, increasing to 78.0% (266/341) and 22.9% (27/118), respectively, in the last five years. The library is a comprehensive, open-access tool that can be used by the malaria community to explore the collective knowledge on antimalarial efficacy (available at https://www.wwarn.org/tools-resources/literature-reviews/wwarn-clinical-trials-publication-library). It is the first of its kind in the field of global infectious diseases, and lessons learnt in its creation can be adapted to other infectious diseases.


Position-effect variegation in Drosophila depends on dose of the gene encoding the E2F transcriptional activator and cell cycle regulator.
A dominant mutation due to the insertion of a P-element at 93E on the third chromosome of Drosophila melanogaster enhances position-effect variegation. The corresponding gene was cloned by transposon tagging and the sequence of the transcript revealed that it corresponds to the gene encoding the transcriptional activator and cell cycle regulator dE2F. The transposon-tagged allele is homozygous viable, and the insertion of the transposon in an intron correlates with a strong reduction in the amount of transcript. A homozygous lethal null allele was found to behave as a strong enhancer when heterozygous. Overexpression of the gene in transgenic flies has the opposite effect of suppressing variegation. A link is established here, and discussed, between the dose of a transcriptional activator, which controls the cell cycle, and epigenetic silencing of chromosomal domains in Drosophila.


Health status and nutritional development of adopted Ethiopian children living in southern Spain: A prospective cohort study.
The first aim of this study was to evaluate the health status and anthropometrical development of adopted children from Ethiopia living in southern Spain. A second aim was to evaluate the association between these parameters and adherence to the Mediterranean dietary pattern. The study sample included 53 adopted children from Ethiopia and a matched sample of 54 native-born children. A physical examination of the children, including height and weight, was conducted in Ethiopia at the time of entry into the adoption process. Height and weight were re-measured at the first day of adoption and 6, 12, and 24 mo after adoption. After 2 y of follow-up, another physical examination was performed, including the KIDMED test, to measure adherence to the Mediterranean diet. Skin and digestive conditions were the most prevalent disorders in Ethiopian children before adoption and at the end of follow-up. Baseline anthropometric characteristics indicated a low wasting prevalence (7.5%); however, stunted growth was more prevalent (35.8%). After 6 mo, the weight-for-age of Ethiopian children was restored (change from baseline P < 0.001), and not significantly different from the Spanish children at 1-y after adoption. Height-for-age also increased from baseline (P < 0.001. A higher KIDMED score was associated with increased weight-for-age (r = 0.279; P = 0.045) and height-for-age (r = 0.385; P = 0.004). This prospective study of adopted Ethiopian children confirmed a rapid growth development that occurred from the beginning of the adoption process and continued after the 2-y of follow-up. A higher adherence to the Mediterranean diet was associated with better growth development, which reinforces the importance of a balanced and adequate diet in growing children.


Evanescent photosynthesis: exciting cyanobacteria in a surface-confined light field.
The conversion of solar energy to chemical energy useful for maintaining cellular function in photosynthetic algae and cyanobacteria relies critically on light delivery to the microorganisms. Conventional direct irradiation of a bulk suspension leads to non-uniform light distribution within a strongly absorbing culture, and related inefficiencies. The study of small colonies of cells in controlled microenvironments would benefit from control over wavelength, intensity, and location of light energy on the scale of the microorganism. Here we demonstrate that the evanescent light field, confined near the surface of a waveguide, can be used to direct light into cyanobacteria and successfully drive photosynthesis. The method is enabled by the synergy between the penetration depth of the evanescent field and the size of the photosynthetic bacterium, both on the order of micrometres. Wild type Synechococcus elongatus (ATCC 33912) cells are exposed to evanescent light generated through total internal reflection of red (λ = 633 nm) light on a prism surface. Growth onset is consistently observed at intensity levels of 79 ± 10 W m(-2), as measured 1 μm from the surface, and 60 ± 8 W m(-2) as measured by a 5 μm depthwise average. These threshold values agree well with control experiments and literature values based on direct irradiation with daylight. In contrast, negligible growth is observed with evanescent light penetration depths less than the minor dimension of the rod-like bacterium (achieved at larger light incident angles). Collectively these results indicate that evanescent light waves can be used to tailor and direct light into cyanobacteria, driving photosynthesis.


Progress and challenges in maternal health in western China: a Countdown to 2015 national case study.
China is one of the few Countdown countries to have achieved Millennium Development Goal 5 (75% reduction in maternal mortality ratio between 1990 and 2015). We aimed to examine the health systems and contextual factors that might have contributed to the substantial decline in maternal mortality between 1997 and 2014. We chose to focus on western China because poverty, ethnic diversity, and geographical access represent particular challenges to ensuring universal access to maternal care in the region. In this systematic assessment, we used data from national census reports, National Statistical Yearbooks, the National Maternal and Child Health Routine Reporting System, the China National Health Accounts report, and National Health Statistical Yearbooks to describe changes in policies, health financing, health workforce, health infrastructure, coverage of maternal care, and maternal mortality by region between 1997 and 2014. We used a multivariate linear regression model to examine which contextual and health systems factors contributed to the regional variation in maternal mortality ratio in the same period. Using data from a cross-sectional survey in 2011, we also examined equity in access to maternity care in 42 poor counties in western China. Maternal mortality declined by 8·9% per year between 1997 and 2014 (geometric mean ratio for each year 0·91, 95% CI 0·91-0·92). After adjusting for GDP per capita, length of highways, female illiteracy, the number of licensed doctors per 1000 population, and the proportion of ethnic minorities, the maternal mortality ratio was 118% higher in the western region (2·18, 1·44-3·28) and 41% higher in the central region (1·41, 0·99-2·01) than in the eastern region. In the rural western region, the proportion of births in health facilities rose from 41·9% in 1997 to 98·4% in 2014. Underpinning such progress was the Government's strong commitment to long-term strategies to ensure access to delivery care in health facilities-eg, professionalisation of maternity care in large hospitals, effective referral systems for women medically or socially at high risk, and financial subsidies for antenatal and delivery care. However, in the poor western counties, substantial disparity by education level of the mother existed in access to health facility births (44% of illiterate women vs 100% of those with college or higher education), antenatal care (17% vs 69%) had at least four visits), and caesarean section (8% vs 44%). Despite remarkable progress in maternal survival in China, substantial disparities remain, especially for the poor, less educated, and ethnic minority groups in remote areas in western China. Whether China's highly medicalised model of maternity care will be an answer for these populations is uncertain. A strategy modelled after China's immunisation programme, whereby care is provided close to the women's homes, might need to be explored, with township hospitals taking a more prominent role. Government of Canada, UNICEF, and the Bill & Melinda Gates Foundation.


The accuracy and precision of radiostereometric analysis in upper limb arthroplasty.
Background and purpose - Radiostereometric analysis (RSA) is an accurate method for measurement of early migration of implants. Since a relation has been shown between early migration and future loosening of total knee and hip prostheses, RSA plays an important role in the development and evaluation of prostheses. However, there have been few RSA studies of the upper limb, and the value of RSA of the upper limb is not yet clear. We therefore performed a systematic review to investigate the accuracy and precision of RSA of the upper limb. Patients and methods - PRISMA guidelines were followed and the protocol for this review was published online at PROSPERO under registration number CRD42016042014. A systematic search of the literature was performed in the databases Embase, Medline, Cochrane, Web of Science, Scopus, Cinahl, and Google Scholar on April 25, 2015 based on the keywords radiostereometric analysis, shoulder prosthesis, elbow prosthesis, wrist prosthesis, trapeziometacarpal joint prosthesis, humerus, ulna, radius, carpus. Articles concerning RSA for the analysis of early migration of prostheses of the upper limb were included. Quality assessment was performed using the MINORS score, Downs and Black checklist, and the ISO RSA Results - 23 studies were included. Precision values were in the 0.06-0.88 mm and 0.05-10.7° range for the shoulder, the 0.05-0.34 mm and 0.16-0.76° range for the elbow, and the 0.16-1.83 mm and 11-124° range for the TMC joint. Accuracy data from marker- and model-based RSA were not reported in the studies included. Interpretation - RSA is a highly precise method for measurement of early migration of orthopedic implants in the upper limb. However, the precision of rotation measurement is poor in some components. Challenges with RSA in the upper limb include the symmetrical shape of prostheses and the limited size of surrounding bone, leading to over-projection of the markers by the prosthesis. We recommend higher adherence to RSA guidelines and encourage investigators to publish long-term follow-up RSA studies.


Gout and the risk of epilepsy: A population-based cohort study.
Gout is a chronic disease related to uric acid metabolism. It involves crystals of uric acid accumulating in the joints, causing joint pain and releasing cytokines that trigger inflammation. Inflammation is a key component in the pathogenesis of epilepsy. Thus, we conducted a cohort study to investigate if epilepsy is associated with gout and determine the risk of epilepsy in patients with gout.The gout cohort was obtained from the Registry of Catastrophic Illnesses Patient Database (RCIPD). We identified 104,238 patients who were aged 20 years or more and newly diagnosed with gout between 2000 and 2011 and 3 outpatient visits or history of gout-specific hospitalization between 2000 and 2011. Patients without gout were frequency matched with the gout cohort at a 2:1 ratio according to age, sex, comorbidities, and year of gout diagnosis.The gout cohort showed a 1.27-fold higher overall crude hazard ratio (HR) for epilepsy compared with the control cohort. After we adjusted the analyses by age, sex, and comorbidities the gout patients displayed an increased risk of epilepsy compared with the control group (adjusted HR = 1.25, 95% confidence interval = 1.15-1.36).This study revealed a significantly higher risk of epilepsy in patients with gout. It provides further evidence for the debate around the effect of gout on brain health.


Facilitators and challenges to start-up of the colorectal cancer screening demonstration program.
The Centers for Disease Control and Prevention (CDC) funded the Colorectal Cancer Screening Demonstration Program in 2005. To assess the feasibility of providing community-based colorectal cancer screening, CDC is conducting a multiple-case study as part of a larger evaluation effort. This article highlights key facilitators and challenges common to the five programs studied during the start-up period. The multiple-case study that includes all five program sites is being conducted during the 3-year program as part of process evaluation efforts. Data collection for program start-up occurred during August 2005 through September 2006. Data include approximately 70 interviews with program staff and stakeholders, document review, and observations. Both case-specific and cross-case analyses were conducted. On the basis of the cross-case analysis, we identified four factors that facilitated program start-up and four factors that challenged program start-up. Facilitating factors included 1) pre-existing program infrastructure, 2) partnerships, 3) clinical expertise, and 4) program champions. Factors challenging program start-up included 1) contracts with endoscopists, 2) resources for treating medical complications of screening and for cancer treatment, 3) administrative barriers, and 4) resource limitations. Additionally, preplanning was critical, allowing programs to efficiently initiate activities once funds became available. The most important facilitator identified was the ability to build on pre-existing infrastructure, which provided experienced staff, partnerships, and provider relationships, as well as aided program integration with other chronic disease programs. Results also suggest that substantial planning and partnership development can begin before funds are secured to implement a colorectal cancer screening program.


Passive airborne dust sampling with the electrostatic dustfall collector: optimization of storage and extraction procedures for endotoxin and glucan measurement.
We recently introduced a passive dust sampling method for airborne endotoxin and glucan exposure assessment-the electrostatic dustfall collector (EDC). In this study, we assessed the effects of different storage and extraction procedures on measured endotoxin and glucan levels, using 12 parallel EDC samples from 10 low exposed indoor environments. Additionally, we compared 2- and 4-week sampling with the prospect of reaching higher dust yields. Endotoxin concentrations were highest after extraction with pyrogen-free water (pf water) + Tween. Phosphate-buffered saline (PBS)-Tween yielded significantly (44%) lower levels, and practically no endotoxin was detected after extraction in pf water without Tween. Glucan levels were highest after extraction in PBS-Tween at 120 degrees C, whereas extracts made in NaOH at room temperature or 120 degrees C were completely negative. Direct extraction from the EDC cloth or sequential extraction after a preceding endotoxin extraction yielded comparable glucan levels. Sample storage at different temperatures before extraction did not affect endotoxin and glucan concentrations. Doubling the sampling duration yielded similar endotoxin and only 50% higher glucan levels. In conclusion, of the tested variables, the extraction medium was the predominant factor affecting endotoxin and glucan yields.


Defective Fc gamma RII gene expression in macrophages of NOD mice: genetic linkage with up-regulation of IgG1 and IgG2b in serum.
A quantitative trait locus for increased IgG serum levels in the NOD mouse strain was mapped to distal chromosome 1, close to the fcgr2 locus encoding the low-affinity type II receptor for the Fc portion of IgG (Fc gamma RII). Expression of membrane-inserted (b2) and soluble (b3) isoforms of Fc gamma RII was strongly decreased in macrophages of NOD compared with C57BL/6 (B6) mice. In contrast, B cell-specific (Fc gamma RIIb1) isoform was only slightly decreased and Fc gamma RIII was not altered. This Fc gamma RII regulatory defect was cis-encoded by fcgr2 or by a closely linked locus, occurred at the mRNA level, and was associated with multiple mutations in the fcgr2 gene promoter. In relation with this defect, binding of IgG1- and IgG2b- but not IgG2a-opsonized RBC by macrophages of NOD and congenic B6.NOD-fcgr2 mice was severely impaired, but was normal in macrophages of NOD.B6-fcgr2 congenic mice, indicating that Fc gamma RII plays a nondispensable role in binding of IgG1 and IgG2b isotypes. Likewise, serum levels of IgG1 and IgG2b but not IgG2a were up-regulated in NOD compared with NOD.B6-fcgr2 congenic mice. These findings indicate that macrophage Fc gamma RII may regulate serum IgG1 and IgG2b through their catabolism, and validate the NOD strain as a model to investigate the functions of Fc gamma RII isoforms.


First passage time to allopatric speciation.
Allopatric speciation is a mechanism to evolve reproductive isolation; it is caused by the accumulation of genetic differences between populations while they are geographically isolated. Here, we studied a simple stochastic model for the time until speciation caused by geographical isolation in fragmented populations that experience recurrent but infrequent migration between subpopulations. We assumed that mating incompatibility is controlled by a number of loci that behave as neutral characters in the accumulation of novel mutations within each population. Genetic distance between populations was defined as the number of incompatibility-controlling loci that differ between them. Genetic distance increases through the separate accumulation of mutations in different populations, but decreases after a successful migration event followed by genetic mixing between migrants and residents. We calculated the time to allopatric speciation, which occurs when the genetic distance exceeds a specified threshold. If the number of invasive individuals relative to the resident population is not very large, diffusion approximation provides an accurate prediction. There is an intermediate optimal rate of migration that maximizes the rate of species creation by recurrent invasion and diversification. We also examined cases that involved more than two populations.


Peripheral Leukocyte Migration in Ferrets in Response to Infection with Seasonal Influenza Virus.
In order to better understand inflammation associated with influenza virus infection, we measured cell trafficking, via flow cytometry, to various tissues in the ferret model following infection with an A(H3N2) human seasonal influenza virus (A/Perth/16/2009). Changes in immune cells were observed in the blood, bronchoalveolar lavage fluid, and spleen, as well as lymph nodes associated with the site of infection or distant from the respiratory system. Nevertheless clinical symptoms were mild, with circulating leukocytes exhibiting rapid, dynamic, and profound changes in response to infection. Each of the biological compartments examined responded differently to influenza infection. Two days after infection, when infected ferrets showed peak fever, a marked, transient lymphopenia and granulocytosis were apparent in all infected animals. Both draining and distal lymph nodes demonstrated significant accumulation of T cells, B cells, and granulocytes at days 2 and 5 post-infection. CD8+ T cells significantly increased in spleen at days 2 and 5 post-infection; CD4+ T cells, B cells and granulocytes significantly increased at day 5. We interpret our findings as showing that lymphocytes exit the peripheral blood and differentially home to lymph nodes and tissues based on cell type and proximity to the site of infection. Monitoring leukocyte homing and trafficking will aid in providing a more detailed view of the inflammatory impact of influenza virus infection.


Women's beliefs regarding food restrictions during common childhood illnesses: a hospital based study.
Malnutrition continues to be a major problem in Pakistan. Inadequate nutrition contributes substantially to childhood death and disease. Restriction of diet during common childhood illnesses further compromises the nutritional status of the child. The present study aims to determine the beliefs and practices regarding food restrictions during common childhood illnesses. A cross-sectional survey was conducted at Community Health Centre (CHC), of The Aga Khan University Hospital, Karachi to understand the beliefs and practices regarding food restrictions in common childhood illnesses. Four hundred adult females were interviewed from July-September 2000. A self-administered questionnaire was filled by the respondent. Major sources of information about restriction of various foods during different illnesses were relatives. Sixty five percent of the respondents believed that heavy food should be restricted during diarrhoea and oily food during jaundice. Sixty six percent of the respondents believed that cold food should be restricted during cold/cough and twenty three percent believed that oily food should be restricted during typhoid. Beliefs' regarding food restrictions during illnesses plays a very important role on the nutrition status. Surprisingly, medical doctors and other health care givers were also the source of information for advising food restrictions in certain childhood illnesses. Therefore there is a need for educating the physicians and other health care workers along with the mothers about food concepts and feeding practices during health and diseases. Protein energy malnutrition can be reduced to some extent if wrong dietary beliefs about child feeding practices in a community can be removed with health education programme.