A missense mutation of the gene encoding voltage-dependent sodium channel (Nav1.1) confers susceptibility to febrile seizures in rats.
Although febrile seizures (FSs) are the most common convulsive syndrome in infants and childhood, the etiology of FSs has remained unclarified. Several missense mutations of the Na(v)1.1 channel (SCN1A), which alter channel properties, have been reported in a familial syndrome of GEFS+ (generalized epilepsy with febrile seizures plus). Here, we generated Scn1a-targeted rats carrying a missense mutation (N1417H) in the third pore region of the sodium channel by gene-driven ENU (N-ethyl-N-nitrosourea) mutagenesis. Despite their normal appearance under ordinary circumstances, Scn1a mutant rats exhibited remarkably high susceptibility to hyperthermia-induced seizures, which involve generalized clonic and/or tonic-clonic convulsions with paroxysmal epileptiform discharges. Whole-cell patch-clamp recordings from HEK cells expressing N1417H mutant channels and from hippocampal GABAergic interneurons of N1417H mutant rats revealed a significant shift of the inactivation curve in the hyperpolarizing direction. In addition, clamp recordings clearly showed the reduction in action potential amplitude in the hippocampal interneurons of these rats. These findings suggest that a missense mutation (N1417H) of the Na(v)1.1 channel confers susceptibility to FS and the impaired biophysical properties of inhibitory GABAergic neurons underlie one of the mechanisms of FS.


Thermosensitive biomimetic polyisocyanopeptide hydrogels may facilitate wound repair.
Changing wound dressings inflicts pain and may disrupt wound repair. Novel synthetic thermosensitive hydrogels based on polyisocyanopeptide (PIC) offer a solution. These gels are liquid below 16 °C and form gels beyond room temperature. The architecture and mechanical properties of PIC gels closely resemble collagen and fibrin, and include the characteristic stiffening response at high strains. Considering the reversible thermo-responsive behavior, we postulate that PIC gels are easy to apply and remove, and facilitate healing without eliciting foreign body responses or excessive inflammation. Biocompatibility may be higher in RGD-peptide-functionalized PIC gels due to enhanced cell binding capabilities. Full-thickness dorsal skin wounds in mice were compared to wounds treated with PIC gel and PIC-RGD gel for 3 and 7 days. No foreign body reactions and similar wound closure rates were found in all groups. The level of macrophages, myofibroblasts, epithelial migration, collagen expression, and blood vessels did not significantly differ from controls. Surprisingly, granulocyte populations in the wound decreased significantly in the PIC gel-treated groups, likely because foreign bacteria could not penetrate the gel. RGD-peptides did not further improve any effect observed for PIC. The absence of adverse effects, ease of application, and the possibilities for bio-functionalization make the biomimetic PIC hydrogels suitable for development into wound dressings.


Elevated level of high-sensitivity C-reactive protein is important in determining prognosis in chronic heart failure.
While the adverse prognostic impact of a high level of high sensitivity C-reactive protein (hs-CRP) in coronary artery disease is well known, we lack data about the prognostic importance of hs-CRP in chronic heart failure (CHF) patients. The aim was to investigate the relationship between hs-CRP and prognosis in CHF patients. Patients (n=258, 74 females) with CHF of both ischemic and non-ischemic etiology were followed up for a mean period of 17 + or - 13 months. The primary end-point was cardiac mortality. During the follow-up period, 71 patients died of cardiac causes. Left ventricular ejection fractions of these patients were lower, but not significantly so (23 + or - 10 vs. 26 + or - 10%, p=0.09). hs-CRP (4.57 + or - 5.35 vs. 1.88 + or - 2.75 mg/dl, p<0.001), brain natriuretic peptide (BNP) (1621 + or - 1361 vs. 736 + or - 914 pg/ml, p<0.001), and creatinine (1.7 + or - 1.7 vs. 1.2 + or - 0.6 mg/dl, p=0.015) levels were significantly higher in CHF patients with cardiac mortality. Using stepwise multivariate Cox proportional hazards regression analyses, hs-CRP proved to be a significant independent predictor of cardiac death (hazard ratio: 1.1, 95%CI: 1.05-1.15, p<0.001). hs-CRP can provide additional prognostic information for the risk stratification of CHF patients. These insights might ultimately also affect the treatment of CHF patients.


Double-blind randomized study evaluating regeneration of the rat transected sciatic nerve after suturing and postoperative low-power laser treatment.
This double-blind randomized study evaluated the therapeutic effect of low-power laser irradiation (LPLI) on peripheral nerve regeneration, after complete transection and direct anastomosis of the rat sciatic nerve. After this procedure, 13 of 24 rats received postoperative LPLI, with a wavelength of 780 nm laser, applied transcutaneously, 30 min daily for 21 consecutive days, to corresponding segments of the spinal cord and to the injured sciatic nerve. Positive somatosensory evoked responses were found in 69.2 percent of the irradiated rats (p = 0.019), compared to 18.2 percent of the non-irradiated rats. Immunohistochemical staining in the laser-treated group showed an increased total number of axons (p = 0.026), and better quality of the regeneration process, due to an increased number of large-diameter axons (p = 0.021), compared to the non-irradiated control group. The study suggests that postoperative LPLI enhances the regenerative processes of peripheral nerves after complete transection and anastomosis.


Meta-analysis: effect of monotherapy and combination therapy with inhibitors of the renin angiotensin system on proteinuria in renal disease.
Reduction of proteinuria is associated with delayed progression of chronic kidney disease. Reports suggest that angiotensin-receptor blockers (ARBs) reduce proteinuria, but results are variable. The relative effect of ARBs and angiotensin-converting enzyme (ACE) inhibitors, and their combined administration, remains uncertain. To establish the effect of ARBs versus placebo and alternative treatments, and the effect of combined treatment with ARBs and ACE inhibitors, on proteinuria. English-language studies in MEDLINE and the Cochrane Library Central Register of Controlled Trials (January 1990 to September 2006), reference lists, and expert contacts. Randomized trials of ARBs versus placebo, ACE inhibitors, calcium-channel blockers, or the combination of ARBs and ACE inhibitors in patients with or without diabetes and with microalbuminuria or proteinuria for whom data were available on urinary protein excretion at baseline and at 1 to 12 months. Two investigators independently searched and abstracted studies. Forty-nine studies involving 6181 participants reported results of 72 comparisons with 1 to 4 months of follow-up and 38 comparisons with 5 to 12 months of follow-up. The ARBs reduced proteinuria compared with placebo or calcium-channel blockers over 1 to 4 months (ratio of means, 0.57 [95% CI, 0.47 to 0.68] and 0.69 [CI, 0.62 to 0.77], respectively) and 5 to 12 months (ratio of means, 0.66 [CI, 0.63 to 0.69] and 0.62 [CI, 0.55 to 0.70], respectively). The ARBs and ACE inhibitors reduced proteinuria to a similar degree. The combination of ARBs and ACE inhibitors further reduced proteinuria more than either agent alone: The ratio of means for combination therapy versus ARBs was 0.76 (CI, 0.68 to 0.85) over 1 to 4 months and 0.75 (CI, 0.61 to 0.92) over 5 to 12 months; for combination therapy versus ACE inhibitors, the ratio of means was 0.78 (CI, 0.72 to 0.84) over 1 to 4 months and 0.82 (CI, 0.67 to 1.01) over 5 to 12 months. The antiproteinuric effect was consistent across subgroups. Most studies were small, varied in quality, and did not provide reliable data on adverse drug reactions. Proteinuria reduction is only a surrogate for important progression of renal failure. The ARBs reduce proteinuria, independent of the degree of proteinuria and of underlying disease. The magnitude of effect is similar regardless of whether the comparator is placebo or calcium-channel blocker. Reduction in proteinuria from ARBs and ACE inhibitors is similar, but their combination is more effective than either drug alone. Uncertainty concerning adverse effects and outcomes that are important to patients limits applicability of findings to clinical practice.


Retinal vascular occlusion in patients infected with human immunodeficiency virus.
To describe the clinical features of retinal vascular occlusions (macrovasculopathy) and the proportion of affected patients with human immunodeficiency virus (HIV) infection. Retrospective chart review of all HIV-infected individuals with retinal vascular occlusions seen at a single tertiary care clinic between November 1983 and May 1998. Retinal vascular occlusion was found in 38 eyes of 33 (1.3%) of the 2,484 consecutive patients examined. Of the 33 patients, 16 (48.5%) had central retinal vein occlusion, 9 (27.3%) had branch retinal vein occlusion, 4 (12.1%) had hemiretinal vein occlusion, and 1 each (3.0%) had central retinal arterial occlusion, branch retinal arterial occlusion, and combined hemiretinal vein occlusion and branch retinal artery occlusion. One patient (3.0%) had Purtscher-like retinopathy in one eye. Multiple regression analysis revealed that macrovasculopathy was associated with the presence of noninfectious retinal microvasculopathy (odds ratio, 5.76; 95% confidence interval, 2.59-12.80). Vision at the time of diagnosis ranged between 20/20 and no light perception. Twenty-five (75.8%) of the 33 patients had follow-up examinations for a mean period of 9.1 months. Ocular findings during follow-up included capillary dropout in 3 patients (11.5%), posterior pole neovascularization in 4 (15.4%), vitreous hemorrhage in 4 (15.4%), and optic atrophy in 6 (23.1%). Vision at the last eye examination ranged between 20/20 and no light perception and was 20/200 or worse in 15 (39.5%) of 38 eyes. Patients with HIV infection have a small risk of retinal vascular occlusion, but complications and visual loss are common in affected patients. There is a strong association between noninfectious retinal microvasculopathy and retinal vein occlusions.


Kruppel-like factors regulate the Lama1 gene encoding the laminin alpha1 chain.
Laminin-1 (alpha1beta1gamma1), a basement membrane (BM) constituent, has been associated with differentiation processes and also with malignant progression. In the intestinal tissue, the alpha1 chain is expressed and secreted in the subepithelial BM during the developmental period; in the adult rodent tissue, it is restricted to the BM of the dividing cells. To understand how laminin alpha1 chain expression is regulated, we cloned and characterized a 2-kb promoter region of the Lama1 mouse gene. Analysis of the promoter was conducted in the Caco2-TC7 intestinal epithelial cells by transient transfection of serially deleted and site-directed mutated promoter constructs, by electrophoretic mobility shift assays, and expression of selected transcription factors. We determined that a proximal region, which includes an Sp1-binding GC box and a Krüppel-like element, was important for the promoter activity. This region is conserved between the human and mouse genes. Interestingly, two Krüppel-like factors KLF4 and KLF5 exhibit opposing effects on the Lama1 promoter activity that are decreased and increased, respectively, in the intestinal epithelial cells. These data corroborate the complementary expression of KLF4 and KLF5 along the intestinal crypt-villus axis and the parallel expression of KLF5 and laminin alpha1 chain in the crypt region. Finally, we showed that glucocorticoids stimulate the promoter activity. This study is the first characterization of the Lama1 promoter; we identified regulatory elements that may account for the expression pattern of the endogenous protein in the mouse intestine.


On the mechanism of induction of DNA synthesis in cultured arterial smooth muscle cells by leukotrienes. Possible role of prostaglandin endoperoxide synthase products and platelet-derived growth factor.
The induction of DNA replication in rat aortic smooth muscle cells (SMCs) by leukotrienes (LTs) was studied in order to elucidate the mechanisms of action in further detail. The effect of LTB4 was blocked by the prostaglandin (PG) synthesis inhibitor indomethacin and the effects of LTC4 and LTD4 were blocked by the cysteinyl-containing leukotriene receptor antagonists FPL 55712 and ICI 198615. These observations suggest that LTB4 and the cysteinyl-containing leukotrienes act via distinct receptors and point to a role for prostaglandin endoperoxide synthase products in bringing about the effect of LTB4. Radioimmunological determinations and analyses of [3H]arachidonic acid metabolism showed that the SMCs were able to synthesize PGI2 (measured as the stable metabolite 6-keto-PGF1 alpha), PGE2, PGF2 alpha and 15(S)hydroxy-eicosatetraenoic acid (15(S)HETE). Moreover, picomolar concentrations of arachidonic acid, PGI2, PGE2, PGF2 alpha and 15(S)HETE induced DNA replication in the SMCs under serum-free conditions, whereas linoleic acid, 6-keto-PGF1 alpha and 5(S)HETE were inactive in this respect. Analysis of conditioned media for mitogenic activity (with or without antibodies against platelet-derived growth factor, PDGF) and for the presence of material competing with radioiodinated PDGF for binding to specific cell surface receptors indicated that LTB4 stimulated release of PDGF or a PDGF-like molecule from the cells. These findings suggest that the growth-promoting effect of LTB4 is mediated via a prostaglandin endoperoxide synthase product and/or PDGF produced by the cells themselves.


Iridium(iii) complexes with five-membered heterocyclic ligands for combined photodynamic therapy and photoactivated chemotherapy.
Organometallic iridium complexes have emerged as potent anticancer agents in recent years. In this work, three cyclometalated iridium(iii) complexes Ir1-Ir3 containing monodentate five-membered heterocyclic ligands have been synthesized and characterized. Upon visible light (425 nm) irradiation, the five-membered heterocyclic ligands will dissociate from the metal centre. Moreover, Ir1-Ir3 can also act as effective singlet oxygen photosensitizers. Thus, Ir1-Ir3 can exert their light-mediated activation of anticancer effects by dual modes including ligand exchange reactions and generation of singlet oxygen (1O2) upon visible light irradiation. Notably, Ir1 displays a high phototoxicity index of 61.7 against human cancer cells. Further studies show that light-mediated anticancer properties exerted by Ir1-Ir3 occur through reactive oxygen species (ROS) generation, caspase activation, and eventually apoptosis induction. Our study demonstrates that these complexes can act as novel dual-mode light-mediated anticancer agents.


Adhesion of Phytophthora palmivora zoospores: electron microscopy of cell attachment and cyst wall fibril formation.
Zoospores of Phytophthora palmivora adhered to a plastic film surface were examined by electron microscopy. Three stages of adhesion were compared: (1) non-adhesive, unencysted zoospores, (2) adhered incipient cysts, and (3) adhered mature cysts. Thin sections of incipient cysts revealed cells attached to the film surface through the partially discharged contents of the so-called peripheral vesicles; this seems to be the first step in cell adhesion. In mature cysts, the adhesive appeared to have been compacted into an electron-dense deposit binding the cyst wall to the plastic surface. The adhesion zone was also examined in face view after lysing attached incipient cysts with sodium dodecyl sulphate. Cyst wall microfibrils were seen together with an amorphous substance (presumably the adhesive material). The microfibrils were in various stages of formation. Seemingly, adhesion and microfibril formation take place concurrently. The possibility was considered that the material contained in the peripheral vesicles serves in both cell adhesion and microfibril elaboration.


The surface glycoproteins of Trypanosoma cruzi encode a superfamily of variant T cell epitopes.
Trypanosoma cruzi is an obligate intracellular protozoan parasite. During mammalian infection, extracellular and intracellular parasites simultaneously express multiple members of a polymorphic surface protein superfamily. The effect of this extensive surface protein polymorphism on the mammalian host T cell response is not known. In this report, we identified a surface protein MHC class II-restricted T cell epitope (epitope 1), and cloned 10 surface protein cDNAs that encode epitope 1 variants. All these cDNA variant epitopes were processed and presented to T cells, and some functioned as partial T cell agonists. CD4 T cells primed with epitope 1, and challenged with epitope 1, proliferated and expressed IFN-gamma and IL-4. In contrast, CD4 T cells primed with a mixture of variant epitopes and epitope 1, and challenged with epitope 1, expressed IL-4, but did not proliferate or express IFN-gamma. The data suggest that the simultaneous expression of polymorphic surface proteins that encode variant T cell epitopes may limit the expression of each surface protein epitope below a threshold level required to stimulate a protective IFN-gamma response against that epitope. This system of T cell evasion is unique and contrasts with the sequential antigenic variation used by African trypanosomes to evade Abs directed against their surface proteins.


Molecular mechanisms of HIV latency.
HIV seeds reservoirs of latent proviruses in the earliest phases of infection. These reservoirs are found in many sites, including circulating cells, the lymphoid system, the brain, and other tissues. The "shock and kill" strategy, where HIV transcription is reactivated so that antiretroviral therapy and the immune system clear the infection, has been proposed as one approach to curing AIDS. In addition to many defective viruses, resting hematopoietic cells harbor transcriptionally latent HIV. Understanding basic mechanisms of HIV gene expression provides a road map for this strategy, allowing for manipulation of critical cellular and viral transcription factors in such a way as to maximize HIV gene expression while avoiding global T cell activation. These transcription factors include NF-κB and the HIV transactivator of transcription (Tat) as well as the cyclin-dependent kinases CDK13 and CDK11 and positive transcription elongation factor b (P-TEFb). Possible therapies involve agents that activate these proteins or release P-TEFb from the inactive 7SK small nuclear ribonucleoprotein (snRNP). These proposed therapies include PKC and MAPK agonists as well as histone deacetylase inhibitors (HDACis) and bromodomain and extraterminal (BET) bromodomain inhibitors (BETis), which act synergistically to reactivate HIV in latently infected cells.


Pharmacogenetic assessment of toxicity and outcome in patients with metastatic colorectal cancer treated with LV5FU2, FOLFOX, and FOLFIRI: FFCD 2000-05.
The aim was to investigate whether germline polymorphisms within candidate genes known or suspected to be involved in fluorouracil (FU), oxaliplatin, and irinotecan pathways were associated with toxicity and clinical outcome in patients with metastatic colorectal cancer (mCRC). Blood samples from 349 patients included in the Fédération Francophone de Cancérologie Digestive 2000-05 randomized trial, which compared FU plus leucovorin (LV5FU2) followed by FU, leucovorin, and oxaliplatin (FOLFOX) followed by FU, leucovorin, and irinotecan (FOLFIRI; sequential arm) with FOLFOX followed by FOLFIRI (combination arm) in terms of progression-free survival (PFS) and overall survival, were collected. Twenty polymorphisms within the DPD, TS, MTHFR, ERCC1, ERCC2, GSTP1, GSTM1, GSTT1, and UGT1A1 genes were genotyped. The ERCC2-K751QC allele was independently associated with an increased risk of FOLFOX-induced grade 3 or 4 hematologic toxicity (P = .01). In the sequential arm, TS-5'UTR3RG and GSTT1 alleles were independently associated with response to LV5FU2 (P = .009) and FOLFOX (P = .01), respectively. The effect of oxaliplatin on tumor response increased with the number of MTHFR-1298C alleles (test for trend, P = .008). The PFS benefit from first-line FOLFOX was restricted to patients with 2R/2R (hazard ratio [HR] = 0.39; 95% CI, 0.23 to 0.68) or 2R/3R (HR = 0.59; 95% CI, 0.42 to 0.82) TS-5'UTR genotypes, respectively. Conversely, patients with the TS-5'UTR 3R/3R genotype did not seem to benefit from the adjunction of oxaliplatin (HR = 0.96; 95% CI, 0.66 to 1.40; trend between the three HRs, P = .006). A pharmacogenetic approach may be a useful strategy for personalizing and optimizing chemotherapy in mCRC patients and deserves confirmation in additional prospective studies.


Vitamin D status and community-acquired pneumonia: results from the third National Health and Nutrition Examination Survey.
To investigate the association between serum 25-hydroxyvitamin D [25(OH)D] level and history of community-acquired pneumonia (CAP). We identified 16,975 individuals (≥17 years) from the third National Health and Nutrition Examination Survey (NHANES III) with documented 25(OH)D levels. To investigate the association of 25(OH)D with history of CAP in these participants, we developed a multivariable logistic regression model, adjusting for demographic factors (age, sex, race, poverty-to-income ratio, and geographic location), clinical data (body mass index, smoking status, asthma, chronic obstructive pulmonary disease, congestive heart failure, diabetes mellitus, stroke, chronic kidney disease, neutropenia, and alcohol consumption), and season. Locally weighted scatterplot smoothing (LOWESS) was used to depict the relationship between increasing 25(OH)D levels and the cumulative frequency of CAP in the study cohort. The median [interquartile range (IQR)] serum 25(OH)D level was 24 (IQR 18-32) ng/mL. 2.1% [95% confidence interval (CI): 1.9-2.3] of participants reported experiencing a CAP within one year of their participation in the national survey. After adjusting for demographic factors, clinical data, and season, 25(OH)D levels <30 ng/mL were associated with 56% higher odds of CAP [odds ratio 1.56; 95% confidence interval: 1.17-2.07] compared to levels ≥30 ng/mL. LOWESS analysis revealed a near linear relationship between vitamin D status and the cumulative frequency of CAP up to 25(OH)D levels around 30 ng/mL. Among 16,975 participants in NHANES III, 25(OH)D levels were inversely associated with history of CAP. Randomized controlled trials are warranted to determine the effect of optimizing vitamin D status on the risk of CAP.


Intracellular determinants of hippocampal CA1 place and silent cell activity in a novel environment.
For each environment a rodent has explored, its hippocampus contains a map consisting of a unique subset of neurons, called place cells, that have spatially tuned spiking there, with the remaining neurons being essentially silent. Using whole-cell recording in freely moving rats exploring a novel maze, we observed differences in intrinsic cellular properties and input-based subthreshold membrane potential levels underlying this division into place and silent cells. Compared to silent cells, place cells had lower spike thresholds and peaked versus flat subthreshold membrane potentials as a function of animal location. Both differences were evident from the beginning of exploration. Additionally, future place cells exhibited higher burst propensity before exploration. Thus, internal settings appear to predetermine which cells will represent the next novel environment encountered. Furthermore, place cells fired spatially tuned bursts with large, putatively calcium-mediated depolarizations that could trigger plasticity and stabilize the new map for long-term storage. Our results provide new insight into hippocampal memory formation.


Prenatal diagnosis of congenital neuroblastoma. Analysis of 4 cases and review of the literature.
Advances in prenatal diagnostics during the last 10 years have enabled the examiner to detect even rare fetal disorders such as fetal tumours. Congenital neuroblastoma is the most frequent solid neoplasm in infancy, with a retroperitoneal cystic or solid mass being a sonographic sign of the conditions. We present 4 cases of neuroblastoma showing suspicious prenatal ultrasound findings. The investigation comprises detection during pregnancy, typical sonographic signs, as well as the postnatal outcome. In addition, a review of the literature is undertaken with a focus on prenatal sonographic signs of congenital neuroblastomas. In all 4 cases, a cystic tumour was detected during the 3rd trimester of pregnancy by means of B-mode sonography. One boy died of disseminated metastases at the age of 26 months. The other 3 survived after surgery and have remained healthy. The detection of a cystic suprarenal mass is suspicious of a congenital neuroblastoma. The delivery should take place at a perinatal centre.


Kodiak seamount not flat-topped.
Earlier surveys in the Aleutian Trench southeast of Kodiak Island, Alaska, indicated that Kodiak Seamount had a flat top and was a tablemount or guyot. This seamount is of special significance because it has been supposed that its surface was eroded at the same time as those of a line of guyots to the southeast. If so, its present position in the axis of the Aleutian Trench indicates that the line of guyots was formed before the trench. A two-part survey in 1965 showed that Kodiak Seamount is not flat-topped, and should be eliminated from the category of guyots. Reflection profiling records indicate that the seamount was formed before the adjacent sediments were deposited, and that the small trough, or moat, on the south side is a depositional feature probably formed by a scouring effect or by the acceleration of turbidity currents around the base of the mount.


Synergistic blocking of RAS downstream signaling and epigenetic pathway in KRAS mutant pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) is a highly fatal malignancy and lacks effective therapeutic targets. Trametinib is considered to be a promising potential indirectly targeted KRAS inhibitor in PDAC. However, the clinical outcomes were poor. JQ1 displayed a significant synergistic effect when combined with chemotherapy or potential targeted therapy in pancreatic cancer. The impact of Trametinib and JQ1 combination treatment in PDAC remains to be fully elucidated. The efficacy of trametinib and JQ1 on cell proliferation and cytotoxicity was assayed in 7 KRAS mutant pancreatic cancer cell lines. The cytotoxic effects of drugs either alone or in combination were evaluated using a luminescent cell viability assay. Immunoblot analysis was carried out to investigate changes in p62 and autophagy. We found that either trametinib or JQ1 alone inhibited the proliferation of some pancreatic cancer cell lines with KRAS alterations, irrespective of the mutational loci of KRAS and the aberrant status of the other driver genes. The synergistic effects of combination treatment of trametinib and JQ1 were observed in both trametinib-resistant and trametinib-sensitive cells. In trametinib-sensitive PDAC cells, the combined treatment definitely inhibited p62 expression compared with trametinib alone, while LC3 expression at high levels changed little. In trametinib-resistant PDAC cells, the combination of MEK/BET inhibitor dramatically decreased p62 expression compared with single agent, while p62 expression increased after anti-autophagic therapy was added. Blocking RAS downstream signaling and epigenetic pathway synergistically increases the antiproliferative activity in KRAS mutant PDAC cells. Combination therapeutic synergism may induce different cell death modes in different pancreatic cancer subtypes.


RECK--a newly discovered inhibitor of metastasis with prognostic significance in multiple forms of cancer.
The RECK (reversion-inducing cysteine rich protein with Kazal motifs) protein was initially discovered by its ability to induce reversion in ras-activated fibroblasts. The key action of RECK is to inhibit matrix metalloproteinases (MMPs) involved in breakdown of the extracellular matrix (ECM), and angiogenesis-namely MMP-2, MMP-9 and MTP-1. To this effect, it plays important physiological roles in embryogenesis and vasculogenesis. Additionally, it has a significant effect on tumorigenesis by limiting angiogenesis and invasion of tumours through the ECM. RECK has been studied in the context of a number of human tumours including colorectal, breast, pancreas, gastric, hepatocellular, prostate, and non-small cell lung carcinoma. In many of these tumours, RECK is down-regulated most likely as a result of inhibition at the Sp1 promoter site. MMP-2 and MMP-9 generally show an inverse association with RECK expression, but there are exceptions to this rule. Likewise, a reduction in tumour microvascular density (MVD) and VEGF have also been correlated with increased RECK levels, although more studies are required to define this effect. The predominant finding across all human tumour studies is a significantly improved prognosis (due to decreased invasion and metastasis) in tumours with preserved RECK expression. Although further research is required, RECK is a promising prognostic marker and potential therapeutic agent in multiple cancers.


Measuring foot and ankle injury outcomes: common scales and checklists.
Functional health outcome measures are increasingly being used in both clinica trials and practice as measures of patient health. Whilst outcome measures can be generic, there are a number of foot and ankle specific measures available and in use. These are being used as not only region specific but also disease specific measures of patient function. Unfortunately not all of these outcome measures have been completely validated which leads to challenges in applying the results of outcomes research to specific patients. Continued work however is being done in this area and these challenges provide opportunities for further investigation into the role of functional outcome scores specific to the foot and ankle.


Conditioned medium from human dental pulp stem cells treats spinal cord injury by inhibiting microglial pyroptosis.
Human dental pulp stem cell transplantation has been shown to be an effective therapeutic strategy for spinal cord injury. However, whether the human dental pulp stem cell secretome can contribute to functional recovery after spinal cord injury remains unclear. In the present study, we established a rat model of spinal cord injury based on impact injury from a dropped weight and then intraperitoneally injected the rats with conditioned medium from human dental pulp stem cells. We found that the conditioned medium effectively promoted the recovery of sensory and motor functions in rats with spinal cord injury, decreased expression of the microglial pyroptosis markers NLRP3, GSDMD, caspase-1, and interleukin-1β, promoted axonal and myelin regeneration, and inhibited the formation of glial scars. In addition, in a lipopolysaccharide-induced BV2 microglia model, conditioned medium from human dental pulp stem cells protected cells from pyroptosis by inhibiting the NLRP3/caspase-1/interleukin-1β pathway. These results indicate that conditioned medium from human dental pulp stem cells can reduce microglial pyroptosis by inhibiting the NLRP3/caspase-1/interleukin-1β pathway, thereby promoting the recovery of neurological function after spinal cord injury. Therefore, conditioned medium from human dental pulp stem cells may become an alternative therapy for spinal cord injury.


Attentional bias, craving and cannabis use in an inpatient sample of adolescents and young adults diagnosed with cannabis use disorder: The moderating role of cognitive control.
Motivational processes like attentional bias and craving have been related to substance use. However, results are inconclusive. The present cross-sectional study was designed to replicate and extend previous research by investigating the relationships between attentional bias, craving, cognitive control and (severity of) cannabis use in a sample of inpatient adolescents and young adults (aged 18-30) diagnosed with CUD according to DSM-5. Contrary to expectations, our sample did not show attentional bias for cannabis words, neither did attentional bias correlate with craving, cognitive control or (severity of) cannabis use. In line with our hypotheses, however, increased session-induced craving was correlated to more daily cannabis use and reduced cognitive control. Furthermore, participants who displayed reduced cognitive control used more cannabis per day. A bootstrapped hierarchical regression model showed that, contrary to expectations, cognitive control did not modulate the relationships between attentional bias, craving and cannabis use. This study highlights the unique role of craving in relation to cannabis use and extends previous findings that cognitive control appears to have no moderating role regarding cannabis use disorder. Based on our results, it might well be that the underlying mechanisms of cannabis use disorder differ from those in other substance use disorders.


Epithelial-mesenchymal interactions in the alteration of gene expression and morphology following lung injury.
Numerous studies using morphologic techniques have demonstrated the plasticity of pulmonary epithelial cells and the requirement for complex intercellular interactions for regeneration of normal epithelium following lung injury. Recent developments in the molecular characterization of genes expressed in the lung have generated additional tools for evaluation of lung cell phenotypes and interactions. This review discusses the relationship between molecular and morphologic changes in lung cells during injury and repair. Changes in epithelial morphology and their differentiated gene products may be mediated through alterations in expression of numerous factors acting through paracrine and autocrine mechanisms. Tumor necrosis factor alpha, transforming growth factor beta, and retinoic acid, whose roles in the regulation of epithelial cell differentiation have been well documented, will be discussed in the context of normal lung development and repair from injury.


[Changes in vasoactive intestinal polypeptide levels in intestinal obstruction and the influence of da cheng qi decoction].
The role of vasoactive intestinal polypeptide (VIP) in small intestine obstructed rabbits and the therapeutic mechanism of Da Cheng Qi Decoction (DCQD) were studied. VIP concentrations of both blood and intestinal tissue were measured by specific radio-immunoassay in a rabbits' mechanical small intestinal obstruction model using fix of ileum to the right low abdominal wall, the influences of duodenal perfusion of DCQD on VIP levels were also investigated. 24 hours after ligating ileum, VIP of arterial plasma was elevated 7 fold, reaching 138.65 +/- 25.58 pmol/L, that of portal vein plasma was 4 times of the arterial value. VIP immunoactivity was detected in peritoneal fluid. VIP content of duodenal tissue in 16 intestine obstructed rabbits was 2 times high as that in 15 controls, whereas that of colonal tissue was inversed. DCQD caused a 50% decrease of elevated plasma VIP in experimental animals and a 65% increase in controls. The present work demonstrates that VIP might be released into the portal and peripheral circulation and mediate local and systemic pathophysiologic alterations accompanying small intestinal obstruction, such as hyperemia and edema of intestinal wall, accumulation of fluid in the lumen. VIP changes might account for the redistribution of blood flow in the obstructed segment of small intestine and the distal site of obstruction. DCQD has a therapeutic effect. The mechanism of dual modulate action of DCQD on VIP concentration is beyond our knowledge.


Aging and cancer: can mTOR inhibitors kill two birds with one drug?
The main risk factor for a number of diseases, including cancer, is aging. By delaying the effects of aging, many years of research indicate that diseases associated with aging are reduced by prolongevity interventions such as reductions in caloric intake and mice genetically deficient for growth factors. Although studies of dietary and growth factor restriction have been highly informative regarding the aging process, they are both unrealistic for human application. Recent preclinical results with a pharmacological prolongevity agent (rapamycin) provide a proof-of-concept that such an approach is feasible in human populations. Exactly how rapamycin works to extend lifespan is under increasingly intense investigation. In addition, these studies underscore the critical role that the intracellular target of rapamycin (TOR) plays in one of the deepest mysteries of life, aging. How age-associated diseases interface with TOR and its signaling systems, and the tremendous opportunities for discovery of new drugs that target both aging and its associated diseases is one of the most exciting areas of research currently being conducted in this new era of aging research.


Common data elements for clinical research in Friedreich's ataxia.
To reduce study start-up time, increase data sharing, and assist investigators conducting clinical studies, the National Institute of Neurological Disorders and Stroke embarked on an initiative to create common data elements for neuroscience clinical research. The Common Data Element Team developed general common data elements, which are commonly collected in clinical studies regardless of therapeutic area, such as demographics. In the present project, we applied such approaches to data collection in Friedreich's ataxia (FRDA), a neurological disorder that involves multiple organ systems. To develop FRDA common data elements, FRDA experts formed a working group and subgroups to define elements in the following: ataxia and performance measures; biomarkers; cardiac and other clinical outcomes; and demographics, laboratory tests, and medical history. The basic development process included identification of international experts in FRDA clinical research, meeting by teleconference to develop a draft of standardized common data elements recommendations, vetting of recommendations across the subgroups, and dissemination of recommendations to the research community for public comment. The full recommendations were published online in September 2011 at http://www.commondataelements.ninds.nih.gov/FA.aspx. The subgroups' recommendations are classified as core, supplemental, or exploratory. Template case report forms were created for many of the core tests. The present set of data elements should ideally lead to decreased initiation time for clinical research studies and greater ability to compare and analyze data across studies. Their incorporation into new, ongoing studies will be assessed in an ongoing fashion to define their utility in FRDA.


Direct or indirect association in a complex disease: the role of SLC22A4 and SLC22A5 functional variants in Crohn disease.
A common haplotype spanning 250 kb on chromosome 5q31 is strongly associated with Crohn disease (CD). Recently, two functional variants within the SLC22A4 and SLC22A5 genes at this locus (IBD5), L503F (c.1507C > T) and G-207C (c.-207G > C), have been proposed to contribute directly to susceptibility to CD. However, extensive linkage disequilibrium at the IBD5 locus has complicated efforts to distinguish causal variants from association of the general risk haplotype. We genotyped the SLC22A4 and SLC22A5 variants and other polymorphisms across the risk haplotype in four populations of European origin, and applied regression-based haplotype analysis to over 1,200 fully genotyped case-control pairs, modeling case/control status on the presence of one or more SNPs to test for conditional association and to identify risk haplotypes. We found highly significant association of SNPs at the IBD5 locus with Crohn disease in all populations tested. However, the frequencies of L503F and G-207C in individuals who did not carry the general IBD5 risk haplotype were not significantly different in cases and controls, with associated disease odds ratios (ORs) of 0.90 (95% CI, 0.57-1.40) and 0.90 (95% CI, 0.65-1.23), respectively. Haplotype analysis showed that addition of the SLC22A4 and SLC22A5 variants to a null model that included the background risk haplotype did not significantly improve the model fit. In addition to the common risk haplotype, several rare haplotypes had an increased frequency in cases compared to controls. This study suggests that the molecular basis for Crohn disease susceptibility at the IBD5 locus remains to be defined, and highlights the challenge of the identification of causal variants in a complex disease in regions of extensive linkage disequilibrium.


[Cardiotoxicity of 5-fluorouracil. Apropos of a new case].
Five - Fluorouracil (5 FU) is widely used in oncology, especially in tumors of the gastrointestinal tract and in carcinoma of the breast. Its side effects (gastrointestinal, haematological, alopecia) are well known. However, its cardiac effects are not widely appreciated, despite an incidence similar to that observed with the anthracyclines (1.6% vs 2.2%) and a high mortality rate (12.5%). A new case of coronary insufficiency is reported illustrating the absence of a dose-effect relationship and the frequency of recurrence of symptoms when 5 FU is reintroduced even with coronary vasodilator therapy. The mechanism of cardiotoxicity in this case seems to be coronary spasm.


Wolbachia transmission dynamics in Formica wood ants.
The role of Wolbachia endosymbionts in shaping the mitochondrial diversity of their arthropod host depends on the effects they have on host reproduction and on the mode of transmission of the bacteria. We have compared the sequence diversity of wsp (Wolbachia surface protein gene) and the host mtDNA in a group of Formica ant species that have diverged approximately 0.5 million years ago (MYA). The aim was to study the relationship of Wolbachia and its ant hosts in terms of vertical and horizontal transmission of the bacteria. All studied ant species were doubly infected with two Wolbachia strains (wFex1 and wFex4) all over their geographical distribution area in Eurasia. The most common haplotypes of these strains were identical with strains previously described from a more distantly related Formica ant, with an estimated divergence time of 3.5 - 4 MYA. Some strain haplotypes were associated to the same or closely related mtDNA haplotypes as expected under vertical transmission. However, in several cases the wsp haplotypes coexisted with distant mtDNA haplotypes, a pattern which is more compatible with horizontal transmission of the bacteria. Two lines of evidence suggest that the sharing of Wolbachia strains by all F. rufa species is rather due to horizontal than vertical transmission. First, the fact that endosymbiont strains identical to those of F. rufa ants have been found in another species that diverged 3.5-4 MYA strongly suggests that horizontal transfer can and does occur between Formica ants. Second, the frequent sharing of identical Wolbachia strains by distant mitochondrial lineages within the F. rufa group further shows that horizontal transmission has occurred repeatedly. Nevertheless, our dataset also provides some evidence for longer-term persistence of infection, indicating that Wolbachia infection within this host clade has been shaped by both horizontal and vertical transmission of symbionts. The fact that all the ants were infected irrespective of the family structure of their societies gives no support to the proposed hypotheses that the spreading of Wolbachia in ants might be associated to the types of their societies.


Smoking cessation knowledge and perceptions of cancer care providers at six Latin American cancer institutions.
To assess the characteristics, self-reported tobacco use, knowledge, and perceptions about smoking cessation among cancer care providers (CCPs), as well as perceived barriers to inform interventions that can potentially improve quitting rates and the prognosis of cancer patients in Latin America. A cross-sectional study was conducted among 996 CCPs in six cancer institutions located in Argentina, Brazil, Colombia, Mexico, and Peru. An online survey consisting of 28 close-ended questions adapted from the 2012 International Association for the Study of Lung Cancer survey and the Global Adult Tobacco Survey was administered. The majority of CCPs, ranging from 86.1% in Mexico to 95.9% in Brazil, agreed or strongly agreed that smoking cessation should be integrated into cancer treatment. However, inadequate training on smoking cessation was reported by 66.9%, 69.4%, 70.4%, 72.9%, 85.8%, and 86.4% in Mexico, Colombia (Floridablanca), Argentina, Peru, Brazil, and Colombia (Medellín), respectively, and this difference was statistically significant (p < 0.001). Moreover, current cigarette smoking prevalence among CCPs was 2.5% in Brazil, 4.6% in Peru, 6.3% in Colombia (Floridablanca), 10.4% in Colombia (Medellín), 11.5% in Mexico, and 15.1% in Argentina, showing a statistically significant difference (p < 0.001). Efforts in Latin America should be geared toward assisting CCPs with their quitting efforts and training in smoking cessation practices aimed at achieving a better prognosis and improving cancer patients' quality of life.


The Emergence of Integrated Information, Complexity, and 'Consciousness' at Criticality.
Integrated Information Theory (IIT) posits that integrated information ( Φ ) represents the quantity of a conscious experience. Here, the generalized Ising model was used to calculate Φ as a function of temperature in toy models of fully connected neural networks. A Monte-Carlo simulation was run on 159 normalized, random, positively weighted networks analogous to small five-node excitatory neural network motifs. Integrated information generated by this sample of small Ising models was measured across model parameter spaces. It was observed that integrated information, as an order parameter, underwent a phase transition at the critical point in the model. This critical point was demarcated by the peak of the generalized susceptibility (or variance in configuration due to temperature) of integrated information. At this critical point, integrated information was maximally receptive and responsive to perturbations of its own states. The results of this study provide evidence that Φ can capture integrated information in an empirical dataset, and display critical behavior acting as an order parameter from the generalized Ising model.


Next generation pharmaceutical impactor: a new impactor for pharmaceutical inhaler testing. Part III. extension of archival calibration to 15 L/min.
An extension of the archival calibration of the recently developed 30-100-L/min seven-stage impactor, the Next Generation Pharmaceutical Impactor (NGI), has been undertaken at 15 L/min. The NGI stage cut sizes are 0.98-14.1 microm aerodynamic diameter at this flow rate. This 15-L/min calibration was motivated by the desire to sample the entire aerosol produced by a nebulizer when tested in accordance with a new international standard developed by the Comite Européen de Normalisation (CEN), as well as the need to test various types of inhalers at flow rates lower than 30 L/min for pediatric applications. Measurements were undertaken with monodisperse oleic acid droplets in the range of 0.7-22 microm aerodynamic diameter following a procedure established in the original 30-100-L/min calibration study. The NGI was found to be effective for particle size separation at 15 L/min. Users should decide the most applicable configuration that meets their needs, based on the following recommendations: (1) the pre-separator should not normally be used, as its performance is significantly degraded by the influence of gravity, resulting in interference with stage 1; and (2) a filter should be inserted below the micro-orifice collector (MOC), as the size corresponding to 80% collection efficiency of the MOC becomes excessively large with decreasing flow rate, so that this component becomes ineffective as a means of collecting fine particles that penetrate beyond stage 7.


Lumbar bone mass predicts low back pain in males.
Longitudinal study of lumbar bone mass as predictor of low back pain (LBP). To investigate whether low bone mineral content (BMC) and bone mineral density (BMD) values at the age of 36 years are associated with the prevalence of LBP at the age of 42 years among the study population of the Amsterdam Growth and Health Longitudinal Study. Results of epidemiological, clinical, and in vitro studies indicate that spinal injuries, caused by mechanical loading, might be a cause of LBP. BMC and BMD are determinants of spinal strength. We therefore hypothesized that BMC and BMD are associated with LBP. At the age of 36 years, the lumbar BMC and BMD were determined by dual-energy x-ray absorptiometry in 140 men and 152 women. At the age of 42 years, the participants were asked whether they had experienced LBP in the previous 12 months. Logistic regression analyses were performed to determine odds ratios (ORs)-adjusted for stature, body weight, physical activity, and smoking-for the relationship of BMC and BMD with LBP. BMC and BMD at the age of 36 years were significantly associated with the reported 12-month prevalence of LBP at the age of 42 years. This association, however, was observed only for men and not for women. Men within the quartile with the lowest BMC or BMD values had higher odds for LBP with ORs of 4.78 (95% confidence interval, 1.52-15.00) and 3.48 (95% confidence interval, 1.23-9.85), respectively. For a male population that is not characterized by osteoporosis or old age, lower lumbar BMC and BMD values at the age of 36 years are associated with an increased risk of reporting to have had LBP in the previous 12 months at the age of 42 years.


Star-shaped tetrathiafulvalene-fused coronene with large pi-extended conjugation.
A tristar shaped, planar TTF-fused coronene 1 was synthesized. Its electronic properties have been studied experimentally by the combination of electrochemistry and UV-vis-NIR spectroscopy. Thereby, a nanosized graphite fragment is largely extended in its size, supplemented with a multielectron donor functionality, and shaped to a strongly chromophoric species absorbing intensely in the visible part of the optical spectrum.


[The organ specificity of the expression of a number of genetic-biochemical systems in sheep].
The analysis of isozyme spectrum of lactate (LDH), malate (ME), isocytrate (IDH) dehydrogenases, and esterases (EST) in sheep lung, kidney, heart, liver, spleen, skeletal muscles, skin, and erythrocytes was carried out. It was revealed, that the expression of ME and IDH loci was related with histogenesis of investigated organs. Activity of IDH was maximal and of ME was minimal in epithelial organs. The minimal activity of IDH and maximal one of ME were revealed in organs of mesenchymal histogenesis. New data suggesting LDH regulation activity system in skin independent from LDH system in erythrocytes were obtained. Spectrum of esterases in skin proved to be associated with the group differences of wool production characters.


MDL1 is a high copy suppressor of ATM1: evidence for a role in resistance to oxidative stress.
The yeast ATM1 gene is essential for normal cellular iron homeostasis. Deletion of ATM1 results in mitochondrial iron accumulation and increased sensitivity to oxidative stress and transition metal toxicity. Atm1p is an ATP-binding cassette (ABC) transporter localized to the mitochondrial inner membrane. The specific function of Atm1p has not been determined, though roles in both mitochondrial iron export and cytosolic Fe-S cluster assembly have been proposed. We undertook a screen for yeast genes capable of suppressing the abnormalities of cellular iron metabolism demonstrated by Deltaatm1 cells. One of the genes we identified was MDL1, which like ATM1, encodes a mitochondrial inner membrane ABC transporter. Mdl1p has previously been shown to function in the export of peptides from the mitochondrial matrix. We demonstrate that over-expression of MDL1 in Deltaatm1 cells results in a reduction of mitochondrial iron content, and decreased sensitivity to H(2)O(2) and transition metal toxicity. Additionally, in studies of the effect of over-expression and deletion of MDL1, we have identified a novel role for Mdl1p in the regulation of cellular resistance to oxidative stress.


Heart Rate Variability and Sleep-Related Breathing Disorders in the General Population.
Obstructive sleep apnea seems to have an important influence on the autonomic nervous system. In this study, we assessed the relations of sleep apnea-related parameters with 24-hour heart rate variability (HRV) in a large population of young and healthy adults. Participants aged 25 to 41 years with a body mass index <35 kg/m(2) and without known obstructive sleep apnea were included in a prospective population-based cohort study. HRV was assessed using 24-hour electrocardiographic monitoring. The SD of all normal RR intervals (SDNN) was used as the main HRV variable. Apnea-Hypopnea Index (AHI) and oxygen desaturation index (ODI) were obtained from nighttime pulse oximetry with nasal airflow measurements. We defined sleep-related breathing disorders as an AHI ≥5 or an ODI ≥5. Multivariable regression models were constructed to assess the relation of HRV with either AHI or ODI. Median age of the 1,255 participants was 37 years, 47% were men, and 9.6% had an AHI ≥5. Linear inverse associations of SDNN across AHI and ODI groups were found (p for trend = 0.006 and 0.0004, respectively). The β coefficients (95% CI) for the relation between SDNN and elevated AHI were -0.20 (-0.40 to -0.11), p = 0.04 and -0.29 (-0.47 to -0.11), p = 0.002 for elevated ODI. After adjustment for 24-hour heart rate, the same β coefficients (95% CI) were -0.06 (-0.22 to 0.11), p = 0.51 and -0.14 (-0.30 to 0.01), p = 0.07, respectively. In conclusion, even early stages of sleep-related breathing disorders are inversely associated with HRV in young and healthy adults, suggesting that they are tightly linked with autonomic dysfunction. However, HRV and 24-hour heart rate seem to have common information.


Abortive infection of the baculovirus Autographa californica nuclear polyhedrosis virus in Sf-9 cells after mutation of the putative DNA helicase gene.
Homologous recombination between the Autographa californica nuclear polyhedrosis virus (AcNPV) genome and a 0.6-kbp-long DNA fragment derived from the putative DNA helicase gene of Bombyx mori nuclear polyhedrosis virus generates eh2-AcNPV, an expanded-host-range AcNPV mutant (S. Maeda, S.G. Kamita, and A. Kondo, J. Virol. 67:6234-6238, 1993). After inoculation at a high multiplicity of infection (MOI), eh2-AcNPV replicates efficiently in both the Sf-9 (AcNPV-permissive) and BmN (non-AcNPV-permissive) cell lines. In this study, we found that after the inoculation of Sf-9 cells at a low MOI (i.e., 1 and 0.1 PFU per cell), the release of eh2-AcNPV virions was dramatically reduced (approximately 900- and 10,000-fold, respectively, at 72 h postinoculation) compared with that of wild-type AcNPV. In addition, the titer of eh2-AcNPV determined by plaque assay on Sf-9 cells was approximately 200-fold lower than that determined by plaque assay on BmN cells. Analyses of gene expression and viral DNA replication after low-MOI eh2-AcNPV inoculation of Sf-9 cells indicated that viral early genes were expressed normally. However, DNA replication and late-gene expression were significantly reduced. These findings suggested that abortive infection occurred at the stage of viral DNA replication in nearly all low-MOI eh2-AcNPV-infected Sf-9 cells. In the larvae of Spodoptera frugiperda, the organism from which Sf-9 cells are derived, the infectivity of eh2-AcNPV was lower than that of AcNPV; however, abortive infection was not found.


General principles of management of the paraplegic.
The management of patients with traumatic paraplegia and tetraplegia has improved markedly in recent years. Survival now approaches the normal life expectancy, due to modern intensive care and understanding of potential complications of the spinal paralysis. Rehabilitation techniques lead to excellent physical and psychological adjustment, and community-based follow up with cooperation from the specialized spinal units has ensured continued good care after discharge.


Contrasting heterozygosity-fitness correlations across life in a long-lived seabird.
Selection is a central force underlying evolutionary change and can vary in strength and direction, for example across time and space. The fitness consequences of individual genetic diversity have often been investigated by testing for multilocus heterozygosity-fitness correlations (HFCs), but few studies have been able to assess HFCs across life stages and in both sexes. Here, we test for HFCs using a 26-year longitudinal individual-based data set from a large population of a long-lived seabird (the common tern, Sterna hirundo), where 7,974 chicks and breeders of known age were genotyped at 15 microsatellite loci and sampled for life-history traits over the complete life cycle. Heterozygosity was not correlated with fledging or post-fledging prospecting probabilities, but was positively correlated with recruitment probability. For breeders, annual survival was not correlated with heterozygosity, but annual fledgling production was negatively correlated with heterozygosity in males and highest in intermediately heterozygous females. The contrasting HFCs among life stages and sexes indicate differential selective processes and emphasize the importance of assessing fitness consequences of traits over complete life histories.


A human monoclonal autoantibody to breast cancer identifies the PDZ domain containing protein GIPC1 as a novel breast cancer-associated antigen.
We have been studying the native autoimmune response to cancer through the isolation of human monoclonal antibodies that are cancer specific from cancer patients. To facilitate this work we previously developed a fusion partner cell line for human lymphocytes, MFP-2, that fuses efficiently with both human lymph node lymphocytes and peripheral blood lymphocytes. Using this unique trioma fusion partner cell line we isolated a panel of autologous human monoclonal antibodies, from both peripheral blood and lymph node lymphocytes, which are representative of the native repertoire of anti-cancer specific antibodies from breast cancer patients. The current study employs immunocytochemistry, immunohistochemistry, Western blot analysis as well as Northern blots, Scatchard binding studies and finally SEREX analysis for target antigen identification. By application of an expression cloning technique known as SEREX, we determined that the target antigen for two monoclonal antibodies, 27.B1 and 27.F7, derived from lymph node B-cells of a breast cancer patient, is the PDZ domain-containing protein known as GIPC1. This protein is highly expressed not only in cultured human breast cancer cells, but also in primary and metastatic tumor tissues and its overexpression appears to be cancer cell specific. Confocal microscopy revealed cell membrane and cytoplasmic localization of the target protein, which is consistent with previous studies of this protein. We have determined that GIPC1 is a novel breast cancer-associated immunogenic antigen that is overexpressed in breast cancer. Its role, however, in the initiation and/or progression of breast cancer remains unclear and needs further clarification.


Acute fatal toxoplasmosis in squirrels (Sciurus carolensis) with bradyzoites in visceral tissues.
Acute toxoplasmosis was diagnosed in 3 gray squirrels (Sciurus carolensis) from Louisiana and Pennsylvania. The predominant lesion was multifocal necrosis in several organs, especially of the lymph nodes. Numerous Toxoplasma gondii tachyzoites were seen in lesions, and the diagnosis was confirmed immunohistochemically by reaction with polyclonal T. gondii-specific antibodies. Tissue cysts were seen in several organs, including lung alveoli. The presence of tissue cysts in alveoli of pet squirrels maybe of public health concern if tissue cysts excreted in nasal secretions are swallowed by children.


Unsuspected fungal endophthalmitis diagnosed in vitrectomy specimen.
A vitrectomy specimen obtained with the roto-extractor from a diabetic patient with vitreous haemorrhage revealed fungal endophthalmitis on microscopical examination. Endophthalmitis was not suspected before the procedure. Extensive medical examinations failed to reveal any evidence of systemic fungal disease. The case exemplifies endogenous mycotic endophthalmitis of occult origin, the first of its kind, we believe, to be diagnosed in this manner.


Induced defences alter the strength and direction of natural selection on reproductive traits in common milkweed.
Evolutionary biologists have long sought to understand the ecological processes that generate plant reproductive diversity. Recent evidence indicates that constitutive antiherbivore defences can alter natural selection on reproductive traits, but it is unclear whether induced defences will have the same effect and whether reduced foliar damage in defended plants is the cause of this pattern. In a factorial field experiment using common milkweed, Asclepias syriaca L., we induced plant defences using jasmonic acid (JA) and imposed foliar damage using scissors. We found that JA-induced plants experienced selection for more inflorescences that were smaller in size (fewer flowers), whereas control plants only experienced a trend towards selection for larger inflorescences (more flowers); all effects were independent of foliar damage. Our results demonstrate that induced defences can alter both the strength and direction of selection on reproductive traits, and suggest that antiherbivore defences may promote the evolution of plant reproductive diversity.


TRIF Differentially Regulates Hepatic Steatosis and Inflammation/Fibrosis in Mice.
Toll-like receptor 4 (TLR4) signaling is activated through 2 adaptor proteins: MyD88 and TIR-domain containing adaptor-inducing interferon-β (TRIF). TLR4 and MyD88 are crucial in nonalcoholic steatohepatitis (NASH) and fibrosis. However, the role of TRIF in TLR4-mediated NASH and fibrosis has been elusive. This study investigated the differential roles of TRIF in hepatic steatosis and inflammation/fibrosis. A choline-deficient amino acid defined (CDAA) diet was used for the mouse NASH model. On this diet, the mice develop hepatic steatosis, inflammation, and fibrosis. TLR4 wild-type and TLR4-/- bone marrow chimeric mice and TRIF-/- mice were fed CDAA or a control diet for 22 weeks. Hepatic steatosis, inflammation, and fibrosis were examined. In the CDAA diet-induced NASH, the mice with wild-type bone marrow had higher alanine aminotransferase and hepatic tumor necrosis factor levels than the mice with TLR4-/- bone marrow. The nonalcoholic fatty liver disease activity score showed that both wild-type and TLR4-/- bone marrow chimeras had reduced hepatic steatosis, and that both types of chimeras had similar levels of inflammation and hepatocyte ballooning to whole-body wild-type mice. Notably, wild-type recipients showed more liver fibrosis than TLR4-/- recipients. Although TRIF-/- mice showed reduced hepatic steatosis, these mice showed more liver injury, inflammation, and fibrosis than wild-type mice. TRIF-/- stellate cells and hepatocytes produced more C-X-C motif chemokine ligand 1 (CXCL1) and C-C motif chemokine ligand than wild-type cells in response to lipopolysaccharide. Consistently, TRIF-/- mice showed increased CXCL1 and CCL3 expression along with neutrophil and macrophage infiltration, which promotes liver inflammation and injury. In TLR4-mediated NASH, different liver cells have distinct roles in hepatic steatosis, inflammation, and fibrosis. TRIF promotes hepatic steatosis but it inhibits injury, inflammation, and fibrosis.


Mobile phone use among motorcyclists and electric bike riders: A case study of Hanoi, Vietnam.
Motorcyclist injuries and fatalities are a major concern of many developing countries. In Vietnam, motorcycles are involved in more than 70% of all road traffic crashes. This paper aims to explore the prevalence and factors associated with mobile phone use among motorcyclists and electric bike riders, using a case study of Hanoi, Vietnam. A cross-sectional observation survey was undertaken at 12 sites, in which each site was surveyed during a two-hour peak period from 16:30 to 18:30 for two weekdays and one weekend day. A total of 26,360 riders were observed, consisting of 24,759 motorcyclists and 1601 electric bike riders. The overall prevalence of mobile phone use while riding was 8.4% (95% CI: 8.06-8.74%) with calling having higher prevalence than screen operation: 4.64% (95% CI: 4.39-4.90%) vs. 3.76% (95% CI: 3.52-3.99%) respectively. Moreover, the prevalence of mobile phone use was higher among motorcyclists than electric bike riders: 8.66% (95%CI: 8.30-9.01%) vs. 4.43% (95% CI: 3.40-5.47%) respectively. Logistic regression analyses revealed that mobile phone use while riding was associated with vehicle type, age, gender, riding alone, weather, day of week, proximity to city centre, number of lanes, separate car lanes, red traffic light duration, and police presence. Combining greater enforcement of existing legislations with extensive education and publicity programs is recommended to reduce potential deaths and injuries related to the use of mobile phones while riding.


Social support and unmet needs among older trans and gender non-conforming people during the COVID-19 'lockdown' in the UK.
Previous research has suggested that older trans and gender non-conforming (TGNC) people may face particular challenges related to stigma, social exclusion and discrimination in later life. However, direct data on social support and needs in older TGNC population both internationally and within the UK is limited due to the small, dispersed nature of this population, and the absence of specific data collection on aging TGNC populations. During the UK COVID-19 lockdown in summer 2020, older people and those with long-term health conditions were advised to adopt particular precautions. This study aimed to explore older TGNC people's social support networks, key concerns and unmet needs during the COVID-19 'lockdown'. A UK cross-sectional survey of LGBT + people aged 60+ (n = 375) was undertaken during the lockdown. This paper analyses responses from the subset (n = 38) of TGNC participants. The majority of TGNC respondents described diverse social networks, often centered around friends and non-kin social networks, although partners and adult children were also significant for some. In most cases, those with existing strong networks continued to maintain social connections during lockdown, albeit with some regrets about loss of activities and face-to-face connection. However, a minority of respondents had experienced greater challenges prior to lockdown, and may have been at increased vulnerability during the pandemic, for example indicating that they had no-one to call on for practical support in an emergency. When asked about unmet needs and challenges, social isolation was repeatedly raised as the most frequent concern. Several respondents also mentioned issues specifically affecting TGNC communities, including access to gender affirming care and a perceived rise in social intolerance. Health and social care providers should be aware of the diversity of support networks within TGNC communities. There may also be benefits in community sector interventions to help older TGNC build and maintain strong social networks.


Physical and psychological improvements after phase II cardiac rehabilitation in patients with myocardial infarction.
We have designed a new 4-week hospitalized phase II cardiac rehabilitation program. The purpose of the present study is to clarify whether the physical and psychological status of patients with myocardial infarction (MI) improves after participation in our program. Twenty-nine patients (27 males, two females) with acute MI who enrolled in the 4-week hospitalized phase II rehabilitation program were assessed. All patients enrolled in this study had received coronary interventions. The rehabilitation consisted of exercise training, education and counseling. We evaluated the physical and psychological status of the patients before and just after the program, and at a 6-month follow up. The physical status was assessed by exercise tolerance measured by the peak oxygen consumption and anaerobic threshold, frequency of exercise, and serum concentrations of triglyceride, total cholesterol, high-density lipoprotein-cholesterol, and low-density lipoprotein-cholesterol. The psychological status was assessed by the Spielberger state-trait anxiety inventory questionnaire (STAI) and the self-rating questionnaire for depression (SRQ-D). Thirty-four patients (27 men, seven women) with MI who did not participate in our rehabilitation program served as a control group. After participation in our rehabilitation program, exercise tolerance and the serum lipid profiles of the patients were improved compared with those before rehabilitation. These parameters had improved significantly 6 months after rehabilitation. The STAI anxiety score was improved significantly and the SRQ-D depression score tended to be improved just after the rehabilitation program. Regular physical activity was continued even 6 months after the completion of the program. Our hospitalized phase II cardiac rehabilitation program improved the management of cardiac risk factors and the psychological status in patients with MI. This comprehensive program may contribute to the secondary prevention of MI as well as the recovery of physical and psychological activities.


Relationship between oxygen concentration and temperature in an exothermic warming device.
Actively warming hypothermic patients or preventing hypothermia is critical in optimising outcomes in patients with traumatic injuries. Our aim was to investigate the effect of ambient oxygen concentration on the rate and change in temperature of the TechTrade Ready-Heat II exothermic (oxygen-activated) warming blanket, to evaluate safety and ascertain the risk of thermal injury. A mannequin covered with an exothermic blanket was placed in a sealed oxygen tent. An ambulance blanket was placed between the TechTrade Ready-Heat II exothermic blanket and the mannequin. Two temperature probes were placed directly on the surface of the mannequin; one on the torso away from the heating packs and the other directly beneath the exothermic heating pack. The mannequin was exposed to increasing oxygen concentrations at 10% increments, starting at 21%. The experiment was conducted nine times, each time using a new blanket. Maximum temperature of the mannequin 'skin' and rate of rise were recorded from both temperature sensors. In room air (21% oxygen), the mannequin surface reached 52°C after 60 min, matching manufacturer specifications. At 30% oxygen concentration, the temperature directly beneath the exothermic heating pack exceeded the 65°C threshold at which rapid thermal burns occur, reaching 72.5°C, with minimal change in overall torso temperature. The supplemental use of oxygen in patients with traumatic injuries that increases ambient oxygen levels in the presence of exothermic warming devices may represent a significant risk to the patient. We suggest that prehospital care providers remain highly vigilant of heat when using high-flow oxygen and the subsequent fire risk, while manufacturers of exothermic blankets should consider ways to improve safety.


Predominantly sigma-bonded metal carbonyl cations (sigma-carbonyls): new synthetic, structural, and bonding concepts in metal carbonyl chemistry.
Predominantly sigma-bonded metal carbonyl cations (sigma-carbonyls) are conveniently generated in the Lewis superacid SbF(5) or the conjugate Brønsted-Lewis superacid HFbondSbF(5), primarily by solvolytic or reductive carbonylations. Thermally stable salts are formed with the fluoroantimonate(V) ions [SbF(6)](-) and [Sb(2)F(11)](-). The salts are characterised by analytical, structural, spectroscopic and computational methods. Most homoleptic carbonyl cations have very regular geometries, comensurate with their d-electron configurations: linear (d(10)), square planar (d(8)) or octahedral(d(6)). The cations with metals in oxidation states of +2 or +3 are termed "superelectrophilic". Extended molecular structures form by significant interionic C-F contacts with electrophilic carbon as acceptor. To account for all experimental observations, a conceptually simple synergetic bonding model is proposed. An outlook at anticipated future developments based on very recent results is provided.


Lack of immunotoxic effects of repeated exposure to atrazine associated with the adaptation of adrenal gland activation.
T cell-dependent IgM antibody production and natural killer cell (NKC) activity were assessed in SD rats orally administered atrazine for 28 days to males (0, 6.5, 25, or 100 mg/kg/day) or females (0, 3, 6, or 50 mg/kg/day), or 30 or 500 ppm in diet (3 or 51 mg/kg/day). Anti-asialo GM1 antibodies (NKC) and cyclophosphamide (antibody-forming cell assay [AFC]) served as positive controls. Pituitary (ACTH, prolactin), adrenal (corticosterone, progesterone, aldosterone), and gonadal (androgens, estrogens) hormones were assessed after 1, 7, and/or 28 days of treatment. Food intake and body weights were significantly reduced in the highest dosed males, and transiently affected in females. Urinary corticosterone levels were not increased in atrazine-treated groups in either sex at any time point measured (10, 22, or 24 days). Corticosterone and progesterone were elevated in males after a single atrazine dose ≥6.5 mg/kg/day, but not after 7, 14, or 28 doses. There were no effects on adrenal, pituitary, or gonadal hormones in females. Atrazine did not suppress the AFC response or decrease NKC function after 28 days in males or females. Atrazine had no effect on spleen weights or spleen cell numbers in males or females, although thymus weights were elevated in males receiving the highest dose. The lack of immunotoxic effect of atrazine was associated with diminished adrenal activation over time in males, and no effects on adrenal hormones in females.


Upper-body kinematics in team-handball throw, tennis serve, and volleyball spike.
Overarm movements are essential skills in many different sport games; however, the adaptations to different sports are not well understood. The aim of the study was to analyze upper-body kinematics in the team-handball throw, tennis serve, and volleyball spike, and to calculate differences in the proximal-to-distal sequencing and joint movements. Three-dimensional kinematic data were analyzed via the Vicon motion capturing system. The subjects (elite players) were instructed to perform a team-handball jump throw, tennis serve, and volleyball spike with a maximal ball velocity and to hit a specific target. Significant differences (P < 0.05) between the three overarm movements were found in 17 of 24 variables. The order of the proximal-to-distal sequencing was equal in the three analyzed overarm movements. Equal order of the proximal-to-distal sequencing and similar angles in the acceleration phase suggest there is a general motor pattern in overarm movements. However, overarm movements appear to be modifiable in situations such as for throwing or hitting a ball with or without a racket, and due to differences at takeoff (with one or two legs).


Impact of out-of-hours admission on patient mortality: longitudinal analysis in a tertiary acute hospital.
Emergency hospital admission on weekends is associated with an increased risk of mortality. Previous studies have been limited to examining single years and assessing day-not time-of admission. We used an enhanced longitudinal data set to estimate the 'weekend effect' over time and the effect of night-time admission on all-cause mortality rates. We examined 246 350 emergency spells from a large teaching hospital in England between April 2004 and March 2014. Outcomes included 7-day, 30-day and in-hospital mortality rates. We conducted probit regressions to estimate the impact on the absolute difference in the risk of mortality of two key predictors: (1) admission on weekends (19:00 Friday to 06:59 Monday); and (2) night-time admission (19:00 to 06:59). Logistic regressions were used to estimate ORs for relative mortality risk differences. Crude 30-day mortality rate decreased from 6.6% in 2004/2005 to 5.2% in 2013/2014. Adjusted mortality risk was elevated for all out-of-hours periods. The highest risk was associated with admission on weekend night-times: 30-day mortality increased by 0.6 percentage points (adjusted OR: 1.17, 95% CI 1.10 to 1.25), 7-day mortality by 0.5 percentage points (adjusted OR: 1.23, 95% CI 1.12 to 1.34) and in-hospital mortality by 0.5 percentage points (adjusted OR: 1.14, 95% CI 1.08 to 1.21) compared with admission on weekday daytimes. Weekend night-time admission was associated with increased mortality risk in 9 out of 10 years, but this was only statistically significant (p<0.05) in 5 out of 10 years. There is an increased risk of mortality for patients admitted as emergencies both on weekends and during the night-time. These effects are additive, so that the greatest risk of mortality occurs in patients admitted during the night on weekends. This increased risk appears to be consistent over time, but the effects are small and are not statistically significant in individual hospitals in every year.


Toward the defined and xeno-free differentiation of functional human pluripotent stem cell-derived retinal pigment epithelial cells.
The production of functional retinal pigment epithelium (RPE) cells from human embryonic (hESCs) and human induced pluripotent stem cells (hiPSCs) in defined and xeno-free conditions is highly desirable, especially for their use in cell therapy for retinal diseases. In addition, differentiated RPE cells provide an individualized disease model and drug discovery tool. In this study, we report the differentiation of functional RPE-like cells from several hESC lines and one hiPSC line in culture conditions, enabling easy translation to clinical quality cell production under Good Manufacturing Practice regulations. Pluripotent stem cells were cultured on human fibroblast feeder cells in serum-free medium. The differentiation toward RPE was induced by removing basic fibroblast growth factor and feeder cells from the serum-free conditions. RPE differentiation was also achieved using xeno-free and defined culture conditions. The RPE cell morphology and pigmentation of the cells were analyzed and the expression of genes and proteins characteristic for RPE cells was evaluated. In vitro functionality of the cells was analyzed using ELISA measurements for pigment epithelium derived factor (PEDF) secretion and phagocytosis of photoreceptor outer segments (POS). The integrity of the generated RPE layers was analyzed using transepithelial electric resistance measurements. We generated putative RPE cells with typical pigmented cobblestone-like morphology. The expression of RPE-specific markers was confirmed at the gene and protein level. The differentiated cells were able to phagocytose POS and secrete PEDF characteristic of native RPE cells. In addition, cultured cells formed a polarized epithelium with high integrity and exhibited excellent transepithelial electric resistance values, indicating well established, tight junctions. Moreover, we introduced an improved method to generate functional putative RPE cells without xeno-components under defined conditions. We have developed a progressive differentiation protocol for the production of functional RPE-like cells from hESCs and hiPSCs. Our results demonstrate that putative hESC-RPE and hiPSC-RPE express genes and proteins characteristic for RPE cells, as well as being able to phagocytose POS and secrete PEDF. Furthermore, our results show that RPE-like cells can be differentiated in xeno-free and defined culture conditions, which is mandatory for Good Manufacturing Practice-production of these cells for clinical use.


Prophylactic gene therapy with human tissue kallikrein ameliorates limb ischemia recovery in type 1 diabetic mice.
Diabetes macro- and microvascular disease causes tissue hypoperfusion. This deficit, together with a failure to mount an adequate angiogenic response, might explain why vascular occlusion evolves more severely among diabetic patients. The present study investigated whether prophylactic gene therapy with human tissue kallikrein (hTK) may protect diabetic limbs from the consequences of supervening ischemia. Vehicle (saline) or an adenovirus carrying the gene for either hTK (Ad.hTK) or luciferase (Ad.Luc) was injected into left adductor muscles of streptozotocin-induced type 1 diabetic mice 2 weeks before operative occlusion of the ipsilateral femoral artery. Saline-injected nondiabetic mice served as controls. Hindlimb blood flow recovery was analyzed sequentially over the 2 weeks after ischemia induction. At necroscopy, microvessel density and endothelial cell proliferation and apoptosis were quantified in skeletal muscles. We found that limb perfusion recovery of saline-injected type 1 diabetic mice is delayed because of insufficient reparative neovascularization and excessive activation of endothelial cell apoptosis. By contrast, prophylactic Ad.hTK renewed the ability to mount an appropriate neovascularization response to ischemia, suppressed apoptosis, and upregulated endothelial nitric oxide synthase expression. Ultimately, correction of diabetic endotheliopathy by Ad.hTK allowed proper perfusion recovery as seen in nondiabetic mice. These discoveries disclose new therapeutic options for the treatment of diabetic complications.


Seasonal Variations of Locomotor Activity Rhythms in Melatonin-Proficient and -Deficient Mice under Seminatural Outdoor Conditions.
Locomotor activity patterns of laboratory mice are widely used to analyze circadian mechanisms, but most investigations have been performed under standardized laboratory conditions. Outdoors, animals are exposed to daily changes in photoperiod and other abiotic cues that might influence their circadian system. To investigate how the locomotor activity patterns under outdoor conditions compare to controlled laboratory conditions, we placed 2 laboratory mouse strains (melatonin-deficient C57Bl and melatonin-proficient C3H) in the garden of the Dr. Senckenbergische Anatomie in Frankfurt am Main. The mice were kept singly in cages equipped with an infrared locomotion detector, a hiding box, nesting material, and with food and water ad libitum. The locomotor activity of each mouse was recorded for 1 year, together with data on ambient temperature, light, and humidity. Chronotype, chronotype stability, total daily activity, duration of the activity period, and daily diurnality indices were determined from the actograms. C3H mice showed clear seasonal differences in the chronotype, its stability, the total daily activity, and the duration of the activity period. These pronounced seasonal differences were not observed in the C57Bl. In both strains, the onset of the main activity period was mainly determined by the evening dusk, whereas the offset was influenced by the ambient temperature. The actograms did not reveal infra-, ultradian, or lunar rhythms or a weekday/weekend pattern. Under outdoor conditions, the 2 strains retained their nocturnal locomotor identity as observed in the laboratory. Our results indicate that the chronotype displays a seasonal plasticity that may depend on the melatoninergic system. Photoperiod and ambient temperature are the most potent abiotic entraining cues. The timing of the evening dusk mainly affects the onset of the activity period; the ambient temperature during this period influences the latter's duration. Humidity, overall light intensities, and human activities do not affect the locomotor behavior.


Ebola GP-specific monoclonal antibodies protect mice and guinea pigs from lethal Ebola virus infection.
Ebola virus (EBOV) causes acute hemorrhagic fever in humans and non-human primates with mortality rates up to 90%. So far there are no effective treatments available. This study evaluates the protective efficacy of 8 monoclonal antibodies (MAbs) against Ebola glycoprotein in mice and guinea pigs. Immunocompetent mice or guinea pigs were given MAbs i.p. in various doses individually or as pools of 3-4 MAbs to test their protection against a lethal challenge with mouse- or guinea pig-adapted EBOV. Each of the 8 MAbs (100 µg) protected mice from a lethal EBOV challenge when administered 1 day before or after challenge. Seven MAbs were effective 2 days post-infection (dpi), with 1 MAb demonstrating partial protection 3 dpi. In the guinea pigs each MAb showed partial protection at 1 dpi, however the mean time to death was significantly prolonged compared to the control group. Moreover, treatment with pools of 3-4 MAbs completely protected the majority of animals, while administration at 2-3 dpi achieved 50-100% protection. This data suggests that the MAbs generated are capable of protecting both animal species against lethal Ebola virus challenge. These results indicate that MAbs particularly when used as an oligoclonal set are a potential therapeutic for post-exposure treatment of EBOV infection.


Denmark: the elderly living in style.
In the next 20 years every country in Western Europe will see a rise in its total numbers of people aged over 60, and since old people are now living longer the proportion aged over 75 will also rise. Both physical and mental disorders become much more common between 60 and 75, so that if these old people are to be provided with accommodation and medical services at contemporary standards many more places will be needed in sheltered flats, nursing homes, and geriatric hospitals. Yet in the current economic recession most European countries are trying to cut back on public expenditure. How do politicians and administrators reconcile the conflict between the demands created by demographic change and the freeze on public spending? How will the care of the elderly change in response to this conflict? Tony Smith is examining the ways in which old people are looked after in several European countries and their plans for the future. This first article deals with Denmark.


MicroRNA-449a overexpression, reduced NOTCH1 signals and scarce goblet cells characterize the small intestine of celiac patients.
MiRNAs play a relevant role in regulating gene expression in a variety of physiological and pathological conditions including autoimmune disorders. MiRNAs are also important in the differentiation and function of the mouse intestinal epithelium. Our study was aimed to look for miRNA-based modulation of gene expression in celiac small intestine, and particularly for genes involved in cell intestinal differentiation/proliferation mechanisms. A cohort of 40 children (20 with active CD, 9 on a gluten-free diet (GFD), and 11 controls), were recruited at the Paediatrics Department (University of Naples Federico II). The expression of 365 human miRNAs was quantified by TaqMan low-density arrays. We used bioinformatics to predict putative target genes of miRNAs and to select biological pathways. The presence of NOTCH1, HES1, KLF4, MUC-2, Ki67 and beta-catenin proteins in the small intestine of CD and control children was tested by immunohistochemistry. The expression of about 20% of the miRNAs tested differed between CD and control children. We found that high miR-449a levels targeted and reduced both NOTCH1 and KLF4 in HEK-293 cells. NOTCH1, KLF4 signals and the number of goblet cells were lower in small intestine of children with active CD and in those on a GFD than in controls, whereas more nuclear beta-catenin staining, as a sign of the WNT pathway activation, and more Ki67 staining, as sign of proliferation, were present in crypts from CD patients than in controls. In conclusion we first demonstrate a miRNA mediated gene regulation in small intestine of CD patients. We also highlighted a reduced NOTCH1 pathway in our patients, irrespective of whether the disease was active or not. We suggest that NOTCH pathway could be constitutively altered in the celiac small intestine and could drive the increased proliferation and the decreased differentiation of intestinal cells towards the secretory goblet cell lineage.


Global patterns of care in advanced stage mycosis fungoides/Sezary syndrome: a multicenter retrospective follow-up study from the Cutaneous Lymphoma International Consortium.
Advanced-stage mycosis fungoides (MF)/Sézary syndrome (SS) patients are weighted by an unfavorable prognosis and share an unmet clinical need of effective treatments. International guidelines are available detailing treatment options for the different stages but without recommending treatments in any particular order due to lack of comparative trials. The aims of this second CLIC study were to retrospectively analyze the pattern of care worldwide for advanced-stage MF/SS patients, the distribution of treatments according to geographical areas (USA versus non-USA), and whether the heterogeneity of approaches has potential impact on survival. This study included 853 patients from 21 specialist centers (14 European, 4 USA, 1 each Australian, Brazilian, and Japanese). Heterogeneity of treatment approaches was found, with up to 24 different modalities or combinations used as first-line and 36% of patients receiving four or more treatments. Stage IIB disease was most frequently treated by total-skin-electron-beam radiotherapy, bexarotene and gemcitabine; erythrodermic and SS patients by extracorporeal photochemotherapy, and stage IVA2 by polychemotherapy. Significant differences were found between USA and non-USA centers, with bexarotene, photopheresis and histone deacetylase inhibitors most frequently prescribed for first-line treatment in USA while phototherapy, interferon, chlorambucil and gemcitabine in non-USA centers. These differences did not significantly impact on survival. However, when considering death and therapy change as competing risk events and the impact of first treatment line on both events, both monochemotherapy (SHR = 2.07) and polychemotherapy (SHR = 1.69) showed elevated relative risks. This large multicenter retrospective study shows that there exist a large treatment heterogeneity in advanced MF/SS and differences between USA and non-USA centers but these were not related to survival, while our data reveal that chemotherapy as first treatment is associated with a higher risk of death and/or change of therapy and thus other therapeutic options should be preferable as first treatment approach.


Toughening and Crack Healing Mechanisms in Nanotwinned Diamond Composites with Various Polytypes.
As an emerging ceramic material, recently synthesized nanotwinned diamond composites with various polytypes embedded in nanoscale twins exhibit unprecedented fracture toughness without sacrificing hardness. However, the toughening and crack healing mechanisms at the atomic scale and the associated crack propagation process of nanotwinned diamond composites remain mysterious. Here, we perform large-scale atomistic simulations of crack propagation in nanotwinned diamond composites to explore the underlying toughening and crack healing mechanisms in nanotwinned diamond composites. Our simulation results show that nanotwinned diamond composites have a higher fracture energy than single-crystalline and nanotwinned diamonds, which originates from multiple toughening mechanisms, including twin boundary and phase boundary impeding crack propagation, crack deflection and zigzag paths in nanotwins and sinuous paths in polytypes, and the formation of disordered atom clusters. More remarkably, our simulations reproduce more detailed crack propagation processes at the atomic scale, which is inaccessible by experiments. Moreover, our simulations reveal that crack healing occurs due to the rebonding of atoms on fracture surfaces during unloading and that the extent of crack healing is associated with whether the crack surfaces are clean. Our current study provides mechanistic insights into a fundamental understanding of toughening and crack healing mechanisms in nanotwinned diamond composites.


[Circulation of the influenza virus on poultry-raising farms].
The results of examinations for immunity to influenza in chickens and the personnel handling them at the permanently observed farm are presented. Antibodies mainly to the same antigenic variants of human influenza viruses were found in chicken and human blood sera. Alongside with antihemagglutinins to influenza A (H3N2) virus strains, chicken in 1983-1984 were found to have antibodies to influenza B virus epidemically important in those years. Thus, in poultry farms human influenza viruses are introduced into poultry populations. The strains circulating among fowls may probably be potentially dangerous for man.


Relative importance of the enzymic hydrolysis of sexamethonium in plasma and tissues: studies in Rhesus monkeys.
1. Suxamethonium was given by intravenous infusion to anaesthetized Rhesus monkeys and the infusion rate required to produce after 10 min a 50% reduction in twitch tension of the indirectly stimulated gastrocnemius or tibialis anterior muscle (IR50) was calculated from results obtained with paired infusions of two concentrations, repeated at intervals. The IR50 values in three monkeys were (388, 958 and 1,010 nmol/kg)/minute.2. In one experiment the cholinesterase activity in plasma was increased to 125 and then to 182% of the initial value by infusions of purified human cholinesterase (usual enzyme). This increased the IR50 by 34 and then by 60%, respectively.3. Inhibition by iso-OMPA of cholinesterase activity in plasma and tissues (four monkeys) by more than 90% lowered the mean IR50 value to (69 +/- 11 (S.E.M.) nmol/kg)/minute. Restoration of cholinesterase activity in the plasma of these animals by infusion of purified human cholinesterase raised the IR50's to values within the normal range.4. It is concluded that in the Rhesus monkey most of the infused suxamethonium is hydrolyzed by the cholinesterase in plasma before reaching the motor endplates. This agrees with observations in man with usual cholinesterase and contrasts with experiments in the cat where the hydrolysis of suxamethonium by cholinesterase in tissues was found to be more important than the hydrolysis by cholinesterase in plasma.


Cocaine blocks extraneuronal uptake of norepinephrine by the pregnant human uterus.
Premature labor is one of the most common complications associated with cocaine abuse during pregnancy. Still, the effect of cocaine on the pregnant uterus is largely unknown. Although inhibition of neuronal uptake is the most important effect of cocaine in most tissues, after mid-pregnancy, the uterus has few functioning adrenergic nerve endings. To determine whether cocaine has any effect on uptake during pregnancy, we evaluated the ability of the term pregnant human uterus to take up [3H]-norepinephrine (9 x 10(-8) mol/L) and the ability of cocaine (10(-6)-10(-8) mol/L) to block this uptake. Because d-propranolol has been shown to block the direct effects of cocaine on the pregnant rabbit uterus, we also evaluated the ability of d-propranolol (2 x 10(-6) mol/L) to block the effect of cocaine on catecholamine uptake. The ability of the Uptake 2 inhibitor hydrocortisone (2 x 10(-5) mol/L) to block catecholamine uptake was also studied. We found that [3H]-norepinephrine was taken up by both the pregnant myometrium and endometrium, and that cocaine blocked this uptake by up to 55% at concentrations as low as 10(-7) mol/L. D-propranolol had no effect on the ability of cocaine to block catecholamine uptake. Hydrocortisone blocked uptake by the endometrium by 15% but did not block uptake by the myometrium. We conclude that the pregnant human uterus at term retains the ability to take up catecholamines and that cocaine blocks this extraneuronal uptake. This may explain, in part, the association of cocaine use with premature labor.


The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations.
Dengue fever is the most important mosquito-borne viral disease of humans with more than 50 million cases estimated annually in more than 100 countries. Disturbingly, the geographic range of dengue is currently expanding and the severity of outbreaks is increasing. Control options for dengue are very limited and currently focus on reducing population abundance of the major mosquito vector, Aedes aegypti. These strategies are failing to reduce dengue incidence in tropical communities and there is an urgent need for effective alternatives. It has been proposed that endosymbiotic bacterial Wolbachia infections of insects might be used in novel strategies for dengue control. For example, the wMelPop-CLA Wolbachia strain reduces the lifespan of adult A. aegypti mosquitoes in stably transinfected lines. This life-shortening phenotype was predicted to reduce the potential for dengue transmission. The recent discovery that several Wolbachia infections, including wMelPop-CLA, can also directly influence the susceptibility of insects to infection with a range of insect and human pathogens has markedly changed the potential for Wolbachia infections to control human diseases. Here we describe the successful transinfection of A. aegypti with the avirulent wMel strain of Wolbachia, which induces the reproductive phenotype cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, providing optimal phenotypic effects for invasion. Under semi-field conditions, the wMel strain increased from an initial starting frequency of 0.65 to near fixation within a few generations, invading A. aegypti populations at an accelerated rate relative to trials with the wMelPop-CLA strain. We also show that wMel and wMelPop-CLA strains block transmission of dengue serotype 2 (DENV-2) in A. aegypti, forming the basis of a practical approach to dengue suppression.