Lyme arthritis synovial gammadelta T cells instruct dendritic cells via fas ligand.
gammadelta T cells participate in the innate immune response to a variety of infectious microorganisms. They also link to the adaptive immune response through their induction of maturation of dendritic cells (DC) during the early phase of an immune response when the frequency of Ag-specific T cells is very low. We observe that in the presence of Borrelia burgdorferi, synovial Vdelta1 T cells from Lyme arthritis synovial fluid potently induce maturation of DC, including production of IL-12, and increased surface expression of CD40 and CD86. The activated DC are then able to stimulate the Vdelta1 T cells to up-regulate CD25. Both of these processes are initiated primarily by Fas stimulation rather than CD40 activation of DC via high expression of Fas ligand by the Vdelta1 T cells. DC are resistant to Fas-induced death due to expression of high levels of the Fas inhibitor c-FLIP. This effect serves to divert Fas-mediated signals from the caspase cascade to the ERK MAPK and NF-kappaB pathways. The findings affirm the importance of the interaction of certain T cell populations with DC during the early phases of the innate immune response. They also underscore the view that as levels of c-FLIP increase, Fas signaling can be diverted from induction of apoptosis to pathways leading to cell effector function.


Characteristics in the epidemiology of bovine viral diarrhea virus (BVDV) of relevance to control.
An understanding of the driving forces of BVDV transmission can be gained by considering the reproductive rate, between individuals and between herds. The former determines the prospects for eliminating the infection from herds, and the latter is the key to persistence at the population level. In this paper, the relation between these two characteristics, their underlying parameters and measures and priorities for BVDV control are discussed. A general model for BVDV control is outlined, with bio-security, virus elimination and monitoring as three necessary consecutive elements, and with immunization as an optional step. A distinction is made between systematic and non-systematic approaches to BVDV control (where the former refers to a monitored and goal-oriented reduction in the incidence and prevalence of BVDV infection and the latter to where measures are implemented on a herd-to-herd decision basis and without systematic monitoring in place). Predictors of progress for systematic control approaches in general are discussed in terms of the abilities: to prevent new infections, to rapidly detect new cases of infection, to take action in infected herds and to gain acceptance by stakeholders. We conclude that an understanding not only of the biology, but also of the social factors - human behavior, the motives that makes stakeholders follow advice and the cultural differences in this respect - are important factors in forming recommendations on alternative strategies for BVDV control.


[Leather dusts and cancerogenic effects].
Work exposure to carcinogenic chemicals in the shoe industry is still debated owing to continuous technological developments. A possible causal role in sino-nasal cancer development was attributed to the leather dusts basing on epidemiologic studies. Nevertheless, convincing conclusions regarding toxicodinamic-involved processes are actually missing. An hypothesis pointing on tannins toxic action shows insufficient and contrasting data. A cancerogenic risk seems to be attributable for workers engaged in the whole process of shoe industry.


Beetles, ants, wasps, or flies? An ethnobiological study of edible insects among the Awajún Amerindians in Amazonas, Peru.
Insects are known to be able to provide valuable nutrients to indigenous populations across the Amazon. However, studies on traditional insect use in the Peruvian Amazon are scarce. This study documents edible insect diversity and characterizes their food and collection patterns in eight Awajún communities in the Peruvian Amazon. Additionally, we reviewed what has been known to date about the nutrient composition of the documented species. The survey was conducted among the Awajún populations living in the Huampami, Paisa, Achu, and Tseasim communities in the Cenepa district and the Shijap, San Mateo, Kusu, and Listra communities in the Imaza district. Data collection was conducted through a freelisting exercise complemented by a semi-structured inquiry form in the Awajún language. In total, 104 informants (72 men and 32 women) aged between 16 to 73 years were interviewed. The Awajún people use at least 12 insect species, with Rhynchophorus palmarum, Atta cephalotes, and Rhinostomus barbirostris being the most important ones. Beetles of the family Curculionidae represent the culturally most salient taxon. In the more accessible and developed Imaza district, the Awajún tend to eat almost exclusively R. palmarum, while in the more isolated and preserved Cenepa district, the community's preferences are linked with more species. Although men are the main insect collectors, women cited more edible insects on average. The insects are eaten mainly roasted or raw. Further use patterns and differences between the districts are discussed. Traditional knowledge related to edible insects and the ecosystems they occur in is widespread among the Awajún populations, and insects still represent an important part of the indigenous food system. This ethnobiological survey discovered five species that are newly recorded as edible insects. Chemical composition of insects deemed edible by the Awajún ought to be analyzed in the future and awareness about their nutritional importance should be raised to harness the potential of this underutilized yet nutrient-rich traditional food.


Malaligned bioprosthetic valve causing left ventricular outflow tract obstruction.
Left ventricular outflow tract obstruction resulting from strut impingement upon the interventricular septum is a rare complication of bioprosthetic mitral valve insertion. Obstruction is more likely to develop when a small, high profile prosthetic valve is inserted into a patient with a small outflow tract. The likelihood of this complication may be reduced by appropriate modification of surgical technique.


New insights on the systematics, palaeoecology and palaeobiology of a plesiosaurian with soft tissue preservation from the Toarcian of Holzmaden, Germany.
The Posidonienschiefer Formation (Toarcian) of Holzmaden, Baden-Württemberg in southwestern Germany has yielded several excellently preserved plesiosaurian specimens and received considerable research attention. The plesiosaurians found within these deposits are always significantly outnumbered by ichthyosaurs, and close examination of these rare specimens is crucial to a better understanding of the diversity and palaeoecology of Plesiosauria in this very peculiar ecosystem. The plesiosaurian specimen SMNS 51945 found in this area is a juvenile individual consisting of a partial, crushed skull and an exquisitely preserved post-cranial skeleton. Its anatomical characters seem to differ from the long-necked plesiosauroids Microcleidus brachypterygius and Seeleyosaurus guilelmiimperatoris that are the most abundant taxa within the plesiosaurian assemblage. The post-cranial skeleton preserves very likely soft tissues composed of buff-coloured and dark-coloured structures around the vertebral column and hindlimb of the animal. A network of buff-coloured fibres located posterior to the hindlimb most likely represents phosphatised collagen fibres as already found in some ichthyosaur specimens, confirming that wing area in plesiosaurians was much larger than that suggested by skeletal remains alone. The specimen also contains gastroliths (sand-sized grains mainly composed of quartz) in the stomach cavity suggesting the animal spent at least some of its time in shallow coastal waters, tens or hundreds of kilometres from the final place of burial.


[Effects of leucine on adipogenesis in 3T3-L1 preadipocytes during and after differentiation].
To observe the effects of leucine on adipogenesis in 3T3-L1 preadipocyte during and after differentiation, and to investigate possible mechanisms. Respectively, 0.0 (control), 0.5, 1.0 and 2.0 mmol/L leucine was added in 3T3-L1 cells and cell proliferation was measured by MTT. Then, 3T3-L1 preadipocyte was induced to differentiate. Leucine was added during whole differentiation period, or after differentiation for 4 days. The cells were stained with Oil Red O dye to observe lipid droplet. The culture media were collected and used to determine glycerol contents. Meanwhile, protein expressions related to lypolytic enzymes, leptin signaling pathway were determined by Western blot. MTT result showed that cell viabilities were (100.00±12.10)%, (102.73±12.38)%, (103.94±14.65)%, (108.70±5.05)% in 0.0, 0.5, 1.0 and 2.0 mmol/L leucine groups, respectively, there were no significant differences in cell proliferation among 4 groups (F=1.07, P=0.383). When 0.0, 0.5, 1.0 and 2.0 mmol/L leucine was added during differentiation, the relative number of lipid droplet was 1.00±0.06, 0.94±0.09, 0.82±0.08 and 0.79±0.04, respectively (F=11.74, P<0.001), and it was significantly lower in 1.0 and 2.0 mmol/L leucine groups than in control group (P=0.002 and P<0.001, respectively). There was no significant difference in lipid droplet when leucine was added after differentiation (F=0.16, P=0.924). When leucine was added during differentiation, the increment of glyceride contents in medium was (65.04 ± 11.75), (71.45 ± 23.71), (79.37 ± 17.63) and (110.32 ± 25.36) μmol/L, respectively (F=2.92, P=0.100). And it was significantly higher in 2.0 mmol/L leucine group (110.32 ± 25.36) μmol/L than in control group (65.04 ± 11.75) μmol/L (t=2.73, P=0.026). No significant difference of the increment of glyceride contents among 4 groups was observed when leucine was added after differentiation (F=0.80, P=0.528). Western blot results showed that leucine treatment during differentiation upregulated expression level of hormone-sensitive lipase phosphorylation (after 0.0 and 2.0 mmol/L leucine treatment,the protein levels were 1.00 ± 0.08 vs. 2.54 ± 0.27, P<0.001) , and downregulated the protein expression levels of perilipin A, leptin and leptin-related pathway, such as leptin receptor, Janus kinase 2 and suppressor of cytokine signaling-3 (after 0.0 and 2.0 mmol/L leucine was added, the protein levels were (1.00 ± 0.03) vs. (0.31 ± 0.07) , (1.00 ± 0.08) vs. (0.22±0.07) , (1.00±0.07) vs. (0.21 ± 0.04) , (1.00 ± 0.03) vs. (0.35 ± 0.05) , (1.00 ± 0.06) vs. (0.34 ± 0.05) , P<0.001). Leucine treatment after differentiation had no effects on these protein expressions (all P>0.05). Leucine inhibits adipogenesis during 3T3-L1 preadipocyte differentiation by the regulation of lypolytic enzymes and leptin signaling pathway; however, leucine has no effect on adipogenesis when differentiation completed.


Formation of homocytotropic antibodies with penicilloyl specificity in the rabbit.
The formation of homocytotropic antibodies (IgE) as determined by immunochemical characteristics against penicilloyl in rabbits was shown. The production of such antibodies to azidocilloyl-human serum albumin (AzO10-HSA) in alum was found optimal using 1 mg of antigen in the tested range of 0.01-8 mg. At the lowest dose (0.01 mg) only haemagglutinating but no IgE antibodies were formed. Immunization with azidocilloyl-bovine gamma-globulin (AzO9-BGG) resulted in a slower increase in antibody levels than was caused by AzO10-HSA. Univalent benzylpenicilloyl-epsilon-aminocaproate or ampicillin given together with the antigen upon immunization decreased the levels of penicllloyl specific IgE and haemagglutinating antibodies, but induced the formation of IgE antibodies against the carrier molecule. Further, the administration of penicilloyl specific IgG antibodies diminished formation of both IgE and haemagglutinating antibodies, but no antibodies specific for the carrier were formed. The usefulness of this animal model for the experimental study of penicillin allergy is discussed.


Comparison of conventional and liquid-based cytology, and human papillomavirus testing using SurePath preparation in Japan.
We compared the detection rate of cervical neoplasias between a liquid-based cytology (LBC) method using SurePath and the conventional method. We also studied the feasibility of human papillomavirus (HPV) typing by linear array assay. Cytological specimens from 1551 Japanese women were prepared using the conventional and SurePath methods; the cytological and histological results from biopsy samples were compared. HPV typing using an HPV linear array assay was carried out on residual specimens using the SurePath method. The cytodiagnostic results showed a concordance rate of 85.3% (Κ= 0.46) between the two methods. The sensitivity of lesions histopathologically diagnosed as CIN1 or above was not significantly different between the two methods (P = 0.575-1.000). The receiver operating characteristic curve analysis of the detectability in CIN2 or above revealed no significant difference between the two methods (P = 0.096). Among the 44 patients who underwent HPV typing using a linear array assay, 33 samples were eligible for HPV testing and were stored at ambient temperature. In conclusion, the SurePath and conventional methods have equivalent abilities for detecting cervical lesions. After preparation for cytological diagnosis, use of the remaining cells from the SurePath specimens to perform HPV typing using the linear array method could be feasible.


Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension.
BACKGROUND Establishing a shunt-induced pulmonary arterial hypertension (PAH) model in mice would be of great scientific value, but no such models have been reported to date. Here, we established a shunt-associated PAH in mice to investigate the characteristics of pulmonary vascular remodeling, which provides a new platform for the in-depth study of PAH associated with congenital heart disease (CHD). MATERIAL AND METHODS Eighty mice were randomly divided into the heavy shunt group (n=32), the small shunt group (n=32), the sham operation group (n=8), and the control group (n=8). The septum of the abdominal aorta and inferior vena cava was cut directly to create a heavy abdominal aortocaval shunt. Pulmonary artery pressure, right ventricular hypertrophy index, and lung tissue morphology were evaluated in the 4th, 6th, 8th, and 12th weeks in the shunt groups. RESULTS Shunt-associated PAH by abdominal aortocaval shunt in mice was successfully established. The shunt patency rate was significantly higher in the heavy shunt group. Significant differences were observed between the heavy shunt group and other groups in terms of pulmonary artery pressure and the right ventricular hypertrophy index. Tissue sections revealed a thickened pulmonary intimal layer and muscular layer and stenosis of the lumen in the shunt groups. Immunofluorescent assay results showed significant proliferations of PAH smooth muscle cells and endothelial cells, consistent with the clinical pulmonary vascular remodeling seen in human patients with severe PAH. CONCLUSIONS Shunt-associated PAH established by directly cutting the septum between the abdominal aorta and inferior vena cava is a stable and reliable model for research on PAH associated with CHD.


Effect of cell density and epidermal growth factor on the inducible expression of CYP3A and CYP1A genes in human hepatocytes in primary culture.
The influence of cell density and epidermal growth factor (EGF) on the expression and inducibility of cytochrome P450 (CYP) genes of the CYP3A and CYP1A families in adult human hepatocytes in primary culture has been evaluated. Only when cultured at subconfluence and in the presence of EGF did hepatocytes exhibit a proliferative response, assessed by measuring DNA synthesis and cyclin A accumulation. In the absence of EGF, the accumulation of CYP3A4 and CYP1A2 messenger RNAs (mRNAs) in response to their respective inducers (rifampicin and dioxin) was dramatically decreased in subconfluent culture with respect to confluent cultures. The presence of EGF only slightly decreased the accumulation of these mRNAs in both confluent and subconfluent cultures. The accumulation of CYP2D6 and CYP2E1 proteins, which are constitutively expressed in confluent cultures, and the production of fibrinogen and apolipoprotein (Apo) B100 exhibited similar behavior, while nicotinamide adenine dinucleotide phosphate cytochrome c reductase activity was affected neither by cell density nor by EGF. In contrast, the accumulation of CYP1A1 mRNA in response to dioxin was similar in confluent and subconfluent cultures, irrespective of the presence of EGF. Interestingly, CYP3A7, a gene that is preferentially expressed in the fetal liver, was expressed constitutively neither in confluent nor in subconfluent cultures, irrespective of the presence of EGF. It is concluded that the loss of cell-cell contacts rather than the proliferative status of cells per se is responsible for the dramatic decrease in the expression of CYP genes, normally expressed in the adult human liver.


Maternal experience influences the establishment of visual/auditory, but not olfactory recognition of the newborn lamb by ewes at parturition.
Maternal behavior in sheep is characterized by the rapid establishment of individual recognition of the lamb through the use of different sensory modalities. Olfactory recognition mediates acceptance at suckling whereas visual/auditory cues are involved in recognition from a distance. This study investigates (a) the timing of both types of recognition and (b) whether they can be influenced by maternal experience. Olfactory recognition was assessed at lambing, 30 min, 1, 2, or 4 hr postpartum by presenting successively an alien and the familiar lamb. Recognition at a distance was assessed at 6, 8, and 12 hr postpartum by using a two-choice test between the familiar and an alien lamb. A majority (33/51) of ewes showed selectivity at suckling as early as 30 min after parturition, and no differences due to maternal experience were observed. By contrast, in the two-choice recognition test, multiparous ewes showed a preference for their familiar lamb at 6 hr whereas primiparous mothers did so only after 24 hr of mother-young contact: The performance of biparous mothers was intermediate. Thus, both types of recognition can be rapidly established after parturition, and maternal experience has a differential effect on the dynamics of these learning processes.


Effects of normobaric oxygen and melatonin on reperfusion injury: role of cerebral microcirculation.
In order to protect the brain before an irreversible injury occurs, penumbral oxygenation is the primary goal of current acute ischemic stroke treatment. However, hyperoxia treatment remains controversial due to the risk of free radical generation and vasoconstriction. Melatonin is a highly potent free radical scavenger that protects against ischemic stroke. Considering its anti-oxidant activity, we hypothesized that melatonin may augment the survival-promoting action of normobaric oxygen (NBO) and prevent brain infarction. Herein, we exposed mice to 30 or 90 min of intraluminal middle cerebral artery occlusion (MCAo) and evaluated the effects of NBO (70% or 100% over 90 min), administered either alone or in combination with melatonin (4 mg/kg, i.p.), on disseminate neuronal injury, neurological deficits, infarct volume, blood-brain barrier (BBB) permeability, cerebral blood flow (CBF) and cell signaling. Both NBO and particularly melatonin alone reduced neuronal injury, neurological deficits, infarct volume and BBB permeability, and increased post-ischemic CBF, evaluated by laser speckle imaging (LSI). They also improved CBF significantly in the ischemic- core and penumbra, which was associated with reduced IgG extravasation, DNA fragmentation, infarct volume, brain swelling and neurological scores. Levels of phosphorylated Akt, anti-apoptotic Bcl-xL, pro-apoptotic Bax and endothelial nitric oxide synthase (NOS) were re-regulated after combined oxygen and melatonin delivery, whereas neuronal and inducible NOS, which were increased by oxygen treatment, were not influenced by melatonin. Our present data suggest that melatonin and NBO are promising approaches for the treatment of acute-ischemic stroke, which encourage proof-of-concept studies in human stroke patients.


A TiSe monolayer as a superior anode for applications of Li/Na/K-ion batteries.
Using density functional theory (DFT), we investigated the energy-storage capabilities of a two-dimensional TiSe monolayer for applications of the anode material of Li/Na/K-ion batteries. The TiSe monolayer showed high thermodynamic stability at 800 K according to ab initio molecular dynamics (AIMD) simulation. The ion-diffusion barrier was estimated to be 0.29/0.36/0.33 eV for Li/Na/K, respectively, indicating the high-rate capacity of this material. The theoretical specific capacity was 422.63 mA h g-1 for Li/Na/K, with an energy density of 1000.19, 802.30, and 802.41 mW h g-1, respectively. Fully charged TiSe was mechanically stable according to the calculated elastic constants. Our results show that the TiSe monolayer could be used as an excellent anode material for Li/Na/K-ion batteries.


Prognostic Value of the Preoperative Neutrophil to Lymphocyte Ratio in Resectable Gastric Cancer.
This study aimed to investigate the prognostic value of the preoperative neutrophil to lymphocyte ratio (NLR) in resectable gastric cancer (GC). This was a retrospective review of 1030 patients with resectable GC managed between 2005 and 2011. Patients were stratified into 2 groups, those with a preoperative NLR >3.44 and those with a preoperative NLR ≤ 3.44. Clinicopathological data affecting patient prognosis were collected prospectively and analyzed. The high NLR (> 3.44) group had a higher proportion of a platelet to lymphocyte ratio > 132, tumor size > 4.8 cm, T4 lesions, metastatic tumors, a ratio of metastatic to examined lymph nodes >0.18, positive resection margins, and presence of vascular or lymphatic invasion than the low NLR (≤ 3.44) group. Patients with a high preoperative NLR had significantly lower 3- and 5-year overall survival rates than those with a low preoperative NLR (55.1% vs 71.0% and 47.2% vs 64.1%, respectively; P < 0.001). Preoperative NLR was a prognostic factor for resectable GC in multivariate analysis. More aggressive tumor behavior was observed in patients with resectable GC with a high preoperative NLR than in those with a low preoperative NLR. High preoperative NLR was an independent unfavorable prognostic factor. Measurement of this ratio may serve as a clinically accessible and useful biomarker for patient outcomes.


William Marsden's Yorkshire family, 1749-1922.
William Marsden (1796-1867), a Yorkshireman, founded two major London hospitals. We know little of his family background, about which his only biography gives a highly inaccurate account. It suggests his family were landowners and gentlemen farmers. Although their origins were more humble, some at least of his Yorkshire relatives prospered-through gardening, banking, manufacturing and even medicine. This paper presents information on four generations of Yorkshire Marsdens.


A double-blind placebo-controlled investigation of the psychomotor profile of clopidogrel in healthy volunteers.
Clopidogrel is a thienopyridine antiplatelet agent that inhibits adenosine diphosphate-dependent platelet activation and aggregation and is currently one of the most widely prescribed antiplatelet drugs for the treatment of symptomatic coronary artery disease. Several large clinical trials have demonstrated that it has a potent protective effect against adverse vascular events, including coronary, cerebral, and peripheral arterial disease. These clinical trials have reported among other clopidogrel's adverse effects, the adverse effects related to the central nervous system including headache and dizziness occurring in 7.6% and 6.2% of patients, respectively. Aim of this study is to detect possible effect of clopidogrel on human psychomotor performance in healthy volunteers. Using a double-blind, placebo-controlled, balanced, between-subject design at laboratories of the pharmacology department in Al-Mustansiryiah University Medical College, Baghdad, Iraq, during the academic year 2007-2008. Fifty-four young healthy volunteers were enrolled, they received either single oral doses of clopidogrel 37.5 mg (Plavix; Sanofi-Synthelabo) or placebo. Each treatment was given to 27 subjects. Measurements of psychomotor performance including the choice reaction time and its 2 components, recognition reaction time and movement reaction time, and the critical flicker fusion threshold using the computerized Leed psychomotor tester were performed just before medication and 2 hours afterward. No statistically significant difference in the parameters measured was detected among the two treatment groups. In this study, we found that single oral doses of clopidogrel in our group of normal healthy volunteers did not affect their psychomotor performance.


Chikungunya Virus Seroprevalence and Associated Factors among Hospital Attendees in Two States of Southwest Nigeria: A Preliminary Assessment.
Chikungunya virus (CHIKV) is a re-emerging pathogen causing long-term polyarthritis and encephalitis. In conducting a preliminary investigation, we hypothesized that there is no serologic evidence of CHIKV infection among attendees of selected hospitals in Lagos and Osun States, Nigeria. Sera from 304 consecutively selected participants were screened for CHIKV IgG and IgM using ELISA. Findings were analyzed vis-à-vis participants' demographic and clinical data. Over 90.0% of the participants had never heard of CHIKV despite the fact that a large proportion of them (88.8%) had secondary/tertiary education. Overall, 41.8% were positive for, at least, one antibody type (IgG or IgM), while about 16.0% of the participants had dual seropositivity (CHIKV IgG and IgM) with gender as associated factor (odds ratio [OR]: 2.8, p = 0.03). Prevalence rates were 31.8% and 38.4% for CHIKV IgG and IgM, respectively. Only hospital location (Osogbo) was associated with CHIKV IgG (OR: 2.2, p = 0.009), while gender alone was associated with CHIKV IgM (OR: 3.0, p = 0.001). Participants seropositive for CHIKV antibodies were mostly adults (18-59 yrs) belonging to the active work-force; five (22.7%) and three (20.0%) of the pregnant participants had CHIKV IgG and IgM, respectively. Detection of CHIKV IgM in some participants might make them potentially infectious to the newborn and mosquito vectors. Importantly, participants positive for either IgG or IgM had fever (72.8%, 67.2%) and general body pains (61.7%, 57.6%), respectively. This ELISA-based study revealed serologic evidence of CHIKV infection among hospital attendees in Lagos and Osun states with the group-specific prevalence rates being considerably high. Chikungunya virus (CHIKV); Chikungunya (CHIK); enzyme-linked immunosorbent assay (ELISA); immunoglobulin G or M (IgG/IgM); odds ratio (OR); non-structural proteins (nsP); hemagglutination inhibiting (HI); complement fixing (CF); neutralization test (NT); immunofluorescence assay (IFA); plaque reduction neutralization test (PRNT); confidence interval (CI); analysis of variance (ANOVA); body temperature (BT); Building Nigeria's Response to Climate Change (BNRCC).


Retinoblastoma and Hirschsprung disease with a 13q14 to 22 deletion.
Hirschsprung disease, bilateral retinoblastoma, and craniofacial dysmorphism with an interstitial deletion of chromosome 13 is a rare gene deletion syndrome that has twice before been described in white children. The authors present the third report of such a case in a South African child of Indian-Asian descent.


Experimental and Computational Studies of Dinitrogen Activation and Hydrogenation at a Tetranuclear Titanium Imide/Hydride Framework.
The activation of N2 by a tetranuclear titanium(III) diimide/tetrahydride complex, [(Cp'Ti)4(μ3-NH)2(μ-H)4] (1) (Cp' = C5Me4SiMe3), which was obtained by the reaction of the Cp'-ligated titanium trialkyl complex Cp'Ti(CH2SiMe3)3 with H2 and N2, was investigated in detail by experimental and density functional theory studies. The reaction of 1 in the solid state with N2 (1 atm) at 180 °C gave the dinitride/diimide complex [(Cp'Ti)4(μ3-N)2(μ3-NH)2] (2) through the incorporation, cleavage, and partial hydrogenation of one molecule of N2 and release of two molecules of H2. At 130 °C, the formation of 2 was not observed, but instead, dehydrogenation of 1 took place through cleavage of the N-H bond in an imide ligand, followed by deprotonation of the other imide ligand with a hydride ligand, affording the dinitride/tetrahydride complex [(Cp'Ti)4(μ3-N)2(μ-H)4] (3). Upon heating under N2 (1 atm) at 180 °C, 3 was quantitatively converted to the dinitride/diimide complex 2. This transformation was initiated by migration of a hydride ligand to a nitride ligand to give one imide unit, followed by N2 coordination to a Ti atom and H2 release through the reductive elimination of two hydride ligands. The other imide ligand in 2 was formed by hydride migration to one of the two nitride ligands generated through the cleavage of the newly incorporated N2 unit. The hydrogenation of 2 with H2 (100 atm) at 180 °C afforded the tetraimide complex [(Cp'Ti)4(μ3-NH)4] (4). This reaction was initiated by σ-bond metathesis between H2 and a titanium-nitride bond, followed by migration of the resulting hydride ligand to the remaining nitride ligand. In all of these transformations, the interplay among the hydride, imide, and nitride ligands, including the reversible dehydrogenation/hydrogenation of imide and nitride species, at the multimetallic titanium framework has a critically important role.


Platinum-based chemotherapy: gastrointestinal immunomodulation and enteric nervous system toxicity.
The efficacy of chemotherapeutic treatment of colorectal cancer is challenged by severe gastrointestinal side effects, which include nausea, vomiting, constipation, and diarrhea. These symptoms can persist long after the treatment has been ceased. An emerging concept is the ability of platinum-based drugs to stimulate immunity, which is in contrast to conventional chemotherapeutic agents that are immunosuppressive. Here, we review the immunomodulatory aspects of platinum-based anticancer chemotherapeutics and their impact on gastrointestinal innervation. Given the bidirectional communication between the enteric nervous system and gastrointestinal immune system; exploring the consequences of platinum-induced immunogenicity will facilitate better understanding of gut dysfunction caused by chemotherapeutic agents. We propose that the development of future successful chemotherapeutics should rely on targeting the mechanisms underlying long-term gastrointestinal side effects.


Intact recognition of facial emotion in Parkinson's disease.
Although the basal ganglia have been shown to be critical for the expression of emotion in prosody and facial expressions, it is unclear whether they are also critical for recognition of emotions. Selective pathology of parts of the basal ganglia is a hallmark of individuals with Parkinson's disease, and such patients have been examined in several studies of emotion. We examined 18 patients with Parkinson's disease (11 men, 7 women) and 13 age-, education-, gender ratio-, and IQ-matched normal controls on their ability to recognize emotions signaled by facial expressions. Parkinson's patients performed entirely normally on a quantitative task of recognizing emotional facial expressions. The findings do not support the notion that the sectors of basal ganglia that are dysfunctional in Parkinson's disease are essential for recognizing emotion in facial expressions.


Contributions of the histone arginine methyltransferase PRMT6 to the epigenetic function of RUNX1.
Hematopoietic differentiation is directed by transcription factors such as RUNX1. RUNX1 binds to specific DNA binding sites in regulatory elements of genes and recruits epigenetic cofactors to target loci. In this way histone modification patterns and the chromatin environment are altered, which results in adjusted gene expression. The process of transcription factor binding and cofactor recruitment is dynamic and strongly influenced by specific posttranslational modifications, which are triggered by signaling. In this way cellular signaling is integrated at the epigenetic level by transcription factors. The identification of epigenetic cofactors and the study of their epigenetic influence on transcription is crucial for the understanding of transcription factor function in differentiation and disease. In this article, the recent observation that RUNX1 is associated with the protein arginine methyltransferase 6 will be reviewed. PRMT6 triggers H3R2me2a at RUNX1 target genes; this histone modification negatively influences the positive H3K4me3 mark and this way acts repressive. The RUNX1/PRMT6 association has an impact on bivalent histone marks. Upon differentiation, a RUNX1 corepressor complex with PRMT6 is exchanged with a RUNX1 coactivator complex. Furthermore, the potential cross talk of transcription factors and epigenetic cofactors with histone marks will be discussed.


A novel role for Arabidopsis CBL1 in affecting plant responses to glucose and gibberellin during germination and seedling development.
Glucose and phytohormones such as abscisic acid (ABA), ethylene, and gibberellin (GA) coordinately regulate germination and seedling development. However, there is still inadequate evidence to link their molecular roles in affecting plant responses. Calcium acts as a second messenger in a diverse range of signal transduction pathways. As calcium sensors unique to plants, calcineurin B-like (CBL) proteins are well known to modulate abiotic stress responses. In this study, it was found that CBL1 was induced by glucose in Arabidopsis. Loss-of-function mutant cbl1 exhibited hypersensitivity to glucose and paclobutrazol, a GA biosynthetic inhibitor. Several sugar-responsive and GA biosynthetic gene expressions were altered in the cbl1 mutant. CBL1 protein physically interacted with AKINβ1, the regulatory β subunit of the SnRK1 complex which has a central role in sugar signaling. Our results indicate a novel role for CBL1 in modulating responses to glucose and GA signals.


Functional and comparative genome analysis of novel virulent actinophages belonging to Streptomyces flavovirens.
Next Generation Sequencing (NGS) technologies provide exciting possibilities for whole genome sequencing of a plethora of organisms including bacterial strains and phages, with many possible applications in research and diagnostics. No Streptomyces flavovirens phages have been sequenced to date; there is therefore a lack in available information about S. flavovirens phage genomics. We report biological and physiochemical features and use NGS to provide the complete annotated genomes for two new strains (Sf1 and Sf3) of the virulent phage Streptomyces flavovirens, isolated from Egyptian soil samples. The S. flavovirens phages (Sf1 and Sf3) examined in this study show higher adsorption rates (82 and 85%, respectively) than other actinophages, indicating a strong specificity to their host, and latent periods (15 and 30 min.), followed by rise periods of 45 and 30 min. As expected for actinophages, their burst sizes were 1.95 and 2.49 virions per mL. Both phages were stable and, as reported in previous experiments, showed a significant increase in their activity after sodium chloride (NaCl) and magnesium chloride (MgCl2.6H2O) treatments, whereas after zinc chloride (ZnCl2) application both phages showed a significant decrease in infection. The sequenced phage genomes are parts of a singleton cluster with sizes of 43,150 bp and 60,934 bp, respectively. Bioinformatics analyses and functional characterizations enabled the assignment of possible functions to 19 and 28 putative identified ORFs, which included phage structural proteins, lysis components and metabolic proteins. Thirty phams were identified in both phages, 10 (33.3%) of them with known function, which can be used in cluster prediction. Comparative genomic analysis revealed significant homology between the two phages, showing the highest hits among Sf1, Sf3 and the closest Streptomyces phage (VWB phages) in a specific 13Kb region. However, the phylogenetic analysis using the Major Capsid Protein (MCP) sequences highlighted that the isolated phages belong to the BG Streptomyces phage group but are clearly separated, representing a novel sub-cluster. The results of this study provide the first physiological and genomic information for S. flavovirens phages and will be useful for pharmaceutical industries based on S. flavovirens and future phage evolution studies.


[Adenosquamous cell carcinoma of the lung with multiple cystic metastases in the liver].
A 70-year-old man was admitted to the hospital because of mild dyspnea, a cough, and hemoptysis. A chest X-ray film and a computed tomographic scan showed a mass in the S1.2 region of the left lung, and swollen mediastinal lymph noes. Cytologic examination of sputum sample resulted in the diagnosis of lung cancer. The tumor did not respond to chemotherapy, and the patient died after seven months. Autopsy disclosed a solid tumor of left lung and many cystic lesions in the liver. Histological examination of the lung lesion revealed adenosquamous cell carcinoma. Metastatic lesions in the liver consisted of adenosquamous cell carcinoma, with predominantly squamous cell carcinoma. Cases of lung cancer in which hepatic metastases have many cystic cavities are rare.


Postneurulation rapid brain growth represents a critical time for encephalocele formation: a chick model.
Current theory regarding the pathogenesis of encephaloceles suggests that the defect occurs after neurulation is completed in which brain tissue herniates through defective mesodermal elements necessary for skull modeling. To better delineate the mechanisms of encephalocele development, we performed a variety of microsurgical manipulations in the developing cranium of the chick embryo during the postneurulation period of early rapid brain growth. The results of the study revealed that encephaloceles could be induced in 78% of chick embryos manipulated at stage 26 that had evidence of marked decompression of the primitive ventricle. On the other hand, control embryos or embryos manipulated without ventricular decompression at stage 26 did not develop encephaloceles (0 of 32). Embryos with decompression of the primitive ventricle treated at earlier stages (20-24) showed only a 5% incidence of encephaloceles. These findings suggest that there is both a critical time point during postneurulation rapid brain growth when encephaloceles are prone to occur and a mesenchymal defect associated with decompression of the primitive ventricle that must be present to induce encephaloceles.


Effectiveness of a risk-based visitor-prioritizing system at a sexually transmitted infection outpatient clinic.
The objective of this study was to study the efficacy/effectiveness of a risk-based visitor-prioritizing system at a sexually transmitted infection (STI) clinic aimed to improve screening capacity by providing tailored service. In April 2004, a prioritizing system was implemented that classifies visitors as high or low risk depending on reported sexual behavior and previous STI events. The high- and low-risk groups are assigned to standard and short screening protocols, respectively. Both protocols include diagnostic testing for syphilis, urogenital gonorrhea, chlamydia, and optional for HIV. To assess the effectiveness of the system, differences in prevalence of STI diagnoses in the standard and short protocol were analyzed by chi test. In total, 14,391 visitors (64%) received standard screening and 8,056 visitors (36%) received short screening. The STI prevalence in both groups was 18.1% and 7.6%, respectively (P <0.001); prevalence of HIV was 1.8% and 0.3%, respectively (P <0.001). The sensitivity of the prioritizing system was 74%. Specificity was substantially lower (42%). This prioritizing system is effective in differentiating between visitors at high and low risk for STI, contributing to provision of tailored STI service, increasing efficiency, and client access to STI service.


A clinicopathologic study on SS18 fusion positive head and neck synovial sarcomas.
To determine clinicopathologic factors on survival in patients with head and neck synovial sarcoma. We retrospectively identified patients with molecularly confirmed synovial sarcomas of the head and neck (SS-HN), either by the presence of SS18-SSX fusion transcript by RT-PCR or SS18 gene rearrangement by FISH, who were managed at our institution over a 20-year period (1996-2015). Kaplan-Meier survival analysis and log-rank test were performed to evaluate variables related to disease specific survival (DSS). Fisher exact test was performed to evaluate variables related to local recurrence. Thirty-four patients (20 males and 14 females, mean of 31years) with SS18-SSX fusion-positive SS-HN were identified. The parapharyngeal region of the neck was the most common site. The mean tumor size was 4.8cm (0.8-10cm). Two-thirds (n=23) of cases had a monophasic histology. The 2, 5 and 10-year DSS rates were 97%, 79% and 68%. The 5-year DSS rates for the adult/pediatric cohort were 74%/88%. Recurrence showed significant effect on DSS (p=0.021). There was no significant effect on DSS with age, therapy modality, tumor site, surgical margin, tumor size (⩽5cm vs. >5cm) and histopathologic subtype. Tumor site (i.e. skull base/paranasal sinus region) was associated with local recurrence (p=0.003). In our cohort DSS rate was associated with recurrence. Tumors located in the skull base/paranasal sinus region were associated with a higher rate of local recurrence. Thus appropriate selection of high risk patients who can benefit from multimodality therapies might improve survival.


Radiolabeled and native antibodies and the prospect of cure of follicular lymphoma.
Advanced-stage follicular lymphoma is incurable by conventional treatment. Rituximab has been introduced in various combinations with chemotherapy and has resulted in a significantly superior treatment outcome compared with chemotherapy alone. Multiple studies have also shown the efficacy of radioimmunotherapy (RIT) both as a single agent and in combination with chemotherapy. Rituximab and RIT have clearly distinct mechanisms of action, the first acting exclusively as a biological treatment, while the second acts by a combination of biologic mechanisms and radiation effects. Despite the therapeutic efficacy of both approaches, the potential exists to further improve both modalities. Repeat administrations of RIT using appropriate radioisotopes for treatment of residual disease or new targeting strategies might afford additional benefits. Unlabeled antibody treatment could potentially benefit from the combination of antibodies directed against different target antigens or combination therapy with cytokines capable of further mobilizing patients' cellular defenses. In this review, we hypothesize that the combination of an optimized biological treatment together with radiolabeled antibodies and chemotherapy early in the disease course of advanced-stage follicular lymphoma may represent the best approach to achieve prolonged disease-free survival and eventually cure.


Heart rate measures from the Apple Watch, Fitbit Charge HR 2, and electrocardiogram across different exercise intensities.
This study compared heart rate (HR) measurements for the Fitbit Charge HR 2 (Fitbit) and the Apple Watch devices with HR measurements for electrocardiogram (ECG). Thirty young adults (15/15 females/males, age 23.5 ± 3.0 years) completed the Bruce Protocol. HR measurements were recorded from the ECG and both devices every minute. Average HR for each participant was calculated for very light, light, moderate, vigorous and very vigorous intensities based on ECG-measured HR. A concordance correlation coefficient (CCC) was calculated to examine the strength of the relationship between ECG measured HR and HR measured by each device. Relative error rates (RER) were also calculated to indicate the difference between each device and ECG. An equivalence test was conducted to examine the equivalence of HRs measured by devices and ECG. The Apple Watch showed lower RER (2.4-5.1%) compared with the Fitbit (3.9-13.5%) for all exercise intensities. For both devices, the strongest relationship with ECG-measured HR was found for very light PA with very high CCC (>.90) and equivalence. The strength of the relationship declined as exercise intensity increased for both devices. These findings indicate that the accuracy of real-time HR monitoring by the Apple Watch and Fitbit Charge HR2 is reduced as exercise intensity increases.


[Birth Cohort Technical Specification Part 3: Participants Information System (T/CPMA 015.3-2020)].
Birth cohort participants information system is a software system with multiple functions including recode basic information, automatic administration, paper-less questionnaire survey, clinical data integration, data visualization. The efficiency and quality of cohort data were improved dramatically by using this system. In this technical specification we described the basic functional requirement and detail steps of the development process of this information system.


Intestinal helminthiasis, nutritional status, and their relationship; a cross-sectional study in urban slum school children in Indonesia.
In a study of urban slum school children (276 boys and 231 girls), in Ujung Pandang, Indonesia, parasitological and anthropometric exams were cross-sectionally performed to assess prevalence and intensity of helminth infections and nutritional status. Prevalence of Ascaris, Trichuris and hookworm was 92%, 98%, and 1.4%, respectively. 91% children had both Ascaris and Trichuris infections. About half of the Ascaris- and Trichuris-infected children (46% and 58%, respectively) had moderate infections. Stunting was seen in 55% of the children, while wasting was observed in 10%. Boys had lower nutritional status than girls (p < 0.001), based on weight-for-age (WA) and height-for-age (HA) Z-scores. Age had an inverse relationship with WA and HA Z-scores (p < 0.0004). A relationship between helminth infections and nutritional status was observed between log Trichuris egg count and WA and HA Z-scores after controlling for age, sex, and log Ascaris egg count (p = 0.048 for HA Z-score, and p = 0.058 for WA Z-score). The relationship was also found when Trichuris infection was categorized into mild, moderate, and severe infections (p = 0.017 and p = 0.001 for HA and WA Z-scores, respectively). Scheffe's test for multiple mean comparisons showed that Trichuris-infected children with above 1,000 eggs per gram feces had significantly lower nutritional status than lower epg or non-infected children (p < 0.001 and p < 0.05 for HA and WA Z-scores respectively).


Sulfuryl transfer: the catalytic mechanism of human estrogen sulfotransferase.
Estrogen sulfotransferase (EST) catalyzes the transfer of the sulfuryl group from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to 17beta-estradiol (E2). The sulfation of E2 prevents it from binding to, and thereby activating, the estrogen receptor. The regulation of EST appears to be causally linked to tumorigenesis in the breast and endometrium. In this study, recombinant human EST is characterized, and the catalytic mechanism of the transfer reaction is investigated in ligand binding and initial rate experiments. The native enzyme is a dimer of 35-kDa subunits. The apparent equilibrium constant for transfer to E2 is (4.5 +/- 0.2) x 10(3) at pH 6.3 and T = 25 +/- 2 degrees C. Initial rate studies provide the kinetic constants for the reaction and suggest a sequential mechanism. E2 is a partial substrate inhibitor (Ki = 80 +/- 5 nM). The binding of two E2 per EST subunit suggests that the partial inhibition occurs through binding at an allosteric site. In addition to providing the dissociation constants for the ligand-enzyme complexes, binding studies demonstrate that each substrate binds independently to the enzyme and that both the E.PAP.E2S and E.PAP.E2 dead-end complexes form. These results strongly suggest a Random Bi Bi mechanism with two dead-end complexes.


Extraction of venom and venom gland microdissections from spiders for proteomic and transcriptomic analyses.
Venoms are chemically complex secretions typically comprising numerous proteins and peptides with varied physiological activities. Functional characterization of venom proteins has important biomedical applications, including the identification of drug leads or probes for cellular receptors. Spiders are the most species rich clade of venomous organisms, but the venoms of only a few species are well-understood, in part due to the difficulty associated with collecting minute quantities of venom from small animals. This paper presents a protocol for the collection of venom from spiders using electrical stimulation, demonstrating the procedure on the Western black widow (Latrodectus hesperus). The collected venom is useful for varied downstream analyses including direct protein identification via mass spectrometry, functional assays, and stimulation of venom gene expression for transcriptomic studies. This technique has the advantage over protocols that isolate venom from whole gland homogenates, which do not separate genuine venom components from cellular proteins that are not secreted as part of the venom. Representative results demonstrate the detection of known venom peptides from the collected sample using mass spectrometry. The venom collection procedure is followed by a protocol for dissecting spider venom glands, with results demonstrating that this leads to the characterization of venom-expressed proteins and peptides at the sequence level.


Machine Learning-Based Early Warning Systems for Acute Care Utilization During Systemic Therapy for Cancer.
Emergency department visits and hospitalizations frequently occur during systemic therapy for cancer. We developed and evaluated a longitudinal warning system for acute care use. Using a retrospective population-based cohort of patients who started intravenous systemic therapy for nonhematologic cancers between July 1, 2014, and June 30, 2020, we randomly separated patients into cohorts for model training, hyperparameter tuning and model selection, and system testing. Predictive features included static features, such as demographics, cancer type, and treatment regimens, and dynamic features, such as patient-reported symptoms and laboratory values. The longitudinal warning system predicted the probability of acute care utilization within 30 days after each treatment session. Machine learning systems were developed in the training and tuning cohorts and evaluated in the testing cohort. Sensitivity analyses considered feature importance, other acute care endpoints, and performance within subgroups. The cohort included 105,129 patients who received 1,216,385 treatment sessions. Acute care followed 182,444 (15.0%) treatments within 30 days. The ensemble model achieved an area under the receiver operating characteristic curve of 0.742 (95% CI, 0.739-0.745) and was well calibrated in the test cohort. Important predictive features included prior acute care use, treatment regimen, and laboratory tests. If the system was set to alarm approximately once every 15 treatments, 25.5% of acute care events would be preceded by an alarm, and 47.4% of patients would experience acute care after an alarm. The system underestimated risk for some treatment regimens and potentially underserved populations such as females and non-English speakers. Machine learning warning systems can detect patients at risk for acute care utilization, which can aid in preventive intervention and facilitate tailored treatment. Future research should address potential biases and prospectively evaluate impact after system deployment.


Simulation of abdomen sonography. Evaluation of a new ultrasound simulator.
We developed and evaluated a simulator for the sonography of the abdomen in order to improve the teaching quality in sonography training. Eleven medicine residents who had received 4 to 12 months full time sonography training performed ultrasound examinations of the right upper quadrant in 5 consecutive patients and in 5 simulator cases. The correctness of their findings and the time required for the examinations were measured. The subjective confidence in their findings and the handling of the ultrasound machines were rated on a visual analogue scale. During patient ultrasound examination 75 % (SEM 9%) of all pathologic findings were recognized by the residents, whereas 71 % (SEM 8%) of the pathologies of the simulator cases were found. This minimal difference was not significant in the paired, two sided t-test (p = 0.15). Severe pathologies did not escape detection. The time required for patient examination(10.57 min, SEM 3.25 min) was not significantly different (p = 0.53) to the time required for the simulator cases (9.59 min, SEM 2.98 min). The subjective confidence in the sonographic findings did not differ significantly (p = 0.39) between the real patient situation (68%, SEM 6%) and the simulation (64%, SEM 12 %). Only the handling of the ultrasound machines was judged to be significantly better (p=0.008) than the simulator (74%,SEM 7% vs. 61 %, SEM 12%). In this first direct crossover comparison between real patient sonography and simulator based scanning we proved that the simulator we developed simulates the real patient examination reliably and reproducibly.


Environment and air pollution like gun and bullet for low-income countries: war for better health and wealth.
The objective of the study is to examine the impact of environmental indicators and air pollution on "health" and "wealth" for the low-income countries. The study used a number of promising variables including arable land, fossil fuel energy consumption, population density, and carbon dioxide emissions that simultaneously affect the health (i.e., health expenditures per capita) and wealth (i.e., GDP per capita) of the low-income countries. The general representation for low-income countries has shown by aggregate data that consist of 39 observations from the period of 1975-2013. The study decomposes the data set from different econometric tests for managing robust inferences. The study uses temporal forecasting for the health and wealth model by a vector error correction model (VECM) and an innovation accounting technique. The results show that environment and air pollution is the menace for low-income countries' health and wealth. Among environmental indicators, arable land has the largest variance to affect health and wealth for the next 10-year period, while air pollution exerts the least contribution to change health and wealth of low-income countries. These results indicate the prevalence of war situation, where environment and air pollution become visible like "gun" and "bullet" for low-income countries. There are required sound and effective macroeconomic policies to combat with the environmental evils that affect the health and wealth of the low-income countries.


Depression in dialysis patients.
Many psychiatric disorders can be seen in patients with chronic renal failure (CRF). Haemodialysis (HD), which is a renal replacement treatment, causes various psychiatric and psychosocial problems. Patients are dependent on treatment and the illness causes various problems. In addition, strict diet and continuous treatment are other stress factors (1,2). Various studies have been published in different regions and countries about the prevalence of depression and the relation between sociodemographic factors and depression in patients treated by continuous ambulatory peritoneal dialysis (CAPD), which has gradually become common in Turkey. However studies, which reflect the authors' region, have become necessary (2,3).


Understanding Performance and Verbal-Communication of Children with ASD in a Collaborative Virtual Environment.
Collaborative virtual environments (CVEs), which allow naturalistic communication between two or more individuals in a shared virtual environment, hold promise as a tool for measuring and promoting social communication between peers. In this work, a CVE platform and a set of CVE-based collaborative games are designed for children with autism spectrum disorder (ASD). Two groups (7 ASD/TD pairs; 7 TD/TD pairs) participated in a pilot study to establish system feasibility and tolerability. We also designed a methodology for capturing meaningful metrics of social communication. Based on these metrics, we found improved game performance and trends in communication of these participants over time. Although preliminary, these results provide important insights on CVE-based interaction for ASD intervention.


SPINK5 is a Tumor-Suppressor Gene Involved in the Progression of Nonsmall Cell Lung Carcinoma through Negatively Regulating PSIP1.
This study aimed to elucidate how SPINK5 affects the malignant phenotypes of NSCLC and the molecular mechanism. NSCLC and adjacent normal tissues were collected to detect the differential level of SPINK5. The influence of SPINK5 on pathological indicators of NSCLC was analyzed. Cellular functions of NSCLC cells overexpressing SPINK5 were assessed by CCK-8, EdU, and transwell assay. By confirming the downstream target of SPINK5, its molecular mechanism on regulating NSCLC was finally explored through rescue experiments. SPINK5 was lowly expressed in NSCLC tissues, and it predicted tumor staging and lymphatic metastasis. In vitro overexpression of SPINK5 declined proliferative and migratory rates in NSCLC cells. PSIP1 was verified as the target gene binding SPINK5, and they displayed a negative correlation in NSCLC tissues. Overexpression of PSIP1 was able to reverse the inhibited proliferative and migratory potentials in NSCLC cells overexpressing SPINK5. SPINK5 level has a close relation to tumor staging and lymphatic metastasis in NSCLC. It serves as a tumor-suppressor gene that inhibits proliferation and migration of NSCLC through negatively regulating PSIP1.


Huntington Disease: Molecular Diagnostics Approach.
Huntington disease (HD) is caused by expansion of a CAG trinucleotide repeat in the first exon of the Huntingtin (HTT) gene. Molecular testing of Huntington disease for diagnostic confirmation and disease prediction requires detection of the CAG repeat expansion. There are three main types of HD genetic testing: (1) diagnostic testing to confirm or rule out disease, (2) presymptomatic testing to determine whether an at-risk individual inherited the expanded allele, and (3) prenatal testing to determine whether the fetus has inherited the expanded allele. This unit includes protocols that describe the complementary use of polymerase chain reactions (PCR) and Southern blot hybridization to accurately measure the CAG trinucleotide repeat size and interpret the test results. In addition, an indirect linkage analysis that does not reveal the unwanted parental HD status in a prenatal testing will also be discussed.


Pervasive developmental disorder in the children of immigrant parents: comparison of different assessment instruments.
The objective of this study was to describe how the Childhood Autism Rating Scale (CARS) behaves in relation to the Autism Diagnostic Observation Schedule (ADOS) and to clinical diagnosis based on the criteria defined in the Diagnostic and Statistical Manual of Mental Disorders, 4 th Edition (DSM-IV) for children of immigrant parents. Forty-nine children of parents who had immigrated to Canada were evaluated. In this sample, the ADOS and the DSM-IV showed complete agreement. Using the standard cut-off point of 30, the CARS showed high specificity and poor sensitivity. The study proposes a cut-off point for the CARS that would include pervasive developmental disorder - not otherwise specified (PDD-NOS). Reducing the cut-off point to 20/21 increased the specificity of the instrument for this group of children without significantly reducing its sensitivity.


Reversed Cherenkov-transition radiation by a charge crossing a left-handed medium boundary.
We analyze the radiation from a charged particle crossing the boundary between an ordinary medium and a "left-handed" metamaterial. We obtain exact and approximate expressions for the field components and develop algorithms for their computation. The spatial radiation in this system can be separated into three distinct components, corresponding to ordinary transition radiation having a relatively large magnitude, Cherenkov radiation, and reversed Cherenkov-transition radiation (RCTR). The last one is explained by reflection and refraction of reversed Cherenkov radiation at the interface. Conditions for generating of RCTR are obtained. We note properties of this radiation that have potential applications in the detection of charged particles and accelerator beams and for the characterization of metamaterial macroscopic parameters (epsilon, mu).


[Evaluation on quality of large sample clinical trials of acupuncture in foreign countries].
Following retrieving articles about foreign large sample clinical trials of acupuncture (more than 500 cases) from MEDLINE and EMBASE databases (1996-2012) by using keywords of "acupuncture" and "clinical trial", a quality analysis was carried out independently by two researchers. A total of 1373 papers were collected and 37 were considered to meet our preformulated standards in accordance with the principles and methods of evidence-based medicine. Majority of these 37 articles were from some advanced countries including Germany, USA, Japan, Demark, Australia, et al. Their paper quality and research quality are varied including the methodology, standards for diagnosis, inclusive and exclusive criteria, follow-up survey, economic effectiveness, adverse effects, acupoint application and syndrome differentiation of traditional Chinese medicine. Of the 37 papers, 17 are multiple center clinical trials, 18 are one center clinical trials, and 2 are not clear. Our Chinese researchers should pay more attention to foreign well-designed, large example, randomized controlled clinical trials and draw their matured experience and strong points to compensate our weak points and to improve our levels in clinical study on acupuncture treatment of clinical disorders.


Typing of Bacillus sporothermodurans and other Bacillus species isolated from milk by repetitive element sequence based PCR.
When analysing raw milk for the presence of Bacillus sporothermodurans, 11 Bacillus strains were isolated which could be differentiated from known Bacillus, Brevibacillus and Paenibacillus spp. by different primer specificity in PCR experiments, indicating that they probably belong to Bacillus species as yet undescribed. Using repetitive element sequence based PCR (REP-PCR), these 11 strains could be clearly distinguished from B. sporothermodurans as well as from each other. Eighty-five B. sporothermodurans strains were characterized by a typical REP-pattern. Using REP-PCR combined with separation on non-denaturing polyacrylamide gel electrophoresis and silver staining, individual B. sporothermodurans strains could be discriminated, which was not possible by methods previously published.


Controllable Microfluidic Production of Drug-Loaded PLGA Nanoparticles Using Partially Water-Miscible Mixed Solvent Microdroplets as a Precursor.
We present a versatile continuous microfluidic flow-focusing method for the production of Doxorubicin (DOX) or Tamoxifen (TAM)-loaded poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs). We use a partially water-miscible solvent mixture (dimethyl sulfoxide DMSO+ dichloromethane DCM) as precursor drug/polymer solution for NPs nucleation. We extrude this partially water-miscible solution into an aqueous medium and synthesized uniform PLGA NPs with higher drug loading ability and longer sustained-release ability than conventional microfluidic or batch preparation methods. The size of NPs could be precisely tuned by changing the flow rate ratios, polymer concentration, and volume ratio of DCM to DMSO (VDCM/VDMSO) in the precursor emulsion. We investigated the mechanism of the formation of NPs and the effect of VDCM/VDMSO on drug release kinetics. Our work suggests that this original, rapid, facile, efficient and low-cost method is a promising technology for high throughput NP fabrication. For the two tested drugs, one hydrophilic (Doxorubicin) the other one hydrophobic (Tamoxifen), encapsulation efficiency (EE) as high as 88% and mass loading content (LC) higher than 25% were achieved. This new process could be extended as an efficient and large scale NP production method to benefit to fields like controlled drug release and nanomedicine.


Conserved intermolecular salt bridge required for activation of protein kinases PKR, GCN2, and PERK.
The protein kinases PKR, GCN2, and PERK phosphorylate translation initiation factor eIF2alpha to regulate general and genespecific protein synthesis under various cellular stress conditions. Recent x-ray crystallographic structures of PKR and GCN2 revealed distinct dimeric configurations of the kinase domains. Whereas PKR kinase domains dimerized in a back-to-back and parallel orientation, the GCN2 kinase domains displayed an antiparallel orientation. The dimerization interfaces on PKR and GCN2 were localized to overlapping surfaces on the N-terminal lobes of the kinase domains but utilized different intermolecular contacts. A key feature of the PKR dimerization interface is a salt bridge interaction between Arg(262) from one protomer and Asp(266) from the second protomer. Interestingly, these two residues are conserved in all eIF2alpha kinases, although in the GCN2 structure, the two residues are too remote to interact. To test the importance of this potential salt bridge interaction in PKR, GCN2, and PERK, the residues constituting the salt bridge were mutated either independently or together to residues with the opposite charge. Single mutations of the Asp (or Glu) and Arg residues blocked kinase function both in yeast cells and in vitro. However, for all three kinases, the double mutation designed to restore the salt bridge interaction with opposite polarity resulted in a functional kinase. Thus, the salt bridge interaction and dimer interface observed in the PKR structure is critical for the activity of all three eIF2alpha kinases. These results are consistent with the notion that the PKR structure represents the active state of the eIF2alpha kinase domain, whereas the GCN2 structure may represent an inactive state of the kinase.


The Ste-Justine Adolescent Idiopathic Scoliosis Cohort Study. Part I: Description of the study.
This study determined the impact of adolescent idiopathic scoliosis (AIS) on health and well being in adulthood by comparing AIS subjects, more than 10 years after referral, to non-AIS subjects according to perceived health status, physical activity, fitness, reproductive health, respiratory condition, and back and neck pain. This paper describes the cohort, the design of the study, and response rates. The study was designed as a comparative retrospective cohort study. A total of 2,092 patients referred for AIS to Ste-Justine Hospital, Montreal, Quebec, between 1960 and 1979 were identified. A population-based control group was selected in 1989-90 by randomly telephoning persons in Quebec. The control group was approximately frequency-matched to the AIS cohort according to age group and broad geographic region of current residence. Patients with adolescent idiopathic scoliosis were traced, and a self-administered questionnaire was sent by post in 1989 to subjects whose addresses were found. Identical questionnaires were sent to subjects in the control group. Seventy-one percent of the cohort (1,476 subjects) returned questionnaires in 1989. Subjects who did not respond to the questionnaires were similar to those who did complete the questionnaires on a number of key factors, except that lower response rates were observed for subjects with scoliotic curves under 20 degrees. The control group consisted of 1,755 subjects (55% response rate). It was similar to the general Quebec population and to the AIS cohort on a number of important sociodemographic factors. Comparisons on health outcomes between the two study groups should not be affected by differential patterns of selection or response.


Children understand subjective (undesirable) desires before they understand subjective (false) beliefs.
Our folk psychology is built around the ascription of beliefs (and related cognitive states) and desires (and related conative states). How and when children develop a concept of these different kinds of propositional attitudes has been the subject of a long-standing debate. Asymmetry accounts assume that children develop a conception of desires earlier than they develop a concept of beliefs. In contrast, the symmetry account assumes that conceptions of both kinds of attitudes are based on the same underlying capacity to ascribe subjective perspectives. Accordingly, a genuine subjective understanding of desires develops in tandem with subjective belief understanding. So far, existing evidence that tested these two accounts remains inconclusive, with inconsistent findings resulting from diverging methods. Therefore, the current study tested between the two accounts in a more systematic way. First, we used a particularly clear test case-value-incompatible (wicked) desires. Such desires are strongly subjective because they are desirable only from the agent's perspective but not from an objective perspective. Second, we probed children's ascription of such desires in the most direct and simplified ways. Third, we directly compared children's desire understanding with their ascription of subjective beliefs. Results revealed that young children were better in reasoning about subjective desires than about subjective beliefs. Desire reasoning was not correlated with subjective belief reasoning, and children did not have more difficulties in reasoning about strongly subjective wicked desires than about neutral desires. All in all, these findings are not in line with the predictions of the symmetry account but speak in favor of the asymmetry account.


Caraway oil inhibits skin tumors in female BALB/c mice.
Caraway oil, supplemented in diet or painted on the skin, inhibited 7,12-dimethylbenz[a]anthracene- (DMBA) and croton oil-induced skin tumors in female BALB/c mice. The inhibition was manifested by disappearance of carcinomas, reduced incidence and number of papillomas, delay of their appearance, retardation of their development, and regression of already established papillomas. Caraway oil is more effective when topically applied than when supplemented in the diet.


Cavernous hemangioma of rhinopharynx: our experience and review of literature.
Hemangiomas are benign tumors originating in the vascular tissues of skin, mucosa, muscles, glands, and bones. Although these tumors are common lesions of the head and neck, they rarely occur in the nasal cavity and paranasal sinuses. Cavernous haemangioma of the lateral wall of the nasopharynx has not previously been reported. We examined the clinical, radiological and therapeutic management of cavernous haemangioma of nasopharynx starting from a clinical case of a 26-year-old woman with a history of recurrent and conspicuous epistaxis and left-sided nasal associated severe obstruction. Nasopharynx examination, by flexible endoscopy, showed a cystic mass borne by the left side wall of the nasopharynx, in contact with the soft palate, covered by intact and regular mucosa. Contrast-enhanced computed tomography (CT) scan, confirmed these findings and showed contextual lamellar calcifications and inhomogeneous enhancement. The nasal endoscopic approach (FESS), under general anesthesia, allowed removal of the mass, without complications, after careful hemostasis of arterial branches. It was possible to establish the precise site of origin of the tumor only during the surgical procedure. Histopathological study showed mucosa with extensive vascular proliferation, with framework of lacunar/cavernous haemangioma, also present at lamellar bone tissue level. An unusual site and an unspecific clinical appearance can make diagnosis and treatment of a cavernous hemangioma of the nasopharynx difficult. The nasal endoscopic technique proved to be reliable in terms of adequate exposure and visualization of the lesion, control of bleeding, and complete removal of the mass.


Hydrothermal Synthesis of Nanoclusters of ZnS Comprised on Nanowires.
Cetyltrimethyl ammonium bromide cationic (CTAB) surfactant was used as template for the synthesis of nanoclusters of ZnS composed of nanowires, by hydrothermal method. The structural and morphological studies were performed by using X-ray diffraction (XRD), scanning electron microscopy (SEM) and high resolution transmission electron microscopy (HRTEM) techniques. The synthesized ZnS nanoclusters are composed of nanowires and high yield on the substrate was observed. The ZnS nanocrystalline consists of hexagonal phase and polycrystalline in nature. The chemical composition of ZnS nanoclusters composed of nanowires was studied by X-ray photo electron microscopy (XPS). This investigation has shown that the ZnS nanoclusters are composed of Zn and S atoms.


[Study on relationship between VEGF-C and lymphangiogenesis and lymph node metastasis in non-small cell lung cancer].
Increasing evidences have shown that vascular endothelial growth factor C (VEGF-C) is involved in the tumor lymphangiogenesis via combining with its ligand, vascular endothelial growth factor receptor-3 (VEGFR-3), which is the most important factor contributing to lymph node metastasis. Futhermore, lymph node metastasis is the most familiar metastatic pathway in non-small cell lung cancer (NSCLC). Based on this knowledge, the present study aims to explore the relationship between VEGF-C and lymphangiogenesis and lymph node metastasis in NSCLC with the use of podoplanin, a novel lymphatic vessel endothelium marker to detect lymphatic vessel. The expression of VEGF-C and podoplanin were detected in 66 paraffin sections of NSCLC and 8 inflammatory pseudotumor tissues by immunohistochemistry (SP). Assessments of podoplanin+ lymphatic vessel density (LVD) and positive rates of VEGF-C were performed and their relationship was analysed. The positive rate of VEGF-C in the inflammatory pseudotumor group (12.5%) was significantly lower than that in the NSCLC group (75.8%) (P < 0.01), the positive rate of VEGF-C in the positive lymph node group was significantly higher than that in the negative lymph node metastasis group (86.5% vs 62.1%, P < 0.05) and there was no difference between the well differentiated group and the poor differentiated group (76.3% vs 75.0%, P > 0.05). LVD in the positive VEGF-C group was significantly higher than that in the negative VEGF-C group (21.3±6.0 vs 17.7±5.1, P < 0.05), LVD in the inflammatory pseudotumor group (10.9±4.9) was distinctly lower than that in the NSCLC group (20.4±5.9) (P < 0.01) and compared with the negative lymph node metastasis group (18.5± 5.5), LVD (21.9±5.9) in the positive lymph node metastasis group increased significantly (P < 0.05). Lymphangiogenesis is a significant factor for tumor lymphatic metastasis, VEGF-C may mediate lymphatic metastasis of NSCLC by the way of inducing lymphangiogenesis.


Effect of inoculants and storage temperature on the microbial, chemical and mycotoxin composition of corn silage.
To evaluate the effect of lactic acid bacteria and storage temperature on the microbial, chemical and mycotoxin composition of corn silage. Corn was harvested at 32.8% dry matter, and chopped to 1 to 2 cm. The chopped material was subjected to three treatments: i) control (distilled water); ii) 1×106 colony forming units (cfu)/g of Lactobacillus plantarum; iii) 1×106 cfu/g of Pediococcus pentosaceus. Treatments in triplicate were ensiled for 55 d at 20°C, 28°C, and 37°C in 1-L polythene jars following packing to a density of approximately 800 kg/m3 of fresh matter, respectively. At silo opening, microbial populations, fermentation characteristics, nutritive value and mycotoxins of corn silage were determined. L. plantarum significantly increased yeast number, water soluble carbohydrates, nitrate and deoxynivalenol content, and significantly decreased the ammonia N value in corn silage compared with the control (p<0.05). P. pentosaceus significantly increased lactic acid bacteria and yeast number and content of deoxynivalenol, nivalenol, T-2 toxin and zearalenone, while decreasing mold population and content of nitrate and 3-acetyl-deoxynivalneol in corn silage when stored at 20°C compared to the control (p<0.05). Storage temperature had a significant effect on deoxynivalenol, nivalenol, ochratoxin A, and zearalenone level in corn silage (p<0.05). Lactobacillus plantarum and Pediococcus pentosaceus did not decrease the contents of mycotoxins or nitrate in corn silage stored at three temperatures.


[Effects of diet and physical activity factors on blood pressure in nine provinces of China: a longitudinal analysis].
To study the effects of diet and physical activity factors on blood pressure in nine provinces, using the multilevel model. Data was collected in the China Health and Nutrition Survey (CHNS). A total of 6706 men and 7140 women aged above 18 who attended at least one of the surveys in the year of 1997, 2000, 2004 and 2006 were selected, and a two-level male and female random intercept-slope growth models were applied to estimate the relationship between the intake of daily salt, vegetable, fruit, fat, protein as well as the time spent on the moderate or heavy physical activity and blood pressure. After controlling for age, education, BMI, drinks and total energy intake, mean of the daily salt intake per person was positively associated with systolic blood pressure in women (β = 0.0481, s(x(-)) = 0.0178, P < 0.01). Mean of the daily vegetable intake per person was negatively associated with systolic blood pressure in men and women, with the regression coefficients as -0.0063, -0.0068 respectively, indicating that if mean of the daily vegetable intake per person increased by 100 g, the systolic blood pressure would decrease by 0.6 mm Hg (1 mm Hg = 0.133 kPa) or more. In addition, the daily vegetable intake was also negatively associated with diastolic blood pressure (P < 0.01). Daily fruit intake was negatively associated with systolic blood pressure both in men and women, with regression coefficients as -0.0029 and -0.0031 respectively. Time spent on the moderate or heavy physical activity was also negatively associated with systolic blood pressure in both men and women, and diastolic blood pressure in women (P < 0.05). No relationship was found between daily fat, protein intake and blood pressure. Daily salt, vegetable, fruit intake, time spent on the moderate or heavy physical activity were associated with blood pressure in both men and women. Programs on integrated lifestyle modifications including dietary salt reduction, eating more vegetable and fruits, increasing physical activity level, plus weight control were critical for the control of high blood pressure.


A cost analysis of implementing mobile health facilitated tuberculosis contact investigation in a low-income setting.
Mobile health (mHealth) applications may improve timely access to health services and improve patient-provider communication, but the upfront costs of implementation may be prohibitive, especially in resource-limited settings. We measured the costs of developing and implementing an mHealth-facilitated, home-based strategy for tuberculosis (TB) contact investigation in Kampala, Uganda, between February 2014 and July 2017. We compared routine implementation involving community health workers (CHWs) screening and referring household contacts to clinics for TB evaluation to home-based HIV testing and sputum collection and transport with test results delivered by automated short messaging services (SMS). We carried out key informant interviews with CHWs and asked them to complete time-and-motion surveys. We estimated program costs from the perspective of the Ugandan health system, using top-down and bottom-up (components-based) approaches. We estimated total costs per contact investigated and per TB-positive contact identified in 2018 US dollars, one and five years after program implementation. The total top-down cost was $472,327, including $358,504 (76%) for program development and $108,584 (24%) for program implementation. This corresponded to $320-$348 per household contact investigated and $8,873-$9,652 per contact diagnosed with active TB over a 5-year period. CHW time was spent primarily evaluating household contacts who returned to the clinic for evaluation (median 30 minutes per contact investigated, interquartile range [IQR]: 30-70), collecting sputum samples (median 29 minutes, IQR: 25-30) and offering HIV testing services (median 28 minutes, IQR: 17-43). Cost estimates were sensitive to infrastructural capacity needs, program reach, and the epidemiological yield of contact investigation. Over 75% of all costs of the mHealth-facilitated TB contact investigation strategy were dedicated to establishing mHealth infrastructure and capacity. Implementing the mHealth strategy at scale and maintaining it over a longer time horizon could help decrease development costs as a proportion of total costs.


Growth hormone refractory interval to levodopa stimulation.
The GH response to repeat l-dopa stimulation was studied in three phases. In phase I and phase II, 16 normal adults divided into four groups were restimulated at 3, 4, 5, and 6 hours with 500 mg l-dopa and 1000 mg l-dopa, respectively. No response could be elicited at 3 hours and only at 6 hours (500 mg l-dopa) and 5 hours (1000 mg l-dopa) did the repeat responses return in magnitude to that of the initial. The period of decreased responsiveness to GH was terned the "refractory interval". In phase III, addition of an inhibitor of peripheral L-aromatic amino acid decarboxylase (50 mg MK-486) to the restimulation dose shortened but did not eliminate the refractoriness. Our results suggest that the refractory interval is not due to a peripherally located mechanism.


Predictive Potential of Head and Neck Squamous Cell Carcinoma Organoids.
In this issue of Cancer Discovery, Driehuis and colleagues establish culture conditions for the rapid, efficient, and long-term expansion of healthy human oral mucosa and head and neck squamous cell carcinoma (HNSCC) tumor organoids. The HNSCC tumor organoids provide a functional platform for analyzing tumor cell phenotype, tumorigenic potential, and drug and radiotherapy response, and they have a potential role in clinical decision-making.See related article by Driehuis et al., p. 852.


Insulin secretion in health and disease: nutrients dictate the pace.
Insulin is a key hormone controlling metabolic homeostasis. Loss or dysfunction of pancreatic β-cells lead to the release of insufficient insulin to cover the organism needs, promoting diabetes development. Since dietary nutrients influence the activity of β-cells, their inadequate intake, absorption and/or utilisation can be detrimental. This review will highlight the physiological and pathological effects of nutrients on insulin secretion and discuss the underlying mechanisms. Glucose uptake and metabolism in β-cells trigger insulin secretion. This effect of glucose is potentiated by amino acids and fatty acids, as well as by entero-endocrine hormones and neuropeptides released by the digestive tract in response to nutrients. Glucose controls also basal and compensatory β-cell proliferation and, along with fatty acids, regulates insulin biosynthesis. If in the short-term nutrients promote β-cell activities, chronic exposure to nutrients can be detrimental to β-cells and causes reduced insulin transcription, increased basal secretion and impaired insulin release in response to stimulatory glucose concentrations, with a consequent increase in diabetes risk. Likewise, suboptimal early-life nutrition (e.g. parental high-fat or low-protein diet) causes altered β-cell mass and function in adulthood. The mechanisms mediating nutrient-induced β-cell dysfunction include transcriptional, post-transcriptional and translational modifications of genes involved in insulin biosynthesis and secretion, carbohydrate and lipid metabolism, cell differentiation, proliferation and survival. Altered expression of these genes is partly caused by changes in non-coding RNA transcripts induced by unbalanced nutrient uptake. A better understanding of the mechanisms leading to β-cell dysfunction will be critical to improve treatment and find a cure for diabetes.


Synthesis and toxicity assessment of 3-oxobutanamides against human lymphocytes and isolated mitochondria.
To reduce costly late-phase compound scrubbing, there has been an increased focus on assessing compounds within in vitro assays that predict properties of human safety liabilities, before preclinical in vivo studies. The aim of our study was to answer the questions that whether the toxicity risk of a series of 3-oxobutanamide derivatives could be predicted by using of human lymphocytes and their isolated mitochondria. Using biochemical and flow cytometry assessments, we demonstrated that exposure of lymphocytes and isolated mitochondria to five 3-oxobutanamide derivatives (1-5) did not exhibit remarkable toxicity at low concentrations (50-500μM) but toxicity could be observed at high concentrations (1000 and 2000μM), particularly for N-(5-(4-bromophenyl)-3-isoxazolyl)-3-oxobutanamide (4) and N-(2-benzothiazolyl)-3-oxo butanamide (5). Compounds 4, 5 and partly N-(5-methyl-3-isoxazol yl)-3-oxo butanamide (1) also showed a marked cellular and mitochondrial toxicity while compound 5 displayed superior toxicity. Compound 5 induced cytotoxicity on human blood lymphocytes which was associated with the generation of intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP) collapse, lysosomal membrane injury, lipid peroxidation and depletion of glutathione. Our results suggested that among assessed compounds, increased toxicity of compound 5 compared to other compounds could be likely attributed to the presence of bromine substituent in 5. Finally our findings proposed that using of antioxidants and mitochondrial/lysosomal protective agents could be beneficial in decreasing the toxicity of 5.


Use of small-diameter capillaries for increasing peptide and protein detection sensitivity in capillary electrophoresis-mass spectrometry.
The use of small ID capillaries is shown to provide a substantial increase in sensitivity for capillary electrophoresis-electrospray ionization/mass spectrometry (CE-ESI/MS). In a comparison using capillaries ranging from either 100 to 10 microns or 50 to 5 microns ID and chemically modified with aminopropylsilane, a 25- to 50-fold increase in sensitivity was observed for both peptide and protein mixtures. This enhanced solute sensitivity allowed the detection of approximately 150 attomoles of melittin (2845 Da) with selected ion monitoring and 600 attomoles of carbonic anhydrase (29,157 Da) while scanning for CE-MS with a quadrupole mass spectrometer. For the protein mixture, mass spectra of sufficient quality for precise molecular weight determination (< or = 0.05%) were obtained for 600 attomole injections using a 5 microns ID capillary. The increase in sensitivity with small capillary diameters can be primarily attributed to a reduced mass flow rate of buffer and other background constituents into the electrospray source, which allows for greater sample ionization efficiency. A model that qualitatively accounts for the results is presented, but quantitative agreement is precluded due to difficulties in accounting for contributions due to a liquid sheath flow used with the electrospray source. The model accounts for the observation that the ESI/MS appears to function as a concentration-sensitive detector under many conditions using large-diameter capillaries. A transition occurs, however, to a regime where the ESI/MS functions as a mass flow-sensitive detector for small-diameter capillaries, where the ESI current is limited by the rate of delivery to the ESI source of charge carrying species in solution. These results suggest peptide and protein analysis at low attomole and subattomole levels should be obtainable with alternative types of mass spectrometers.


Bovine lactoferrin has a nitric oxide-dependent hypotensive effect in rats.
Lactoferrin (LF) is a multifunctional protein that is found in milk, neutrophils, and other biological fluids. Under inflammatory conditions, LF production is increased in the periphery by neutrophils. However, the cardiovascular function of LF is still unknown. In the present study, we investigated the effect of bovine LF (BLF) on the mean blood pressure (MBP) and heart rate (HR) in urethane-anesthetized rats and the vascular function of BLF in the rat thoracic aorta. Intravenous injection of BLF produced dose-dependent decreases in MBP but did not affect HR, while the opioid agonist morphine decreased both MBP and HR. The hypotensive effect of BLF was not altered by naloxone methiodide, which cannot pass through the blood-brain barrier, but was significantly reduced by naloxone hydrochloride, which does pass through the blood-brain barrier. BLF-induced hypotension was completely blocked by the NO synthase inhibitor NG-nitro-l-arginine methyl ester (l-NAME) but not by the inactive enantiomer of l-NAME, NG-nitro-d-arginine methyl ester (d-NAME). BLF-induced hypotension was not altered by the muscarinic ACh receptor antagonist atropine or the cyclooxygenase inhibitor diclofenac. BLF produced relaxation in endothelium-intact but not endothelium-denuded aortic rings precontracted with phenylephrine. The relaxation evoked by BLF was completely blocked by l-NAME but not by d-NAME or the ATP-sensitive potassium channel blocker glibenclamide. These results suggest that BLF causes hypotension via an endothelium-dependent vasodilation that is strongly mediated by NO production and that BLF-induced hypotension also may be mediated by the central opioidergic system.


Exercise-deficit disorder in children: are we ready to make this diagnosis?
Primary care physicians, particularly pediatricians, are expected to promote physical activity and prescribe exercise to patients. Unfortunately, many health care professionals do not have adequate skills or experience in this area, making exercise intervention logistically and psychologically challenging. Exercise-deficit disorder (EDD), which is a new term to the literature, is defined as reduced levels of moderate-to-vigorous physical activity, inconsistent with public health recommendations. Physicians are in a unique position to identify children who are deficient in exercise and initiate prevention strategies. To aid physicians in the diagnosis and treatment of children with EDD, we propose integrating exercise assessment into primary care practice, and, when appropriate, referral to a pediatric exercise specialist. This approach should ensure adequate development of muscle strength and motor skills that are required for successful physical activity participation. The timely identification of children with EDD is essential in preventing resistance to intervention. Connecting pediatric health care professionals with pediatric exercise specialists, culminating in an established professional collaborative system, is critical for promoting healthy behaviors throughout childhood and adolescence.


Plasma concentrations of IFN-gamma and TNF-alpha in psoriatic patients before and after local treatment with dithranol ointment.
IFN-gamma and TNF-alpha plasma levels were measured before and after local treatment in 27 patients. Twenty healthy subjects served as controls. Plasma concentrations of IFN-gamma and TNF-alpha were significantly higher before treatment (178.7 +/- 11.9 pg/ml and 31.9 +/- 11.6 pg/ml, respectively) compared to the control group (139.6 +/- 7.86 pg/ml and 17.1 +/- 7.7 pg/ml, respectively). After treatment IFN-gamma levels were significantly decreased (151.3 +/- 8.3 pg/ml) toward the control group values and TNF-alpha levels were observed even lower than in the controls (11.48 +/- 6.8 pg/ml). No correlations were found between age, duration of psoriasis and plasma levels of cytokines. However, IFN-gamma levels were related, although not significantly, to disease severity (evidenced by the PASI score). The data support the important proinflammatory role of IFN-gamma and TNF-alpha in the clinical manifestation of psoriasis.


Development of self-assembling mixed protein micelles with temperature-modulated avidities.
Elastin-like polypeptides (ELPs) are polypentapeptides that undergo hydrophobic collapse and aggregation above a specific transition temperature, Tt . ELP diblocks sharing a common "core" block (I60) but varying "outer" blocks (A80, P40) were designed, where Tt,I < Tt,A < Tt,P . The formation of ∼55 nm diameter mixed micelles from these ELP diblocks was verified using dynamic light scattering (DLS), multiangle light scattering (MALS) and fluorescence resonance energy transfer (FRET). To confer affinity to the blood circulating protein fibrinogen, a fibrinogen-binding tetrapeptide sequence (GPRP) was fused to A80-I60, while P40-I60 was fused to a non-binding control (GPSP). The self-assembling, peptide-displaying, mixed micelles exhibit temperature-modulated avidities for immobilized and soluble fibrinogen at 32 °C and 42 °C. In this initial proof-of-concept design, the engineered mixed micelles were shown to disengage fibrinogen at elevated temperatures. The modular nature of this system can be used for developing in vivo depot systems that will only be triggered to release in situ upon specific stimuli.


The "Cryptic" Escherichia.
In 2009, five monophyletic Escherichia clades were described and referred to as "cryptic" based on the inability to distinguish them from representative E. coli isolates using diagnostic biochemical reactions. Since this original publication, a number of studies have explored the genomic, transcriptomic, and phenotypic diversity of cryptic clade isolates to better understand their phylogenetic, physiological, and ecological distinctiveness with respect to previously named Escherichia species. This chapter reviews the original discovery of the cryptic clades, discusses available evidence that some are environmentally adapted, and evaluates current support for taxonomic designations of these microorganisms. The importance of these clades to clinical research, epidemiology, population genetics, and microbial speciation is also discussed.


Alexithymia is associated with mood disorders, impairment in quality of life and disability in women with fibromyalgia.
Alexithymia is a personality trait related to the quality of life of women with fibromyalgia (FM). It is still unknown whether alexithymia is associated with the clinical manifestations of FM. The present study describes the relationship between alexithymia and the domains included in the core set recommended by the Outcome Measures in Rheumatology (OMERACT) for FM evaluation. One hundred two women with FM were enrolled in the cross-sectional study. The domains evaluated were alexithymia, pain, fatigue, health-related quality of life, sleep quality, depression, anxiety, and disability. Univariate and multivariate (Kernel Regularized Least Squares method) analyses were performed to assess the relationship between alexithymia and the domains included in the core set recommended by the OMERACT. Alexithymia prevalence was 64.5% (95% Confidence Interval [CI], 54.6%-73.9%) and higher in women with depression (76.1%; 95%CI, 63.8%-86%). Female patients with FM and alexithymia showed higher pain intensity, anxiety and depression levels, and disability perception and lower quality of life, as compared to those with FM without alexithymia. Size effect differences ranged from medium to large and all of them were statistically significant (p<0.05). Using multivariate analysis, alexithymia was significantly associated with worse perceptions of quality of life (except physical health domain) and more disability perception, independently of other variables. However, alexithymia was not significantly associated with pain intensity. Alexithymia plays an important role in clinical manifestations of FM, mainly in the psychological and social dimensions of quality of life and the degree of perceived disability.


Subsequent physical and mental development in infants with positive contraction stress tests.
Little knowledge exists concerning long-term sequelae in children born of pregnancies complicated by chronic uteroplacental insufficiency. This follow-up study evaluates 12 children with positive contraction stress tests (CSTs) 5.5 months to 4.75 years following birth. Subsequent physical growth was uniformly normal in the four infants classified as appropriate for gestational age at birth; however, continued postnatal growth deficiency was present in 50% of the eight infants who suffered from intrauterine growth retardation. No major neurologic abnormalities were detected, and 10 of the 12 displayed normal mental and motor function, as determined by the Denver Developmental Screening Test, Bayley Scales of Infant Development and Wechsler Preschool and Primary Scale of Intelligence. Two of the 12 infants were depressed at birth and had serious neonatal complications. These were the only two children who scored poorly on the Bayley test. Although no conclusions regarding minimal brain dysfunction can be made, the majority of infants with positive CSTs demonstrate grossly normal neurologic and psychologic development.