Significant association of high-grade inflammation and thick lining epithelium with the increased number of Langerhans cells in dentigerous cysts.
Langerhans cells (LCs) are antigen-presenting cells. This study assessed the LC counts in 80 dentigerous cysts (DCs). The CD1a-positive LC numbers in the lining epithelia and subepithelial connective tissues were counted at 80 DC sites without inflammation, 33 DC sites with mild/moderate inflammation, and 9 DC sites with severe inflammation from 80 DC specimens. The mean LC counts in the lining epithelia and subepithelial connective tissues increased significantly from no inflammation (0.5 ± 0.5 and 0.2 ± 0.3 cell/high-power field or HPF, respectively) through mild/moderate inflammation (6.8 ± 1.8 and 2.4 ± 2.0 cells/HPF, respectively) to severe inflammation DC sites (18.9 ± 7.0 and 6.7 ± 5.8 cells/HPF, respectively; all P-values < 0.001). DC sites with inflammation had thicker lining epithelia than those without inflammation. Moreover, the mean LC counts in the lining epithelia and subepithelial connective tissues of DCs were significantly higher in the thicker lining epithelium (>50 μm) group (7.4 ± 6.5 and 2.6 ± 3.4 cells/HPF, respectively) than in the thinner lining epithelium (≦ 50 μm) group (0.5 ± 0.5 and 0.2 ± 0.3 cells/HPF, respectively; both P-values < 0.001). A significant association of high-grade inflammation and thick lining epithelium with the increased LC number in DCs is found. The finding of few LCs in the lining epithelia of DCs without inflammation indicates the reduced immunosurveillance ability against DC lining epithelial cells in DC patients. It needs further studies to confirm the role of reduced immunosurveillance in the enlargement of the DC.


Inhibition of autophagy reduces the rate of fluoride-induced LS8 apoptosis via regulating ATG5 and ATG7.
Excessive fluoride affects ameloblast differentiation and tooth development. The fate of fluorinated ameloblasts is determined by multiple signaling pathways in response to a range of stimuli. Both autophagy and apoptosis are involved in the regulation of dental fluorosis as well as in protein synthesis and enamel mineralization. Emerging evidence suggests that autophagy and apoptosis are interconnected and that their interaction greatly influences cell death. However, the effect of autophagy on apoptosis in fluoride-treated ameloblasts is unclear. Here, we employed an in vitro cellular model of fluorosis in mouse ameloblast-like LS8 cells and induced autophagy using sodium fluoride (NaF). Our findings suggest that NaF treatment induces autophagy in LS8 cells, and ATG5 and ATG7 are important molecules involved in this process. We also showed that NaF treatment reduced cell viability in Atg5/7 siRNA and autophagy inhibitor-treated LS8 cells. More importantly, NaF-induced apoptosis can be reversed by inhibiting early stage of autophagy. In conclusion, our study shows that autophagy is closely related to dental fluorosis, and inhibition of autophagy, especially ATG5/7, reduces fluoride-induced cell death and apoptosis.


[A morphological evaluation of the tissue status of the oral mucosa after anesthesia applied with dicaine and pyromecaine solutions].
Effects of aqueous dicaine and pyromecaine solutions on mucosal tissue structures were studied in 40 syrian hamsters. The results evidence that when dicaine, an ether anesthetic, is applied, injuries prevail over reactive phenomena: a manifest cytotoxic effect of the drug can be seen at all phases of the follow-up, this effect being observed both in the mucosal epithelium and in the deeper layers of the mucosa. An amide anesthetic, pyromecaine, is not characterized by a cytotoxic action on mucosal epithelium cells, though in later periods of the experiment it also may induce side effects presenting as an acute inflammatory reaction developing in the subepithelial [correction of polyepithelial] connective tissue stroma and muscular layer.


Expression of the Escherichia coli yfiD gene responds to intracellular pH and reduces the accumulation of acidic metabolic end products.
The YfiD protein of Escherichia coli has been reported to be an acid-inducible protein. Here it is shown that expression of a yfiD::lac reporter fusion is enhanced up to 3 small middle dot5-fold during acidic growth. The anaerobic transcription factor FNR was confirmed as the major regulator of yfiD expression, and ArcA was found to enhance anaerobic yfiD expression, probably by displacing a repressing FNR dimer in the -93 small middle dot5 region of the promoter. Moreover, the pyruvate sensor PdhR was shown to act as a minor anaerobic repressor of yfiD expression. On the basis of its strong homology to the C-terminal region of pyruvate formate-lyase (PFL) it was predicted that YfiD would be a radical-containing enzyme. The YfiD radical was found to be introduced by the PFL-activase enzyme, but unlike PFL, AdhE did not deactivate radicalized YfiD. The extent of radical activation of YfiD was enhanced by low intracellular pH, and thus it was concluded that both yfiD expression and YfiD activity are affected by growth at low pH. The yfiD mutant strain JRG4033 excreted increased levels of organic acids compared to the parental strain when grown in chemostat culture under oxygen-starved conditions, consistent with the acid-inducibility of yfiD expression and the recently reported ability of YfiD to rescue the activity of oxygenolytically cleaved PFL.


Identification and Characterization of Breakpoints and Mutations on Drosophila melanogaster Balancer Chromosomes.
Balancers are rearranged chromosomes used in Drosophila melanogaster to maintain deleterious mutations in stable populations, preserve sets of linked genetic elements and construct complex experimental stocks. Here, we assess the phenotypes associated with breakpoint-induced mutations on commonly used third chromosome balancers and show remarkably few deleterious effects. We demonstrate that a breakpoint in p53 causes loss of radiation-induced apoptosis and a breakpoint in Fucosyltransferase A causes loss of fucosylation in nervous and intestinal tissue-the latter study providing new markers for intestinal cell identity and challenging previous conclusions about the regulation of fucosylation. We also describe thousands of potentially harmful mutations shared among X or third chromosome balancers, or unique to specific balancers, including an Ankyrin2 mutation present on most TM3 balancers, and reiterate the risks of using balancers as experimental controls. We used long-read sequencing to confirm or refine the positions of two inversions with breakpoints lying in repetitive sequences and provide evidence that one of the inversions, In(2L)Cy, arose by ectopic recombination between foldback transposon insertions and the other, In(3R)C, cleanly separates subtelomeric and telomeric sequences and moves the subtelomeric sequences to an internal chromosome position. In addition, our characterization of In(3R)C shows that balancers may be polymorphic for terminal deletions. Finally, we present evidence that extremely distal mutations on balancers can add to the stability of stocks whose purpose is to maintain homologous chromosomes carrying mutations in distal genes. Overall, these studies add to our understanding of the structure, diversity and effectiveness of balancer chromosomes.


Impact of critical care-trained flight paramedics on casualty survival during helicopter evacuation in the current war in Afghanistan.
The US Army pioneered medical evacuation (MEDEVAC) by helicopter, yet its system remains essentially unchanged since the Vietnam era. Care is provided by a single combat medic credentialed at the Emergency Medical Technician - Basic level. Treatment protocols, documentation, medical direction, and quality improvement processes are not standardized and vary significantly across US Army helicopter evacuation units. This is in contrast to helicopter emergency medical services that operate within the United States. Current civilian helicopter evacuation platforms are routinely staffed by critical care-trained flight paramedics (CCFP) or comparably trained flight nurses who operate under trained EMS physician medical direction using formalized protocols, standardized patient care documentation, and rigorous quality improvement processes. This study compares mortality of patients with injury from trauma between the US Army's standard helicopter evacuation system staffed with medics at the Emergency Medical Technician - Basic level (standard MEDEVAC) and one staffed with experienced CCFP using adopted civilian helicopter emergency medical services practices. This is a retrospective study of a natural experiment. Using data from the Joint Theater Trauma Registry, 48-hour mortality for severely injured patients (injury severity score ≥ 16) was compared between patients transported by standard MEDEVAC units and CCFP air ambulance units. The 48-hour mortality for the CCFP-treated patients was 8% compared to 15% for the standard MEDEVAC patients. After adjustment for covariates, the CCFP system was associated with a 66% lower estimated risk of 48-hour mortality compared to the standard MEDEVAC system. These findings demonstrate that using an air ambulance system based on modern civilian helicopter EMS practice was associated with a lower estimated risk of 48-hour mortality among severely injured patients in a combat setting.


Allosteric MAPKAPK2 inhibitors improve plaque stability in advanced atherosclerosis.
Atherosclerotic vascular disease resulting from unstable plaques is the leading cause of morbidity and mortality in subjects with type 2 diabetes (T2D), and thus a major therapeutic goal is to discover T2D drugs that can also promote atherosclerotic plaque stability. Genetic or pharmacologic inhibition of mitogen-activated protein kinase-activated protein kinase-2 (MAPKAPK2 or MK2) in obese mice improves glucose homeostasis and enhances insulin sensitivity. We developed two novel orally active small-molecule inhibitors of MK2, TBX-1 and TBX-2, and tested their effects on metabolism and atherosclerosis in high-fat Western diet (WD)-fed Ldlr-/- mice. Ldlr-/- mice were first fed the WD to allow atherosclerotic lesions to become established, and the mice were then treated with TBX-1 or TBX-2. Both compounds improved glucose metabolism and lowered plasma cholesterol and triglyceride, without an effect on body weight. Most importantly, the compounds decreased lesion area, lessened plaque necrosis, and increased fibrous cap thickness in the aortic root lesions of the mice. Thus, in a preclinical model of high-fat feeding and established atherosclerosis, MK2 inhibitors improved metabolism and also enhanced atherosclerotic plaque stability, suggesting potential for further clinical development to address the epidemic of T2D associated with atherosclerotic vascular disease.


[Variants of serum cholinesterase (EC 184.108.40.206) in mentally retarded children].
Using screening, electrophoretic and spectrophotometric examinations, the authors studied the polymorphism of serum cholinesterase (CE) (CP 126.96.36.199) in children with undifferentiated oligophrenia. The rates of the identified phenotypes were as follows: CHE1U--0.769; CHE1UD--0.039; CHE1UF--0.192; CHE2(5-)-1.000. The incidence of the CHE1UF variant of the enzyme considerably exceeded that in the control groups (u = 2.44; u = 3.23) while the frequency of the CHE1U variant detection was significantly below that in the control groups (u = 2.70; u = 3.01; u = 3.46). The author discusses the advisability of the identification of CE variants in patients with psychoneurological diseases in order to prevent the development of side effects of myorelaxants and local anesthetics.


Spectrum of asthma in children. 3. Psychological and social components.
Behavioural disturbances in the child, the mother-child and family relationships, and the family social structure were studied in a representative sample of the whole range of asthmatic children and compared with a control group of normal children. Behavioural disturbances occurred more often and at a statistically significant level only in the small group of children with severe and continuing asthma. These children were those with severe chronic airways obstruction as assessed physiologically and also with the most severe allergic manifestations.Predominant in the mother-child relations was an over-concern to protect the child's health in those children with continuing asthma at 14 years of age. The families of the very severely affected group of children showed evidence of more stress than other families. Socioeconomic conditions were not significantly different in any group of asthmatic children compared with the control group.


Congener-specific tissue distribution of aroclor 1254 and a highly chlorinated environmental PCB mixture in rats.
Polychlorinated biphenyl (PCB) mixtures were synthesized and marketed in Eastern European countries, but little is known abouttheir composition, distribution, ortoxicity. PCB-contaminated soil from the former production site of the Polish PCB mixture Chlorofen was collected, and the PCBs were extracted. An in vivo study was performed to investigate the PCB tissue distribution and biochemical effects of this soil extract in immature male Sprague-Dawley rats. Rats were administered 0.05 mmol/kg soil-extracted PCBs or Aroclor 1254 and sacrificed 7 days later, and congener-specific PCB profiles in selected tissues were determined. Distribution of SigmaPCBs (sum of 120 congeners) in tissues was primarily a function of lipid content of the tissues, except for the spleen, which retained more PCB than other tissues. Multivariate analysis of the PCB congener data showed that (a) congener profiles in tissues had changed, as compared to the parent mixture; (b) disposition and redistribution of individual congeners in vivo differed between both mixtures; and (c) more highly chlorinated congeners were retained in the spleens of both treatment groups. Differences in the induction of cytochrome P-450 1A and 2B subfamilies reflected the homologue composition of the respective mixtures and predict a different toxicity profile for Chlorofen than for Aroclor 1254.


Evaluation of state-of-the-art segmentation algorithms for left ventricle infarct from late Gadolinium enhancement MR images.
Studies have demonstrated the feasibility of late Gadolinium enhancement (LGE) cardiovascular magnetic resonance (CMR) imaging for guiding the management of patients with sequelae to myocardial infarction, such as ventricular tachycardia and heart failure. Clinical implementation of these developments necessitates a reproducible and reliable segmentation of the infarcted regions. It is challenging to compare new algorithms for infarct segmentation in the left ventricle (LV) with existing algorithms. Benchmarking datasets with evaluation strategies are much needed to facilitate comparison. This manuscript presents a benchmarking evaluation framework for future algorithms that segment infarct from LGE CMR of the LV. The image database consists of 30 LGE CMR images of both humans and pigs that were acquired from two separate imaging centres. A consensus ground truth was obtained for all data using maximum likelihood estimation. Six widely-used fixed-thresholding methods and five recently developed algorithms are tested on the benchmarking framework. Results demonstrate that the algorithms have better overlap with the consensus ground truth than most of the n-SD fixed-thresholding methods, with the exception of the Full-Width-at-Half-Maximum (FWHM) fixed-thresholding method. Some of the pitfalls of fixed thresholding methods are demonstrated in this work. The benchmarking evaluation framework, which is a contribution of this work, can be used to test and benchmark future algorithms that detect and quantify infarct in LGE CMR images of the LV. The datasets, ground truth and evaluation code have been made publicly available through the website: https://www.cardiacatlas.org/web/guest/challenges.


Purification and amino acid sequence of melanocyte-stimulating hormone from the dogfish Squalus acanthias.
A melanocyte-stimulating hormone (MSH) was isolated by gel filtration and ion-exchange chromatography from extracts of the pituitary glands of dogfish. Sequence studies were carried out on the hormone and its enzymically and chemically cleaved fragments. The sequence of the hormone, Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Met, shows that ten of its 11 residues are the same as ten of the 13 residues of mammalian alpha-MSH. About half of its molecules have the carboxyl group at the C-terminus free and about half are amidated; about a fifth have an extra tyrosine residue on the N-terminus, thereby making 11 residues the same as in mammalian alpha-MSH. Unlike the mammalian hormone, however, none of it was found to be N-acetylated.


Standardization of denaturing gradient gel electrophoresis (DGGE) for discrimination of Victoria and Yamagata lineages of influenza B.
In this study, a PCR-DGGE protocol was standardized in order to distinguish Victoria and Yamagata influenza B lineages directly from clinical samples. After routine multiplex PCR characterization, amplicons of the haemagglutinin gene bearing a 40bp-length GC clamp were generated by nested-PCR and analyzed by electrophoresis in 6% polyacrylamide gel with a 25-45% urea-formamide gradient. The results showed a perfect correlation between DGGE and phylogenetic analyses for all compared samples, besides some distinct profiles in Victoria and Yamagata groups that could be used to infer variability inside these groups. In summary, this DGGE protocol for the haemagglutinin gene is rapid, useful and efficient, being an alternative for discrimination between the influenza B lineages.


Interleukin 7 induces CD4+ T cell-dependent tumor rejection.
The potential of interleukin 7 (IL-7) to induce an antitumor response in vivo was analyzed. Therefore, the IL-7 gene was expressed in the plasmacytoma cell line J558L. Although the growth of IL-7-producing cells was not retarded in vitro, the IL-7-producing cells were completely rejected upon injection into mice. Tumor rejection was observed only in syngeneic but not in nude mice. The tumor-suppressive effect could be abolished by the parallel injection of an anti-IL-7 monoclonal antibody. Immunohistochemical analysis revealed IL-7-dependent infiltration of the tumor tissue by CD4+ and CD8+ T lymphocytes, and also type 3 complement receptor-positive (CR3+) cells, predominantly macrophages. Depletion of T cell subsets in tumor-bearing mice showed the absolute dependence of the antitumor response on CD4+ cells, whereas tumor rejection was unaffected by depletion of CD8+ cells. In addition to CD4+ cells, CR3+ cells were also needed for tumor rejection. The antitumor effect of IL-7 was confirmed by expression of the IL-7 gene in a second tumor cell line of different cellular origin. Together, our results demonstrate that a high local IL-7 concentration at the tumor site obtained by tumor cell-targeted gene transfer leads to tumor rejection involving a cellular mechanism that seems to be different from the ones observed in analogous experiments with other cytokines.


Complement evasion of pathogens: common strategies are shared by diverse organisms.
Infectious diseases represent a major health problem. Based on the limited efficacy of existing drugs and vaccines and the increasing antibiotic resistance new strategies are needed to fight infectious diseases. A better understanding of pathogen-host interaction is one important aspect to identify new virulence factors and antimicrobial and anti-inflammatory compounds utilized by pathogens represent an additional source for effective anti-inflammatory compounds. Complement forms a major defense line against invading microbes, and pathogens have learned during evolution to breach this defense line. The characterization of how pathogens evade complement attack is a rapidly developing field of current research. Pathogens mimic host surfaces and bind host complement regulators. Similarly pathogens utilize a number of complement inhibitory molecules which help to evade complement attack and which display anti-inflammatory activity. The molecular identification of these molecules, as well as the functional characterization of their roles at the pathogen-host interface is an important and emerging field of infection biology. In addition, pathogens utilize multiple sets of such regulators as redundancy and multiplicity is important for immune and complement evasion. Here we summarize the current scenarios of this emerging field which identifies multiple virulence factors and complement evasion strategies, but which at the same time reveals common mechanisms for immune and complement defense.


Socioeconomic status and prevalence of obesity and diabetes in a Mexican American community, Cameron County, Texas, 2004-2007.
Mexican Americans are at increased risk for obesity and diabetes. We established a cohort on the United States-Mexico border to determine the prevalence of obesity and diabetes in this Mexican American population and to see whether minor economic advantages had any effect on health. We randomly selected and extensively documented 810 people aged 35 to 64 years. Weighted data were analyzed to establish prevalence of obesity and diabetes and other markers of poor health such as elevated glycated hemoglobin levels. Rates of obesity (body mass index > or = 30 kg/m(2)) were 57% in the first (lower) of 4 socioeconomic strata by income and were 55.5% in the third (higher). People in the higher socioeconomic stratum were significantly less likely to have undiagnosed diabetes (2% vs 9%). Among people aged 55 to 64 years, rates of diabetes were significantly higher among those in the lower socioeconomic stratum than among those in the higher stratum. Rates of undiagnosed diabetes had similar differences. Approximately three-fourths of the respondents reported having no health insurance, and we found no difference between people in different socioeconomic strata. Rates of obesity and diabetes in this border community are among the highest in the United States. Belonging to the lower socioeconomic stratum significantly increased the likelihood of having undiagnosed diabetes and, in patients too young to be eligible for Medicare, the overall risk of developing diabetes. Modest improvement in income has a beneficial effect on health in this racial/ethnic minority community.


Review of cellular mechanisms of tumor osteolysis.
The cellular and biochemical mechanisms that direct the destruction of bone at sites of tumor osteolysis are unknown. To better understand the mechanisms through which tumors direct bone resorption, research has focused on developing in vivo and in vitro experimental models that are useful for studying this process. In vivo experimental systems have been developed that permit study of tumor osteolysis from human and murine tumors, and that permit the study of tumors that arise from (sarcoma) or can metastasize (breast cancer) to bone. Recent research has focused on three questions: (1) Are osteoclasts or tumor cells responsible for bone resorption during tumor osteolysis? (2) What are the cellular mechanisms that are responsible for bone resorption during tumor osteolysis, and (3) what are the tumor cell products that regulate the cellular mechanisms that are responsible for tumor osteolysis? It has been determined that osteoclasts are responsible for bone resorption at sites of tumor osteolysis by enhancing the binding of osteoclast to bone, by inducing osteoclastic bone resorption, and by stimulating osteoclast formation. Attempts to identify tumor cell products that regulate these cellular mechanisms are in progress, and findings suggest that production of macrophage colony stimulating factor may be required for tumor osteolysis to occur with some tumors.


Conserved L464 in p97 D1-D2 linker is critical for p97 cofactor regulated ATPase activity.
p97 protein is a highly conserved, abundant, functionally diverse, structurally dynamic homohexameric AAA enzyme-containing N, D1, and D2 domains. A truncated p97 protein containing the N and D1 domains and the D1-D2 linker (ND1L) exhibits 79% of wild-type (WT) ATPase activity whereas the ND1 domain alone without the linker only has 2% of WT activity. To investigate the relationship between the D1-D2 linker and the D1 domain, we produced p97 ND1L mutants and demonstrated that this 22-residue linker region is essential for D1 ATPase activity. The conserved amino acid leucine 464 (L464) is critical for regulating D1 and D2 ATPase activity by p97 cofactors p37, p47, and Npl4-Ufd1 (NU). Changing leucine to alanine, proline, or glutamate increased the maximum rate of ATP turnover (kcat) of p47-regulated ATPase activities for these mutants, but not for WT. p37 and p47 increased the kcat of the proline substituted linker, suggesting that they induced linker conformations facilitating ATP hydrolysis. NU inhibited D1 ATPase activities of WT and mutant ND1L proteins, but activated D2 ATPase activity of full-length p97. To further understand the mutant mechanism, we used single-particle cryo-EM to visualize the full-length p97L464P and revealed the conformational change of the D1-D2 linker, resulting in a movement of the helix-turn-helix motif (543-569). Taken together with the biochemical and structural results we conclude that the linker helps maintain D1 in a competent conformation and relays the communication to/from the N-domain to the D1 and D2 ATPase domains, which are ∼50 Å away.


Toll-like receptor 4 expression in the epithelium of inflammatory periapical lesions. An immunohistochemical study.
Toll-like receptors (TLR) are essential for the innate immune response against invading pathogens and have been described in immunocompetent cells of areas affected by periapical disease. Besides initiating the inflammatory response, they also directly regulate epithelial cell proliferation and survival in a variety of settings. This study evaluates the in situ expression of TLR4 in periapical granulomas (PG) and radicular cysts, focusing on the epithelial compartment. Twenty-one periapical cysts (PC) and 10 PG were analyzed; 7 dentigerous non-inflamed follicular cyst (DC) served as control. TLR4 expression was assessed by immunohistochemistry. TLR4 immunoreaction products were detected in the epithelium of all specimens, with a higher percentage of immunostained cells in PG. Although TLR4 overexpression was detected in both PG and PC, there were differences that seemed to be related to the nature of the lesion, since in PG all epithelial cells of strands, islands and trabeculae were strongly immunoreactive for TLR4, whereas in PC only some areas of the basal and suprabasal epithelial layers were immunostained. This staining pattern is consistent with the action of TLR4: in PG it could promote formation of epithelial cell rests of Malassez and in epithelial strands and islands the enhancement of cell survival, proliferation and migration, whereas in PC TLR4 could protect the lining epithelium from extensive apoptosis. These findings go some way towards answering the intriguing question of why many epithelial strands or islands in PG and the lining epithelium of apical cysts regress after non-surgical endodontic therapy, and suggest that TLR4 plays a key role in the pathobiology of the inflammatory process related to periapical disease.


Response to selection for fertility in a fall-lambing sheep flock.
Selection for fertility in a fall lambing system was applied for 5 yr (1.45 generations) in a crossbred sheep population. Three flocks were involved: a fall-lambing selected (S) flock of 125 ewes and 10 rams, a fall-lambing environmental control (EC) flock of 55 ewes and five rams, and a spring-lambing genetic control flock of 45 ewes and five rams used to produce replacements for EC. Estimated breeding values for fertility (ewes lambing per ewe exposed) were calculated for animals mated in each year of the study using a single-trait animal model and increased more rapidly in S (.0138/yr) than in EC (.0067/yr). After accounting for genetic drift, EBV of ewes mated in S in the final year of the study averaged .070 +/- .032 (P = .08). A correlated response of .0087 lambs/yr was observed in S for fall litter size Genetic trends in all body weights were positive in EC, likely because of supplemental selection on body size in ram lambs used in EC. Ninety-day weight, maternal effects on weaning weight, and 90-d scrotal circumference increased in S, but direct effects on birth and weaning weights declined. In order to focus on realized selection response in fertility, EBV for fertility of 330 replacement ewe lambs from S and EC were calculated using only data collected after their birth. These realized EBV were then regressed on pedigree EBV calculated at the time the ewe lambs were born. The regression was significant when all relationships were used to calculate both EBV (.20 +/- .08) but only approached significance (.13 +/- .07) when only records of the ewe lambs themselves, their descendants, and their paternal half-sibs were used to calculate realized EBV. Also, EBV for all ewes were calculated at the start of each breeding season and were higher (P < .01) for ewes that subsequently lambed than for open ewes. These results suggest that selection can be used to improve fertility in fall-lambing sheep flocks.


Intersecting pathways in cell biology.
The endocytic pathway is involved in activation and inhibition of cellular signaling. Thus, defining the regulatory mechanisms that link endocytosis and cellular signaling is of interest. An emerging link between these processes is a family of proteins called intersectins (ITSNs). These multidomain proteins serve as scaffolds in the assembly of endocytic vesicles and also regulate components of various signaling pathways, including kinases, guanosine triphosphatases, and ubiquitin ligases. This review summarizes research on the role of ITSNs in regulating both endocytic and signal transduction pathways, discusses the link between ITSNs and human disease, and highlights future directions in the study of ITSNs.


The mRNA structure has potent regulatory effects on type 2 iodothyronine deiodinase expression.
Type 2 deiodinase (D2) is a selenoenzyme catalyzing the activation of T(4) to T(3). D2 activity/mRNA ratios are often low, suggesting that there is significant posttranscriptional regulation. The D2 mRNA in higher vertebrates is more than 6 kb, containing long 5' and 3' untranslated regions (UTRs). The D2 5'UTRs are greater than 600 nucleotides and contain 3-5 short open reading frames. These full-length 5'UTRs reduce the D2 translation efficiency approximately 5-fold. The inhibition by human D2 5'UTR is localized to a region containing the first short open reading frame encoding a tripeptide-MKG. This inhibition was abolished by mutating the AUG start codon and weakened by modification of the essential purine of the Kozak consensus. Deletion of the 3.7-kb 3'UTR of the chicken D2 mRNA increased D2 activity approximately 3.8-fold due to an increase in D2 mRNA half-life. In addition, alternatively spliced D2 mRNA transcripts similar in size to the major 6- to 7-kb D2 mRNAs but not encoding an active enzyme are present in both human and chicken tissues. Our results indicate that a number of factors reduce the D2 protein levels. These mechanisms, together with the short half-life of the protein, ensure limited expression of this key regulator of T(4) activation.


Performance Characteristics of the Digital Biograph Vision PET/CT System.
This study evaluated the performance of the Biograph Vision digital PET/CT system according to the NEMA NU 2-2012 standard (published by the National Electrical Manufacturers Association [NEMA]) to allow for a reliable, reproducible, and intersystem-comparable performance measurement. Methods: The new digital PET/CT system features silicon photomultiplier-based detectors with 3.2-mm lutetium oxyorthosilicate crystals and full coverage of the scintillator area. The PET components incorporate 8 rings of 38 detector blocks, and each block contains 4 × 2 mini blocks. Each mini block consists of a 5 × 5 lutetium oxyorthosilicate array of 3.2 × 3.2 × 20 mm crystals coupled to a silicon photomultiplier array of 16 × 16 mm, resulting in an axial field of view of 26.1 cm. In this study, PET/CT system performance was evaluated for conformation with the NEMA NU 2-2012 standard, with additional measurements described in the new NEMA NU 2-2018 standard. Spatial resolution, sensitivity, count-rate performance, accuracy of attenuation and scatter correction, image quality, coregistration accuracy, and time-of-flight performance were determined. Measurements were directly compared with results from its predecessor, the Biograph mCT Flow, using existing literature. Moreover, feasibility to comply with the European Association of Nuclear Medicine Research Ltd. (EARL) criteria was evaluated, and some illustrative patient PET images were obtained. Results: The Biograph Vision showed a transverse and axial spatial resolution of 3.6 and 3.5 mm, respectively, in full width at half maximum at a 1-cm offset from the center of the field of view (measured with a 22Na 0.25-mm point source), a NEMA sensitivity of 16.4 kcps/MBq, and a NEMA peak noise-equivalent count-rate of 306 kcps at 32 kBq/mL. Time-of-flight resolution varied from 210 to 215 as count-rate increased up to the peak noise-equivalent count-rate. The overall image contrast seen with the NEMA image quality phantom ranged from 77.2% to 89.8%. Furthermore, the system was able to comply with the current and future EARL performance criteria. Conclusion: The Biograph Vision outperforms the analog Biograph mCT Flow, and the system is able to meet European harmonizing performance standards.


Duration of virus shedding after trivalent intranasal live attenuated influenza vaccination in adults.
To characterize the probability and duration of viral shedding among adults given trivalent live attenuated influenza vaccine (LAIV). Prospective surveillance study. Nasal wash samples were collected from adult volunteers at baseline and on days 3, 7, and 10 and between days 17 and 21 following intranasal LAIV vaccination. The presence, titer, and identification of each specific strain of influenza virus shed were determined by standard methodology. Twenty subjects received LAIV. No samples were positive for influenza virus at baseline. After LAIV vaccination, influenza virus was recovered from 10 of 20 vaccinees on day 3, from 1 of 18 vaccinees on day 7, and from none of the samples on days 10 or 17 through 21. Vaccinees who shed vaccine virus were significantly younger than those who did not (mean age, 26.4 vs 38.6 years; P < .01). Although the presence of specific mucosal immunoglobulin A to influenza B was associated with significantly less shedding of influenza B after vaccination (P = .02), associations of shedding with other measures of immunity were not detected. The duration of shedding of vaccine virus after LAIV in adults i s limited and may be associatedwith an individual's prior influenza vaccination history.


Prediction of Lymph Node Maximum Standardized Uptake Value in Patients With Cancer Using a 3D Convolutional Neural Network: A Proof-of-Concept Study.
The purpose of this study is to determine whether a convolutional neural network (CNN) can predict the maximum standardized uptake value (SUVmax) of lymph nodes in patients with cancer using the unenhanced CT images from a PET/CT examination, thus providing a proof of concept for potentially using deep learning to diagnose nodal involvement. Consecutive initial staging PET/CT scans obtained in 2017 for patients with pathologically proven malignancy were collected. Two blinded radiologists selected one to 10 lymph nodes from the unenhanced CT portion of each PET/CT examination. The SUVmax of the lymph nodes was recorded. Lymph nodes were cropped and used with the primary tumor histology type as input for a novel 3D CNN with predicted SUVmax as the output. The CNN was trained using one cohort and tested using a separate cohort. An SUVmax of 2.5 or greater was defined as FDG avid. Two blinded radiologists separately classified lymph nodes as FDG avid or not FDG avid on the basis of unenhanced CT images and separately using a short-axis measurement cutoff of 1 cm. Logistic regression analysis was performed. A total of 400 lymph nodes (median SUVmax, 6.8 [interquartile range {IQR}, 2.7-11.6]; median short-axis, 1.1 cm [IQR, 0.9-1.6 cm]) in 136 patients were used for training. A total of 164 lymph nodes (median SUVmax, 3.5 [IQR, 1.9-8.6]; median short-axis, 1.0 cm [IQR, 0.7-1.4 cm]) in 49 patients were used for testing. The predicted SUVmax was associated with the real SUVmax (β estimate = 0.83, p < 0.0001). The predicted SUVmax was associated with FDG avidity (p < 0.0001), with an ROC AUC value of 0.85, and it improved when combined with radiologist qualitative assessment and short-axis criteria. A CNN is able to predict with moderate accuracy the SUVmax of lymph nodes, as determined from the unenhanced CT images and tumor histology subtype for patients with cancer.


Novel role for inhibitor of differentiation 2 in the genesis of angiotensin II-induced hypertension.
Angiotensin (Ang) II-induced target-organ damage involves innate and acquired immunity. Mice deficient for the helix-loop-helix transcription factor inhibitor of differentiation (Id2(-/-)) lack Langerhans and splenic CD8a+ dendritic cells, have reduced natural killer cells, and have altered CD8 T-cell memory. We tested the hypothesis that an alteration in the number and quality of circulating blood cells caused by Id2 deletion would ameliorate Ang II-induced target-organ damage. We used gene-deleted and transgenic mice. We conducted kidney and bone marrow transplants. In contrast to Ang II-infused Id2(+/-), Id2(-/-) mice infused with Ang II remained normotensive and failed to develop albuminuria or renal damage. Bone marrow transplant of Id2(+/-) bone marrow to Id2(-/-) mice did not restore the blunted blood pressure response to Ang II. Transplantation of Id2(-/-) kidneys to Id2(+/-) mice also could not prevent Ang II-induced hypertension and renal damage. We verified the Ang II resistance in Id2(-/-) mice in a model of local tissue Ang II production by crossing hypertensive mice transgenic for rat angiotensinogen with Id2(-/-) or Id2(+/-) mice. Angiotensinogen-transgenic Id2(+/-) mice developed hypertension, albuminuria, and renal injury, whereas angiotensinogen-transgenic Id2(-/-) mice did not. We also found that vascular smooth muscle cells from Id2(-/-) mice showed an antisenescence phenotype. Our bone marrow and kidney transplant experiments suggest that alterations in circulating immune cells or Id2 in the kidney are not responsible for Ang II resistance. The present studies identify a previously undefined role for Id2 in the pathogenesis of Ang II-induced hypertension.


Indole-3-carbinol (I3C)-induced apoptosis in nasopharyngeal cancer cells through Fas/FasL and MAPK pathway.
The apoptotic effects of indole-3-carbinol (I3C) were exhibited in many human cancer cells. However, the apoptotic effects of I3C on nasopharyngeal cancer cells were still unknown. In this study, we first examined the potential effects of I3C on the induction of apoptosis in human nasopharyngeal cancer cells CNE-2 and further characterized the mechanism(s) involved in the effects. Apoptotic cells after treatment with I3C were analyzed by flow cytometry. The change in relative mitochondrial transmembrane potential in the CNE-2 cells was analyzed with rhodamine 123 staining. The protein levels of MAPK family and Fas/FasL were examined by Western blot. Our results indicated that I3C could induce apoptosis of CNE-2 cells in a dose- and time-dependent manner accompanied with increased mitochondrial membrane potential. Treatment with I3C significantly suppressed XIAP, c-IAP1 and Survivin protein, while elevated the expression of Omi, Smac and Cyto-c. Fas/FasL and MAPK pathway were involved in the induction of apoptosis. Taken together, these results demonstrated that I3C may induce mitochondria-mediated apoptosis via the Fas death receptor in CNE-2 cells. This molecular mechanism for apoptotic effect of I3C on nasopharyngeal cancer cells suggested that I3C might become a preventive and therapeutic agent against nasopharyngeal cancer.


Individual and social determinants of multiple chronic disease behavioral risk factors among youth.
Behavioral risk factors are known to co-occur among youth, and to increase risks of chronic diseases morbidity and mortality later in life. However, little is known about determinants of multiple chronic disease behavioral risk factors, particularly among youth. Previous studies have been cross-sectional and carried out without a sound theoretical framework. Using longitudinal data (n = 1135) from Cycle 4 (2000-2001), Cycle 5 (2002-2003) and Cycle 6 (2004-2005) of the National Longitudinal Survey of Children and Youth, a nationally representative sample of Canadian children who are followed biennially, the present study examines the influence of a set of conceptually-related individual/social distal variables (variables situated at an intermediate distance from behaviors), and individual/social ultimate variables (variables situated at an utmost distance from behaviors) on the rate of occurrence of multiple behavioral risk factors (physical inactivity, sedentary behavior, tobacco smoking, alcohol drinking, and high body mass index) in a sample of children aged 10-11 years at baseline. Multiple behavioral risk factors were assessed using a multiple risk factor score. All statistical analyses were performed using SAS, version 9.1, and SUDAAN, version 9.01. Multivariate longitudinal Poisson models showed that social distal variables including parental/peer smoking and peer drinking (Log-likelihood ratio (LLR) = 187.86, degrees of freedom (DF) = 8, p < .001), as well as individual distal variables including low self-esteem (LLR = 76.94, DF = 4, p < .001) increased the rate of occurrence of multiple behavioral risk factors. Individual ultimate variables including age, sex, and anxiety (LLR = 9.34, DF = 3, p < .05), as well as social ultimate variables including family socioeconomic status, and family structure (LLR = 10.93, DF = 5, p = .05) contributed minimally to the rate of co-occurrence of behavioral risk factors. The results suggest targeting individual/social distal variables in prevention programs of multiple chronic disease behavioral risk factors among youth.


The physiological responses to repeated upper-body sprint exercise in highly trained athletes.
To study performance, physiological and biomechanical responses during repeated upper-body sprint exercise. Twelve male elite cross-country skiers performed eight 8-s maximal poling sprints with a 22-s recovery while sitting on a modified SkiErg poling ergometer. Force, movement velocity, cycle rate, work per cycle, oxygen saturation in working muscles and pulmonary oxygen uptake were measured continuously. A 3-min all-out ergometer poling test determined VO2peak, and 1 repetition maximum (1RM) strength was determined in a movement-specific pull-down. Average sprint power was 281 ± 48 W, with the highest power on the first sprint, a progressive decline in power output over the following four sprints, and a sprint decrement of 11.7 ± 4.1 %. Cycle rate remained unchanged, whereas work per cycle progressively decreased (P < 0.05). m. triceps brachii and m. latissimus dorsi were highly desaturated already after the first sprint (all P < 0.05), whereas the response was delayed for m. biceps brachii and m. vastus lateralis. Correspondingly, increases in VO2 mainly occurred over the first two sprints (P < 0.05) and plateaued at approximately 75 % of VO2peak. 1RM correlated with power during the first four sprints and with average sprint power (r = 0.71-0.80, all P < 0.05), whereas VO2peak correlated with power in the last three sprints (r = 0.60-0.71, all P < 0.05). The main decrement in upper-body sprint performance was evident in the first five sprints, followed by highly desaturated muscles and a plateau in pulmonary oxygen uptake already after the first 2-3 sprints. While high maximal strength seems important for producing power, aerobic capacity correlates with power in the last sprints.


Diethylstilbestrol selectively modulates plasma coagulation protein synthesis by the perfused rat liver.
The combined effects of orchiectomy and estrogen administration on synthesis of selected hepatic secretory proteins--antithrombin (AT) III, plasminogen, fibrinogen, factor II (prothrombin), factor VII, fibronectin, and albumin--were studied using the isolated rat liver perfused in vitro for ten hours. Male rat liver donors underwent orchiectomy under ether anesthesia and then received 5.0 mg of diethylstilbestrol (DES) by subcutaneous pellet implantation or a placebo pellet; 14 days later the livers were extracted and perfused in vitro for ten hours. In DES experiments, 1.0 mg of DES was also added directly to the liver perfusate at the outset. Pretreatment with DES resulted in significant increases in cumulative synthesis of factors II (65%) and VII (76%) and significant reduction in cumulative synthesis of both antithrombin III (20%; P = .03) and plasminogen (27%; P less than .01) compared to control perfusions, but synthesis of fibrinogen, fibronectin, and albumin was not significantly affected by addition of the hormone. Plasma samples collected from rat liver donors that had received DES showed similar effects on protein concentrations: significant decreases in concentration of plasminogen and antithrombin III were apparent with no significant changes in concentrations of fibrinogen, fibronectin, or albumin. In additional perfusions, "dose-response" experiments were conducted in which rat liver donors received a subcutaneous DES pellet of 0.5, 5.0, or 50 mg. Synthesis of plasminogen in this group of perfusions was progressively decreased as the concentration of DES administered to the rat liver donor increased. Synthesis of AT III was reduced to the same degree by 5.0 or 50 mg of DES, both being substantially lower than the 0.5-mg experiments. Concentrations of these two proteins in plasma samples from rat liver donors showed changes quite similar to those seen in perfusion experiments; however, plasma fibrinogen concentrations were not different among the three groups of rats.


Intracranial Penetration During Temporal Soft Tissue Filler Injection-Is It Possible?
Treating temporal volume loss for aesthetic and reconstructive purposes can be achieved by superficial or deep injections of soft tissue fillers into the temples. The latter is performed with bone contact that can lead to intracranial penetration when the bone is accidentally penetrated. Based on a clinical case, the potential risk of accidental intracranial penetration was investigated. Twenty fresh-frozen hemi-faces (all Caucasian ethnicity, 10 women, 10 men, mean age 72.8 ± 11.2 years) were investigated. Shape of pterion and bone-stability parameters of the temporal fossa were investigated. Bone stability was tested using uniaxial mechanical indentation (18-G, 1.25-mm diameter, 15-mm length blunt-tip device) until intracranial perforation occurred. Variations in the shape of the pterion, bone thickness, and density correlates were detected, however, without statistical significant differences in side symmetry. Minimum force necessary to penetrate intracranially was 40.4 N. Maximum force generated by an 18-g, 70-mm length blunt-tip cannula was 32.1 ± 4.2 N in 70 mm length and 75.3 ± 10.2 N in 15 mm length. Based on the results of this investigation, it can be concluded that there is a risk for intracranial penetration performing the deep temple injection technique with direct pressure on the bone.


Dichotic listening deficits in children with dyslexia.
Several auditory processing deficits have been reported in children with dyslexia. In order to assess for the presence of a binaural integration type of auditory processing deficit, dichotic listening tests with digits, words and consonant-vowel (CV) pairs were administered to two groups of right-handed 11-year-old children, one group diagnosed with developmental dyslexia and an age-matched control group. Dyslexic children performed more poorly than controls from their left ears when listening to digits and words and from their right ears when listening to CVs. Direction of ear advantage varied across individuals in both groups when tested with digits and CVs, but ear advantage was stable with words. Several factors that may have contributed to inconsistencies in direction of ear advantage are discussed. When the children were tested in a directed response mode, degree of ear advantage differed significantly between groups with both words and digits. More dyslexic than control children demonstrated clinically significant reductions in dichotic listening performance, but no uniform pattern of deficit emerged. Only the double correct score and the left ear score with CV pairs were predictive of word recognition performance in dyslexic children. Binaural integration deficits are present in some children with dyslexia. Auditory processing disorder assessment may help delineate factors that underlie or are associated with reading impairment in this population.


Detection of hemoglobin alterations in a sample population in northwest Mexico. Preliminary report.
Blood samples from 1,000 individuals seen at the Instituto Mexicano del Seguro Social in northwestern Mexico were assayed by multiple systems (acid, alkaline and neutral electrophoresis, heat, freeze and isopropanol stability and erythrocytic morphology under hypoxia) to detect hemoglobin (Hb) abnormalities. Two beta-thalassemia heterozygotes (0.2 per cent), five Hb S heterozygotes (0.5 per cent) and three probable new Hb variants (0.3 per cent) were identified. These preliminary results give a general idea of the gene frequencies in this population, but do not yet permit the evaluation of the relative advantage of multiple methodology for the detection of abnormal Hbs.


Brain awareness week and beyond: encouraging the next generation.
The field of neuroscience is generating increased public appetite for information about exciting brain research and discoveries. As stewards of the discipline, together with FUN and others, the Society for Neuroscience (SfN) embraces public outreach and education as essential to its mission of promoting understanding of the brain and nervous system. The Society looks to its members, particularly the younger generation of neuroscientists, to inspire, inform and engage citizens of all ages, and most importantly our youth, in this important endeavor. Here we review SfN programs and resources that support public outreach efforts to inform, educate and tell the story of neuroscience. We describe the important role the Brain Awareness campaign has played in achieving this goal and highlight opportunities for FUN members and students to contribute to this growing effort. We discuss specific programs that provide additional opportunities for neuroscientists to get involved with K-12 teachers and students in ways that inspire youth to pursue further studies and possible careers in science. We draw attention to SfN resources that support outreach to broader audiences. Through ongoing partnerships such as that between SfN and FUN, the neuroscience community is well positioned to pursue novel approaches and resources, including harnessing the power of the Internet. These efforts will increase science literacy among our citizens and garner more robust support for scientific research.


Searching for a minimal set of behaviors for autism detection through feature selection-based machine learning.
Although the prevalence of autism spectrum disorder (ASD) has risen sharply in the last few years reaching 1 in 68, the average age of diagnosis in the United States remains close to 4--well past the developmental window when early intervention has the largest gains. This emphasizes the importance of developing accurate methods to detect risk faster than the current standards of care. In the present study, we used machine learning to evaluate one of the best and most widely used instruments for clinical assessment of ASD, the Autism Diagnostic Observation Schedule (ADOS) to test whether only a subset of behaviors can differentiate between children on and off the autism spectrum. ADOS relies on behavioral observation in a clinical setting and consists of four modules, with module 2 reserved for individuals with some vocabulary and module 3 for higher levels of cognitive functioning. We ran eight machine learning algorithms using stepwise backward feature selection on score sheets from modules 2 and 3 from 4540 individuals. We found that 9 of the 28 behaviors captured by items from module 2, and 12 of the 28 behaviors captured by module 3 are sufficient to detect ASD risk with 98.27% and 97.66% accuracy, respectively. A greater than 55% reduction in the number of behaviorals with negligible loss of accuracy across both modules suggests a role for computational and statistical methods to streamline ASD risk detection and screening. These results may help enable development of mobile and parent-directed methods for preliminary risk evaluation and/or clinical triage that reach a larger percentage of the population and help to lower the average age of detection and diagnosis.


A 5' regulatory sequence containing two Ets motifs controls the expression of the Wiskott-Aldrich syndrome protein (WASP) gene in human hematopoietic cells.
The recently-identified Wiskott-Aldrich syndrome protein gene (WASP) is responsible for the Wiskott-Aldrich X-linked immunodeficiency as well as for isolated X-linked thrombocytopenia (XLT). To characterize the regulatory sequences of the WASP gene, we have isolated, sequenced and functionally analyzed a 1.6-Kb DNA fragment upstream of the WASP coding sequence. Transfection experiments showed that this fragment is capable of directing efficient expression of the reporter chloramphenicol acetyltransferase (CAT) gene in all human hematopoietic cell lines tested. Progressive 5' deletions showed that the minimal sequence required for hematopoietic-specific expression consists of 137 bp upstream of the transcription start site. This contains potential binding sites for several hematopoietic transcription factors and, in particular, two Ets-1 consensus that proved able to specifically bind to proteins present in nuclear extracts of Jurkat cells. Overexpression of Ets-1 in HeLa resulted in transactivation of the CAT reporter gene under the control of WASP regulatory sequences. Disruption of the Ets-binding sequences by side-directed mutagenesis abolished CAT expression in Jurkat cells, indicating that transcription factors of the Ets family play a key role in the control of WASP transcription.


The Experience of Cognitive Intrusion of Pain: scale development and validation.
Patients with chronic pain often report their cognition to be impaired by pain, and this observation has been supported by numerous studies measuring the effects of pain on cognitive task performance. Furthermore, cognitive intrusion by pain has been identified as one of 3 components of pain anxiety, alongside general distress and fear of pain. Although cognitive intrusion is a critical characteristic of pain, no specific measure designed to capture its effects exists. In 3 studies, we describe the initial development and validation of a new measure of pain interruption: the Experience of Cognitive Intrusion of Pain (ECIP) scale. In study 1, the ECIP scale was administered to a general population sample to assess its structure and construct validity. In study 2, the factor structure of the ECIP scale was confirmed in a large general population sample experiencing no pain, acute pain, or chronic pain. In study 3, we examined the predictive value of the ECIP scale in pain-related disability in fibromyalgia patients. The ECIP scale scores followed a normal distribution with good variance in a general population sample. The scale had high internal reliability and a clear 1-component structure. It differentiated between chronic pain and control groups, and it was a significant predictor of pain-related disability over and above pain intensity. Repairing attentional interruption from pain may become a novel target for pain management interventions, both pharmacologic and nonpharmacologic.


Upregulation of matrix metalloproteinase triggers transdifferentiation of retinal pigmented epithelial cells in Xenopus laevis: A Link between inflammatory response and regeneration.
In adult Xenopus eyes, when the whole retina is removed, retinal pigmented epithelial (RPE) cells become activated to be retinal stem cells and regenerate the whole retina. In the present study, using a tissue culture model, it was examined whether upregulation of matrix metalloproteinases (Mmps) triggers retinal regeneration. Soon after retinal removal, Xmmp9 and Xmmp18 were strongly upregulated in the tissues of the RPE and the choroid. In the culture, Mmp expression in the RPE cells corresponded with their migration from the choroid. A potent MMP inhibitor, 1,10-PNTL, suppressed RPE cell migration, proliferation, and formation of an epithelial structure in vitro. The mechanism involved in upregulation of Mmps was further investigated. After retinal removal, inflammatory cytokine genes, IL-1β and TNF-α, were upregulated both in vivo and in vitro. When the inflammation inhibitors dexamethasone or Withaferin A were applied in vitro, RPE cell migration was severely affected, suppressing transdifferentiation. These results demonstrate that Mmps play a pivotal role in retinal regeneration, and suggest that inflammatory cytokines trigger Mmp upregulation, indicating a direct link between the inflammatory reaction and retinal regeneration. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1086-1100, 2017.


Dynamic thiol/disulphide homeostasis as a novel indicator of oxidative stress in obese children and its relationship with inflammatory-cardiovascular markers.
Childhood obesity is an important cause of cardiovascular risk with chronic inflammation. Oxidative stress may contribute to the pathogenesis of obesity-related cardiovascular pathologies. We aimed to evaluate thiol/disulphide homeostasis as a novel and sensitive marker of oxidative stress and to evaluate its relationship with some inflammatory and cardiovascular markers in obese children. In this case-controlled study, 65 children with exogenous obesity and 64 healthy children, as a control group, were included. In both groups, thiol/disulphide homeostasis parameters and inflammatory (white blood cells, platelets, mean corpuscular volume, neutrophil/lymphocyte ratio, and high-sensitivity C-reactive protein) and cardiovascular (epicardial adipose tissue thickness and left ventricular mass index) markers were studied. Correlation analyses of thiol/disulphide homeostasis parameters with body mass index standard deviation scores (BMI SDS) and inflammatory and cardiovascular markers were performed. Receiver-operating characteristic analysis was performed to determine the sensitivity, specificity, and optimal cut-off values of thiol/disulphide homeostasis parameters. Native thiol, total thiol, and native thiol/total thiol ratios (antioxidant parameters) were lower (p<0.05) and disulphide/native thiol and disulphide/total thiol ratios (oxidant parameters) were higher in the obese group than in the control group (p<0.01). A positive correlation of oxidant parameters with BMI SDS and inflammatory markers was found. However, a negative correlation of antioxidant parameters with BMI SDS and inflammatory markers was found. The specificities of disulphide/native thiol and disulphide/total thiol ratios were higher in the obese group. The impairment in thiol/disulphide homeostasis, which is indicative of oxidative stress, is associated with inflammation in obesity. In addition, cardiovascular involvement may also contribute to this impairment.


Zinc: requirements, bioavailabilities and recommended dietary allowances.
This review has approached zinc requirements, bioavailabilities and recommended dietary allowances from a historical view. For example, a requirement for zinc was first demonstrated for the microorganism Aspergillus niger more than a century ago, although zinc has been recognized as a dietary nutrient for humans only within the last decade. Dietary requirement was defined as that quantity of zinc which must be provided daily in order to meet the metabolic requirement. The degree of bioavailability determines the total dietary zinc which must be consumed by humans to remain in metabolic equilibrium. In regard to recommended dietary allowances (RDA), they are recommended average daily intakes of nutrients that population groups should consume. RDA are not synonymous with requirements. Metabolic requirements (the quantity that must be provided to the circulating blood daily) have been determined previously using radio-isotope studies and have indicated a calculated need of approximately 6 mg for an adult per day. More recently a similar amount (4-6 mg) has been found to be required daily (intravenously) in order to maintain plasma zinc concentrations and daily urinary zinc excretion within normal range in hospitalized subjects. Average zinc intakes of large segments of the U.S. population are receiving levels approaching one-half or less the RDA with no apparent deleterious effects. These subjects include pregnant and lactating women. In addition, the zinc content of breast milk is lower than previous studies indicated. Thus, it appears that the majority of breast-fed infants are receiving no more than 70% of the recommended intake, with a recent study indicating less than 50%. Factors reported to affect bioavailability of zinc from foods are discussed. These include fiber and phytate. It was concluded that no definite conclusion can be reached regarding the overall effect of food fiber on zinc balance. Many of the studies were of short duration, with the longest being 32 days. It is probable that different sources of food fiber may have different effects on zinc balance. The apparent discrepancy in the literature regarding the effect of soy protein on zinc requirement was cited. Lastly, the phytate:zinc molar ratio concept was designed to test it as a predictor of zinc bioavailability to humans. The concept must be expanded to recognize the relevance of the total daily dietary zinc intake. Specifically, impairment of absorption of zinc by phytate would be more critical if the total dietary intake was low since the metabolic requirements might not be met.(ABSTRACT TRUNCATED AT 400 WORDS)


Ultrasensitive Room Temperature Infrared Photodetection Using a Narrow Bandgap Conjugated Polymer.
Photodetectors operating across the short-, mid-, and long-wave infrared (SWIR-LWIR, λ = 1-14 µm) underpin modern science, technology, and society in profound ways. Narrow bandgap semiconductors that form the basis for these devices require complex manufacturing, high costs, cooling, and lack compatibility with silicon electronics, attributes that remain prohibitive for their widespread usage and the development of emerging technologies. Here, a photoconductive detector, fabricated using a solution-processed narrow bandgap conjugated polymer is demonstrated that enables charge carrier generation in the infrared and ultrasensitive SWIR-LWIR photodetection at room temperature. Devices demonstrate an ultralow electronic noise that enables outstanding performance from a simple, monolithic device enabling a high detectivity (D*, the figure of merit for detector sensitivity) >2.44 × 109 Jones (cm Hz1/2 W-1 ) using the ultralow flux of a blackbody that mirrors the background emission of objects. These attributes, ease of fabrication, low dark current characteristics, and highly sensitive operation overcome major limitations inherent within modern narrow-bandgap semiconductors, demonstrate practical utility, and suggest that uncooled detectivities superior to many inorganic devices can be achieved at high operating temperatures.


Neurovascular Compression in Patients With Trigeminal Neuralgia May Be Associated With Worse Outcomes After Primary Percutaneous Rhizotomy.
Percutaneous rhizotomy may be an effective primary intervention in patients with trigeminal neuralgia who are poor candidates for microvascular decompression or those who desire a less invasive approach. However, the influence of neurovascular compression on pain-free survival after primary percutaneous rhizotomy is not well understood. We retrospectively reviewed all patients undergoing percutaneous rhizotomy at our institution from 1995 to 2022. Patients were included if they had no history of surgical intervention, available preoperative MRI imaging, and postoperative follow-up data. Barrow Neurological Institute pain scores were assigned at various time points. We collected baseline patient information, pain characteristics, and perioperative complications for each patient. In addition, we recorded evidence of pain recurrence. Patients were dichotomized into those with evidence of neurovascular compression on preoperative MRI vs those without. The effect of neurovascular compression on pain-free survival was assessed using Kaplan-Meier Cox proportional hazards analyses. Of the 2726 patients reviewed, 298 met our inclusion criteria. Our study comprised 261 patients with no evidence of neurovascular compression on preoperative MRI vs 37 patients with evidence of neurovascular compression on preoperative MRI. Patients in the compression group had a shorter median duration to recurrence compared with those in the no compression group, P = .01. Kaplan-Meier survival analysis revealed that patients with preoperative evidence of neurovascular compression on MRI imaging demonstrated shorter pain-free survival compared with those without such evidence [hazard ratio = 1.57 (1.03-2.4), P = .037]. Cox proportional hazards analysis demonstrated that evidence of neurovascular compression was associated with poor pain-free survival [hazard ratio = 1.64 (1.06-2.53), P = .03]. Patients with neurovascular compression on preoperative MRI may experience reduced time to recurrence compared with those without after percutaneous rhizotomy. These patients should be counseled on potential reduced efficacy of percutaneous rhizotomy as a primary intervention for their pain.


Haematological aspect of malaria in a population based hospital, Saudi Arabia.
In Saudi Arabia, although the malaria is nearly extinct from urban areas, anophelines breeding however still exists in the coastal and oasis regions of the Kingdom. Imported cases of malaria are also seen as the country continues to employ a large expatriate work force from malarious countries. Data were analysed on 69 malaria patients (35 Saudis; 34 non-Saudis) to assess the haematological and clinical manifestations. Two types of malaria, P. falciparum and P. vivax were mainly present. Anemia was the significant finding in patients with P. falciparum (P = 0.0060). Approximately (32/69) half of the patients had Hb < or = 12 g/dl and among these included 15 cases (22%) with Hb < than 9 g/dl. White blood counts were near normal or mildly decreased despite of high fever. Moderate to severe thrombocytopenia (platelets < or = 160 x 10(9)/L) was seen in 57% of the patients and in 10 cases the counts were appreciably low (< 60 x 10(9)/L). Thrombocytopenia correlated with splenomegaly. The palpable spleen was felt in approximately half of patients with P. falciparum. Findings of this study suggest that in Saudi Arabia we will continue to see malaria cases occasionally, however it is important to become aware of the treatment, as many patients who originated from the third world malarious zones, failed to respond to chloroquine therapy.


The mouse eugenol odorant receptor: structural and functional plasticity of a broadly tuned odorant binding pocket.
Molecular interactions of odorants with their olfactory receptors (ORs) are of central importance for the ability of the mammalian olfactory system to detect and discriminate a vast variety of odors with a limited set of receptors. How a particular OR binds and distinguishes different odorant molecules remains largely unknown on a structural basis. Here we investigated this question for the mouse eugenol receptor (mOR-EG). By screening a large odorant library, we discovered a wide range of chemical structures activating the receptor in heterologous mammalian cells. Potent agonists comprise (i) benzene, (ii) cyclohexane, or (iii) polycyclic structures substituted with alcohol, aldehyde, keto, ether, or esterified carboxylic groups. To detect those amino acids within the receptor that are in contact with a particular bound odorant molecule, we investigated how distinct mOR-EG point mutants were activated by the different odorant agonists found for the wild-type receptor. We identified 11 amino acids as a part of the receptor's ligand binding pocket. Molecular modeling predicted 10 of these residues in transmembrane helices TM3-TM6 and one in the extracellular loop between TM2 and TM3. These amino acids participate in odorant binding with variable importance depending on the type of odorant, revealing functional "fingerprints" of ligand-receptor interactions.


The Crawford variant as a cause of RhD typing discrepancies in blood banks: A case report.
We present the case of a 55-year-old Colombian male who showed a discrepancy in the serological typing of the RhD antigen in his first platelet donation. The discrepancy persisted after a serological investigation with multiple Anti-D monoclonal reagents (IgG and IgM) under different conditions (22°C and 37°C, saline, and LISS/Coombs). Furthermore, partial RhD typing was performed, obtaining negative results with a commercially available panel of six Anti-D reagents. Molecular analysis showed a homozygous deletion of RHD and heterozygosity for the Crawford variant (RHCE*ce, RHCE*ceCF), with a predicted phenotype of C-, c+, E-, e+, Vs+, V+. Following the investigation of this case, this man has made 14 platelet donations showing variable reactivity, with agglutinations ranging from - to 2+. Since Crawford red blood cells express some RhD antigen epitopes, they could cause alloimmunization in RhD negative receptors. Likewise, Anti-D alloantibodies have been documented in Crawford variant carriers. Therefore, it is recommended that carriers of this variant be classified as RhD positive if they are blood donors and RhD negative if they are transfusion recipients. Also, in pregnant women carrying a Crawford variant, Anti-D immunoprophylaxis is recommended.


Expression of aquaporins in inner ear disease.
The inner ear is responsible for hearing and balance and consists of a membranous labyrinth within a bony labyrinth. The balance structure is divided into the otolith organ that recognizes linear acceleration and the semicircular canal that is responsible for rotational movement. The cochlea is the hearing organ. The external and middle ear are covered with skin and mucosa, respectively, and the space is filled with air, whereas the inner ear is composed of endolymph and perilymph. The inner ear is a fluid-filled sensory organ composed of hair cells with cilia on the upper part of the cells that convert changes in sound energy and balance into electric energy through the hair cells to transmit signals to the auditory nerve through synapses. Aquaporins (AQPs) are a family of transmembrane proteins present in all species that can be roughly divided into three subfamilies according to structure and function: 1) classical AQP, 2) aquaglyceroporin, and 3) superaquaporin. Currently, the subfamily of mammalian species is known to include 13 AQP members (AQP0-AQP12). AQPs have a variety of functions depending on their structure and are related to inner ear diseases such as Meniere's disease, sensorineural hearing loss, and presbycusis. Additional studies on the relationship between the inner ear and AQPs may be helpful in the diagnosis and treatment of inner ear disease. Laryngoscope, 130:1532-1539, 2020.


Validity of the Empatica E4 Wristband to Measure Heart Rate Variability (HRV) Parameters: a Comparison to Electrocardiography (ECG).
Wearable monitoring devices are an innovative way to measure heart rate (HR) and heart rate variability (HRV), however, there is still debate about the validity of these wearables. This study aimed to validate the accuracy and predictive value of the Empatica E4 wristband against the VU University Ambulatory Monitoring System (VU-AMS) in a clinical population of traumatized adolescents in residential care. A sample of 345 recordings of both the Empatica E4 wristband and the VU-AMS was derived from a feasibility study that included fifteen participants. They wore both devices during two experimental testing and twelve intervention sessions. We used correlations, cross-correlations, Mann-Whitney tests, difference factors, Bland-Altman plots, and Limits of Agreement to evaluate differences in outcomes between devices. Significant correlations were found between Empatica E4 and VU-AMS recordings for HR, SDNN, RMSSD, and HF recordings. There was a significant difference between the devices for all parameters but HR, although effect sizes were small for SDNN, LF, and HF. For all parameters but RMSSD, testing outcomes of the two devices led to the same conclusions regarding significance. The Empatica E4 wristband provides a new opportunity to measure HRV in an unobtrusive way. Results of this study indicate the potential of the Empatica E4 as a practical and valid tool for research on HR and HRV under non-movement conditions. While more research needs to be conducted, this study could be considered as a first step to support the use of HRV recordings provided by wearables.


MR findings in hereditary isolated growth hormone deficiency.
To describe the MR characteristics by which patients with hereditary isolated growth hormone deficiency (GHD) can be distinguished from patients with other types of GHD. A total of 51 patients with GHD were examined prospectively with MR imaging. On the basis of familial occurrence of GHD and genetic analysis, 10 patients met the criteria for hereditary deficiency. In each case, the height of the pituitary gland, the presence and location of the posterior neurohypophysis, and the completeness of the stalk were recorded. The findings in the hereditary group were compared with those in the rest of the patients. In all 10 patients with hereditary GHD, the adenohypophysis, the neurohypophysis, and the stalk were normal. Of the other 41 patients, the height of the gland was normal in three (7%), the neurohypophysis was abnormal in all, and the stalk was truncated in all but two patients (95%). The subgroup of patients with hereditary GHD exhibited an anatomically normal pituitary-hypothalamic region. This is in contrast to the majority of patients with idiopathic GHD. MR imaging can contribute to the classification of patients with GHD.


The G Protein-Coupled Estrogen Receptor (GPER) Expression Correlates with Pro-Metastatic Pathways in ER-Negative Breast Cancer: A Bioinformatics Analysis.
The G protein-coupled estrogen receptor (GPER, formerly known as GPR30) is a seven-transmembrane receptor that mediates estrogen signals in both normal and malignant cells. In particular, GPER has been involved in the activation of diverse signaling pathways toward transcriptional and biological responses that characterize the progression of breast cancer (BC). In this context, a correlation between GPER expression and worse clinical-pathological features of BC has been suggested, although controversial data have also been reported. In order to better assess the biological significance of GPER in the aggressive estrogen receptor (ER)-negative BC, we performed a bioinformatics analysis using the information provided by The Invasive Breast Cancer Cohort of The Cancer Genome Atlas (TCGA) project and Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) datasets. Gene expression correlation and the statistical analysis were carried out with R studio base functions and the tidyverse package. Pathway enrichment analysis was evaluated with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway on the Database for Annotation, Visualization and Integrated Discovery (DAVID) website, whereas gene set enrichment analysis (GSEA) was performed with the R package phenoTest. The survival analysis was determined with the R package survivALL. Analyzing the expression data of more than 2500 primary BC, we ascertained that GPER levels are associated with pro-migratory and metastatic genes belonging to cell adhesion molecules (CAMs), extracellular matrix (ECM)-receptor interaction, and focal adhesion (FA) signaling pathways. Thereafter, evaluating the disease-free interval (DFI) in ER-negative BC patients, we found that the subjects expressing high GPER levels exhibited a shorter DFI in respect to those exhibiting low GPER levels. Overall, our results may pave the way to further dissect the network triggered by GPER in the breast malignancies lacking ER toward a better assessment of its prognostic significance and the action elicited in mediating the aggressive features of the aforementioned BC subtype.


PdumBase: a transcriptome database and research tool for Platynereis dumerilii and early development of other metazoans.
The marine polychaete annelid Platynereis dumerilii has recently emerged as a prominent organism for the study of development, evolution, stem cells, regeneration, marine ecology, chronobiology and neurobiology within metazoans. Its phylogenetic position within the spiralian/ lophotrochozoan clade, the comparatively high conservation of ancestral features in the Platynereis genome, and experimental access to any stage within its life cycle, make Platynereis an important model for elucidating the complex regulatory and functional molecular mechanisms governing early development, later organogenesis, and various features of its larval and adult life. High resolution RNA-seq gene expression data obtained from specific developmental stages can be used to dissect early developmental mechanisms. However, the potential for discovery of these mechanisms relies on tools to search, retrieve, and compare genome-wide information within Platynereis, and across other metazoan taxa. To facilitate exploration and discovery by the broader scientific community, we have developed a web-based, searchable online research tool, PdumBase, featuring the first comprehensive transcriptome database for Platynereis dumerilii during early stages of development (2 h ~ 14 h). Our database also includes additional stages over the P. dumerilii life cycle and provides access to the expression data of 17,213 genes (31,806 transcripts) along with annotation information sourced from Swiss-Prot, Gene Ontology, KEGG pathways, Pfam domains, TmHMM, SingleP, and EggNOG orthology. Expression data for each gene includes the stage, the normalized FPKM, the raw read counts, and information that can be leveraged for statistical analyses of differential gene expression and the construction of genome-wide co-expression networks. In addition, PdumBase offers early stage transcriptome expression data from five further species as a valuable resource for investigators interested in comparing early development in different organisms. To understand conservation of Platynereis gene models and to validate gene annotation, most Platynereis gene models include a comprehensive phylogenetic analysis across 18 species representing diverse metazoan taxa. PdumBase represents the first online resource for the early developmental transcriptome of Platynereis dumerilii. It serves as a research platform for discovery and exploration of gene expression during early stages, throughout the Platynereis life cycle, and enables comparison to other model organisms. PdumBase is freely available at http://pdumbase.gdcb.iastate.edu .


Relation of race, ethnicity and cardiac surgeons to operative mortality rates in primary coronary artery bypass grafting in California.
The aim of this study was to evaluate whether African American, Hispanic, and Asian patients in California were more likely to undergo coronary artery bypass grafting (CABG) by cardiac surgeons with higher risk-adjusted mortality rates (RAMRs). Clinical data from the California CABG Outcomes Reporting Program were analyzed for all patients who underwent isolated CABG from 2003 to 2006 by surgeons who performed ≥ 10 operations. Surgeons were divided into quintiles on the basis of their RAMRs, with the top-performing surgeons in the first quintile and the lowest performing surgeons in the fifth quintile. There were 72,845 isolated CABG procedures performed by 303 surgeons, including 49,886 in white, 9,380 in Hispanic, 6,867 in Asian, and 2,750 in African American patients. African American and Asian patients underwent CABG by surgeons with higher mean RAMRs (2.90% and 2.99%, respectively) compared with the state average of 2.65% (p <0.001). Compared to white patients, Asian and Hispanic patients were more likely to be treated by surgeons in the lowest quintile (odds ratio [OR] 1.2, 95% confidence interval [CI] 1.11 to 1.3, and OR 1.38, 95% CI 1.30 to 1.48, respectively). African Americans and Hispanics were less likely to be operated on by surgeons in the top quintile compared to white patients (OR 0.80, 95% CI 0.71 to 0.90, and OR 0.81, 95% CI 0.76 to 0.87, respectively). Hispanics were less likely to be treated by surgeons in the top quintile than by surgeons in the lowest quintile (OR 0.65, 95% CI 0.60 to 0.75). In conclusion, racial and ethnic minority patients who undergo isolated CABG in California may be more likely to be operated on by cardiac surgeons with higher RAMRs.


Vitamin D3 and cardiovascular function in rats.
We have previously identified a receptor for 1,25-dihydroxyvitamin D3 in myocardial cells (Simpson, R.U. 1983. Circulation. 68:239.). To establish the relevance of this observation, we evaluated the role of the prohormone vitamin D3 in regulating cardiovascular function. In rats maintained on a vitamin D3-deficient diet for nine weeks, increases in systolic blood pressure (BP) and serum creatine phosphokinase (CPK) were observed. These increases coincided with a reduction of serum calcium from 10.3 to 5.6 mg/dl. However, while serum calcium remained depressed throughout the study, increases in BP and serum CPK were transient. After nine weeks of vitamin D3-depletion, but not after six weeks, ventricular and vascular muscle contractile function were also markedly enhanced. The increase in ventricular contractile function could not be prevented by maintaining serum calcium at 9.0 mg/dl during the period of D3-depletion. These observations suggest a primary role for the vitamin D3-endocrine system in regulating cardiovascular function.


Psychological factors, including alexithymia, in the prediction of cardiovascular risk in HIV infected patients: results of a cohort study.
Psychological factors are known predictors of cardiovascular disease in many clinical settings, but data are lacking for HIV infection. We carried out a prospective cohort study to evaluate potential psychological predictors of preclinical and clinical vascular disease in HIV patients. HIV patients were consecutively enrolled. Demographics, viral and immune parameters and traditional cardiovascular predictors were considered; Intima-Media Thickness (c-IMT, continuous measure) and Carotid Plaques (CPs, focal thickening ≥1.5 mm) were investigated by B-mode ultrasonography; depressive symptoms by the Beck Depression Inventory (BDI-II), Type D personality (Distressed Personality or Type D) by the DS14, alexithymia by the Toronto Alexithymia Scale (TAS-20). Vascular outcomes included transient ischemic attacks or stroke, acute coronary syndrome, myocardial or other organ infarction. We enrolled 232 HIV subjects, 73.9% males, aged 44.5±9.9 y, 38.2% with AIDS diagnosis, 18.3% untreated. Mean Nadir CD4 T-cell counts were 237.5±186.2/mmc. Of them, 224 (96.5%) attended IMT measurements; 201 (86.6%) attended both IMT assessment and psychological profiling. Mean follow-up was 782±308 days. Fifty-nine patients (29.4%) had CPs at baseline. Nineteen patients (9.5%) had ≥1 vascular event; 12 (6.0%) died due to such events (n = 4) or any cause. At baseline cross-sectional multivariate analysis, increasing age, total cholesterol, current smoking and Alexithymia score≥50 were significantly associated with both increased cIMT (linear regression) and CPs (logistic regression). At follow-up analysis, log-rank tests and Cox's regression revealed that only older age (p = 0.001), current smoking (p = 0.019) and alexithymia score≥50 (p = 0.013) were independently associated with vascular events. In HIV-infected subjects, the Alexithymic trait emerges as a strong predictor of increased IMT, presence of CPs and vascular events. Such results are preliminary and require confirmation from studies with larger sample size and longer follow-up.


Identification of IDO-positive and IDO-negative human dendritic cells after activation by various proinflammatory stimuli.
Dendritic cells (DCs) can induce tolerance or immunity. We identified and characterized an IDO-expressing and an IDO-negative human DC population after stimulation by various proinflammatory stimuli. IDO expression was strongly dependent on the maturation status of the cells (CD83-positive cells only). The two DC subpopulations remained IDO positive and IDO negative, respectively, over a time period of at least 48 h. IDO enzyme activity of human DCs was highest during stimulation by strongly maturation-inducing TLR ligands such as highly purified LPS (TLR4 ligand) or polyriboinosinic-polyribocytidilic acid (TLR3 ligand); factors of the adaptive immune system such as IFN-γ, a mixture of cytokines, and IFN-α had lesser stimulatory capacity for IDO induction and activity. After stimulation with CD40L, IDO-positive DCs expressed significantly increased levels of B7 family molecules such as CD40, CD80, CD86, ICOS ligand, as well as PD-L1 (B7-H1) and PD-L2 (B7-DC) compared with the IDO-negative DC subset. At the same time, the inhibitory receptors Ig-like transcripts 3 and 4 were significantly downregulated on IDO-positive cells. Functionally, IDO-positive DCs produced significantly more IL-1β and IL-15 and less IL-10 and IL-6 than the IDO-negative subset after CD40L stimulation. These results show that IDO expression is associated with a distinctive phenotype and functional capacity in mature DCs. It seems likely that the IDO-positive DC subset possesses a regulatory function and might skew a T cell response toward tolerance.


"I knew what was going to happen if I did nothing and so I was going to do something": faith, hope, and trust in the decisions of Canadians with multiple sclerosis to seek unproven interventions abroad.
Chronic cerebrospinal venous insufficiency (CCSVI) treatment is an unproven intervention aimed at relieving some of the symptoms of multiple sclerosis (MS). Despite limited evidence of the efficacy and safety of this intervention, Canadians diagnosed with MS have been traveling abroad to access this procedure as it is not available domestically outside of limited clinical trials. This paper discusses the experiences of Canadians with MS seeking CCSVI treatment abroad. This paper presents a secondary analysis of 15 interviews with participants who had gone abroad for CCSVI treatment. Interviews were conducted over the phone between October 2012 and December 2012. All interviews were digitally recorded and transcribed verbatim. Transcripts were hand coded for: 1) why CCSVI treatment was sought and where it was obtained; 2) the role of having hope for a cure in seeking CCSVI; 3) the impact of MS on everyday life; and 4) the role other people played in the decision to go abroad. The authors identified loss of faith, hope, and trust as themes emerging from the transcripts. The participants experienced a loss of faith with the Canadian health system and especially the neurologists who were responsible for their care and the classification of MS as a neurological disease. Access to CCSVI treatment abroad generated hope in these participants, but they were cautious in their expectations, focusing on symptom management rather than a cure. Trust in their caregivers abroad was generated through the recommendations of other MS sufferers and the credentials of their caregivers abroad. By deciding to seek an unproven intervention abroad, these individuals took on responsibility for their care from the Canadian health system. While evidence of the efficacy of CCSVI treatment is limited, the participants felt that they were making a rational care decision, focusing on the empowerment and renewed hope generated by seeking this intervention. Health professionals and policy makers globally should consider the causes of loss of faith in their domestic care systems and balance the benefits of empowerment and renewed hope against concerns that unproven interventions may create new health risks.


Improving reproducibility and sensitivity in identifying human proteins by shotgun proteomics.
Identifying proteins in cell extracts by shotgun proteomics involves digesting the proteins, sequencing the resulting peptides by data-dependent mass spectrometry (MS/MS), and searching protein databases to identify the proteins from which the peptides are derived. Manual analysis and direct spectral comparison reveal that scores from two commonly used search programs (Sequest and Mascot) validate less than half of potentially identifiable MS/MS spectra (class positive) from shotgun analyses of the human erythroleukemia K562 cell line. Here we demonstrate increased sensitivity and accuracy using a focused search strategy along with a peptide sequence validation script that does not rely exclusively on XCorr or Mowse scores generated by Sequest or Mascot, but uses consensus between the search programs, along with chemical properties and scores describing the nature of the fragmentation spectrum (ion score and RSP). The approach yielded 4.2% false positive and 8% false negative frequencies in peptide assignments. The protein profile is then assembled from peptide assignments using a novel peptide-centric protein nomenclature that more accurately reports protein variants that contain identical peptide sequences. An Isoform Resolver algorithm ensures that the protein count is not inflated by variants in the protein database, eliminating approximately 25% of redundant proteins. Analysis of soluble proteins from a human K562 cells identified 5130 unique proteins, with approximately 100 false positive protein assignments.


The Caspase Inhibitor Z-VAD-FMK Alleviates Endotoxic Shock via Inducing Macrophages Necroptosis and Promoting MDSCs-Mediated Inhibition of Macrophages Activation.
Macrophages play a critical role in the pathogenesis of endotoxin shock by producing excessive amounts of pro-inflammatory cytokines. A pan-caspase inhibitor, zVAD, can be used to induce necroptosis under certain stimuli. The role of zVAD in both regulating the survival and activation of macrophages, and the pathogenesis of endotoxin shock remains not entirely clear. Here, we found that treatment of mice with zVAD could significantly reduce mortality and alleviate disease after lipopolysaccharide (LPS) challenge. Notably, in LPS-challenged mice, treatment with zVAD could also reduce the percentage of peritoneal macrophages by promoting necroptosis and inhibiting pro-inflammatory responses in macrophages. In vitro studies showed that pretreatment with zVAD promoted LPS-induced nitric oxide-mediated necroptosis of bone marrow-derived macrophages (BMDMs), leading to reduced pro-inflammatory cytokine secretion. Interestingly, zVAD treatment promoted the accumulation of myeloid-derived suppressor cells (MDSCs) in a mouse model of endotoxin shock, and this process inhibited LPS-induced pro-inflammatory responses in macrophages. Based on these findings, we conclude that treatment with zVAD alleviates LPS-induced endotoxic shock by inducing macrophage necroptosis and promoting MDSC-mediated inhibition of macrophage activation. Thus, this study provides insights into the effects of zVAD treatment in inflammatory diseases, especially endotoxic shock.


Structural and Kinetic Studies of Formate Dehydrogenase from Candida boidinii.
The structure of formate dehydrogenase from Candida boidinii (CbFDH) is of both academic and practical interests. First, this enzyme represents a unique model system for studies on the role of protein dynamics in catalysis, but so far these studies have been limited by the availability of structural information. Second, CbFDH and its mutants can be used in various industrial applications (e.g., CO2 fixation or nicotinamide recycling systems), and the lack of structural information has been a limiting factor in commercial development. Here, we report the crystallization and structural determination of both holo- and apo-CbFDH. The free-energy barrier for the catalyzed reaction was computed and indicates that this structure indeed represents a catalytically competent form of the enzyme. Complementing kinetic examinations demonstrate that the recombinant CbFDH has a well-organized reactive state. Finally, a fortuitous observation has been made: the apoenzyme crystal was obtained under cocrystallization conditions with a saturating concentration of both the cofactor (NAD(+)) and inhibitor (azide), which has a nanomolar dissociation constant. It was found that the fraction of the apoenzyme present in the solution is less than 1.7 × 10(-7) (i.e., the solution is 99.9999% holoenzyme). This is an extreme case where the crystal structure represents an insignificant fraction of the enzyme in solution, and a mechanism rationalizing this phenomenon is presented.


Growth hormone: a reproductive endocrine-paracrine regulator?
Growth hormone (GH) is not classically considered as a reproductive hormone, although a vast literature indicates that it has roles in reproductive function. It is required for sexual differentiation and pubertal maturation and it participates in gonadal steroidogenesis, gametogenesis and ovulation. GH is also required for fetal nutrition and growth during pregnancy and for mammary development and lactation. Although some of these roles reflect the action of GH on the secretion and action of LH and FSH (Chandrashekar and Bartke, 1998), they also reflect direct actions of GH and indirect actions mediated through the local production of insulin-like growth factor I. Moreover, as GH is produced in gonadal and mammary tissues, these actions may reflect local autocrine or paracrine actions of extrapituitary GH, as well as the endocrine actions of pituitary GH. The roles of GH in reproductive function are considered in this review.


Luminescent imidazolium carboxylate supported aggregate and infinite coordination networks of copper and zinc.
The new copper dimer [LCu(DMF)]2(NO3)4(H2O)(DMF)2 (4), where L = [{1,1'-(CH2)2-C14H8)-3,3'-(CH2CO2)2}{(HCN)2CH}], and porous coordination polymers [{L2Cu(OH2)2}2Br2]x (5) and [{L2Zn(OH2)2}2Br2]x (6) have been isolated from reactions of luminescent imidazolium carboxylate ligand, LH2Br2 (3) and the corresponding metal precursors. The reaction between Cu(NO3)2·3H2O and LH2Br2 (3) in DMF at 100 °C yielded bluish green crystals of tetracationic discrete copper dimer 4, the structure of which contains a rare tetracationic [(DMF)Cu(ii)]2 dimer unit that is bridged by four carboxylates of two L in a "paddle-wheel" structure. When the reaction was carried out in the presence of a water-ethanol-methanol mixture, light green crystals of 5 were obtained. Molecule 5 comprises two-dimensional (2D) porous coordination polymeric sheets consisting of unique symmetrical dinuclear [(C(O)O)Cu(OH2)2(O(O)C)2]2 building blocks, which are connected by imidazolium anthracene spacers. The infinite 2D porous coordination polymeric sheets are further linked by significant intermolecular hydrogen-bonding interactions by bromide anions to form a three-dimensional supramolecular framework. Interestingly, the reaction between zinc dust and LH2Br2 (3) in H2O at room temperature gave similar structural features to those in 5, though they differ in terms of C-O bond distances and M-O-C angles. The solution-state UV-visible absorption spectra of 2-6 in water exhibits the comparable absorption pattern with decrease in the intensity of absorption from 5, 4, 3, 6 and 2, while the solid-state UV-visible absorption spectra of 2-6 are significantly different from the solution-state UV-visible absorption spectra. The considerable change in the fluorescent emission was observed upon complexation of 3 with corresponding metal precursors and the fluorescent emission was shifted towards the red region in the order of 2, 3, 6, 4 and 5 in water.


Myosin-driven actin-microtubule networks exhibit self-organized contractile dynamics.
The cytoskeleton is a dynamic network of proteins, including actin, microtubules, and their associated motor proteins, that enables essential cellular processes such as motility, division, and growth. While actomyosin networks are extensively studied, how interactions between actin and microtubules, ubiquitous in the cytoskeleton, influence actomyosin activity remains an open question. Here, we create a network of co-entangled actin and microtubules driven by myosin II. We combine dynamic differential microscopy, particle image velocimetry, and particle tracking to show that both actin and microtubules undergo ballistic contraction with unexpectedly indistinguishable characteristics. This contractility is distinct from faster disordered motion and rupturing that active actin networks exhibit. Our results suggest that microtubules enable self-organized myosin-driven contraction by providing flexural rigidity and enhanced connectivity to actin networks. Beyond the immediate relevance to cytoskeletal dynamics, our results shed light on the design of active materials that can be precisely tuned by the network composition.