Bioenergetic analysis of ovarian cancer cell lines: profiling of histological subtypes and identification of a mitochondria-defective cell line.
Epithelial ovarian cancer (EOC) is the most lethal of all gynecological cancers, and encompasses distinct histological subtypes that have specific genetic and tissues-of-origin differences. Ovarian clear cell carcinoma (OCCC) represents approximately 10% of cases and has been termed a stress responsive cancer. OCCC is characterized by increased expression of oxidative stress and glycolysis-related genes. In the present study, we hypothesized that bioenergetic profiling might uniquely distinguish OCCC from other EOC histological subtypes. Using an extracellular flux analyzer, OCCC lines (ES-2, TOV-21-G) were shown to be highly metabolically active, with high oxygen consumption rate (OCR) and high extracellular acidification rate (ECAR), indicative of enhanced mitochondrial oxidative phosphorylation and glycolytic rate, respectively. A high bioenergetics profile was associated with the cell lines' ability to form anchorage independent spheroids. Given their high glycolytic and mitochondrial activity, OCCC cells displayed strong sensitivity to 2-deoxy-D-glucose and Rotenone growth inhibition, although this chemosensitivity profile was not specific to only OCCC cells. Bioenergetic profiling also identified a non-OCCC cell line, OVCA420, to have severely compromised mitochondrial function, based on low OCR and a lack of stimulation of maximal respiration following application of the uncoupler FCCP. This was accompanied by mitochondrial morphology changes indicative of enhanced fission, increased expression of the mitochondrial fission protein Drp1, a loss of mitochondrial membrane potential and dependence on glycolysis. Importantly, this loss of mitochondrial function was accompanied by the inability of OVCA420 cells to cope with hypoxic stress, and a compromised ability to stabilize HIF-1α in response to 1% O2 hypoxia. This knowledge may be imperative for researchers planning to utilize this cell line for further studies of metabolism and hypoxia, and suggests that altered mitochondrial fission dynamics represents a phenotype of a subpopulation of EOCs.


Observed characteristics of suicidal hangings: an 11-year retrospective review.
Many studies have been published regarding suicidal hanging deaths, and most forensic pathologists and coroners are very familiar with such causes of death. Forensic pathologists are challenged over their rulings regarding manner of death in part because the general public has a limited scope of knowledge. One such challenge centers on the question of whether a hanging can be a suicide if the individual is not fully suspended. The authors designed a retrospective study to review suspension in hangings and to analyze other criteria used to help in deciding manner of death. We examined 229 suicidal hanging deaths over an 11-year period (1997 through early 2009) using the data from two separate jurisdictions in Ohio. In conclusion, we found that the vast majority (83.4%) of people who hanged themselves were found partially suspended. Among other criteria analyzed, only the presence of petechial hemorrhages and acute neck injury was statistically significant.


A patient with acute myocardial infarction with acute lower extremity arterial embolization underwent amputation under general anesthesia.
Acute peripheral and coronary artery embolism are common complications of diabetes mellitus and greatly affect the clinical outcome of patients with diabetes; however, there are few reports about the symptoms and prognosis of patients with acute myocardial infarction (AMI) and concurrent acute lower extremity arterial embolism (ALEAE). A 44-year-old man with a history of 4 years of type 1 diabetes was admitted to hospital after suddenly experiencing severe pain in his right lower limb and feeling tightness in the left anterior chest area. Ultrasonography revealed distal occlusion of the right superficial femoral artery, and an electrocardiogram showed acute anterior interstitial myocardial infarction. After conservative treatment for 2 days, the patient had severe necrosis of the lower limbs and secondary injury of multiple organs. Haemodialysis and heparin anticoagulant therapy were performed before amputation. Twelve days after the operation, the patient's condition was stable, and he was transferred out of the intensive care unit. If patients with ALEAE miss the opportunity for early treatment, even with AMI, emergency amputation under general anaesthesia is the right strategy to save lives.


Synovial sarcoma of the spine: A report of three cases and review of the literature.
Synovial sarcoma (SS) is a rare sarcoma with distinct morphologic and genetic features, which, despite its name, does not arise from synovium. While most SSs (>80%) arise in the deep soft tissue of the extremities, up to 5% of these tumors are encountered in the body axis including the spine, mediastinum, retroperitoneum, and head/neck regions. Reports of SS located within the spinal axis have been rare to date. We searched the medical records at our institution and found three patients who were diagnosed and treated for SSs involving the spine. We also performed an exhaustive literature search using PubMed to identify all reported cases in the literature. In this study, we report on three SS cases involving the spine. All three cases involved the paraspinal muscles and spinal nerve roots, with one case having a significant leptomeningeal involvement. In two cases, "smaller operations" were performed first because the lesions were thought to be benign, however, when the final pathology identified them as SSs, more radical procedures were performed. Additionally, we identified 14 cases of SSs involving the spine published in the literature and all cases are reviewed here. Due to limited numbers of cases, spine SS long-term outcomes are hard to quantify. The currently accepted standard of treatment for SSs starts with wide surgical excision with negative margins followed by chemotherapy and radiation. We summarize the available literature on spinal SSs and review the current treatment options available for these tumors.


[Effects of combined radioactive and pesticide loads on the population health].
Relationships of the magnitude of environmental radioactive and pesticide factors to morbidity were studied. Comparative investigations showed that there was a correlation between the above environmental factors. Hygienic preventive measure for health promotion in ecologically poor areas (exposed to radiation or pesticides) were developed.


Phylogenetic distribution of compatible solute synthesis genes support a freshwater origin for cyanobacteria.
Previous work using ancestral state reconstruction of habitat salinity preference proposed that the early cyanobacteria likely lived in a freshwater environment. The aim of this study was to test that hypothesis by performing phylogenetic analyses of the genes underlying salinity preferences in the cyanobacteria. Phylogenetic analysis of compatible solute genes shows that sucrose synthesis genes were likely ancestral in the cyanobacteria, and were also likely inherited during the cyanobacterial endosymbiosis and into the photosynthetic algae and land plants. In addition, the genes for the synthesis of compatible solutes that are necessary for survival in marine and hypersaline environments (such as glucosylglycerol, glucosylglycerate, and glycine betaine) were likely acquired independently high up (i.e., more recently) in the cyanobacterial tree. Because sucrose synthesis is strongly associated with growth in a low salinity environment, this independently supports a freshwater origin for the cyanobacteria. It is also consistent with geologic evidence showing that the early oceans were much warmer and saltier than modern oceans-sucrose synthesis alone would have been insufficient for early cyanobacteria to have colonized early Precambrian oceans that had a higher ionic strength. Indeed, the acquisition of an expanded set of new compatible solute genes may have enabled the historical colonization of marine and hypersaline environments by cyanobacteria, midway through their evolutionary history.


MiR-126 Mediates Brain Endothelial Cell Exosome Treatment-Induced Neurorestorative Effects After Stroke in Type 2 Diabetes Mellitus Mice.
Background and Purpose- Stroke patients with type 2 diabetes mellitus (T2DM) exhibit increased vascular and white matter damage and have worse prognosis compared with nondiabetic stroke patients. We investigated the neurorestorative effects of exosomes derived from mouse brain endothelial cells (EC-Exo) as treatment for stroke in T2DM mice and investigated the role of miR-126 in mediating EC-Exo-derived therapeutic benefits in T2DM-stroke mice. Methods- Adult, male BKS.Cg-m+/+Leprdb/J (T2DM) mice were subjected to photothrombotic stroke model. T2DM mice were intravenously injected at 3 days after stroke with (1) PBS; (2) liposome mimic (vehicle control, 3×1010); (3) EC-Exo (3×1010); (4) knockdown of miR-126 in EC-Exo (miR-126-/- EC-Exo, 3×1010). Behavioral and cognitive tests were performed, and mice were sacrificed at 28 days after stroke. Results- Compared with non-DM stroke mice, T2DM-stroke mice exhibit significantly decreased serum and brain tissue miR-126 expression. Endothelial cells and EC-Exo contain high levels of miR-126 compared with other cell types or exosomes derived from other types of cells, respectively (smooth muscle cells, astrocytes, and marrow stromal cells). Compared with PBS or liposome mimic treatment, EC-Exo treatment of T2DM-stroke mice significantly improves neurological and cognitive function, increases axon density, myelin density, vascular density, arterial diameter, as well as induces M2 macrophage polarization in the ischemic boundary zone. MiR-126-/- EC-Exo treatment significantly decreases miR-126 expression in serum and brain, as well as attentuates EC-Exo treatment-induced functional improvement and does not significantly increase axon and myelin density, vascular density, arterial diameter or induce M2 macrophage polarization in T2DM-stroke mice. In vitro, EC-Exo treatment significantly increases primary cortical neuron axonal outgrowth and increases endothelial capillary tube formation whereas miR-126-/- EC-Exo attentuates EC-Exo induced capillary tube formation and axonal outgrowth. Conclusions- EC-Exo treatment of stroke promotes neurorestorative effects in T2DM mice. MiR-126 may mediate EC-Exo-induced neurorestorative effects in T2DM mice. Visual Overview- An online visual overview is available for this article.


Factorial and construct validity of the revised short form integrative psychotherapy alliance scales for family, couple, and individual therapy.
The Integrative Psychotherapy Alliance model brought an interpersonal and systemic perspective to bear on theory, research, and practice on the psychotherapeutic alliance. Questions have been raised about the independence of the theoretical factors in the model and their operationalization in the Individual, Couple, and Family Therapy Alliance Scales. This paper presents results of a confirmatory factor analysis of the scales that delineated at least three distinct interpersonal factors as well as shorter versions of the three scales to facilitate their use in research and practice. The paper also presents the results of a study testing each factor's association with client retention and progress over the first eight sessions in individual and couple therapy. At least two of the interpersonal factors were uniquely associated with progress in individual and couple functioning. Implications of the results for theory, research, practice, and training in individual, couple, and family therapy are elaborated.


Predictors of failure after fecal microbiota transplantation for recurrent Clostridioides difficile infection: a systematic review and meta-analysis.
Fecal microbiota transplantation (FMT) is a highly effective therapy for recurrent Clostridioides difficile infection (CDI), with ~15% 1-year recurrence rate. Small studies have identified variable risk factors associated with FMT failure. We, therefore, performed a systematic review and meta-analysis to evaluate the predictors of FMT failure. A systematic search of Medline, Embase, and Web of Science was performed from January 2013 up to June 2020. Meta-analyses were performed using random-effects models and pooled adjusted odds ratios for risk factors reported in ≥2 studies were calculated. Overall, 2671 patients with recurrent CDI who underwent FMT in 12 studies were included. FMT failure occurred in 454 patients (16.9%) with median follow-up of 3 months (range 2-7.7 months). A total of 9 risk factors were identified in ≥2 studies. Meta-analysis showed that use of non- CDI antibiotics, presence of inflammatory bowel disease, poor quality of bowel preparation, CDI-related hospitalization before FMT, inpatient FMT, and severe CDI were associated with statistically significant increased risk of failure after FMT. Increasing age, female gender, and immunocompromised status were not associated with increased risk for FMT failure. Several risk factors (both modifiable and non-modifiable) are associated with FMT failure. Lower use of antibiotics in the post-FMT period and good bowel preparation at the time of FMT are associated with lower risk of failure after FMT. Additionally, patients with non-modifiable risk factors should be counseled to be particularly alert about recurrent symptoms after FMT.


Purification and partial characterization of high-molecular-weight forms of ectopic calcitonin from a human bronchial carcinoma cell line.
Studies on the high-molecular-weight immunoreactive calcitonin produced ectopically in culture by an epidermoid bronchial carcinoma cell line are reported. In cell-exposed medium, the principal component has a molecular weight of 40000 and molecules of mol.wts. 13000 and 10000 also occur. Only a trace amount of material co-eluting with 35000-mol.wt. human calcitonin is detectable. None of the calcitonins show cross-reactivity with anti-corticotropin serum. The 40000-mol.wt. immunoreactive calcitonin is readily proteolysed to the 13000- and 10000-mol.wt. components, but the 10000-mol.wt. component behaves as a comparatively stable 'core' molecule. By using immunoprecipitation and high-pressure liquid chromatography (h.p.l.c.), it is possible to prepare radiochemically homogeneous 10000-mol.wt. immunoreactive calcitonin from cells grown in the presence of individual 35S- or 3H-labelled amino acids. Peptide mapping of enzymic digests of this material by h.p.l.c. shows that it contains peptides in common with synthetic human calcitonin.


Implicit co-activation of American Sign Language in deaf readers: An ERP study.
In an implicit phonological priming paradigm, deaf bimodal bilinguals made semantic relatedness decisions for pairs of English words. Half of the semantically unrelated pairs had phonologically related translations in American Sign Language (ASL). As in previous studies with unimodal bilinguals, targets in pairs with phonologically related translations elicited smaller negativities than targets in pairs with phonologically unrelated translations within the N400 window. This suggests that the same lexicosemantic mechanism underlies implicit co-activation of a non-target language, irrespective of language modality. In contrast to unimodal bilingual studies that find no behavioral effects, we observed phonological interference, indicating that bimodal bilinguals may not suppress the non-target language as robustly. Further, there was a subset of bilinguals who were aware of the ASL manipulation (determined by debrief), and they exhibited an effect of ASL phonology in a later time window (700-900ms). Overall, these results indicate modality-independent language co-activation that persists longer for bimodal bilinguals.


Exposure of human peripheral blood mononuclear cells (PBMC) to hydrostatic pressure increases their proliferative response to phytohemagglutinin A (PHA) and anti-CD3 antibody.
In the present study we show that a brief exposure of human PBMC to hydrostatic pressure (HyP) increased their proliferative response to PHA and anti-CD3 antibody, assessed by DNA synthesis. The effect of HyP was most prominent at 400 atmospheres of HyP followed by 600 and 200 atmospheres. At any pressure level, the highest effect of HyP was noted when employing PHA and anti-CD3 antibody at 10(-2) dilution. When PBMC were exposed to 400 atmospheres HyP, maximal effect was achieved at 20 minutes of exposure. The highest effect of HyP on DNA synthesis was noted at 48 and 72 hours of incubation with PHA, when exposing cells to pressure for 20 minutes at 400 atmospheres. Exposure of PBMC under similar conditions for 40 minutes, caused an increase in DNA synthesis only at 48 hours incubation with PHA. These results demonstrate that exposure of human PBMC to HyP increases their proliferative response to different polyclonal activators. The possible mechanisms involved in this phenomenon are discussed.


Effects of glucose starvation on the oxidation of fatty acids by maize root tip mitochondria and peroxisomes: evidence for mitochondrial fatty acid beta-oxidation and acyl-CoA dehydrogenase activity in a higher plant.
Fatty acid beta-oxidation was studied in organellar fractions from maize root tips by h.p.l.c. and radiometric analysis of the products of incubations with [1-14C]octanoate and [1-14C]palmitate. In crude organellar fractions containing both mitochondria and peroxisomes, octanoate and palmitate beta-oxidation, as determined by the production of acetyl-CoA, was functional and, for palmitate, was activated 4-12-fold after subjecting the root tips to 48 h of glucose starvation. The sensitivity to a 'cocktail' of respiratory-chain inhibitors containing cyanide, azide and salicylhydroxamate depended on the conditions of incubation, with no inhibition in a medium facilitating peroxisomal beta-oxidation and a significant inhibition in a medium potentially facilitating mitochondrial beta-oxidation. Indeed, preparations of highly purified mitochondria from glucose-starved root tips were able to oxidize octanoate and palmitate to give organic acids of the tricarboxylic acid cycle. This activity was inhibited 5-10-fold by the above cocktail of respiratory-chain inhibitors, with no parallel accumulation of acetyl-CoA, thus showing that the inhibition affected beta-oxidation rather than the pathway from acetyl-CoA to the organic acids. This provides the first evidence that the complete beta-oxidation pathway from fatty acids to citrate was functional in mitochondria from a higher plant. Moreover, an acyl-CoA dehydrogenase activity was shown to be present in the purified mitochondria. In contrast with the peroxisomal activity, mitochondrial beta-oxidation showed the same efficiency with octanoate and palmitate and was strictly dependent on glucose starvation.


Absence of risk factor change in young adults after family heart attack or stroke: the CARDIA Study.
Health behavior theories suggest that occurrence of heart attack or stroke in an immediate family member should increase one's perceived susceptibility to these conditions, which might lead to improved risk factor behavior and control. Changes in measures of smoking, physical activity, lipids/lipoproteins, body weight, and blood pressure were investigated over two consecutive 5-year follow-up periods among 3950 participants (aged 18 to 30 years) in the Coronary Artery Risk Development in Young Adults (CARDIA) study, who either did or did not have an immediate family member experience a heart attack or stroke. Recruitment and examinations for Years 0, 5, and 10 took place in 1985-1986, 1990-1991, and 1995-1996, respectively. After adjustment for baseline demographics and risk factors, young adults who experienced a change in family history of heart attack or stroke over a 5-year period were no more likely than those who did not to quit smoking, or to experience more positive changes in weight, physical activity, LDL cholesterol, HDL cholesterol, triglycerides, or systolic or diastolic blood pressure. These findings persisted among the few study participants with one or more established risk factors at baseline. The occurrence of a heart attack or stroke in an immediate family member does not appear to lead to self-initiated, sustained change in modifiable risk factors in young adults. Since family history of heart attack and stroke is associated with known risk factors and is an independent risk factor for incident development of these conditions, interventions should be developed and tested to motivate sustained risk-factor control following occurrence of a severe vascular event in a family member.


Phylogenetic analyses of ray-finned fishes (Actinopterygii) using collagen type I protein sequences.
Ray-finned fishes (Actinopterygii) are the largest and most diverse group of vertebrates, comprising over half of all living vertebrate species. Phylogenetic relationships between ray-finned fishes have historically pivoted on the study of morphology, which has notoriously failed to resolve higher order relationships, such as within the percomorphs. More recently, comprehensive genomic analyses have provided further resolution of actinopterygian phylogeny, including higher order relationships. Such analyses are rightfully regarded as the 'gold standard' for phylogenetics. However, DNA retrieval requires modern or well-preserved tissue and is less likely to be preserved in archaeological or fossil specimens. By contrast, some proteins, such as collagen, are phylogenetically informative and can survive into deep time. Here, we test the utility of collagen type I amino acid sequences for phylogenetic estimation of ray-finned fishes. We estimate topology using Bayesian approaches and compare the congruence of our estimated trees with published genomic phylogenies. Furthermore, we apply a Bayesian molecular clock approach and compare estimated divergence dates with previously published genomic clock analyses. Our collagen-derived trees exhibit 77% of node positions as congruent with recent genomic-derived trees, with the majority of discrepancies occurring in higher order node positions, almost exclusively within the Percomorpha. Our molecular clock trees present divergence times that are fairly comparable with genomic-based phylogenetic analyses. We estimate the mean node age of Actinopteri at ∼293 million years (Ma), the base of Teleostei at ∼211 Ma and the radiation of percomorphs beginning at ∼141 Ma (∼350 Ma, ∼250-283 Ma and ∼120-133 Ma in genomic trees, respectively). Finally, we show that the average rate of collagen (I) sequence evolution is 0.9 amino acid substitutions for every million years of divergence, with the α3 (I) sequence evolving the fastest, followed by the α2 (I) chain. This is the quickest rate known for any vertebrate group. We demonstrate that phylogenetic analyses using collagen type I amino acid sequences generate tangible signals for actinopterygians that are highly congruent with recent genomic-level studies. However, there is limited congruence within percomorphs, perhaps due to clade-specific functional constraints acting upon collagen sequences. Our results provide important insights for future phylogenetic analyses incorporating extinct actinopterygian species via collagen (I) sequencing.


Single Viral Load Measurements Overestimate Stable Viral Suppression Among HIV Patients in Care: Clinical and Public Health Implications.
The HIV continuum of care paradigm uses a single viral load test per patient to estimate the prevalence of viral suppression. We compared this single-value approach with approaches that used multiple viral load tests to examine the stability of suppression. The retrospective analysis included HIV patients who had at least 2 viral load tests during a 12-month observation period. We assessed the (1) percent with suppressed viral load (<200 copies/mL) based on a single test during observation, (2) percent with suppressed viral loads on all tests during observation, (3) percent who maintained viral suppression among patients whose first observed viral load was suppressed, and (4) change in viral suppression status comparing first with last measurement occasions. Prevalence ratios compared demographic and clinical subgroups. Of 10,942 patients, 78.5% had a suppressed viral load based on a single test, whereas 65.9% were virally suppressed on all tests during observation. Of patients whose first observed viral load was suppressed, 87.5% were suppressed on all subsequent tests in the next 12 months. More patients exhibited improving status (13.3% went from unsuppressed to suppressed) than worsening status (5.6% went from suppressed to unsuppressed). Stable suppression was less likely among women, younger patients, black patients, those recently diagnosed with HIV, and those who missed ≥1 scheduled clinic visits. Using single viral load measurements overestimated the percent of HIV patients with stable suppressed viral load by 16% (relative difference). Targeted clinical interventions are needed to increase the percent of patients with stable suppression.


Investigation of neurotrophic tyrosine kinase receptor 1 fusions and neurotrophic tyrosine kinase receptor family expression in non-small-cell lung cancer and sensitivity to AZD7451 in vitro.
Advances in the molecular segmentation of lung cancer has raised the possibility that neurotrophic tyrosine kinase receptor (NTRK) 1 fusions and NTRK1-3 expression may be promising molecular targets for future therapeutic interventions. We investigated the antitumor effects of a selective pan-NTRK inhibitor, AZD7451, by evaluating its antiproliferative effects on the KM12 cell line (a colorectal cancer cell line harboring a tropomyosin-NTRK1 fusion) and the H460 and H810 cell lines [large-cell neuroendocrine carcinoma (LCNEC) cell lines expressing NTRK2]. Relative quantitative polymerase chain reaction (qPCR) was performed to measure the mRNA levels of the NTRK1-3 tyrosine kinase domain using cDNA extracted from KM12, H460 and H810 cells. The cultures were grown in 6-well plates at a density of 1.0×106 cell/well and treated with AZD7451 at different doses (1, 2.5, 4, 5, 7.5 and 10 nM) using dimethyl sulfoxide as a control. Following a 24-h incubation, the number of surviving cells was measured using a hemocytometer. Furthermore, we performed western blotting of the high-affinity nerve growth factor receptor (TRKA) and NTRK2 (TRKB) proteins and monitored the effects on the downstream signaling pathways Akt and ERK in these cell lines following treatment with AZD7451 (KM12 and H460: 0, 1 and 5 nM; H810: 0 and 5 nM). Immunohistochemical analyses of the surgically resected samples were also performed, using anti-NTRK1,2 antibodies. We performed reverse-transcription PCR and direct sequencing to investigate NTRK fusions in 268 patients; however, were unable to confirm the presence of NTRK fusions in this cohort. Further immunohistochemical analyses of the primary patient samples demonstrated that none of 61 tumors had NTRK1 overexpression and 7 of 39 samples exhibited NTRK2 expression, including 1 LCNEC sample. The qPCR results from the KM12 cell line revealed an apparent increase and overexpression of NTRK1 mRNA levels, while H460 cells exhibited a modest increase and the H810 cell line showed no apparent increase in the expression of any NTRK1-3 isoforms. There were no increases in the NTRK2 mRNA levels in any of the three cell lines, although KM12 and H460 cells exhibited low levels of NTRK2 expression. In vitro growth and proliferation of the KM12 cell line harboring the NTRK1-fusion was found to be potently inhibited by AZD7451 at a concentration of 2 nM. The proliferation of H460 cells was also found to be inhibited at a concentration of 5 nM, while there was no apparent inhibitory effect of AZD7451 on the growth or proliferation of H810 cells. Western blotting of KM12 cells treated with AZD7451 also revealed a potent inhibition of TRKA phosphorylation following AZD7451 treatment. Analysis of H460 cells confirmed the expression and inhibition of phosphorylation of NTRK2, whereas there was little to no expression of TRKA/B in H810 cells. Subsequent in vitro analysis of cell lines treated with the pan-TRK inhibitor AZD7451 suggested that the proliferation of KM12 and H460 cells was significantly inhibited by AZD7451, while H810 cells expressing low levels of wild-type NTRK1-3 were not inhibited. Based on these results, there is potential for a NTRK-dependent proliferation driver in a subpopulation of lung cancer patients with NTRK expression. In addition, pharmacological inhibition with a NTRK inhibitor, such as AZD7451, in cells harboring NTRK1 fusions, may be associated with beneficial antitumor effects.


Adenosine-induced activation of esophageal nociceptors.
Clinical studies implicate adenosine acting on esophageal nociceptive pathways in the pathogenesis of noncardiac chest pain originating from the esophagus. However, the effect of adenosine on esophageal afferent nerve subtypes is incompletely understood. We addressed the hypothesis that adenosine selectively activates esophageal nociceptors. Whole cell perforated patch-clamp recordings and single-cell RT-PCR analysis were performed on the primary afferent neurons retrogradely labeled from the esophagus in the guinea pig. Extracellular recordings were made from the isolated innervated esophagus. In patch-clamp studies, adenosine evoked activation (inward current) in a majority of putative nociceptive (capsaicin-sensitive) vagal nodose, vagal jugular, and spinal dorsal root ganglia (DRG) neurons innervating the esophagus. Single-cell RT-PCR analysis indicated that the majority of the putative nociceptive (transient receptor potential V1-positive) neurons innervating the esophagus express the adenosine receptors. The neural crest-derived (spinal DRG and vagal jugular) esophageal nociceptors expressed predominantly the adenosine A(1) receptor while the placodes-derived vagal nodose nociceptors expressed the adenosine A(1) and/or A(2A) receptors. Consistent with the studies in the cell bodies, adenosine evoked activation (overt action potential discharge) in esophageal nociceptive nerve terminals. Furthermore, the neural crest-derived jugular nociceptors were activated by the selective A(1) receptor agonist CCPA, and the placodes-derived nodose nociceptors were activated by CCPA and/or the selective adenosine A(2A) receptor CGS-21680. In contrast to esophageal nociceptors, adenosine failed to stimulate the vagal esophageal low-threshold (tension) mechanosensors. We conclude that adenosine selectively activates esophageal nociceptors. Our data indicate that the esophageal neural crest-derived nociceptors can be activated via the adenosine A(1) receptor while the placodes-derived esophageal nociceptors can be activated via A(1) and/or A(2A) receptors. Direct activation of esophageal nociceptors via adenosine receptors may contribute to the symptoms in esophageal diseases.


Ethical considerations in human experimentation.
An accepted practice in medical research has been the subordination of individual rights to the benefit of mankind. The Nazi experiments, and other recent abuses, have focused increased attention on the dignity and rights of human research subjects and have led to new regulations to control human experimentation. Central to establishing what constitutes permissible human experimentation are the issues of risk-benefit ratio and informed consent. Evaluating the risk-benefit ratio requires consideration of the nature of the experiment, the likelihood of its success, and the type of subjects to be utilized. Participation in a study requires voluntary "educated" informed consent. Certain categories of subjects, such as children and the mentally handicapped, require special protection in this regard. Finally, formal education in medical ethics and prohibiting publication of unethical studies are likely to prove more effective in curbing unethical experimentation than increasing government regulation.


Ginkgolide B reduces atherogenesis and vascular inflammation in ApoE(-/-) mice.
To investigate whether ginkgolide B (a platelet-activating factor inhibitor) affects vascular inflammation in atherosclerosis-prone apolipoprotein E-deficient (ApoE(-/-)) mice. Human platelets were used to evaluate the effects of ginkgolide B on platelet aggregation and signal transduction. Ginkgolide B attenuated platelet aggregation and inhibited phosphatidylinositol 3 kinase (PI3K) activation and Akt phosphorylation in thrombin- and collagen-activated platelets. ApoE(-/-) mice were administered a high-cholesterol diet for 8 weeks. Plasma platelet factor 4 (PF4) and RANTES (regulated upon activation, normal T-cell expressed, and secreted protein) were then measured using an enzyme-linked immunosorbent assay. Scanning electron microscopy and immunohistochemistry were used to determine atherosclerotic lesions. Ginkgolide B decreased plasma PF4 and RANTES levels in ApoE(-/-) mice. Scanning electron microscopic examination showed that ginkgolide B reduced aortic plaque in ApoE(-/-) mice. Immunohistochemistry analysis demonstrated that ginkgolide B diminished P-selectin, PF4, RANTES, and CD40L expression in aortic plaque in ApoE(-/-) mice. Moreover, ginkgolide B suppressed macrophage and vascular cell adhesion protein 1 (VCAM-1) expression in aorta lesions in ApoE(-/-) mice. Similar effects were observed in aspirin-treated ApoE(-/-) mice. Ginkgolide B significantly reduced atherosclerotic lesions and P-selectin, PF4, RANTES, and CD40L expression in aortic plaque in ApoE-/- mice. The efficacy of ginkgolide B was similar to aspirin. These results provide direct evidence that ginkgolide B inhibits atherosclerosis, which may be associated with inhibition of the PI3K/Akt pathway in activated platelets.


Role of ribosomal protein RPS2 in controlling let-7a expression in human prostate cancer.
We discovered that an inverse relationship exists in the expression of ras/c-myc and ribosomal protein RPS2 with pre-let-7a-1/let-7a/let-7f miRNA and prostate tumor cell malignancy. Nonmalignant IBC-10a cells expressed low levels of ras/RPS2 and elevated pre-let-7a-1/let-7a/let-7f miRNA, whereas the reverse occurred in malignant PCa-20a and PC-3ML cells. Stable transfection of IBC-10a cells with pBABE.ras and pBABE.RPS2 induced ras, c-myc, and RPS2 expression, whereas the levels of let-7a/let-7f miRNA dropped to near zero. Conversely, in pBABE.pre-let-7a-1 transfected PCa-20a and PC-3ML clones, let-7a/let-7f increased whereas ras, RPS2, and c-myc dropped greater than 5-fold. Electrophoretic mobility shift assays, antibody "supershift" assays and immunoprecipitation assays revealed that RPS2 specifically binds pre-let-7a-1 to block RNA processing. Immunoflourescent studies and Northern blots confirmed that RPS2 complexes with pre-let-7a-1 (i.e., in episomal structures) to block processing to let-7a/let-7f, indicating RPS2 may prevent let-7a miRNA expression to indirectly promote oncogene expression. Functional studies further showed that the colony-forming ability (CFA) and invasive activities of IBC-10a cells were significantly enhanced in pBABE-ras.IBC-10a and pBABE-RPS2-IBC-10a clones. Conversely, with the "knockdown" of ras and RPS2 in malignant PC-3ML cells (i.e., in pLKO.TRC.shRNA.ras.PC3-ML, pLKO.TRC.shRNA.RPS2.PC-3ML transfected cells), there was both a loss of these functions and a loss of tumorigenesis in SCID mice. Likewise, with the overexpression of let-7a/let-7f in pBABE.pre-let-7a-1.PC-3ML clones (and PCa-20a clones), CFAs, invasive activities in vitro, and tumorigenesis in vivo were significantly reduced. These results show for the first time that RPS2 blocks pre-let-7a-1 processing to enable ras and c-myc expression and the transformation of primary tumor cells.


Healing of the guinea pig incisor after partial pulp removal.
A histological and microradiographical study was made of eruption and healing of guinea pig lower incisors after endodontic treatment compared with intact incisors. One incisor per animal was cut perpendicularly at the level of the gingiva. One third to one half of the pulp was removed. The experimental periods were 0, 4, 7, 16, 21 and 31 days. Unstained, HE or Brown and Brenn stained sections were studied microscopically. Some teeth were sectioned undecalcified and studied microradiographically. In intact incisors the dentin matrix deposition and mineralization were described. In the dentin, incisally to the pulp proper, narrow canals, the walls of which consisted of mineralized material, were found opening at the occlusal surface. Histologically, in the treated teeth, a beginning of cell-rich fibrodentin deposition at the original dentin wall was present at Day 4. At Day 7 a substantial walling off of the pulp is achieved by cell-rich and some cell-poor fibrodentin. After 16 days large amounts of cell-rich and cell-poor fibrodentin were covered by some tertiary dentin. After 31 days the amount of tertiary dentin was larger than after 16 days and also covered the incisal part of the root canal walls. Further apically some vasodentin and newly formed normal, secondary, dentin was observed. In course of this period the cells of the cell-rich and cell-poor fibrodentin first showed necrobiosis, then necrosis. The cell-rich fibrodentin appeared not fully mineralized. All blood vessels and strands of soft tissue which ran in canals in the fibrodentin came to a dead end. Thus the root canal was fully obturated. Although the teeth appeared (nearly) normal to the naked eye and radiographically after 31 days, histologically they were not. Large amounts of fibrodentin still being present incisally which showed the dual potential of the guinea pig pulp to produce both fibrodentin and orthodentin.


Evaluation of the role of in vivo proteolysis (fibrinogenolysis) in prolonging the thrombin time of human umbilical cord fibrinogen.
These studies have been directed at evaluating the role played by proteolysis (fibrinogenolysis) in vivo in prolonging the thrombin time of human umbilical cord ("fetal") fibrinogen. The aggregation rate of cord fibrin compared with that from adult plasma is always delayed when the reaction is carried out under conditions of relatively high ionic strength (e.g., 0.29); this difference is not apparent at relatively low ionic strength (e.g., 0.09). In addition, as assessed by turbidimetric techniques, the maximum absorbance attained by cord fibrin is considerably less than that attained by adult fibrin. Coagulable fibrinogen catabolites (i.e., fraction I-5) are present in cord plasma and, like their counterparts from adult plasma, lack various portions of the COOH-terminal region of the A alpha chain. However, their presence in plasma does not explain the behavioral differences between cord and adult fibrin. Moreover, differences revealed by turbidimetric comparison of cord and adult fibrin from plasma fraction I-2 persist in fibrin from fraction I-5; it therefore appears that the COOH-terminal region of the A alpha chain does not contain the structure(s) accounting for the unique behavior of "fetal" fibrinogen.


Outcomes of Selective Arthrodesis Based on Joints Affected in 33 Feet with Müller-Weiss Disease.
No consensus exists regarding operative treatment of Müller-Weiss disease (MWD). Its only classification is based solely on Méary's angle and serves neither as guide to management nor prognosis. We report on 33 feet that underwent surgery following failed conservative management. Treatment was directed towards joint(s) involved, as determined by clinical examination, plain radiography and SPECT-CT. Thus, surgery consisted of isolated talonavicular in 6 feet, triple in 8, subtalar and talonavicular in 7, talonaviculocuneiform in 4, talonaviculocuneiform with interpositional tricortical iliac crest graft in 6 and pantalar arthrodesis in 2. PROMIS scores for pain interference and depression decreased significantly (P < 0.001) with significant accompanying increase in physical function (P = 0.003). Union occurred in 31 of 33 feet (94%) with complete resolution of pain at an average follow-up of 84 months. Of the 2 non-unions, one had fracture through the lateral navicular, and the other marked sclerosis and avascularity of the lateral navicular. We describe our pathways for selecting arthrodesis based on the joints affected. Isolated talonavicular arthrodesis was performed in early stages of MWD, which begins at the talonavicular articulation. When disease extended to both sides of the navicular, we performed talonaviculocuneiform arthrodesis. When considering isolated talonavicular, double medial or triple arthrodesis, there should be adequate cancellous bone stock remaining in the lateral part of the navicular, as determined on medial oblique radiographs and CT scan. In case of inadequate bone stock or fracture through the lateral navicular, talonaviculocuneiform arthrodesis with interpositional iliac crest bone graft is recommended.


Pooling samples: the key to sensitive, specific and cost-effective genetic diagnosis of Chlamydia trachomatis in low-resource countries.
The aims of this study were to compare the performance characteristics and cost-effectiveness of pooling endocervical samples for screening and diagnosis of Chlamydia trachomatis, and to investigate the prevalence of C. trachomatis infection in women in Leningrad Oblast, Russia. A total of 1500 endocervical samples were tested individually and when pooled in groups of 5 and 10 samples, respectively. A previously evaluated in-house diagnostic polymerase chain reaction (PCR) assay was utilized. The sensitivity and specificity of the PCR were not affected by either pooling strategy. The estimated prevalence of genital C. trachomatis infection was 6.6%, 6.1% and 6.0% based on individually tested samples, and pools of 5 and 10, respectively. For diagnosis of individual samples, the pooling strategies resulted in cost savings of 53.3% (5 samples per pool) and 44.0% (10 samples per pool). Pooling samples for PCR detection of C. trachomatis is an accurate and cost-saving approach for diagnosis and large-scale prevalence studies in St Petersburg, Russia.


Conformational Dynamics of Poly(T) Single-Stranded DNA at the Single-Molecule Level.
The conformational dynamics of single-stranded DNA (ss-DNA) are implicated in the mechanisms of several key biological processes such as DNA replication and damage repair and have been modeled with those of semiflexible or flexible polymer. The high flexibility and customizability of ss-DNA also make it an excellent polymeric material for materials engineering. Polythymidine (poly(T)) is an excellent model ss-DNA as a flexible polymer since it does not form any secondary structure. However, only limited experimental results have been reported of poly(T) conformational dynamics with a very short length that is not relevant to the aforementioned processes and applications. Here, we provide the first experimental results of the conformational dynamics of poly(T) with lengths in the range of 130-170 nucleotides at the single-molecule level. Our experiments are based on single-molecule FRET and a DNA hairpin structure of which the folding kinetics are governed by the conformational dynamics of poly(T). We found that the folding kinetics deviate far from those of a flexible polymer model with a harmonic bending potential. To this end, we derived a simple model for the kinetics of DNA hairpin folding from the self-avoiding-walk (SAW). Our model describes the conformational dynamics of poly(T) very well and enables estimation of the conformational dimensionality. The estimated dimensionalities suggest that ss-DNA is completely flexible at 100 mM or a higher NaCl concentration, but not at 50 mM. These results will help understand the conformational dynamics of ss-DNA implicated in several key biological processes and maximize the utility of ss-DNA for materials engineering. Also, our system and model provide an excellent platform to investigate the conformational dynamics of ss-DNA.


Controlled Payload Release by Magnetic Field Triggered Neural Stem Cell Destruction for Malignant Glioma Treatment.
Stem cells have recently garnered attention as drug and particle carriers to sites of tumors, due to their natural ability to track to the site of interest. Specifically, neural stem cells (NSCs) have demonstrated to be a promising candidate for delivering therapeutics to malignant glioma, a primary brain tumor that is not curable by current treatments, and inevitably fatal. In this article, we demonstrate that NSCs are able to internalize 2 μm magnetic discs (SD), without affecting the health of the cells. The SD can then be remotely triggered in an applied 1 T rotating magnetic field to deliver a payload. Furthermore, we use this NSC-SD delivery system to deliver the SD themselves as a therapeutic agent to mechanically destroy glioma cells. NSCs were incubated with the SD overnight before treatment with a 1T rotating magnetic field to trigger the SD release. The potential timed release effects of the magnetic particles were tested with migration assays, confocal microscopy and immunohistochemistry for apoptosis. After the magnetic field triggered SD release, glioma cells were added and allowed to internalize the particles. Once internalized, another dose of the magnetic field treatment was administered to trigger mechanically induced apoptotic cell death of the glioma cells by the rotating SD. We are able to determine that NSC-SD and magnetic field treatment can achieve over 50% glioma cell death when loaded at 50 SD/cell, making this a promising therapeutic for the treatment of glioma.


Substrate-mediated stabilization of a tetrameric drug target reveals Achilles heel in anthrax.
Bacillus anthracis is a gram-positive spore-forming bacterium that causes anthrax. With the increased threat of anthrax in biowarfare, there is an urgent need to characterize new antimicrobial targets from B. anthracis. One such target is dihydrodipicolinate synthase (DHDPS), which catalyzes the committed step in the pathway yielding meso-diaminopimelate and lysine. In this study, we employed CD spectroscopy to demonstrate that the thermostability of DHDPS from B. anthracis (Ba-DHDPS) is significantly enhanced in the presence of the substrate, pyruvate. Analytical ultracentrifugation studies show that the tetramer-dimer dissociation constant of the enzyme is 3-fold tighter in the presence of pyruvate compared with the apo form. To examine the significance of this substrate-mediated stabilization phenomenon, a dimeric mutant of Ba-DHDPS (L170E/G191E) was generated and shown to have markedly reduced activity compared with the wild-type tetramer. This demonstrates that the substrate, pyruvate, stabilizes the active form of the enzyme. We next determined the high resolution (2.15 A) crystal structure of Ba-DHDPS in complex with pyruvate (3HIJ) and compared this to the apo structure (1XL9). Structural analyses show that there is a significant (91 A(2)) increase in buried surface area at the tetramerization interface of the pyruvate-bound structure. This study describes a new mechanism for stabilization of the active oligomeric form of an antibiotic target from B. anthracis and reveals an "Achilles heel" that can be exploited in structure-based drug design.


How do aging and age-related hearing loss affect the ability to communicate effectively in challenging communicative conditions?
This study investigated the relation between the intelligibility of conversational and clear speech produced by older and younger adults and (a) the acoustic profile of their speech (b) communication effectiveness. Speech samples from 30 talkers from the elderLUCID corpus were used: 10 young adults (YA), 10 older adults with normal hearing (OANH) and 10 older adults with presbycusis (OAHL). Samples were extracted from recordings made while participants completed a problem-solving cooperative task (diapix) with a conversational partner who could either hear them easily (NORM) or via a simulated hearing loss (HLS), which led talkers to naturally adopt a clear speaking style. In speech-in-noise listening experiments involving 21 young adult listeners, speech samples by OANH and OAHL were rated and perceived as less intelligible than those of YA talkers. HLS samples were more intelligible than NORM samples, with greater improvements in intelligibility across conditions seen for OA speech. The presence of presbycusis affected (a) the clear speech strategies adopted by OAHL talkers and (b) task effectiveness: OAHL talkers showed some adaptations consistent with an increase in vocal effort, and it took them significantly longer than the YA group to complete the diapix task. The relative energy in the 1-3 kHz frequency region of the long-term average spectrum was the feature that best predicted: (a) the intelligibility of speech samples, and (b) task transaction time in the HLS condition. Overall, our study suggests that spontaneous speech produced by older adults is less intelligible in babble noise, probably due to less energy present in the 1-3 kHz frequency range rich in acoustic cues. Even mild presbycusis in 'healthy aged' adults can affect the dynamic adaptations in speech that are beneficial for effective communication.


Long-latency auditory-evoked potentials in severe traumatic brain injury.
To detect the effects of different deviant stimuli on long-latency auditory-evoked potentials (LLAEPs) in patients with severe impairment of consciousness from traumatic brain injury (TBI) and to define their prognostic value for late functional outcome. Correlational study on a prospective cohort. Brain injury rehabilitation center. Eleven volunteers and 21 consecutively sampled patients with severe TBI referred to the inpatient intensive rehabilitation unit of primary care in a university-based system. The LLAEPs recorded with different paradigms; and the Glasgow Outcome Scale (GOS), Disability Rating Scale (DRS), FIMtrade mark instrument, and Neurobehavioural Rating Scale (NBHRS). N100-P150 complex showed high reliability. Patients with good outcomes showed N100 and P150 mean latencies similar to those of unimpaired patients and shorter than patients with unfavorable outcomes. When the deviant stimulus was the patient's name, N100 latency showed high correlations with DRS (p <.007), FIM (p <.01), and NBHRS (p <.009). P250 and P300 showed a low percentage of occurrence with passive paradigms in both patients and controls. Their scores were inversely correlated to the Glasgow Coma Scale (p <.03) and the Innsbruck Coma Scale (p <.003), but no significant correlations were found with functional and behavioral outcomes. Patients with GOS score 1-2 1 year posttrauma had significantly longer latency and lower amplitude of N100 and P150 than those with GOS score 4-5. LLAEPs can be recorded in patients with severe impairment of consciousness by means of passive paradigms. The use of a stimulus that is relevant for the patient can enhance the accuracy of the test and its relationship with functional outcome.


Opportunity costs of gastrointestinal endoscopic training in Canada.
No data exist to define the opportunity costs related to instruction in endoscopic procedures in Royal College of Physicians and Surgeons of Canada-accredited teaching centres. Academic and institutional administrators expect staff to achieve acceptable performance standards. There is a need to measure some of the effects of training activity in the establishment of such standards. To measure the effect of resident training in colonoscopy on real procedure times and, as a secondary goal, to estimate procedural losses related to the process of training. Real procedure times for ambulatory colonoscopy in a single academic, hospital-based endoscopy unit were documented. Times for certified endoscopy instructors functioning solo were compared with times for procedures involving trainees at several levels of colonoscopic experience. Procedural reductions associated with resident training were estimated based on the parameters derived from the results. The analysis was executed retrospectively using prospectively collected data. Resident training prolonged procedure times for ambulatory colonoscopy by 50%. The trainee effect was consistent, although variable in degree, among a variety of endoscopy instructors. Such increased procedure times have the potential to reduce case throughput and endoscopist remuneration. Resident training in colonoscopy in a Canadian certified training program has significant negative effects on case throughput and endoscopist billings. These factors should be considered in any assessment of performance in similar training environments.


[Clinical problems of extended intestinal resections].
Extended intestinal resections constitute a primary surgical problem, and in numerous cases there is the necessity of programming an appropriate nutrition, including parenteral nutrition at patient's home. The aim of the work was to examine treatment problems: surgery indications, complications and the after-effects of extended intestinal resections as well as the procedure rules. The group of 51 patients with extended intestinal resections who were subjected to treatment in the years 1988-2002. The group was comprised of 25 female patients with the average age 64.2 years (34-86) and 26 male patients at the age of 56.3 (21-72). Extended intestinal resections resulted from: cancer--30, ischemia (embolism, thrombosis)--12, injury--3, inflammatory bowel diseases--2, intestinal polyposis--2, diverticulosis with hemorrhages--1, toxic necrosis--1.9 deaths were recorded within the post-operative period: 7 (that is 7 of 12) due to ischemia, 1 due to cancer, 1 due to colon toxicum. The overall mortality rate was 17% (that is 9 deaths of all the 51 patients); the highest rate reached as much as 58.3% in the ischemia cases. The most frequent reason of the deaths following extended intestinal resections performed due to ischemia of intestine was: the impossibility of hemodynamic stabilization--3, the escalation of intestinal ischemia and septic shock--2, simultaneously both causes--2. The average length of time between admittance to hospital and surgical intervention was 5.1 days (0-31); the average length of stay in hospital amounted to 20.8 days (2-102). Extended intestinal resections were the most frequently performed due to neoplastic or vascular reasons. Such surgical interventions are fraught with a high risk of complications and deaths. Furthermore, the foregoing surgeries demand a very expensive therapeutic procedure including the post-operative parenteral nutrition.


In situ distal saphenous vein bypass using the intraluminal valve-disruption technique.
In the last few years there has been a resurgence of interest in in situ saphenous vein bypass for lower extremity revascularization because of improved patency rates. We performed 28 in situ bypass operations in 26 patients with threatened limbs using the intraluminal Hall valve disrupter. Seventy-five percent of these bypasses were to tibial vessels and had a 93% early patency rate. Three late failures were salvaged before thrombosis of the bypass, resulting in a cumulative patency rate of 93% with a mean follow-up of 17 months. In situ saphenous vein bypass has become our procedure of choice for distal reconstruction in severely ischemic limbs because of improved long-term patency compared with reversed-saphenous vein bypass.


[Bone marrow cells transplant in the treatment of chronic chagasic cardiomyopathy].
Chronic chagasic cardiomyopathy remains a major cause of death due to heart failure in Latin American countries and for which there is currently no effective treatment. While chagasic patients wait for the development of more efficient and less toxic chemotherapeutics for the elimination of Trypanosoma cruzi, a new strategy has appeared in an attempt to repair or ameliorate the damage caused to the myocardium of patients with chronic chagasic cardiomyopathy. This therapy, involving the transplant of bone marrow cells obtained from the patient to be treated, may lead to improvement in heart function and in life quality of patients with severe chagasic cardiopathy, similar to that obtained with this approach in the treatment of patients with heart failure of ischemic etiology. The possible effects of cell therapies and its applications in chronic chagasic cardiopaths are discussed in the present report.


Evaluation of novel nutraceuticals based on the combination of oat beta-glucans and a green coffee phenolic extract to combat obesity and its comorbidities. A randomized, dose-response, parallel trial.
Obesity and its associated comorbidities are a major public health concern worldwide. Reduced energy intake and increased physical activity interventions have limited success in the long term. Nutraceuticals might be an alternative means to help lose weight and reduce obesity-associated cardiometabolic risk factors without changes in the habitual diet. The objective of the present study was to comparatively evaluate the efficiency of nutraceuticals based on the combination of a decaffeinated green coffee bean extract (GCBE) and two types of oat beta-glucans (BG) with different physiochemical properties on obesity related biomarkers in overweight/obese subjects. A randomized, dose-response, parallel, blind study was carried out in four groups of subjects (n = 15 each) who consumed for 6 weeks, twice a day, a nutraceutical containing 3 g d-1 or 5 g d-1 doses of 35% or 70% BG and a fixed amount of GCBE providing 600 mg d-1 of phenols. 35% BG showed a 10 and 100 times higher molecular weight and viscosity, respectively, compared to 70% BG. Food intake, anthropometry and different cardiometabolic markers were assessed at the beginning and end of the intervention. According to the general model of variance with repeated measure analysis, the intervention caused positive changes in the levels of total cholesterol, LDL cholesterol, VLDL cholesterol, triglycerides, alanine aminotransferase, aspartate aminotransferase, haemoglobin A1c, insulin, systolic blood pressure (SBP), total body fat percentage (TBF%), visceral fat percentage, and waist and hip circumferences without differences among the treatments, except for SBP and TBF%. Looking into the rates of change [(end value - beginning value)/beginning value] of these parameters, 5 g - 70% BG was the treatment that lowered TBF% the most. In conclusion, 5 g - 70% BG may be more effective in helping to lose weight and additionally, it produced the least bloating according to participants' subjective perception.


Description of Cellulophaga algicola sp. nov., isolated from the surfaces of Antarctic algae, and reclassification of Cytophaga uliginosa (ZoBell and Upham 1944) Reichenbach 1989 as Cellulophaga uliginosa comb. nov.
A group of strains with potent extracellular enzymic activity were isolated from the surfaces of the chain-forming sea-ice diatom Melosira and from an unidentified macrophyte collected from the Eastern Antarctic coastal zone. 16S rDNA sequence analysis indicated that the strains belonged to the genus Cellulophaga and showed greatest similarity to the species Cellulophaga baltica (sequence similarity 97%). Phenotypic characteristics, DNA base composition and DNA-DNA hybridization values clearly separate the Antarctic strains from Cellulophaga baltica and other Cellulophaga species. Thus, the strains form a distinct and novel species and have the proposed name Cellulophaga algicola sp. nov. (type strain IC166T = ACAM 630T). In addition, it was recognized that the species Cytophaga uliginosa (ZoBell and Upham 1944) Reichenbach 1989, a species phylogenetically remote from the type species of the genus Cytophaga, possessed 16S rDNA sequences and phenotypic and chemotaxonomic traits similar to those of other Cellulophaga species. Thus, it was proposed that the species Cytophaga uliginosa be renamed as Cellulophaga uliginosa comb. nov.


Incidence of exercise-induced bronchospasm in Olympic winter sport athletes.
The purpose of this project was to determine the incidence of exercise-induced bronchospasm (EIB) among U.S. Olympic winter sport athletes. Subjects included female and male members of the 1998 U.S. Winter Olympic Team from the following sports: biathlon, cross-country ski, figure skating, ice hockey, Nordic combined, long-track speedskating, and short-track speedskating. Assessment of EIB was conducted in conjunction with an "actual competition" (Olympic Trials, World Team Trials, World Cup Event, U.S. National Championships) or a "simulated competition" (time trial, game), which served as the exercise challenge. Standard spirometry tests were performed preexercise and at 5, 10, and 15 min postexercise. An athlete was considered EIB-positive based on a postexercise decrement in FEV1 > or = 10%. For the seven sports evaluated on the 1998 U.S. Winter Olympic Team, the overall incidence of EIB across all sports and genders was 23%. The highest incidence of EIB was found in cross-country skiers, where 50% of the athletes (female = 57%; male = 43%) were diagnosed with EIB. Across the seven sports evaluated, the prevalence of EIB among the female and male athletes was 26% and 18%, respectively. Among those individuals found to be EIB-positive were athletes who won a team gold medal, one individual silver medal, and one individual bronze medal at the Nagano Winter Olympics. These data suggest that: 1) EIB is prevalent in several Olympic winter sports and affects nearly one of every four elite winter sport athletes; 2) the winter sport with the highest incidence of EIB is cross-country skiing; 3) in general, EIB is more prevalent in female versus male elite winter sport athletes; and 4) athletes may compete successfully at the international level despite having EIB.


The prevalence of xerostomia and salivary gland hypofunction in a cohort of HIV-positive and at-risk women.
The association of xerostomia and salivary gland hypofunction with HIV infection has been established for men but not for women. We investigated the prevalence of these conditions in a national cohort (n = 733) of HIV-positive and at-risk HIV-negative women. Participants in this prospective cross-sectional study were recruited from the Women's Interagency HIV Study (WIHS) at five outpatient USA clinics. Xerostomia was assessed based on "yes" responses to a dry-mouth questionnaire. Samples of unstimulated whole and chewing-stimulated whole saliva were collected under standardized conditions. The major salivary glands were also evaluated clinically. The prevalence of dry-mouth complaint, the absence of saliva upon palpation, and zero unstimulated whole saliva (flow rate = 0 mL/min) were significantly (p = 0.001) higher in HIV-positive women. Adjusted odds of zero unstimulated whole saliva were significantly (p = 0.02) higher in HIV-positive women vs. HIV-negative women (OR = 2.86; 95% CI, 1.23 to 6.63). Significant (p = 0.03) univariate association was found between zero unstimulated whole saliva and CD4 counts. Adjusted odds of zero unstimulated whole saliva were significantly (p = 0.02) higher for HIV-positive women with CD4 < 200 compared with those with CD4 > 500 (OR = 2.61; 95% CI, 1.17 to 5.85). Chewing-stimulated flow rates were not significantly different between seropositive and seronegative women. The prevalence of xerostomia and salivary gland hypofunction appears to be significantly higher in HIV-positive women relative to a comparable group of at-risk seronegative women. Immunosuppression levels measured by CD4 cell counts are significantly associated with xerostomia and salivary gland hypofunction in a population of HIV-positive women.


The Use of Vitamin K2 in Patients With Parkinson's Disease and Mitochondrial Dysfunction (PD-K2): A Theranostic Pilot Study in a Placebo-Controlled Parallel Group Design.
Background: Despite rapid advances in research on Parkinson's disease (PD), in particular in the elucidation of genetic contributions, no disease-modifying therapy has become available to date. Objectives: In the proposed project, we aim to investigate the potential effects of vitamin K2 (long-chain menaquinone 7, MK-7) in genetically determined PD with mitochondrial dysfunction. Methods: A total of 130 study participants (26 biallelic Parkin/PINK1 mutation carriers, 52 sporadic PD patients, and 52 healthy controls) will receive the trial medication (MK-7 or placebo for 1 week). 31P-Magnetic resonance spectroscopy imaging of the forebrain and basal ganglia (31P-MRSI, primary endpoint) as well as other advanced neuroimaging methods, clinical assessment, including quantitative movement analysis, and biomarker sampling will be applied pre- and post-intervention. Innovation: The proposed project is highly translational as it builds on compelling mechanistic data from animal studies as well as on a small preliminary data set in humans. Patients are selected based on their mutation-related mitochondrial dysfunction and compared to disease and a healthy control group in a personalized medicine approach. We will further investigate how neuroimaging and blood-derived biomarkers can predict individual treatment response in sporadic PD. Clinical trial registration: This study was registered at the German Clinical Trial Registry (DRKS, DRKS00019932) on the 19th of December 2019.


Navigating and negotiating: Social processes to describe and explain the student experience of undertaking part-time, taught study on a Master's level degree course in dementia care.
Post graduate education in dementia care is delivered by a number of academic institutions worldwide. Aimed primarily at people in advanced and specialised roles in health and social care, it is designed to develop the knowledge and skills to advance practice and lead to positive changes and benefits to people living with dementia and their social networks. However, undertaking Master's level education can be challenging, particularly when this is undertaken alongside full-time work and family/domestic commitments. To illuminate this student experience further, this paper takes a case study approach to explore the experiences of three students who have successfully completed a Master's programme in dementia care in the North West of England. Their individual and collective experiences highlight the challenges of juggling competing demands and the dynamic processes of navigating the student journey and continually negotiating supports in order to successfully complete the course. The study highlights important implications for the design and supportive structures within post graduate Master's education in dementia care/studies, and potentially in other subject areas also, in order to support students' transition to post graduate study.


Regulation of cytochrome P-450c by glucocorticoids and polycyclic aromatic hydrocarbons in cultured fetal rat hepatocytes.
The actions of polycyclic aromatic hydrocarbons and glucocorticoids to regulate the synthesis of cytochrome P-450c (the major isozyme induced by polycyclic aromatic hydrocarbons) were investigated in fetal rat hepatocytes maintained in primary monolayer culture. Treatment of hepatocytes in culture with 1,2-benzanthracene resulted in a 50-fold increase in 7-ethoxycoumarin O-deethylase activity. The level of P-450c increased in the cells in a time-dependent fashion as determined by immunoelectrophoretic analysis. The inductive effect of BA was potentiated approximately 1.6- to 2.3-fold when 1 microM dexamethasone was included in the culture medium. However, dexamethasone alone had little or no effect on the induction of P-450c. The rate of synthesis of P-450c was examined by immunoisolation of the specific isozyme from total cellular proteins radiolabeled with [35S]methionine and from the protein products formed during in vitro translation of the isolated mRNA. In addition, the amount of mRNA specific for cytochrome P-450c was determined by Northern blot analysis of RNA extracted from cultured cells. The changes in the rates of synthesis and mRNA levels were found to parallel the changes in enzyme activity. The concentration of dexamethasone required to cause a half-maximal increase in P-450c content in the presence of 1,2-benzanthracene was between 10(-8) and 10(-7) M. It is concluded that glucocorticoids act synergistically with polycyclic aromatic hydrocarbons to increase the levels of P-450c expressed in the fetal rat liver, and that this action is likely mediated by the classical type II glucocorticoid receptor.


The development of the characteristic anomalies found in gastroschisis--experimental and clinical data.
Gastroschisis was investigated experimentally as well as clinically. An experimental model developed in the chick embryo demonstrated that the characteristic picture of gastroschisis evolved only if the herniated bowel was exposed to urine components in the allantoic fluid. Remarkable similarities were revealed on comparing changes in human amniotic fluid composition (from 30th week) with changes in chicken allantoic fluid composition (from 15th day of incubation). These changes correlate with progressive fibrotic coating of the exposed bowel loops leading to the characteristic picture of gastroschisis. No fibrous coating was found in human foetuses with a gestational age under 30 weeks, while investigation of these foetuses did confirm that gastroschisis itself occurs at an early developmental stage (6th-8th week). No primary structural defects were found in the nervous system of the bowel wall neither in the experimental gastroschisis model nor in the human cases investigated. The postoperative delay in intestinal motility affecting some gastroschisis patients was found to be secondary to multifocal ischaemic damage of the bowel wall.


Hypertension and disrupted blood pressure circadian rhythm in type 2 diabetic db/db mice.
Human Type 2 diabetes is associated with increased incidence of hypertension and disrupted blood pressure (BP) circadian rhythm. Db/db mice have been used extensively as a model of Type 2 diabetes, but their BP is not well characterized. In this study, we used radiotelemetry to define BP and the circadian rhythm in db/db mice. We found that the systolic, diastolic, and mean arterial pressures were each significantly increased by 11, 8, and 9 mmHg in db/db mice compared with controls. In contrast, no difference was observed in pulse pressure or heart rate. Interestingly, both the length of time db/db mice were active (locomotor) and the intensity of locomotor activity were significantly decreased in db/db mice. In contrast to controls, the 12-h light period average BP in db/db mice did not dip significantly from the 12-h dark period. A partial Fourier analysis of the continuous 72-h BP data revealed that the power and the amplitude of the 24-h period length rhythm were significantly decreased in db/db mice compared with the controls. The acrophase was centered at 0141 in control mice, but became scattered from 1805 to 0236 in db/db mice. In addition to BP, the circadian rhythms of heart rate and locomotor activity were also disrupted in db/db mice. The mean arterial pressure during the light period correlates with plasma glucose, insulin, and body weight. Moreover, the oscillations of the clock genes DBP and Bmal1 but not Per1 were significantly dampened in db/db mouse aorta compared with controls. In summary, our data show that db/db mice are hypertensive with a disrupted BP, heart rate, and locomotor circadian rhythm. Such changes are associated with dampened oscillations of clock genes DBP and Bmal1 in vasculature.


Vitamin A in regulation of insulin responsiveness: mini review.
Vitamin A, retinol, circulates in blood bound to retinol-binding protein (RBP4) which, in turn, associates with another serum protein, transthyretin (TTR), to form a ternary retinol-RBP4-TTR complex. At some tissues, retinol-bound (holo-) RBP4 is recognised by a receptor termed stimulated by retinoic acid 6 (STRA6) which transports retinol into cells. This mini-review summarises evidence demonstrating that, in addition to functioning as a retinol transporter, STRA6 is also a signalling receptor which is activated by holo-RBP4. The data show that STRA6-mediated retinol transport induces receptor phosphorylation, in turn activating a Janus kinases2/signal transducers and activators of transcription (STAT)3/5 cascade that culminates in induction of STAT target genes. STRA6-mediated retinol transport and cell signalling are inter-dependent, and both functions critically rely on intracellular retinol trafficking and metabolism. Hence, STRA6 couples 'sensing' of vitamin A homeostasis and metabolism to cell signalling, allowing it to control important biological functions. For example, by inducing the expression of the STAT target gene suppressor of cytokine signalling 3, STRA6 potently suppresses insulin responses. These observations provide a rationale for understanding the reports that elevation in serum levels of RBP4, often observed in obese mice and human subjects, causes insulin resistance. The observations indicate that the holo-RBP4 /STRA6 signalling cascade may comprise an important link through which obesity leads to insulin resistance and suggest that the pathway may be a novel target for treatment of metabolic diseases.


The biology of Schistosoma japonicum, Philippine strain: a review.
Some important and adequately studied aspects of the biology of Schistosoma japonicum in the Philippines were reviewed and presented. The identity of the species of human blood fluke in the Philippines was established by Tubangui who also was the first to complete the life cycle of the parasite in his laboratory. Successful cross-infections of different strains of S. Japonicum from different mammalian hosts including man suggest that there is only one common strain for all of the definitive hosts in the Philippines. Studies on egg-laying habits, factors affecting hatching of eggs, eruption of cercariae from snails, cercarial survival and undocumented obervations are considered in this review.


DNA bridging and looping by HMO1 provides a mechanism for stabilizing nucleosome-free chromatin.
The regulation of chromatin structure in eukaryotic cells involves abundant architectural factors such as high mobility group B (HMGB) proteins. It is not understood how these factors control the interplay between genome accessibility and compaction. In vivo, HMO1 binds the promoter and coding regions of most ribosomal RNA genes, facilitating transcription and possibly stabilizing chromatin in the absence of histones. To understand how HMO1 performs these functions, we combine single molecule stretching and atomic force microscopy (AFM). By stretching HMO1-bound DNA, we demonstrate a hierarchical organization of interactions, in which HMO1 initially compacts DNA on a timescale of seconds, followed by bridge formation and stabilization of DNA loops on a timescale of minutes. AFM experiments demonstrate DNA bridging between strands as well as looping by HMO1. Our results support a model in which HMO1 maintains the stability of nucleosome-free chromatin regions by forming complex and dynamic DNA structures mediated by protein-protein interactions.


[Staging and neoadjuvant therapy of squamous cell carcinoma of esophagus].
Once the diagnosis of esophageal cancer is established, the decision on treatment will depend on the stage of the disease. Since improvement of prognosis can only be expected in patients with complete removal of their tumor, preoperative staging plays a pivotal role in the decision-making process. Preoperative diagnostic procedures should define the tumor in its relation to the tracheal bifurcation (site), determine the depth of tumor invasion (T status), evaluate regional lymph node metastases (N1 disease) and exclude distant metastases (M1 disease). Endosonography represents currently the most accurate imaging technique for detecting the correct T stage over the correct N stage. A higher accuracy rate may be achieved by combining endosonography with other staging modalities such as computed tomography. Chest x-ray, and percutaneous ultrasonography (abdominal, neck) form the diagnostic basis in staging M1 disease. Computed tomography (neck, chest and abdomen) is currently the best method to detect metastases in the liver and in celiac nodes. Staging laparoscopy when combined with laparoscopic ultrasonography shows a higher sensitivity than ultrasonography and computed tomography in the diagnosis of smaller metastases and peritoneal seedings. En bloc esophagectomy together with the regional lymph nodes remains the treatment of choice in medically fit patients with localized esophageal carcinoma (Stage I-IIB, T1-T2/N0-N1/M0). Due to early involvement of mediastinal structures, curative resection is unlikely to be achieved in patients with locally advanced esophageal carcinoma (Stage III, T3-T4/N0-N1/M0). Most available data indicate that neoadjuvant radiochemotherapy leads in a significant number of patients to downstaging of the tumor, increases the rate of R0 resection, improves local tumor control, and prolongs the recurrence free interval. However, neoadjuvant radiochemotherapy resulted in a marked increase of morbidity and postoperative mortality without improvement of survival. At present, neoadjuvant therapy is still experimental and there is no consensus for an optimal treatment regimen. Its use outside of an investigational setting can not be recommended. Future research must focus on more effective and less toxic neoadjuvant modalities (e.g. new chemotherapy agents, hyperthermia).


Inositol-specific phospholipase C in low and fast proliferating hepatoma cell lines.
Inositol lipid cycle, among the pletora of signalling events, is directly involved in cell growth. It is located both in the cytoplasm and in the nucleus. Disturbances may cause uncontrolled proliferation of the cell and ultimately cancer. The phosphatidyl inositol phospolipase C (PLC) is a key enzyme in the hydrolysis of polyphosphoinositides (PIs) and could be differently involved in the normal and pathological cell growth. We report immunochemical and immunocytochemical demonstrations that the PLC isoforms are present in both cytoplasmic and nuclear compartments of low and fast proliferating hepatoma cells. The PLC activity is increased in fast proliferating cells, in which PLC delta1 and to a greater extent PLC delta4 are more expressed at cytosolic level, suggesting an involvement of PI specific PLCs in the progression of cell cycle and in the control of cell proliferation and possibly of neoplastic cell growth.


Avoided-crossing-based ultrasensitive photonic crystal fiber refractive index sensor.
We propose and demonstrate an ultrasensitive photonic crystal fiber refractive index sensor by introducing the avoided-crossing effect in a bent-controlled fluid-filled photonic-bandgap fiber (FF-PBGF). By controlling the bend radius of the FF-PBGF, resonant couplings between the fundamental core mode and the cladding modes are realized and transmission notches are observed in the transmission band of the FF-PBGF. By changing the refractive index or temperature of the bent FF-PBGF, a sensitivity of 32,400 nm per refractive index unit (or 13.1 nm/degrees C) is achieved, which is the highest for a fiber device to date, to our best knowledge.


Transcranial magnetic stimulation for the treatment of anxiety disorder.
Anxiety is currently one of the main mood changes and can impair the quality of life of the individual when associated with other neurological or psychiatric disorders. Neuromodulation has been highlighted as a form of treatment of several pathologies, including those involving anxiety symptoms. Among the neuromodulatory options with the potential to improve mood changes, we highlight repetitive transcranial magnetic stimulation (rTMS). rTMS is a viable therapeutical option for neuropsychiatric dysfunctions of high prevalence and is important for the understanding of pathological and neuropsychological adaptation processes. Even with this potential, and high relevance of intervention, we observe the scarcity of literature that covers this subject. The objective of this study was to carry out a survey of the current literature, using scientific databases for the last five years. We found 32 studies reporting the effects of rTMS on anxiety, 7 on anxiety disorders and 25 on anxiety symptoms as comorbidities of neurological or psychiatric disorders. This survey suggests the need for further studies using TMS for anxiety in order to seek strategies that minimize these anxiety effects on the quality of life of the victims of this disorder.


Kinesin's processivity results from mechanical and chemical coordination between the ATP hydrolysis cycles of the two motor domains.
Kinesin is a processive motor protein: A single molecule can walk continuously along a microtubule for several micrometers, taking hundreds of 8-nm steps without dissociating. To elucidate the biochemical and structural basis for processivity, we have engineered a heterodimeric one-headed kinesin and compared its biochemical properties to those of the wild-type two-headed molecule. Our construct retains the functionally important neck and tail domains and supports motility in high-density microtubule gliding assays, though it fails to move at the single-molecule level. We find that the ATPase rate of one-headed kinesin is 3-6 s(-1) and that detachment from the microtubule occurs at a similar rate (3 s(-1)). This establishes that one-headed kinesin usually detaches once per ATP hydrolysis cycle. Furthermore, we identify the rate-limiting step in the one-headed hydrolysis cycle as detachment from the microtubule in the ADP.P(i) state. Because the ATPase and detachment rates are roughly an order of magnitude lower than the corresponding rates for two-headed kinesin, the detachment of one head in the homodimer (in the ADP.P(i) state) must be accelerated by the other head. We hypothesize that this results from internal strain generated when the second head binds. This idea accords with a hand-over-hand model for processivity in which the release of the trailing head is contingent on the binding of the forward head. These new results, together with previously published ones, allow us to propose a pathway that defines the chemical and mechanical cycle for two-headed kinesin.


An overview on the current status and future prospects in Aspergillus cellulase production.
Cellulase is a new research point besides glucoamylase, amylase, and protease in the enzyme industry. Cellulase can decompose lignocellulosic biomass into small-molecule sugars, which facilitates microbial utilization; thus, it has a vast market potential in the field of feed, food, energy, and chemistry. The Aspergillus was the first strain used in cellulase preparation because of its safety and non-toxicity, strong growth ability, and high enzyme yield. This review provides the latest research and advances on preparing cellulase from Aspergillus. The metabolic mechanisms of cellulase secretion by Aspergillus, the selection of fermentation substrates, the comparison of the fermentation modes, and the effect of fermentation conditions have been discussed in this review. Also, the subsequent separation and purification techniques of Aspergillus cellulase, including salting out, organic solvent precipitation, ultrafiltration, and chromatography, have been declared. Further, bottlenecks in Aspergillus cellulase preparation and corresponding feasible approaches, such as genetic engineering, mixed culture, and cellulase immobilization, have also been proposed in this review. This paper provides theoretical support for the efficient production and application of Aspergillus cellulase.


Development of a Patient-Specific 3D-Printed Liver Model for Preoperative Planning.
Liver surgery is widely used as a treatment modality for various liver pathologies. Despite significant improvement in clinical care, operative strategies, and technology over the past few decades, liver surgery is still risky, and optimal preoperative planning and anatomical assessment are necessary to minimize risks of serious complications. 3D printing technology is rapidly expanding, and whilst appliactions in medicine are growing, but its applications in liver surgery are still limited. This article describes the development of models of hepatic structures specific to a patient diagnosed with an operable hepatic malignancy. Anatomy data were segmented and extracted from computed tomography and magnetic resonance imaging of the liver of a single patient with a resectable liver tumor. The digital data of the extracted anatomical surfaces was then edited and smoothed, resulting in a set of digital 3D models of the hepatic vein, portal vein with tumor, biliary tree with gallbladder, and hepatic artery. These were then 3D printed. The final models of the liver structures and tumor provided good anatomical detail and representation of the spatial relationships between the liver tumor and adjacent hepatic structures and could be easily manipulated and explored from different angles. A graspable, patient-specific, 3D printed model of liver structures could provide an improved understanding of the complex liver anatomy and better navigation in difficult areas and allow surgeons to anticipate anatomical issues that might arise during the operation. Further research into adequate imaging, liver-specific volumetric software, and segmentation algorithms are worth considering to optimize this application.


Blue Light Potentiates Antibiotics in Bacteria via Parallel Pathways of Hydroxyl Radical Production and Enhanced Antibiotic Uptake.
In the age of antimicrobial resistance, the urgency by which novel therapeutic approaches need to be introduced into the clinical pipeline has reached critical levels. Antimicrobial blue light (aBL), as an alternative approach, has demonstrated promise as a stand-alone therapeutic method, albeit with a limited window of antimicrobial activity. Work by others indicates that treatment with antibiotics increases the production of reactive oxygen species (ROS) which may, in part, contribute to the bactericidal effects of antibiotics. These findings suggest that there may be potential for synergistic interactions with aBL, that similarly generates ROS. Therefore, in this study, the mechanism of aBL is investigated, and the potential for aBL to synergistically promote antibiotic activity is similarly evaluated. Furthermore, the translatability of using aBL and chloramphenicol in combination within a mouse model of Acinetobacter baumanii burn infection is assessed. It is concluded that porphyrins and hydroxyl radicals driven by "free iron" are paramount to the effectiveness of aBL; and aBL is effective at promoting multiple antibiotics in different multidrug-resistant bacteria. Moreover, rROS up-regulation, and promoted antibiotic uptake are observed during aBL+antibiotic exposure. Lastly, aBL combined with chloramphenicol appears to be both effective and safe for the treatment of A. baumannii burn infection. In conclusion, aBL may be a useful adjunct therapy to antibiotics to potentiate their action.


Predictive value of the 8th edition American Joint Commission Cancer (AJCC) nodal staging system for patients with cutaneous squamous cell carcinoma of the head and neck.
The 8th edition American Joint Committee on Cancer (AJCC8) provides the same nodal staging system for mucosal and cutaneous squamous cell carcinoma of the head and neck (HNcSCC) and includes extranodal extension (ENE) as an adverse prognostic criterion. This study evaluates the prognostic efficacy of the AJCC8 pathologic nodal staging system (pN) for HNcSCC. Univariate analysis of 382 patients with metastatic HNcSCC staged according to both the 7th (AJCC7) and the 8th edition staging systems. The AJCC7 pN3 category was associated with reduced disease specific survival (DSS HR 5.49; 95% CI: 1.83-16.53; P = 0.002) and overall survival (OS HR 3.42; 95% CI: 1.54-7.58; P = 0.002) as compared with pN1. However, no difference was observed between pN1, pN2, and pN3 categories as defined by the AJCC8. Also, when comparing Stages III and IV as defined by AJCC8, there was no difference in DSS (HR 0.75; 95% CI: 0.34-1.67; P = 0.478) or OS (HR 0.88; 95% CI: 0.51-1.51; P = 0.648). The AJCC8 performed poorly as a prognostic indicator for patients with metastatic HNcSCC in this cohort. HNcSCC would benefit from a staging system that accounts for its unique biologic characteristics distinct from mucosal SCC.


MAPping the kinase landscape of macrophage activation.
ARL11 is a tumor suppressor gene with established pro-apoptotic properties, but its function beyond this role is poorly understood. A new analysis of macrophage activation has identified ARL11 as a novel regulator of a mitogen-activated protein kinase (MAPK). These findings expand on the function of ARL11 beyond its tumor suppressor activity and highlight a novel role as a regulator of macrophage activation and inflammatory response.


Perception, attitudes, preparedness and experience of chemotherapy-induced alopecia among breast cancer patients: a qualitative study.
Regardless of its negative impact on quality of life, little is known about the importance of alopecia from the patients' perspective. This study aimed to explore the whole experience of chemotherapy- induced alopecia among Korean breast cancer patients including perception, attitudes, preparedness, and changes after alopecia. Patients expected to experience or had experienced alopecia were recruited at a tertiary hospital in Seoul, Korea. Semi-structured in-depth interviews were performed in 21 patients. Recurrent issues were identified and placed into thematic categories. All patients think that appearance is important and they pay attention to how they look like. They had negative perceptions about alopecia. Patients were not well prepared for alopecia, and experienced substantial physical, psychological and social distress. Lack of information and limited social support combined with negative images of cancer made it difficult for patients to overcome the trauma and deterred them from usual daily activities resulting in poor quality of life. Patients were not well prepared for alopecia and negative perceptions, lack of preparedness, and limited social support and resources increased alopecia- related distress. Educational programs for preparing patients to cope with alopecia distress and advocate activities to change people's negative perception about alopecia are needed to reduce the burden imposed by alopecia in cancer patients.


Psychological problems and psychosocial predictors of cigarette smoking behavior among undergraduate students in Malaysia.
Cigarette smokers have their own motivation and justification to smoke. For example, smoking reduces their stress or enhances their pleasure. This study aimed to identify the (a) prevalence of cigarette smokers among undergraduates in Malaysia, (b) gender differences in nicotine dependence among current smokers, (c) differences in psychological problems (depression, anxiety and stress) based on the status of smoking cigarettes (current, former and non-smokers) and (d) extent to which precipitating factors (tension reduction, addiction, automatism, handling, social interaction, pleasure, and stimulation) predict the smoking behavior among current smokers. In this study 780 undergraduate students participated from a private university in Kuala Lumpur and Selangor state in Malaysia. The Depression, Stress and Anxiety Scale, Modified Reason for Smoking Scale and Fagerstrom Nicotine Dependence Test were used to measure psychological problems, predictors of smoking behavior and nicotine dependency among current smokers. The results showed that 14.7%(n=106) of the students were smokers. Current smokers exhibited more psychological problems (depression, anxiety and stress) compared to former and non-smokers. Addiction, tension reduction, pleasure and automatism were predictors of smoking behavior among the current smoking students. Step wise regression analysis showed that smoking behavior was highly predicted by nicotine dependency or addiction. Smoking students were motivated to smoke cigarettes as they believed that it reduced their tension and enhance pleasure. Hence, there is a need for health promotion and anti-tobacco prevention as cigarette smokers experience more psychological problems. Nicotine dependency or addition was one of the major causes for smoking behavior among the student population in Malaysia.


Modulation of Th17 Proliferation and IL-17A Gene Expression by Acetylated Form of Apigenin in Patients with Multiple Sclerosis.
The presence of Th17 cells in CNS lesion of MS patients due to their inflammatory cytokines secretion is in line with the deterioration of the disease. Currently, the use of natural compounds with anti-inflammatory properties such as flavonoids have been considered to reduce inflammation in these patients, but the remaining issue is how deliver these compounds to the site of inflammation. Acetylation is a way to better uptake compound by cells and cross through cellular layers with tight junctions. This study aimed to investigate the in vitro effects of the Apigenin 3-Acetate on Th17 cells of MS patients and compare its efficacy with Apigenin and Methyl Prednisolone Acetate. IC50 for Apigenin 3-Acetate, and Methyl Prednisolone Acetate were determined using three healthy volunteers. The peripheral blood mononuclear cells (PBMCs) of five MS patients were isolated and co-cultured with a selected dose of Apigenin, Apigenin 3-Acetate, and Methyl Prednisolone Acetate for 48 hr, and then theproliferation of Th17 cells in isolated PBMCs was assessed by flow cytometry. The levels of RAR-related orphan receptor (RORC) and IL-17A expression were also determined by quantitative real-time PCR. The results showed that Apigenin 3-Acetate inhibited Th17 cells proliferation (P value: 0.018) at 80 µM concentration after 48 hr. Additionally, IL-17A gene expression significantly (P value≤ 0.0001) inhibited by Apigenin, Apigenin 3-Acetate and Methyl Prednisolone Acetate in 80 µM, 80 µM and 2.5 µM (selected dose in IC50 determination) respectively These results demonstrate that Acetate increases anti-inflammatory effects of Apigenin on Th17 cells.


The regularity of orthosis use and the reasons for disuse in stroke patients.
This is a Retrospective descriptive study. Orthoses and walking aids are used frequently in stroke rehabilitation to facilitate ambulation. The aim was to describe the regularity of orthosis use and the reasons for disuse in stroke after discharge from inpatient rehabilitation. The study included 64 (43 men, 21 women) subacute stroke patients who had been discharged from our clinic in the last 6 months. Demographic and clinical findings, proposed orthoses and walking aids, frequency of usage for the proposed orthoses, reasons for disuse, and the ambulation levels were recorded. A total of 54 (84.4%) patients had an ankle-foot orthosis and 10 (15.6%) patients had a knee-ankle-foot orthosis. The orthosis frequency of use was every day in 38 (59.4%) patients and one to seven times a week in seven (10.9%) patients, whereas 19 (29.7%) did not use them. The reasons for orthosis disuse were finding them unnecessary in seven (27%) patients, usage difficulties in six (23%) patients, pressure sensation in five (19.2%) patients, the belief that they did not make life easier in five (19.2%) patients, lack of a suitable environment in one (3.8%) patient, orthosis wear in one (3.8%) patient, and systemic disease in one (3.8%) patient. Age, sex, residence, the disease duration, and recommended orthosis duration (how long the prescribed orthosis was used), recommended orthosis, range of motion limitation in the lower extremities, presence of spasticity, type of orthosis, and the ambulation level were not statistically significantly associated with the frequency of orthosis use (P>0.05). The only significant clinical factor for the frequency of orthosis use was the lower extremity Brunnstrom neurophysiological recovery stage (P<0.008). Orthosis use had been discontinued by 29% of the patients. The most common reasons reported for discontinuing orthosis use were that the patients found it unnecessary, usage difficulty, pressure sensation, not making life easier, lack of a suitable environment, and orthosis wear. The only significant clinical factor for the frequency of orthosis use was the lower extremity Brunnstrom neurophysiological recovery stage.


Reduction in intensity of Pneumocystis carinii pneumonia in mice by aerosol administration of gamma interferon.
Pneumocystis carinii pneumonia in patients with AIDS may result from impaired local release of gamma interferon from lung lymphocytes and subsequent failure of macrophage activation. We tested this hypothesis with mice rendered immunodeficient by selective depletion of CD4+ lymphocytes and inoculated intratracheally with P. carinii. After aerosol administration of recombinant murine gamma interferon, the intensity of P. carinii infection in these mice was reduced in comparison with that in mice not treated with gamma interferon. Histologic scoring of lung tissue did not reveal additional pulmonary inflammation attributable to gamma interferon. Aerosol administration of gamma interferon may reduce the intensity of P. carinii infection by augmentation of host defense, despite persistent CD4+ lymphocyte depletion.


Assembly of porphyrin-based uranium organic frameworks with (3,4)-connected pto and tbo topologies.
Three (3,4)-connected uranyl-organic frameworks (UOFs) with pto and tbo topologies were synthesized. The UOF with a pto net possesses a 2-fold interpenetrating network and exhibits 2D interconnected channels. The UOF with a tbo net is constructed from two types of ultralarge nanocages. All these compounds can efficiently remove large cationic dye crystal violet (CV) through a cation exchange mechanism.