Systemic RNA Interference Defective (SID) genes modulate dopaminergic neurodegeneration in C. elegans.
The fine-tuning of gene expression is critical for all cellular processes; aberrations in this activity can lead to pathology, and conversely, resilience. As their role in coordinating organismal responses to both internal and external factors have increasingly come into focus, small non-coding RNAs have emerged as an essential component to disease etiology. Using Systemic RNA interference Defective (SID) mutants of the nematode Caenorhabditis elegans, deficient in gene silencing, we examined the potential consequences of dysfunctional epigenomic regulation in the context of Parkinson's disease (PD). Specifically, the loss of either the sid-1 or sid-3 genes, which encode a dsRNA transporter and an endocytic regulatory non-receptor tyrosine kinase, respectively, conferred neuroprotection to dopaminergic (DA) neurons in an established transgenic C. elegans strain wherein overexpression of human α-synuclein (α-syn) from a chromosomally integrated multicopy transgene causes neurodegeneration. We further show that knockout of a specific microRNA, mir-2, attenuates α-syn neurotoxicity; suggesting that the native targets of mir-2-dependent gene silencing represent putative neuroprotective modulators. In support of this, we demonstrated that RNAi knockdown of multiple mir-2 targets enhanced α-syn-induced DA neurodegeneration. Moreover, we demonstrate that mir-2 overexpression originating in the intestine can induce neurodegeneration of DA neurons, an effect that was reversed by pharmacological inhibition of SID-3 activity. Interestingly, sid-1 mutants retained mir-2-induced enhancement of neurodegeneration. Transcriptomic analysis of α-syn animals with and without a sid-1 mutation revealed 27 differentially expressed genes with human orthologs related to a variety of diseases, including PD. Among these was pgp-8, encoding a P-glycoprotein-related ABC transporter. Notably, sid-1; pgp-8 double mutants abolished the neurodegeneration resulting from intestinal mir-2 overexpression. This research positions known regulators of small RNA-dependent gene silencing within a framework that facilitates mechanistic evaluation of epigenetic responses to exogenous and endogenous factors influencing DA neurodegeneration, revealing a path toward new targets for therapeutic intervention of PD.


Involvement of dorsal hippocampal and medial septal nicotinic receptors in cross state-dependent memory between WIN55, 212-2 and nicotine or ethanol in mice.
The present study examined whether nicotinic acetylcholine receptors (nAChRs) of the CA1 regions of the dorsal hippocampus and medial septum (MS) are involved in cross state-dependent memory retrieval between WIN55, 212-2 (WIN, a non-selective CB1/CB2 receptor agonist) and nicotine or ethanol. Memory retrieval was measured in one-trial step-down type passive avoidance apparatus in male adult mice. Pre-training intraperitoneal administration of WIN (0.1-1mg/kg) dose-dependently impaired memory retrieval when it was tested 24h later. Pre-test systemic administration of nicotine (0.6 and 0.7mg/kg, s.c.) or ethanol (0.5g/kg, i.p.) improved WIN-induced memory impairment, suggesting a cross state-dependent memory retrieval between the drugs. Pre-test intra-CA1 microinjection of nicotine (1 and 2μg/mouse) before systemic administration of an ineffective dose of nicotine (0.5mg/kg, s.c.) or ethanol (0.25g/kg) significantly reversed WIN-induced memory impairment. Pre-test intra-CA1 microinjection of mecamylamine (1 and 3μg/mouse) inhibited cross state-dependent memory between WIN and nicotine or ethanol. Moreover, pre-test intra-MS microinjection of nicotine (1 and 2μg/mouse) in combination with systemic administration of a lower dose of nicotine (0.5mg/kg), but not ethanol (0.25g/kg), improved memory impairment induced by pre-training administration of WIN. On the other hand, in the animals that received pre-training WIN and pre-test systemic administration of nicotine (0.7mg/kg), but not ethanol (0.5g/kg), pre-test intra-MS microinjection of mecamylamine (1-5μg/mouse) inhibited WIN-nicotine state-dependent memory retrieval. It should be noted that pre-test intra-CA1 or intra-MS microinjection of nicotine or mecamylamine by itself had no effect on memory retrieval and also could not reverse memory impairment induced by pre-training administration of WIN. It can be concluded that WIN and nicotine or WIN and ethanol can induce state-dependent memory retrieval. In addition, our results showed that a cross-state dependency between these drugs may be mediated through a cholinergic nicotinic mechanism.


Face-Dependent Reconstruction of Bi-Oxyiodides toward Selective Growth of (BiO)2 CO3 Edge Side to Maximize CO2 Conversion Efficiency.
Chemical reconstruction of bismuth oxyiodides using bicarbonates is tried to selectively grow (BiO)2 CO3 edge side. Orthorhombic o-Bi5 O7 I undergoes a total reconstruction process by its phase transformation into tetragonal (BiO)2 CO3 (BOC-o) to form a well-aligned nanosheet array with maximally exposing CO3 2- moiety at the edge side. The post-reconstruction BOC-o catalyst achieved 100 % Faradaic efficiency at -0.86 V vs. RHE for CO2 -to-formate conversion. However, another conservative reconstruction of tetragonal t-BiOI into tetragonal (BiO)2 CO3 (BOC-t) exposed majorly a less reactive [BiO]+ layer. At low overpotential regions, the catalytic cycle of BOC-o begins with the initial conversion of the CO3 2- moiety into formate at the [-OBi-(CO3 )-BiO-] site, but at high overpotential regions, the [BiO]+ layer undergoes reduction to metallic Bi and multi-catalytic species proceed with CO2 reduction. Otherwise, the deactivation of Bi+ site by an organic molecule switched on another catalysis of proton reduction, preventing CO2 reduction.


Diagnostic value of real-time capillary thermal cycler in virus detection.
Molecular-based detection methods such as PCR techniques have had a significant impact on the diagnosis of viral infections because of their superior sensitivity and rapid turnaround time. This review describes the use of real-time PCR on the capillary thermal cycler, the Roche LightCycler trade mark, for early disease detection in diagnostic virology. The advantages of using the LightCycler, the detection processes using SYBR Green I and different hybridization strategies will be discussed in detail, with specific examples drawn from our in-house viral assays. The use of the LightCycler for the investigation of two recent viral outbreaks in Singapore will also be briefly described.


The set of naturally processed peptides displayed by DR molecules is tuned by polymorphism of residue 86.
The response to tetanus toxoid (TT) was studied in immune donors that carry two alleles of DR5 that differ only at DR beta residue 86: DRB1*1101 (G86, abbreviated 1101) and DRB1*1104 (V86, abbreviated 1104). A large number of TT-specific T cell clones was isolated and the epitopes recognized in association with 1101 and 1104 were mapped. We found that two epitopes (p2 and p32) can be recognized in association with both 1101 and 1104 while three epitopes (p23, p27 and p30) are recognized in association with 1101, but never in association with 1104. The sets of naturally processed self peptides displayed by 1101 and 1104 were characterized using alloreactive T cell clones. We found that all 1104 alloreactive clones recognize both 1104 and 1101, while approximately 30% of the alloreactive 1101 clones fail to recognize 1104. Thus it is apparent that both naturally processed TT and self peptides displayed on 1104 molecules represent a fraction of those displayed on 1101 molecules. The mechanism responsible for this differential presentation was investigated by comparing the capacity of 1101 and 1104 antigen-presenting cells to present TT or synthetic peptides to specific T cells and by measuring the binding of these peptides to DR molecules. Three sets of results suggest that V86 acts as a constraint to the binding of naturally processed peptides: (i) all 1104-restricted or alloreactive T cell clones recognize TT- or allo-epitopes presented by 1101 as well, thus ruling out a major effect of V86 as a residue determining T cell restriction specificity; (ii) presentation of naturally processed peptides correlates in general with the capacity of long synthetic peptides to bind to 1101 or 1104 and (iii) while the naturally processed p30 epitope discriminates between 1101 and 1104, a short synthetic peptide binds equally well to and is comparably recognized in association with both 1101 and 1104. Taken together these results suggest that the natural polymorphism at residue 86 might be a molecular switch that finely tunes the complexity of the peptide collection presented on DR molecules.


Genetic and Functional Analysis of GRIN2A in Tumor Samples.
Ionotropic glutamate receptors (iGluRs) are large integral membrane multi-protein complexes that create ion channels in plasma membranes. Upon binding of receptor specific ligands (e.g., glutamate), increased efflux or influx of mono- or divalent cations (e.g., Ca2+) promotes synaptic transmission, cellular migration, and survival. Three classes of iGluRs were originally defined after their respective agonists: AMPA, kainate, and NMDA receptors (NMDARs). Recently, we examined iGluR families at the genetic level using Next-Generation Sequencing (NGS) (whole-exome sequencing (WES)) and discovered a high prevalence of somatic mutations within the gene for one of the NMDAR subunits, GRIN2A, specifically in malignant melanoma. Following confirmation of the somatic mutations, we focused on functional characterization of a subset of the GRIN2A mutants that demonstrated a loss of NMDAR functionality. We used gene expression and protein biochemistry to examine complex formation between GluN1 subunit (encoded by GRIN1) and GluN2A subunit (encoded by GRIN2A), anchorage-independent growth in soft agar and cellular migration. Furthermore, we used shRNA depletion of endogenous GRIN2A in melanoma cells expressing either wild-type GRIN2A or mutant GRIN2A and measured cellular proliferation compared to negative controls. Our data show that somatic mutation of certain residues in GluN2A results in increased survival and is the first such report to demonstrate the functional importance of GRIN2A mutations in melanoma and the significance ionotropic glutamate receptor signaling plays in malignant melanoma.


Progesterone receptor, bc1-2 and bax expression in meningiomas.
Meningiomas are generally benign central nervous system neoplasms, which frequently express progesterone receptor (PR) and only rarely express the estrogen receptor (ER). For breast cancer, a relation between steroid hormone receptors and proteins involved in the apoptotic process has been described. For meningiomas, the exact relation between PR and these proteins is not known. In this study, ER, PR, bcl-2 and bcl-2-associated x protein (Bax) expression levels were determined in meningioma cytosols. As a reference for our experimental conditions, we also determined these proteins in breast cancer cytosols. PR and ER were determined with a ligand-binding assay and scatchard-plot analysis. The expression levels of the anti- and pro-apoptotic proteins, bcl-2 and Bax, respectively, were determined by immunoblotting. In 65% of the meningioma, bcl-2 expression was found in variable amounts. In contrast to breast cancer, a significant negative association between PR and bcl-2 was found (P < 0.01). Bax expression appeared constitutive, not related to PR, and 2.6 times higher than breast cancer. As both PR and bcl-2 appear positively associated with prognosis, the negative relationship between bcl-2 and PR found in this study might have some biological and clinical significance.


Spontaneous bilateral femur neck fracture secondary to grand mal seizure: A case report.
Spontaneous bilateral femur neck fracture is a rare entity in the general population. A 17-year-old immobile, developmentally delayed male with the sequelae of cerebral palsy fractured both femoral necks during a grand mal epileptic seizure. He had been treated with valproic acid as an antiseizure medication for about 10 years; otherwise, he had no history of drug use. The laboratory analysis was normal except a marked vitamin D deficiency. Closed reduction and osteosynthesis with percutaneous cannulated screws were performed. Solid union was observed at 6 mo, and rapid postoperative rehabilitation was started. A femoral neck fracture may occur in a person with epilepsy presenting with hip pain in the emergency department.


Pembrolizumab alone or with chemotherapy for recurrent or metastatic head and neck squamous cell carcinoma: Health-related quality-of-life results from KEYNOTE-048.
To assess health-related quality of life (HRQoL) with first-line pembrolizumab, pembrolizumab-chemotherapy, or cetuximab-chemotherapy in recurrent or metastatic head and neck squamous cell carcinoma (R/M HNSCC) in the phase 3 KEYNOTE-048 trial (NCT02358031). HRQoL was measured using the European Organisation for Research and Treatment of Cancer 30-question quality-of-life (EORTC QLQ-C30), the EORTC 35-question quality-of-life head and neck cancer-specific module (EORTC QLQ-H&N35), and the EuroQol 5-dimension 3-level instruments (EQ-5D-3L). Secondary endpoints included mean change from baseline in EORTC QLQ-C30 global health status/quality of life (GHS/QoL) at week 15 and time to deterioration (TTD) in EORTC QLQ-C30 GHS/QoL and EORTC QLQ-H&N35 pain and swallowing. Of 882 enrolled participants, 844 received ≥ 1 dose of study treatment and completed ≥ 1 HRQoL assessment; adherence was ≥ 79% at week 15 across treatment groups. At week 15, EORTC QLQ-C30 GHS/QoL scores remained stable; no clinically meaningful between-group differences were observed (least squares mean difference, pembrolizumab vs cetuximab-chemotherapy, 0.24; 95% CI, -3.34 to 3.82; pembrolizumab-chemotherapy vs cetuximab-chemotherapy, 0.40; 95% CI, -3.46 to 4.26). Median TTD in EORTC QLQ-C30 GHS/QoL and EORTC QLQ-H&N35 pain and swallowing scores was not reached over 51 weeks across groups, showing stable HRQoL. TTD was similar between groups for EORTC QLQ-C30 GHS/QoL (pembrolizumab vs cetuximab-chemotherapy: HR, 1.38; 95% CI, 0.95-2.00; pembrolizumab-chemotherapy vs cetuximab-chemotherapy: HR, 1.37; 95% CI, 0.94-2.00), as was TTD in EORTC QLQ-H&N35 pain and swallowing scores. Pembrolizumab monotherapy and pembrolizumab-chemotherapy extended OS while maintaining HRQoL, further supporting first-line use for R/M HNSCC.


Molecular epidemiology of KI and WU polyomaviruses in healthy blood donors, south-eastern France.
The epidemiology of human polyomaviruses KI (KIPyV) and WU (WUPyV) in healthy populations is described poorly in the literature. The frequency of KIPyV and WUPyV viraemia was evaluated in a cohort of blood donors from south-eastern France. Plasma samples (n=640) were investigated for the presence of KIPyV/WUPyV DNA using a conserved real-time PCR detection system (VP2 gene). Three plasma samples (3/640; ∼0.5%) exhibited a positive fluorescence signal, with a low viral load (<500 copies/ml plasma); no additional amplicons were identifiable by agarose gel analysis. Sequencing highlighted the KIPyV origin of the three amplified sequences and the occurrence of point mutations. The sustained detection of KIPyV DNA in two serial samples (9 months) was in favor of a possible persistence of the virus in blood of healthy individuals. Further studies will be needed in order to explore both the prevalence and potential clinical impact of KIPyV/WUPyV on infected hosts.


The perfect storm: A quadrifecta of complications with takotsubo cardiomyopathy.
Takotsubo cardiomyopathy is generally considered to have a favorable prognosis. However, several major problems may complicate the clinical course. We present a patient with four simultaneous complications of shock, left ventricular thrombus, left ventricular outflow obstruction, and severe mitral regurgitation. Despite this dramatic presentation, the patient had a complete recovery with resolution of all four complications within weeks.


Acrokeratosis paraneoplastica (Bazex syndrome) associated with primary cutaneous squamous cell carcinoma of the lower leg, vitiligo and alopecia areata.
We report a case of acrokeratosis paraneoplastica (AP; Bazex syndrome), characterized by typical palmoplantar hyperkeratosis and psoriasiform scaly erythema of the acral regions, associated with primary cutaneous squamous cell carcinoma (SCC) on the left lower leg. This 54-year-old Japanese man subsequently developed vitiligo, and alopecia areata of the scalp. Serial monitoring of squamous cell carcinoma antigen (SCC-Ag) demonstrated that the severity of the clinical manifestations of AP paralleled the serum concentrations of SCC-Ag. We suggest that an immune-mediated mechanism underlies the development of AP in this patient.


IGF-1R as an anti-cancer target--trials and tribulations.
Type I insulin-like growth factor receptor (IGF-1R) has long been recognized for its role in tumorigenesis and growth, but only recently have the tools for targeting the IGF pathway become available. More than 10 IGF/IGF-1R inhibitors have entered clinical trials, and these belong to three main classes: (1) monoclonal antibodies against IGF-1R, (2) monoclonal antibodies against IGF-1R ligands (IGF-1 and IGF-2), and (3) IGF-1R tyrosine kinase inhibitors. These IGF-1R-targeting agents share common effects on IGF-1R signaling but differ in mechanisms of action, spectrum of target inhibition, and pharmacological features. Clinical activity of IGF-1R inhibitors has been demonstrated with sustained responses in a small number of patients with select tumor types, such as Ewing sarcoma and thymoma. However, many large clinical trials involving patients with adult tumors, including non-small cell lung cancer, breast cancer, and pancreatic cancer, failed to show clinical benefit in the overall patient population. Possible reasons for failure include the complexity of the IGF-1R/insulin receptor system and parallel growth and survival pathways, as well as a lack of patient selection markers. While IGF-1R remains a valid target for selected tumor types, identification of predictive markers and rational combinations will be critical to success in future development.


Systematic development of a group quantification method using evaporative light scattering detector for relative quantification of ginsenosides in ginseng products.
The determination for the contents of multi-components in ginseng products has come to the fore by demands of in-depth information, but the associated industries confront the high cost of securing pure standards for the continuous quality evaluation of the products. This study aimed to develop a prospective high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method for relative quantification of ginsenosides in ginseng products without a considerable change from the conventional gradient analysis. We investigated the effects of mobile phase composition and elution bandwidth, which are potential variables affecting the ELSD response in the gradient analysis. Similar ELSD response curves of nine major ginsenosides were obtained under the identical flow injection conditions, and the response increased as the percentage of organic solvent increased. The nine ginsenosides were divided into three groups to confirm the effect of elution bandwidth. The ELSD response significantly decreased in case of the late eluted ginsenoside in the individual groups under the isocratic conditions. With the consideration of the two important effects, stepwise changes of the gradient condition were carried out to reach a group quantification method. The inconsistent responses of the nine ginsenosides were reconstituted to three normalized responses by the stepwise changes of the gradient condition, and this result actualized relative quantification in the individual groups. The availability was confirmed by comparing the ginsenoside contents in a base material of ginseng products determined by the direct and group quantification method. The largest difference in the determination results from the two methods was 8.26%, and the difference of total contents was only 0.91%.


[Epitheliocytes of fundal glands and the relative volume of the parietal microflora of the stomach and small intestine mucosa in experimental chronic duodenal ulcer].
The effect of experimental chronic duodenal ulcers and vagotomy on fundal gland epitheliocytes and membrane microflora has been studied in rats using light microscopy and stereometry on semithin sections. It is shown that in ulcers the relative amount of perietal and zymogen cells increases, while the volume of mucocytes decreases. Vagotomy leads to a decrease in the relative amount of parietal and zymogen cells and increases the relative amount of mucocytes. The relative volume of membrane microflora in gastric fundal and pyloroantral regions, duodenum and jejunum diminishes in ulcers and increases in vagotomy, as compared to the control.


Inhibition of Liver Cancer Cell Growth by Triethylsilyl Resveratrol through Targeting Phosphoinositide-3 Kinase Pathway.
Liver cancer is one of the commonly detected malignancy and third highest cause of deaths caused by cancer worldwide. The current study investigated the effect of triethylsilyl resveratrol on HuH7 and hep3B cell proliferation and explored the underlying mechanism. The viability of HuH7 and hep3B cells was suppressed to 23 and 18%, respectively on exposure to 2 µM triethylsilyl resveratrol for 72 h. Triethylsilyl resveratrol treatment of HuH7 cells led to a prominent increase in expression of Atg-5, LC3B-II and Beclin proteins. In triethylsilyl resveratrol treated HuH7 cells expression of activated PI3K and Akt proteins showed a prominent decrease compared to the control cells. Moreover, p-mTOR protein expression was also suppressed in HuH7 cells on treatment with 2 µM triethylsilyl resveratrol. In triethylsilyl resveratrol treated cells a marked increase in ERK1/2 phosphorylation was observed compared to the control cells. Treatment with 2 µM triethylsilyl resveratrol for 72 h led to a significant (p < 0.05) increase in GFP-LC3B labelling in HuH7 cells compared to the control cells. Thus, triethylsilyl resveratrol reduced liver cancer cell viability through increase expression of proteins associated with autophagy. Moreover, it increased p-ERK1/2 expression and targeted activation of PI3K/Akt protein in HuH7 cells. Therefore, triethylsilyl resveratrol may be studied further as a promising therapeutic agent for treatment of liver cancer.


A constitutively expressed 36 kDa exochitinase from Bacillus thuringiensis HD-1.
A 36 kDa chitinase was purified by ion exchange and gel filtration chromatography from the culture supernatant of Bacillus thuringiensis HD-1. The chitinase production was independent of the presence of chitin in the growth medium and was produced even in the presence of glucose. The purified chitinase was active at acidic pH, had an optimal activity at pH 6.5, and showed maximum activity at 65 degrees C. Of the various substrates, the enzyme catalyzed the hydrolysis of the disaccharide 4-MU(GlnAc)(2) most efficiently and was therefore classified as an exochitinase. The sequence of the tryptic peptides showed extensive homology with Bacillus cereus 36 kDa exochitinase. The 1083 bp open reading frame encoding 36 kDa chitinase was amplified with primers based on the gene sequence of B. cereus 36 kDa exochitinase. The deduced amino-acid sequence showed that the protein contained an N-terminal signal peptide and consisted of a single catalytic domain. The two conserved signature sequences characteristic of family 18 chitinases were mapped at positions 105-109 and 138-145 of Chi36. The recombinant chitinase was expressed in a catalytically active form in Escherichia coli in the vector pQE-32. The expressed 36 kDa chitinase potentiated the insecticidal effect of the vegetative insecticidal protein (Vip) when used against neonate larvae of Spodoptera litura.


Increased upper airway cytokines and oxidative stress in severe obstructive sleep apnoea.
Inflammation may contribute to upper airway pathophysiology in obstructive sleep apnoea (OSA). Our objective was to compare upper airway pro-inflammatory cytokine expression, oxidative stress and connective tissue deposition in severe (n = 25) versus mild (n = 17) OSA patients. Upper airway surgical specimens were separated by predominance of either mucosal or muscle tissue. Expression levels of interleukin (IL)-1α, IL-6, interferon-γ, RANTES (regulated on activation, normal T-cell expressed and secreted), transforming growth factor (TGF)-β and l-selectin were measured by ribonuclease protection assay. Oxidative stress was assessed via protein carbonyl group detection by immunoblotting. Histochemistry was employed for immunolocalisation of selected cytokines and connective tissue morphometry. In the severe OSA group, expression of IL-1α, IL-6 and TGF-β was significantly higher in mucosa-predominant tissues, whereas in muscle-predominant specimens, RANTES expression was greater in severe OSA. Increased protein carbonylation was observed in severe OSA within both mucosal and muscle compartments. Immunohistochemistry localised TGF-β to submucosal and perimuscular inflammatory cells, while IL-6 was primarily localised to myocytes. Consistent with the pro-fibrotic cytokine profile observed in mucosa-predominant tissue, morphometric analysis revealed greater submucosal and perimuscular connective tissue in severe OSA subjects. There is increased pro-inflammatory and pro-fibrotic cytokine expression, oxidative stress, and connective tissue deposition in upper airway tissues from severe versus mild OSA patients.


A selective ACAT-1 inhibitor, K-604, stimulates collagen production in cultured smooth muscle cells and alters plaque phenotype in apolipoprotein E-knockout mice.
Acyl-coenzyme A:cholesterol O-acyltransferase-1 (ACAT-1) plays an essential role in macrophage foam cell formation and progression of atherosclerosis. We developed a potent and selective ACAT-1 inhibitor, K-604, and tested its effects in apoE-knockout mice. Administration of K-604 to 8-week-old apoE-knockout mice for 12 weeks at a dose of 60 mg/kg/day significantly reduced macrophage-positive area and increased collagen-positive area in atherosclerotic plaques in the aorta without affecting plasma cholesterol levels or lesion areas, indicating direct plaque-modulating effects of K-604 on vascular walls independent of plasma cholesterol levels. Pactimibe, a nonselective inhibitor of ACAT-1 and ACAT-2, reduced plasma cholesterol levels but did not affect macrophage- or collagen-positive areas. The size of macrophages and cholesteryl ester contents in the aorta were reduced by K-604. Exposure of cultured human aortic smooth muscle cells to K-604 resulted in increased procollagen type 1 contents in the culture supernatant and increased procollagen type 1 mRNA levels. Procollagen production was unaffected by pactimibe even at a concentration that inhibited cholesterol esterification to the basal level. Thus, the plaque-modulating effects of K-604 can be explained by stimulation of procollagen production independent of ACAT inhibition in addition to potent inhibition of macrophage ACAT-1.


Effects and mechanism of acid rain on plant chloroplast ATP synthase.
Acid rain can directly or indirectly affect plant physiological functions, especially photosynthesis. The enzyme ATP synthase is the key in photosynthetic energy conversion, and thus, it affects plant photosynthesis. To clarify the mechanism by which acid rain affects photosynthesis, we studied the effects of acid rain on plant growth, photosynthesis, chloroplast ATP synthase activity and gene expression, chloroplast ultrastructure, intracellular H(+) level, and water content of rice seedlings. Acid rain at pH 4.5 remained the chloroplast structure unchanged but increased the expression of six chloroplast ATP synthase subunits, promoted chloroplast ATP synthase activity, and increased photosynthesis and plant growth. Acid rain at pH 4.0 or less decreased leaf water content, destroyed chloroplast structure, inhibited the expression of six chloroplast ATP synthase subunits, decreased chloroplast ATP synthase activity, and reduced photosynthesis and plant growth. In conclusion, acid rain affected the chloroplast ultrastructure, chloroplast ATPase transcription and activity, and P n by changing the acidity in the cells, and thus influencing the plant growth and development. Finally, the effects of simulated acid rain on the test indices were found to be dose-dependent.


PROBLEM PROFILES OF AT-RISK YOUTH IN TWO SERVICE PROGRAMS: A MULTI-GROUP, EXPLORATORY LATENT CLASS ANALYSIS.
Baseline data collected in two brief intervention projects (BI-Court and Truancy Project) were used to assess similarities and differences in subgroups of at-risk youth. Classifications of these subgroups were based on their psychosocial characteristics (e.g., substance use). Multi-group latent class analysis (LCA) identified two BI-Court subgroups of youth, and three Truant subgroups. These classes can be viewed as differing along two dimensions, substance use involvement and emotional/behavioral issues. Equality tests of means across the latent classes for BI-Court and Truancy Project youths found significant differences that were consistent with their problem group classification. These findings highlight the importance of quality assessments and allocating appropriate services based on problem profiles of at-risk youth.


Immuno-viral therapy of brain tumors by combination of viral therapy with cancer vaccination using a replication-conditional HSV.
Here we developed an effective therapeutic approach using a replication-conditional mutant of herpes simplex virus (HSV), G207, for the treatment of metastatic tumors in the immunologically privileged central nervous system. An experimental model of brain metastasis was developed using BALB/c mice that harbored both intracranial (i.c.) and subcutaneous (s.c.) mouse CT26 colon adenocarcinoma tumors. Intratumoral injections of G207 into s.c. tumors elicited cytotoxic T-cell responses not only to HSV but also to a tumor antigen; however, only a limited antitumor effect was observed on metastatic brain tumors. To improve this antitumor effect, G207 was also injected into the brain tumor. After intratumoral injections of G207 into both i.c. and s.c. CT26 tumors, a significant antitumor effect was observed in the metastatic brain tumors. This therapeutic efficacy was absent in athymic mice, indicating that the antitumor effect could be mediated by T cells. Cytotoxic T-cell responses to HSV and the tumor antigen were induced by injections of G207 into i.c. and s.c. CT26 tumors. These results suggest that HSV-infected brain tumors may be efficiently eliminated by the induced anti-HSV T cells as well as by antitumor T cells. Therefore, this strategy of immuno-viral therapy, involving direct viral oncolytic activities and inducing antitumor and antiviral immune responses, may be useful for the treatment of tumors in the immunologically privileged central nervous system.


LncRNAs and circular RNAs as endothelial cell messengers in hypertension: mechanism insights and therapeutic potential.
Endothelial cells are major constituents in the vasculature, and they act as important players in vascular homeostasis via secretion/release of vasodilators and vasoconstrictors. In healthy arteries, endothelial cells play a key role in the regulation of vascular tone, cellular adhesion, and angiogenesis. A shift in the functions of the blood vessels toward vasoconstriction, proinflammatory state, oxidative stress and deficiency of nitric oxide (NO) might lead to endothelial dysfunction, a key event implicated in the pathophysiology of cardiovascular metabolic diseases, including diabetes, atherosclerosis, arterial hypertension and pulmonary arterial hypertension (PAH). Thus, reversibility of endothelial dysfunction may be beneficial for maintaining vascular homeostasis. In recent years, accumulative evidence has documented that noncoding RNAs (ncRNAs) are critically involved in endothelial homeostasis. Specifically, long noncoding RNAs (lncRNAs) and circular RNAs are highly expressed in endothelial cells where they serve as important mediators in normal endothelial functions. Dysregulation of lncRNAs and circular RNAs has been tightly associated with hypertension-related endothelial dysfunction. In this review, we will summarize the current progression and underlying mechanisms of lncRNA and circular RNA in endothelial cell biology under hypertensive conditions. We will also highlight their potential as biomarkers or therapeutic targets for hypertension and its associated endothelial dysfunction.


Lymphocyte immunophenotyping in dogs with lymphopenia of common causes.
Lymphocyte immunophenotyping can be useful for evaluating immune competence and predicting the disease prognosis. It is essential to gain knowledge about canine lymphocyte immunophenotypes in various conditions. The study deals with the characteristics of lymphopenia in dogs, with an emphasis on lymphocyte immunophenotyping by flow cytometry. Blood samples from 44 dogs with lymphopenia were included in the study. All lymphopenias sent from veterinary clinics to the diagnostic laboratory were analyzed. The hematological and biochemical abnormalities were investigated, as well as the effect of the age. Lymphopenias were classified according to the level of C-reactive protein (CRP). The percentage of T cells, B cells, Th cells and Tc cells, and T/B and Th/Tc ratios were determined by flow cytometry. Lymphopenias often occurred in dogs over 7 years of age (79.5 %). The most common were postoperative lymphopenia (31.8 %) and inflammatory diseases (29.5 %), most commonly affecting the gastrointestinal tract. Frequent abnormalities were monocytosis (56.8 %), increased CRP (72.7 %) and decreased albumin/globulin ratio (50.0 %). The percentage of Th lymphocytes was significantly lower in the group with elevated CRP than in the group with basal CRP (P = 0.0329). A negative correlation was found between the level of CRP and the percentage of Th lymphocytes (r = -0.3278, P = 0.0390). This study provided new insights into the appearance, incidence and classification of canine lymphopenia.


Defining the role of T cell-derived leptin in the modulation of hepatic or intestinal inflammation in mice.
The role of leptin in the immune system has been well established. While adipocytes represent the major source, leptin production by lymphocytes, infiltrating at the site of inflammation, was recently demonstrated. However, the significance of this locally released leptin remains unresolved. In the present study, two models in which absence of leptin-signalling is associated with protection were employed: the model of ConA-induced hepatitis and the CD4(+)CD45Rb(high) transfer model of colitis. For the ConA model, scid mice were reconstituted with either WT or leptin-deficient (ob/ob) CD4(+) T cells. Eight weeks post transfer, ConA was injected and serum ALT, TNFalpha, leptin as well as liver mononuclear cell activation and histological signs of inflammation were evaluated. No difference between recipients of WT or ob/ob cells was observed for any of the parameters evaluated. In the second model, either WT or ob/ob CD4(+)CD45Rb(high) cells were transferred into scid mice. No histological differences were detected, although recipients of ob/ob cells showed higher weight loss compared to recipients of WT cells. Spontaneous production of IL-6 from colon cultures obtained from recipients of ob/ob cells was reduced compared to recipients of WT cells, whereas stimulation of lamina propria lymphocytes with leptin resulted in a higher IFNgamma release in recipients of ob/ob cells compared to recipients of WT cells. In conclusion, the present study provides evidence that T cell-derived leptin does not play a major role in the regulation of the inflammatory process, indicating that the adipose tissue is the critical player in the immune-modulating effects of leptin.


Construction of a flocculent Saccharomyces cerevisiae strain secreting high levels of Aspergillus niger beta-galactosidase.
A flocculent Saccharomyces cerevisiae strain secreting Aspergillus niger beta-galactosidase activity was constructed by transforming S. cerevisiae NCYC869-A3 strain with plasmid pVK1.1 harboring the A. niger beta-galactosidase gene, lacA, under the control of the ADH1 promoter and terminator. Compared to other recombinant S. cerevisiae strains, this recombinant yeast has higher levels of extracellular beta-galactosidase activity. In shake-flask cultures, the beta-galactosidase activity detected in the supernatant was 20 times higher than that obtained with previously constructed strains (Domingues et al. 2000a). In bioreactor culture, with cheese-whey permeate as substrate, a yield of 878.0 nkat/gsubstrate was obtained. The recombinant strain is an attractive alternative to other fungal beta-galactosidase production systems as the enzyme is produced in a rather pure form. Moreover, the use of flocculating yeast cells allows for enzyme production with high productivity in continuous fermentation systems with facilitated downstream processing.


Quantitative perfusion mapping of the human lung using 1H spin labeling.
To evaluate the feasibility and reproducibility of a noninvasive, rapid and quantitative pulmonary perfusion mapping method using a two-compartment tissue model in combination with a (1)H spin labeling technique. Ten healthy volunteers and three patients with cystic fibrosis (CF) were examined on a 1.5-T whole-body scanner. Global and selective lung T(1) maps based on an inversion recovery Snapshot FLASH technique were acquired from each subject with breath-holds at end-expiration. For comparison, corresponding Gd-DTPA-enhanced (1)H MR perfusion images were also obtained from each CF patient. Quantitative perfusion maps were calculated from the global and selective T(1) maps. The measured perfusion rates of the upper right lung in volunteers ranged from 400 to 600 mL/100 g/minute. The method showed a high intra-study reproducibility and low relative errors. In CF-patients, perfusion defects detected using Gd-DTPA-enhanced MR imaging were also detected using the spin labeling method. The perfusion rates of diseased lung tissues were less than 200 mL/100 g/minute. Noninvasive, robust and quantitative (1)H MR mapping of pulmonary perfusion was successfully performed using a rapid lung T(1) mapping in combination with spin labeling within the imaging slice. The proposed method has the potential to provide both important qualitative functional information and quantitative pulmonary perfusion rates in various lung diseases at various stages without the need of contrast agents.


Incidence of trichothecenes and zearalenone in poultry feed mixtures from Slovakia.
A total of 50 samples of poultry feed mixtures of Slovakian origin were analyzed for eight toxicologically significant Fusarium mycotoxins, namely zearalenone (ZON), A-trichothecenes: diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2) and B-trichothecenes: deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON) and nivalenol (NIV). The A-trichothecenes and the B-trichothecenes were detected by means of high pressure liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS) and gas chromatography electron capture detection (GC-ECD), respectively. Reversed phase-high performance liquid chromatography with a fluorescence detector (RP-HPLC-FLD) was used for ZON detection. The most frequent mycotoxin detected was T-2, which was found in 45 samples (90%) in relatively low concentrations ranging from 1 to 130 microg kg(-1) (average 13 microg kg(-1)), followed by ZON that was found in 44 samples (88%) in concentrations ranging from 3 to 86 microg kg(-1) (average 21 microg kg(-1)). HT-2 and DON were detected in 38 (76%) and 28 (56%) samples, respectively, in concentrations of 2 to 173 (average 18 microg kg(-1)) for HT-2 and 64 to 1230 microg kg(-1) sample (average 303 microg kg(-1)) for DON. The acetyl-derivatives of DON were in just four samples, while NIV was not detected in any of the samples investigated. In as many as 22 samples (44%), a combination of four simultaneously co-occurring mycotoxins, i.e. T-2, HT-2, ZON and DON, was revealed. Despite the limited number of samples investigated during this study poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in Central Europe. This is the first reported study dealing with zearalenone and trichothecene contamination of poultry mixed feeds from Slovakia.


Influence of the polyanion on the physico-chemical properties and biological activities of polyanion/DNA/polycation ternary polyplexes.
It was recently reported that polyanion/DNA/polycation ternary polyplexes markedly improve gene transfection activity in comparison with the original DNA/polycation binary polyplexes. In this study to explore the influence of the polyanion on the physico-chemical properties and biological activity of polyanion/pDNA/polycation ternary polyplexes four types of biocompatible polyanions were selected, mainly based on the acid strength of the anionic functional groups and the molecular rigidity on forming ternary polyplexes with 25 kDa polyethyleneimine and DNA. Polyanion loosening of the DNA polyplex, weakening of the adsorption of serum proteins and improving of cellular uptake, which are thought to be important factors leading to a high transfection efficiency of DNA ternary polyplexes, were specifically investigated. Electrophoresis retardation analysis indicated that the loosening capacity of polyanions depended on the pK(a) value of the functional anion groups as well as the flexibility of the polyanion. The low pK(a) and flexible structure of the polyanions tended to loosen the compact DNA polyplexes. Thermodynamic analysis by isothermal titration calorimetry provided direct evidence about the serum protein-DNA ternary polyplex interactions. The polyanion/pDNA/polycation ternary polyplexes exhibited obviously lower binding affinities and less adsorption to serum proteins compared with the original DNA/polycation binary polyplexes. These relatively stable DNA ternary polyplexes maintained high levels of cellular uptake and intracellular accumulation in serum-containing medium that correlated with their high transfection efficiency. In contrast, the original pDNA/polycation binary polyplexes became clustered by strong adsorption of large amounts of serum proteins, leading to a sharp reduction in cellular uptake and intracellular accumulation, and thus low gene transfer efficiency. These results provide a basis for the development of polyanion/DNA/polycation ternary polyplexes for polyfection.


Percutaneous vertebroplasty relieves pain in metastatic cervical fractures.
Percutaneous vertebroplasty is currently an alternative for treating vertebral fractures of the thoracic and lumbar spine, providing both pain control and vertebral stabilization. In the cervical spine, however, percutaneous vertebroplasty is technically challenging because of the complex anatomy of this region. We evaluated the technical feasibility, complication rate, and ability of percutaneous vertebroplasty to provide pain relief in patients with painful metastatic cervical fractures. We retrospectively reviewed 62 patients (24 men) who, between May 2005 and May 2009, underwent vertebroplasty to treat painful metastatic cervical fractures. Each patient was evaluated by a visual analog scale for pain, number of pain analgesics, and CT and MRI before, the day after, and at 3 months after the procedure. Two of the 62 patients had asymptomatic cement leakage in the soft tissues. We observed no delayed complications. Mean pretreatment and 24-hour posttreatment visual analog scale pain scores were 7.9±1.7 and 1.5±2, respectively. Immediately after surgery, the pain completely disappeared in 25 (40%) patients. Administration of analgesics was suspended in 34 (55%) patients whereas in 27 (39%) patients the median analgesics use decreased from two pills per day (range, 0-3) to 0 (range, 0-3). In two (3%) patients, analgesics administration was continued due to the persistence of pain. At 3 months, the patients reported a mean visual analog scale pain score of 1.7±2. Our data suggest, in selected patients, percutaneous vertebroplasty may be performed with a high technical success rate combined with a low complication rate, providing immediate pain relief lasting at least 3 months and a reduction in the use of analgesic drugs. Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence.


Structure and action of heteronemertine polypeptide toxins: importance of amphipathic helix for activity of Cerebratulus lacteus toxin A-III.
The marine heteronemertine Cerebratulus lacteus produces a family of protein cytolysins designated as A-toxins. Limited subtilisin digests of the most abundant homolog, toxin A-III, yield two major products which may be purified by high-performance liquid chromatography. One product is shown to represent residues 1-86 and the other contains the entire toxin sequence (1-95). Both polypeptides are shown to lack internal protease nicks. The 1-95 polypeptide retains full cytolytic activity in comparison to native toxin, whereas 1-86 has an activity that is approximately four times lower. Extensive treatment of A-III with carboxypeptidase Y yields a polypeptide containing residues 1-75 which is totally devoid of hemolytic activity. Residues 63-95 of native A-III have been predicted to form a relatively hydrophobic alpha-helix which is potentially important for activity. The circular dichroism spectrum of 1-95 is in excellent agreement with both experimental and Chou-Fasman-predicted secondary structures of native A-III, while the spectra of 1-86 and 1-75 indicate a loss of helicity quantitatively consistent with the removal of residues 87-95 and 76-95, respectively. Combined with our earlier data on bilayer penetration by N-terminal sequences (K. M. Blumenthal (1982) Biochemistry 21, 4229-4233], the current results indicate a direct involvement of both ends of A-III in lytic activity. The C-terminal region may function by contributing a membrane binding site in the form of an amphipathic helix.


Well-Based Antibody Arrays.
Multiplex immunoassays are important tools in basic research and diagnostics. The ability to accurately quantify the presence of several antigens within an individual sample all at once has been useful in developing a proteomics view of biology. This in turn has enabled the development of disease-associated immunodiagnostic panels for better prognosis and well-being. Moreover, it is well understood that such multiplexing approaches lend themselves to automation, thereby reducing labor while providing the ability to dramatically conserve both reagent and sample all of which will reduce the cost per test. Here we describe various methods to create and use multiplex immunoassays in the wells of microtiter plates or similar formats.


Lack of contribution of P2X receptors to neurally mediated vasoconstriction in the rabbit kidney in vivo.
The contribution of adenosine triphosphate (ATP) to the neural control of regional renal perfusion in vivo remains unknown. We therefore examined whether P2X receptors mediate renal vascular responses to electrical stimulation of the renal nerves (RNS) in pentobarbitone anaesthetized rabbits. Responses to RNS were tested before and during renal arterial infusion of alpha,beta-methylene ATP (alpha,beta-mATP, 7-56 microg kg(-1) min(-1)) to desensitize P2X1 receptors. RNS consisted of 3 min trains at graded frequencies and short trains of RNS (4-32 pulses). Three-minute trains of RNS reduced renal blood flow (RBF), cortical laser Doppler flux (CLDF), and medullary LDF (MLDF) by -90 +/- 3%, -89 +/- 3% and -31 +/- 11%, respectively, at 4 Hz. MLDF was reduced less than CLDF or RBF. During short train RNS, RBF, CLDF and MLDF were reduced by -22 +/- 2%, -15 +/- 2% and -12 +/- 2%, respectively, for 32 s at 1 Hz. CLDF and MLDF were reduced to a similar extent. Infusion of alpha,beta-mATP induced transient reductions in RBF, CLDF and MLDF, but within 5 min these variables had recovered to control levels. Vascular responses to RNS were not significantly altered by alpha,beta-mATP treatment. In the rabbit kidney in vivo, alpha,beta-mATP-sensitive receptors mediate vasoconstriction and reduce perfusion in both cortical and medullary vascular beds. However, these receptors do not mediate neurally induced reductions in renal perfusion.


An evaluation of two approaches to exercise conditioning in pulmonary rehabilitation.
To compare the effectiveness of two forms of exercise training in pulmonary rehabilitation. A prospective, randomized, unblinded, 8-week trial. A hospital-based outpatient pulmonary rehabilitation program. Forty patients (20 patients in each group) with COPD who were referred for pulmonary rehabilitation. We compared the short-term effectiveness of a high-intensity, lower-extremity endurance program with a low-intensity, multicomponent calisthenics program for the rehabilitation of patients with COPD. The high-intensity group trained predominately on the stationary bicycle and treadmill, with a goal of exercising at > or = 80% of maximal level determined from incremental testing for 30 min per session. The low-intensity group performed predominately classroom exercises for approximately 30 min per session. For both groups, twice-weekly sessions were held for 8 weeks. The primary outcome measure was health status, measured using the Chronic Respiratory Disease Questionnaire. Other outcomes included peak oxygen consumption on incremental treadmill exercise testing, exertional dyspnea, treadmill endurance time, the number of sit-to-stand repetitions and arm lifts in 1 min, overall dyspnea, and questionnaire-rated functional status. Both groups showed significant postrehabilitation improvement in exercise variables, exertional and overall dyspnea, functional performance, and health status. Patients in the high-intensity group showed greater increases in treadmill endurance and greater reductions in exertional dyspnea, whereas those in the low-intensity group showed greater increases in arm-endurance testing. Both groups had similar improvements in overall dyspnea, functional performance, and health status. Despite differences in exercise performance, both high-intensity, lower-extremity endurance training and low-intensity calisthenics led to similar short-term improvements in questionnaire-rated dyspnea, functional performance, and health status.


The fatty acid pattern of triglycerides and FFA in serum of spontaneously hypertensive rats (SHR).
The fatty acid patterns of serum triglycerides and FFA in SHR and in normotensive controls aged 4, 8 and 26 weeks were estimated by gas-liquid chromatography. In serum triglycerides of SHR, the percentage of linoleic acid (C18:2) was lower and the content of arachidonic acid (C20:4) higher than in age-matched control animals. A continuous increase in palmitic (C16) and linoleic acids as well as a decrease in arachidonic acid has been found with advancing age, the most striking differences existing between 4- and 8-week-old animals, i.e. before onset of arterial hypertension in SHR. In the pre-hypertensive stage, the percentage of arachidonic acid was about 3 times as high as in later stages in SHR. This gradation was, however, even more pronounced in normotensive control rats. The C18:2/C20:4-ratio of triglycerides was lower in SHR but increased with age in both groups reaching a 5--8-fold level. Similar behavior of the FFA fatty acid pattern was less marked. Alterations in levels of linoleic and arachidonic acids are of interest because of their pathogenic role as precursors of prostaglandins in the development of genetically spontaneous hypertension in rats. The results are discussed in connection with the hypotensive effect of a linoleic acid-rich diet recently reported in hypertensive rats.


BIX01294, an inhibitor of histone methyltransferase, induces autophagy-dependent differentiation of glioma stem-like cells.
Glioblastoma (GBM) contains rare glioma stem-like cells (GSCs) with capacities of self-renewal, multi-lineage differentiation, and resistance to conventional therapy. Drug-induced differentiation of GSCs is recognized as a promising approach of anti-glioma therapy. Accumulating evidence suggests that unique properties of stem cells depend on autophagy. Here we demonstrate that BIX01294, an inhibitor of a G9a histone methyltransferase (introducing H3K9me2 and H3K27me3 repressive marks) triggers autophagy in human glioma cells. Pharmacological or genetic inhibition of autophagy decreased LC3-II accumulation and GFP-LC3 punctation in BIX01294-treated cells. GSCs-enriched spheres originating from glioma cells and GBM patient-derived cultures express lower levels of autophagy related (ATG) genes than the parental glioma cell cultures. Typical differentiation inducers that upregulate neuronal and astrocytic markers in sphere cultures, increase the level of ATG mRNAs. G9a binds to the promoters of autophagy (LC3B, WIPI1) and differentiation-related (GFAP, TUBB3) genes in GSCs. Higher H3K4me3 (an activation mark) and lower H3K9me2 (the repressive mark) levels at the promoters of studied genes were detected in serum-differentiated cells than in sphere cultures. BIX01294 treatment upregulates the expression of autophagy and differentiation-related genes in GSCs. Pharmacological inhibition of autophagy decreases GFAP and TUBB3 expression in BIX01294-treated GSCs suggesting that BIX01294-induced differentiation of GSCs is autophagy-dependent.


Subgroup analysis of the ASPirin in Reducing Events in the Elderly randomized clinical trial suggests aspirin did not improve outcomes in older adults with chronic kidney disease.
The role of aspirin for primary prevention in older adults with chronic kidney disease (CKD) is unclear. Therefore, post hoc analysis of the randomized controlled trial ASPirin in Reducing Events in the Elderly (ASPREE) was undertaken comparing 100 mg of enteric-coated aspirin daily against matching placebo. Participants were community dwelling adults aged 70 years and older in Australia, 65 years and older in the United States, all free of a history of dementia or cardiovascular disease and of any disease expected to lead to death within five years. CKD was defined as present at baseline if either eGFR under 60mL/min/1.73m2 or urine albumin to creatinine ratio 3 mg/mmol or more. In 4758 participants with and 13004 without CKD, the rates of a composite endpoint (dementia, persistent physical disability or death), major adverse cardiovascular events and clinically significant bleeding in the CKD participants were almost double those without CKD. Aspirin's effects as estimated by hazard ratios were generally similar between CKD and non-CKD groups for dementia, persistent physical disability or death, major adverse cardiovascular events and clinically significant bleeding. Thus, in our analysis aspirin did not improve outcomes in older people while increasing the risk of bleeding, with mostly consistent effects in participants with and without CKD.


The HLA-B*38:110 allele identified in a volunteer bone marrow donor.
HLA-B*38:110 allele differs from HLA-B*38:01:01:01 in codon 202 in exon 4.


Hydrogen-bonded structures and low temperature transitions of the confined water in subnano channels.
The interfacial and confined water have long been attractive objects due to their crucial roles in biological, geological processes, etc. In this paper, we investigate the hydrogen-bonded structures of water and their low temperature transitions in the subnano channels of AlPO4-11 for the first time on the basis of infrared spectroscopy. The number of the adsorbed water molecules is estimated to be 8.45 per channel in one unit cell by thermogravimetric analysis. It is found that the confined water molecules are involved in saturated and unsaturated coordination with different hydrogen bond strengths at ambient temperature. The former refers to ice-like four-coordinated water and the latter includes liquid-like structures, Al-coordinated and relatively free water molecules. Unique coordination between water molecules and framework Al sites is responsible for the ice-like structures in the channels above the ice melting point. The appearance of liquid-like structures is closely related to the strong channel confinement, which does not allow the formation of extensive tetrahedral hydrogen-bonded configuration. As temperature decreases, a structural transformation of confined water happens in the channels of AlPO4-11. Isolated small water oligomers and two new components with stronger hydrogen bonds, such as low-density amorphous ice-like structures and a kind of low-density liquid-like structures are preferred. Our results provide important insights into the structural organizations and thermal-dynamic behaviors of confined water in extreme narrow channels.


Reliability of Evaluating Achilles Tendon Vascularization Assessed With Doppler Ultrasound Advanced Dynamic Flow.
The reliability of quantifying intratendinous vascularization by high-sensitivity Doppler ultrasound advanced dynamic flow has not been examined yet. Therefore, this study aimed to investigate the intraobserver and interobserver reliability of evaluating Achilles tendon vascularization by advanced dynamic flow using established scoring systems. Three investigators evaluated vascularization in 67 recordings in a test-retest design, applying the Ohberg score, a modified Ohberg score, and a counting score. Intraobserver and interobserver agreement for the Ohberg score and modified Ohberg score was analyzed by the Cohen κ and Fleiss κ coefficients (absolute), Kendall τ b coefficient, and Kendall coefficient of concordance (W; relative). The reliability of the counting score was analyzed by intraclass correlation coefficients (ICC) 2.1 and 3.1, the standard error of measurement (SEM), and Bland-Altman analysis (bias and limits of agreement [LoA]). Intraobserver and interobserver agreement (absolute/relative) ranged from 0.61 to 0.87/0.87 to 0.95 and 0.11 to 0.66/0.76 to 0.89 for the Ohberg score and from 0.81 to 0.87/0.92 to 0.95 and 0.64 to 0.80/0.88 to 0.93 for the modified Ohberg score, respectively. The counting score revealed an intraobserver ICC of 0.94 to 0.97 (SEM, 1.0-1.5; bias, -1; and LoA, 3-4 vessels). The interobserver ICC for the counting score ranged from 0.91 to 0.98 (SEM, 1.0-1.9; bias, 0; and LoA, 3-5 vessels). The modified Ohberg score and counting score showed excellent reliability and seem convenient for research and clinical practice. The Ohberg score revealed decent intraobserver but unexpected low interobserver reliability and therefore cannot be recommended.


Rho-kinase inhibitor Y-27632 attenuates pulmonary hypertension in hyperoxia-exposed newborn rats.
To test the hypothesis that neonatal hyperoxia induced pulmonary hypertension accompanied by increased Rho-kinase expression in rat lungs and that Rho-kinase inhibitor could attenuate right ventricular hypertrophy and pulmonary arterial remodeling. Newborn rats were exposed to >95% O2 in the first week after birth, then to 60% O2 in the following 2 weeks. Control pups were exposed to room air over the same periods. The pups were injected with either Rho-kinase inhibitor Y-27632 (10 mg·kg(-1)·d(-1), ip) or vehicle from postnatal d 14 to 20. Lung and heart tissues were collected on postnatal d 7 and 21. Rho-kinase activity in lungs was measured using Western blotting and immunohistochemistry. The right ventricular hypertrophy and arterial medial wall thickness (MWT) were assessed morphologically. Rho-kinase activity in lungs was comparable between the hyperoxic and control pups on postnatal d 7, but it had a more than 2-fold increase in the hyperoxic pups on postnatal d 21. Moreover, the hyperoxic exposure induced structural features of pulmonary hypertension, as shown by the right ventricular hypertrophy and significantly increased arterial MWT. Administration with Y-27632 effectively blocked the hyperoxia-induced increase of Rho-kinase activity in lungs, and attenuated the right ventricular hypertrophy. Rho-kinase inhibitor may be a novel therapy for attenuating the hyperoxia-induced structural changes in pulmonary hypertension.


Highly efficient degradation of metronidazole drug using CaFe2O4/PNA nanohybrids as metal-organic catalysts under microwave irradiation.
Catalytic degradation based on microwave irradiation is an emerging technique which promises prompt and efficient catalytic degradation of organic pollutants. Calcium ferrite (CaFe2O4), poly(1-napththylamine) (PNA), and PNA/CaFe2O4 nanohybrids were synthesized via microwave-assisted technique. The properties of the as-prepared CaFe2O4, PNA, and PNA/CaFe2O4 nanohybrids were characterized by the thermogravimetric analysis (TGA), FTIR, XRD, SEM, and ultraviolet-visible spectrophotometry (UV-vis) analyses. The formation of inorganic-organic hybrids was confirmed by the FTIR and XRD studies. Loading of PNA was confirmed to be 8%, 16%, 32%, and 40% in CaFe2O4 which was established by TGA studies and the thermal stability was found to follow the order: CaFe2O4 > 8-PNA/CaFe2O4 > 16-PNA/CaFe2O4 > 32-PNA/CaFe2O4 > 40-PNA/CaFe2O4 > PNA. CaFe2O4 and PNA revealed band gap values of 3.42 eV and 2.60 eV respectively while for the PNA/CaFe2O4 nanohybrids, the values were found to be ranging between 2.46 and 3.00 eV. The PNA modified CaFe2O4 nanohybrids showed higher degradation efficiency towards metronidazole (MTZ) drug as compared with PNA and pure CaFe2O4. MTZ drug showed around 94% degradation within 21 min of microwave irradiation using 40-PNA/CaFe2O4 as catalyst. The enhanced catalytic activity was attributed to the high surface area of the nanohybrid catalyst as well as improved microwave catalytic activity of PNA. The reactive species responsible for degradation were confirmed by scavenger studies which formation of ·OH and O2·- radicals. Recyclability tests showed that the 40-PNA/CaFe2O4 nanohybrid exhibited 86% degradation of MTZ (90 mg/l) even after the third cycle, which reflected higher reusability of the catalyst. The MTZ fragments were identified using liquid chromatography-mass spectrometry (LC-MS).


How do we deliver our findings? Analysis of podium presentations at shoulder meetings.
The aim of the present study was to evaluate the time structure of oral presentations delivered at three shoulder congresses: shoulder sessions at the American Academy of Orthopaedic Surgeons (AAOS) Meeting, European Foundation of National Associations of Orthopaedics and Traumatology (EFORT) Congress, and International Congress of Shoulder and Elbow Surgery (ICSES). A total of 160 oral presentations at the 2016 AAOS Annual Meeting, 17th EFORT Congress, and 13th ICSES were included. Podium presentations were categorized by topic, congress, inclusion of video support, and nationality of the speaker. Total time and time dedicated to each section of the presentation (introduction, methods, results, discussion and conclusions) were collected for all podium presentations. Approximately 34% of speakers exceeded time constraints. No differences were found in the times that presenters used for the introduction, methods, results, and conclusions sections (p > 0.05). However, when extended introductions were delivered, the results and conclusions sections were shortened (r = - 0.2 and r = - 0.21, respectively). Inclusion of video support tended to result in exceedance of time limits (p < 0.01). One third of the shoulder surgeons exceeded time constraints in their conference presentations, and no distinctions were found in time allocations for different sections of the presentations. Longer introductions may lead to time restriction in the results and conclusions sections.


Evaluating the validity of self-reported smoking in Mexican adolescents.
We aimed to evaluate the validity of the self-reported smoking indicator used in the Global Youth Tobacco Survey (GYTS). 43 middle and high-school classrooms from 26 schools were selected from Mexico City and Cuernavaca, Morelos. A total of 1257 students provided both a questionnaire and a urine sample. Sensitivity and specificity of self-reported smoking compared to urinary cotinine. Validity indices were evaluated by subgroups of gender, social acceptability of smoking (ie, smoking parents or friends) and smoking frequency. Sensitivity and specificity for current smoking were 93.2% and 81.7%, respectively. Validity indices remained stable across gender. Parental smoking status moderated the validity of self-report, which had lower sensitivity in adolescents with non-smoking parents (86.7%) than in adolescents with smoking parents (96.6%). Sensitivity and specificity increased with smoking frequency. This first validation study of self-reported current smoking used in the GYTS among Mexican adolescents suggests that self-reported smoking in the past 30 days is a valid and stable indicator of current smoking behaviour. This measure appears suitable for public health research and surveillance.


Automated analysis of living cells through the quantitative use of automated phase contrast microscopy.
A simplified theory of image formation in phase contrast microscopy is presented. It is shown that the phase shift induced in light (related to the refractive index) by the observed object can be reconstructed, point by point, from the phase-contrast digitally sampled image through an appropriate algorithm. This allows one to make quantitative observations on unstained, living cells.


Endobronchial Carcinoid Tumor in a Girl with Initial Histologic Diagnosis of Leiomyoma.
Endobronchial tumors represent the rarest cause of airway obstruction in pediatric population. Due to rarity of the condition, a high index of suspicion is required for early diagnosis. We report a patient in whom diagnostic bronchoscopic biopsy was reported as leiomyoma while post resection histopathology showed an atypical carcinoid.


Quantitative nitric oxide production by rat, bovine and porcine macrophages.
The aim of this work was to compare in vitro nitric oxide (NO) production by rat, bovine and porcine macrophages. NO production was induced by lipopolysaccharide (LPS) or by phorbol 12-myristate 13-acetate (PMA) with ionomycin or recombinant interferon gamma (rIFN-gamma) and was assessed by Griess reaction. NO synthase type II (NOS II) expression was quantified by immunocytochemistry, Western blot and real-time polymerase chain reaction (RT-PCR). There were differences in NO production by pulmonary alveolar macrophages (PAM) in all species tested. The largest amounts of NO were produced by rat PAM. Less NO was produced by bovine PAM. Moreover, PAM in rats and cows differed in their abilities to respond to various stimulators. Neither porcine PAM nor Kupffer cells produced NO. Stimulation of porcine PAM with alternative concentrations of LPS did not lead to inducing NO production. Stimulation of porcine PAM with rIFN-gamma together with LPS led to a significant increase in the expression of NOS II mRNA, albeit without detectable NO production or NOS II expression on the protein level.


Newer cephalosporins: lessons to be learned from clinical trials in intraabdominal infections.
Recent clinical trials testing the efficacy of newer cephalosporins in intraabdominal infections lack a number of the study design criteria proposed by Solomkin et al. Nevertheless, these trials all support the use of these newer cephalosporins in the treatment of intraabdominal infections. These newer agents demonstrate increased in vitro antimicrobial activity against potential aerobic pathogens, which suggests that they may be used in combination with an antianaerobic drug in the treatment of intraabdominal infections. Therefore, agents including cefotaxime, cefoperazone, ceftazidime, cefuroxime, ceftriaxone, and aztreonam may be chosen in place of the standard aminoglycoside. Previous studies have demonstrated the necessity of both aerobic and anaerobic coverage in empiric therapy of intraabdominal infections. Cefotetan, ceftizoxime, cefoxitin, moxalactam, mezlocillin, and piperacillin all have in vitro activity against aerobes and anaerobes (including B. fragilis), thus, these agents have a potential as monotherapy for acute intraabdominal infections in otherwise healthy patients. Directed combination chemotherapy or perhaps imipenem alone is recommended for persistent peritonitis and intraabdominal infections in immunocompromised hosts.


Protein composition of PNS myelin: developmental comparison of control and quaking mice.
Protein compositions were determined for sciatic nerve myelin isolated from young and adult control and quaking (Qk) mice. Age-related changes in the relative amounts of large (Pl) and small (Pr) basic proteins were found. In control animals, the ratio Pr/Pl increased with age, a change similar to that observed for the large (Bl) and small (Bs) CNS myelin basic proteins of adult mice. Pr/Pl also increased with age in the Qk mouse sciatic nerve, but only to the point that the value in the adult Qk mouse was similar to that observed for young control animals, a situation reminiscent of the effect of the Qk mutation on CNS basic proteins. Thus, our data suggest that the Qk mutation has a similar effect on peripheral nervous system (PNS) and CNS basic proteins. Our findings are consistent with recent electrophoretic and immunochemical data showing that PNS and CNS myelin basic proteins in rodents are analogous, and they suggest that the genetic program controlling basic protein expression is common to oligodendroglia and Schwann cells.


Systematic reviews for informing rehabilitation practice: an introduction.
The research literature that rehabilitation clinicians need to be familiar with has become too large for anyone to read, and numerous published studies are too complex for many practitioners to understand and fruitfully use. One method to keep up with new findings is through systematic reviews. Systematic reviews can be effective tools that help guide rehabilitation practice by identifying the best research that provides the evidence for enhanced clinical decision-making. This article describes how systematic reviews are created, indicates where rehabilitation clinicians may find them, and refers to a resource that may be of use in evaluating their quality and applicability.


The cellular distribution of the Wld s chimeric protein and its constituent proteins in the CNS.
The C57BL/Wld s mouse is a mutant strain of mouse that shows greatly slowed Wallerian degeneration both in the central and peripheral nervous system. Using immunohistochemistry, immunofluorescence and Western blotting, we have investigated the distribution of the chimeric Wld s protein and its different components in neurons of the CNS of Wld s mice and wild-type C57BL/6J mice. The expression of the Wld s protein is restricted to the nucleus in Wld s mice. Wld s was not detected in axons. The Wld s mice express both the normal and chimeric forms of ubiquitination factor E4 (Ube 4b) and nicotinamide mononucleotide adenylyltransferase-1 (Nmnat-1). The normal forms were expressed both in the cytoplasm and the nuclei of neurons in Wld s mice and wild-type mice, and were also present in the axon. The normal form of Ube4b, mono- and poly-ubiquitin and IkappaBalpha, a substrate of Ube4b, were not differentially expressed in Wld s mice compared with wild-type mice. However, the expression of both the normal and mutant forms of Nmnat-1 was higher in the nuclei of Wld s mice compared with wild-type mice. Therefore, axon protection in Wld s mice does not appear to be controlled by expression of Wld s protein in the axons per se and also is unlikely to be related to the different activity of Ube4b either in general ubiquitination or toward this particular substrate. The increased Nmnat-1 activity in the nucleus of Wld s mice compared with wild-type mice seems to be a significant factor in the axon protection. It is not known whether the expression of the Nmnat-1 in the axon is significant.


Expression and mutational analysis of Nm23-H1 in liver metastases of colorectal cancer.
It has been proposed that nm23-H1, a candidate suppressor gene for metastasis, plays an important role in metastasis formation of human tumours. In order to investigate its role in the progression of colorectal cancer, we analysed 22 liver metastases of this malignancy with respect to mutational changes, loss of heterozygosity and expression levels of nm23-H1. Although genetic alterations in nm23-H1 have recently been described in those colorectal adenocarcinomas which give rise to distant metastases, we were unable to detect any mutation in the coding sequence of nm23-H1 in the metastatic tissue itself. We further analysed the metastases with respect to allelic deletions at the chromosomal locus of nm23. However, no loss of heterozygosity could be detected in ten informative cases. Moreover, the mRNA expression levels of nm23-H1 in the metastatic tissues were not significantly different from those in normal colon mucosa. Thus, although nm23-H1 might be involved in metastasis suppression of certain tumour types, in colorectal tumour progression its role remains to be determined.


Natural killer cell regulation of murine embryonic pulmonary fibroblast survival in vivo.
We examined the role of the natural killer (NK) cell in controlling the survival of embryonic pulmonary fibroblasts in vivo. In vitro, both primary embryonic fibroblasts and an embryonic fibroblast line (10T1/2) were lysed by syngeneic C3H/HeN splenocytes threefold more efficiently than primary adult fibroblasts. The membrane phenotype of the effector cells was typical of NK cells. It was asialo GM1+, Lyt2.1-, Lyt 1.1-, Thy 1.2-. The cytotoxicity of the effector cell could be enhanced by IFN-alpha/beta but was deficient in the C3H/HeJ bg/bg mutant. Iododeoxyuridine (131I-dUrd)-labeled embryonic fibroblasts were injected intravenously into syngeneic mice with either enhanced or deficient NK function and their survival in the lung was quantitated. Enhanced fibroblast survival was detected in the NK deficient C3H/HeJ beige (bg/bg) mutant strain compared to its normal littermate C3H/HeJ (bg/+). A second method of NK depletion by pretreatment with rabbit anti-asialo GM1 antiserum also produced a striking increase in fibroblast survival. Poly(I:C) significantly enhanced the elimination of pulmonary fibroblasts from the lung between 4 and 24 hr after injection. Poly(I:C) did not enhance clearance of pulmonary fibroblasts in the C3H/HeJ (bg/bg) mutant, but did so in the normal littermate C3H/HeJ (bg/+). In conclusion, we have shown that the survival of embryonic pulmonary fibroblasts was inversely correlated with in vivo NK activity suggesting a possible role for this cytotoxic cell in the control of fibroblast growth in vivo.


Right hemicolectomy does not confer a survival advantage in patients with mucinous carcinoma of the appendix and peritoneal seeding.
Traditionally epithelial malignancies of the appendix with or without carcinomatosis have been treated by right hemicolectomy. Recent accumulation of a large number of patients with this disease has enabled a re-evaluation of this surgical judgement. Clinical data on 501 patients with epithelial malignancy of the appendix were collected prospectively. All patients had peritoneal seeding at the time of referral and were treated by cytoreductive surgery and perioperative intraperitoneal chemotherapy. The main independent variable for statistical analysis was the surgical procedure used to resect the primary cancer (appendicectomy alone versus right hemicolectomy). Nineteen other clinical and pathological variables were considered as control variables. The endpoint for all analyses was survival. Median follow-up after the initial diagnosis was 4 years. The rate of regional lymph node positivity was 5.0 per cent. When the incidence of lymph node metastasis was determined by histological type, it was statistically significantly higher in intestinal (66.7 per cent) than in mucinous (4.2 per cent) tumours (P < 0.001). The presence of lymph node metastases had no influence on prognosis (P = 0.155). The surgical procedure (appendicectomy alone versus right hemicolectomy) had an influence on patient survival by univariate analysis (P < 0.001), but not by multivariate analysis (P = 0.258). Right hemicolectomy does not confer a survival advantage in patients with mucinous appendiceal tumours with peritoneal seeding. These data suggest that right hemicolectomy should be avoided unless metastatic involvement of the appendiceal or distal ileocolic lymph nodes is documented by biopsy, or the resection margin is inadequate.


Aromatherapy for Preoperative Anxiety: A Pilot Study.
The aim of this study was to evaluate the use of a lavender aromatherapy skin patch on anxiety and vital sign variability during the preoperative period in female patients scheduled for breast surgery. This observational pilot study used a one-group, pretest and post-test design. Participants received an aromatherapy patch in addition to standard preoperative care. Anxiety levels were assessed with a 10-cm visual analog scale at baseline and then every 15 minutes after patch placement. Vital sign measurements were recorded at the same interval. There was a statistically significant decrease (P = .03) in the anxiety visual analog scale measurements from baseline to final scores. Findings from this study suggest the use of aromatherapy is beneficial in reducing anxiety experienced by females undergoing breast surgery. Further research is needed to address the experience of preoperative anxiety, aromatherapy use, and the challenges of managing preoperative anxiety.


Moral Motivation and the Basal Forebrain.
Moral motivations drive humans to sacrifice selfish needs to serve the needs of others and internalized sociocultural norms. Over the past two decades, several brain regions have been associated with different aspects of moral cognition and behaviour. Only more recently, however, investigations have highlighted the importance of the basal forebrain for moral motivation. This includes the septo-hypothalamic region, implicated in kinship bonding across mammal species, and the closely connected subgenual frontal cortex. Understanding the neuroanatomy of moral motivation and its impairments will be fundamental for future research aiming to promote prosocial behaviour and mental health.


Ovarian carcinoma initially presenting as breast cancer two years prior to diagnosis: A case report and review of literature.
•Ovarian cancer presenting as a primary breast cancer two years prior•Ovarian cancer with metastases to breast is rare.•Metastases to the breast generally present as a recurrence.•Delay in diagnosis likely due to chemotherapy given for breast disease.


Kinetics of sodium-dependent solute transport by rabbit renal and jejunal brush-border vesicles using a fluorescent dye.
The kinetics of Na-coupled solute transport by renal and jejunal brush-border vesicles in the rabbit were examined using the potential-sensitive fluorescent dye diS-C3-(5). All organic solutes known to be transported across these membranes by Na-coupled mechanisms increase the fluorescence of the dye in the presence of Na, but not K. An increase in fluorescence (delta F) corresponds to a depolarization of the electrical potential difference (5-60 mV) across the brush-border membrane in the intact cell. delta F was independent of the valency of the transported solute. The fluorescence response was saturable, and for twelve solutes the Kf, i.e. the concentration of the substrate generating 50% of the maximal response, agreed quite closely with the Kt values reported from tracer studies. For six solutes increasing the Na concentration decreased Kf, and this agrees with the effect of Na on the kinetics of succinate transport in renal vesicles. We conclude that D-glucose, neutral amino acids and imino acids are co-transported with Na across both renal and jejunal brush-border membranes, and that carboxylic acids, beta-amino acids, and dibasic amino acids are co-transported with Na across the renal, but not jejunal, membranes.


Deficiency of mouse CD4+CD25+Foxp3+ regulatory T cells in xenogeneic pig thymus-grafted nude mice suffering from autoimmune diseases.
Xenogeneic thymus transplantation can efficiently induce specific immune tolerance to donor antigens in athymic recipients. However, many nude mice suffer from autoimmune diseases (AID) for over 10 weeks after xenogeneic thymus transplantation. CD4+CD25+Foxp3+ regulatory T (Treg) cells were recently determined to play a pivotal role in keeping immune tolerance in humans and mice. Thus, we investigated this subpopulation of Treg cells in the periphery of pig thymus-grafted nude mice suffering from AID. Our results showed that the expression of Foxp3, CTLA-4 and GITR on mouse CD4+CD25+ T cells and the ratio of CD4+CD25+Foxp3+ Treg cells to CD4+ T cells were significantly decreased in the periphery of pig thymus-grafted nude mice suffering from AID, compared with healthy pig or mouse thymus-grafted nude mice. Furthermore, mouse CD4+CD25+ T cells in pig thymus-grafted nude mice suffering from AID showed more severe deficiency in immunosuppressive function compared with the counterpart in xenogeneic pig or syngeneic thymus-grafted nude mice without AID. Thus, the decreased frequency, altered phenotype and functional deficiency of mouse CD4+CD25+ Treg cells in pig thymus-grafted nude mice may contribute to the development of AID in this model.


Bioinformatics Analysis of DNA Methylation Through Bisulfite Sequencing Data.
DNA methylation plays an important role in the regulation of gene expression as one of the epigenetic modifications. The bisulfite sequencing is widely used to determine the patterns of genomic methylation as a gold standard technology allowing conversion of the unmethylated cytosines to uracils that are represented as Ts in the sequencing reads. This chapter introduces the methodology for analyzing bisulfite sequencing data using various bioinformatics tools.


Heme synthesis in the lead-intoxicated mouse embryo.
Incorporation of 55Fe and of [14C]glycine was studied in control embryos and mothers and in those which had received lead in the diet from day 7 of pregnancy. Incorporation of Fe into heme of embryonic liver which increases markedly for controls on day 17 of pregnancy was depressed greatly and showed no such increase in lead-intoxicated embryos. These embryos were retarded in growth but had normal heme concentrations in body and liver. Incorporation of glycine into embryonic heme and proteins was not affected. Data on incorporation in the mothers are also presented. It is thought that the impaired synthesis of heme in lead-intoxicated embryos limits their body growth during the late phase of pregnancy.


Ultrastructural characteristics of finger-like membrane protrusions in cell competition.
A small number of oncogenic mutated cells sporadically arise within the epithelial monolayer. Newly emerging Ras- or Src-transformed epithelial cells are often apically eliminated during competitive interactions between normal and transformed cells. Our recent electron microscopy (EM) analyses revealed that characteristic finger-like membrane protrusions are formed at the interface between normal and RasV12-transformed cells via the cdc42-formin-binding protein 17 (FBP17) pathway, potentially playing a positive role in intercellular recognition during apical extrusion. However, the spatial distribution and ultrastructural characteristics of finger-like protrusions remain unknown. In this study, we performed both X-Y and X-Z EM analyses of finger-like protrusions during the apical extrusion of RasV12-transformed cells. Quantification of the distribution and widths of the protrusions showed comparable results between the X-Y and X-Z sections. Finger-like protrusions were observed throughout the cell boundary between normal and RasV12 cells, except for apicalmost tight junctions. In addition, a non-cell-autonomous reduction in protrusion widths was observed between RasV12 cells and surrounding normal cells under the mix culture condition. In the finger-like protrusions, intercellular adhesions via thin electron-dense plaques were observed, implying that immature and transient forms of desmosomes, adherens junctions or unknown weak adhesions were distributed. Interestingly, unlike RasV12-transformed cells, Src-transformed cells form fewer evident protrusions, and FBP17 in Src cells is dispensable for apical extrusion. Collectively, these results suggest that the dynamic reorganization of intercellular adhesions via finger-like protrusions may positively control cell competition between normal and RasV12-transformed cells. Furthermore, our data indicate a cell context-dependent diversity in the modes of apical extrusion.


[Phosphonates and their importance for dental practice].
In the light of a literature review, the author deals with the significance of anorganic pyrophosphate in the mineralization process in the organism. Diphosphonates are synthetic substances which act on mineralisation. The author made experiments with them, the results of which might be of importance in the stomatological sphere with regard to caries reduction and plaque inhibition.


Bone ankylosis. a rare manifestation of gout.
A 44 year old man with neglected tophaceous gout developed progressive bony and fibrous ankylosis of the lower leg articulations extending from a toe through the ankle, in spite of three years of otherwise successful therapy.


The effects of k-space data undersampling and discontinuities in keyhole functional MRI.
The effects of the k-space data undersampling and the discontinuities associated with the keyhole approach in functional magnetic resonance imaging (fMRI) have been addressed. Undersampling of high k-space data increases the high spatial-frequency noise by a factor of square root m, where m is the number of images sharing the high k-space data. However, the effects of the noise can be effectively removed using clustered-pixel statistical analysis. The amplitude and phase discontinuities may result in an increase in the low spatial-frequency noise level, which has no impact on specificity and only a minimal impact on sensitivity. Although dramatically reducing acquisition time and preserving the spatial resolution of functional magnetic resonance imaging, the keyhole technique may have a limited capability in detecting highly focal activations.


Progress in multicellular human cardiac organoids for clinical applications.
Advances in self-organizing cardiac organoids to recapitulate human cardiogenesis have provided a powerful tool for unveiling human cardiac development, studying cardiovascular diseases, testing drugs, and transplantation. Here, we highlight the recent remarkable progress on multicellular cardiac organoids and review the current status for their practical applications. We then introduce key readouts and tools for assessing cardiac organoids for clinical applications, address major challenges, and provide suggestions for each assessment method. Lastly, we discuss the current limitations of cardiac organoids as miniature models of the human heart and suggest a direction for moving forward toward building the mini-heart from cardiac organoids.


Monoclonal antibody against human sperm-specific YLP12 peptide sequence involved in oocyte binding.
The monoclonalantibody (mAb) against YLP12 peptide was raised and its immunobiological properties were examined. In the Western blot procedure, the YLP12 mAb recognized a specific protein band of approximately 50 +/- 5 kD in human sperm extract and approximately 72 +/- 5 kD in human testis extract. The myeloma Ig control did not recognize these specific protein bands. In the immunofluorescence studies, the YLP12 mAb, and not the myeloma Ig, predominantly reacted with the acrosome regions of methanol-fixed human sperm. In the acrosome reaction assay, the YLP12 mAb showed a significant (p < .001) and a concentration-dependent inhibition of acrosome reaction. The myeloma Ig did not affect the acrosome reaction. There was no apparent effect of antibodies on sperm motility. Thus, the monoclonal antibody, if humanized by genetic engineering technology, may provide a useful immunocontraceptive agent.


Histochemical studies on catecholaminergic cells in the carp retina.
Histochemical studies on catecholaminergic cells were conducted with the carp (Cyprinus carpio) retina. Catecholamine (CA)-containing cell bodies appear sparsely distributed among amacrine cells in the innermost cellular row of the inner nuclear layer (INL) and occasionally in the outer half part of the inner plexiform layer (IPL); only exceptionally are they found among ganglion cells. The fluorescent cells interspersed with the amacrine cells and in the IPL send their fiber processes toward both the outer plexiform layer (OPL) and the IPL; the fine fibers form dense networks in the INL and IPL. Pretreatment of the fish with intramuscular injection of reserpine (20 hr prior to enucleation) completely depleted CA from the retina. The fluorescence of catecholaminergic cells was enhanced, and the number of fluorescent cells visible was increased, by intravitreous injection of L-DOPA, DA, and NA (3 prior to enucleation). A combination of pretreatment with intramuscular reserpine and intravitreous NA was particularly effective. These results indicate that catecholamines may play an important role in the modulation of the membrane potential of horizontal cells.


Phanton limb sensation under subarachnoid and epidural analgesia - a comparative clinical study of two hundred cases.
Two hundred patients scheduled for various surgical procedures, under subarachnoid and epidural anaesthesia were divided in two groups of 100 for each technique. Subarachnoid analgesia was obtained with five per cent lidocaine, while epidural analgesia was accomplished with 1.5 per cent or 2 per cent lidocaine with adrenaline. This study shows that phantom sensation is painless and self limiting and that it lasts only for the duration of motor and proprioceptive blockade. It does not require any special treatment except psychotherapy, supplemented if needed by tranquilizers. Patients should be positioned after motor blockade has been established.


Giant pseudo-aneurysm of the right ventricular outflow tract after Tetralogy of Fallot repair.
Images of the diagnosis and correction of a pseudo-aneurysm at the right ventricular outflow tract, one of the rarest complications of Tetralogy of Fallot surgical correction.


Echocardiographic diagnosis of acute pericardial effusion in penetrating chest trauma.
Pericardial tamponade persists as a diagnostic challenge to the clinician. The immediate diagnosis is particularly difficult when the patient is well compensated hemodynamically. In contrast to the deceptively stable appearance, the patient is at increased risk due to delayed onset, recognition, and therapy. A case of penetrating thoracic trauma is presented in which the concern about pericardial tamponade was considerable. However, the elements of clinical assessment were inconclusive. Two-dimensional echocardiography was employed in the emergency department to detect a pericardial effusion and thus avoid delayed management. Pericardial tamponade and associated vascular injuries were confirmed at surgery.


LC-MS/MS determination of Synercid injections.
Synercid is a combination of two semisynthetic pristinamycin derivatives, quinupristin and dalfopristin in 30:70 (w/w) ratio. A rapid and specific high-performance liquid chromatography-mass spectrometry was developed for the determination of quinupristin and dalfopristin using positive electrospray tandem mass spectrometry (+ESI-MS/MS). Multiple reaction monitoring transitions at 1023.05>134.34 and 691.87>166.26 were selected for the quantitation of quinupristin and dalfopristin, respectively. The assay run cycle-time was approximately 2.0 min injection-to-injection. The assay was linear up to concentration of 4000 ng x ml(-1) quinupristin and 1920 ng x ml(-1) dalfopristin. The lowest limits of quantitation of quinupristin and dalfopristin were found to be 1000 and 480 ng x ml(-1), respectively. Quantitation was based on peak area measurement of quinupristin and dalfopristin using weighed linear regression. Linear relationships with correlation coefficients (r>0.99) were automatically computed for both constituents by MASSLYNX quantify program. The ratio of the slopes of the calibration curves of quinupristin and dalfopristin was found to be 0.425, which matches the nominal ratio composition of the antimicrobial compounds in Synercid. The %RSD ranges were 2.3-4.0% for dalfopristin and 1.3-4.2% for quinupristin, whereas the %DEV ranges were (-7.5+3.7) and (-1.2+9.1%), respectively, indicating appropriate precision and accuracy. Recoveries of 99.5-103.8% and 97.8-99.0% of quinupristin and dalfopristin, respectively, were computed from Synercid injection. The described method is recommended for rapid determination of the contents and for tracking the stability and compatibility of quinupristin and dalfopristin in Synercid injection.


Analysis of immediate changes of water-soluble metabolites in alkali-burned rabbit cornea, aqueous humor and lens by high-resolution 1H-NMR spectroscopy.
To investigate immediate changes in water-soluble metabolites of ocular tissue in alkali-burned eyes by using high-resolution 1H-NMR spectroscopy. Adult New Zealand rabbit eyes were burned with 1 M NaOH for 1 min. Normal eyes were used as control. Samples from aqueous humor and perchloric acid extracts of the cornea and lens were analyzed on a NMR spectrometer operating at 500 MHz for protons. Metabolites were quantified by comparing peak area with an added internal standard, TSP (3'-trimethylsilylpropinate-2,2,3,3-d4). Alkali burn of corneal surface causes immediate changes in concentration of many water-soluble metabolites in the anterior segment. Even as far away as the lens a significant increase in lactate was found. Cornea showed a significant increase in glucose and a significant decrease in hypo-taurine concentration. Most changes were observed in aqueous humor, with significant increases in succinate, creatine, scyllo- and myo-inositol and a significant decrease in citrate concentration. Furthermore, a small decrease in ascorbate concentration in aqueous humor was observed. The present study provides a valuable contribution to the knowledge of metabolic alterations in alkali-burned eyes. It shows that 1H-NMR spectroscopy is well suited for simultaneous qualitative and quantitative analysis of changes of metabolite concentrations in damaged tissues. This can help us to better evaluate and understand the biological alterations due to alkali burn.


Double labeling of human leukemic cells using 3H-cytarabine and monoclonal antibody against bromodeoxyuridine.
A new technique using immunofluorescence and autoradiography is described, in which the DNA of cells in S phase are labeled with two different probes. This method makes it possible to study the relationship between DNA synthesis and the uptake and/or incorporation of chemotherapeutic agents into normal or neoplastic cells. An example is provided in which the incorporation of 3H-cytarabine into DNA is demonstrated to occur only in cells which were synthesizing DNA during exposure to 3H-cytarabine. Other radioactively labeled probes can be used as well.


Cohaerins G-K, azaphilone pigments from Annulohypoxylon cohaerens and absolute stereochemistry of cohaerins C-K.
Four azaphilones, for which the trivial names cohaerins G, H, I and K are proposed, were isolated from the methanolic stromatal extract of Annulohypoxylon cohaerens together with the known metabolites cohaerins C-F and 4,5,4',5'-tetrahydroxy-1,1'-binaphthyl (BNT). Their planar structures were determined by NMR spectroscopy and by mass spectrometry. While their core structure is identical with cohaerin C and F, respectively, subgroups 2-hydroxy-6-methylphenyl and (1R,2R,4S)-4-hydroxy-2-methyl-6-oxocyclohexyl account for the structural diversity as substituents at C-3 of the azaphilone core. The absolute stereochemistry was assigned by NOE NMR experiments, CD spectroscopy and derivatisation with Mosher's acid; in addition, the stereochemistry of cohaerins C-F was revised. The metabolites showed cytotoxic effects besides a weak antimicrobial activity.


Leiomyosarcoma of artery and vein.
Five cases of leiomyosarcoma of blood vessels--three venous, two arterial--are reported. Treatment was surgical alone in two patients with involvement of the internal mammary artery and the long saphenous vein. The other three were treated initially by surgery, followed by radiation therapy, and surgery aimed at either total or partial removal of metastases. Two patients are alive and well, free of disease. Two patients have died of their disease despite all modalities of treatment, and one patient has a life-threatening metastases and is not expected to survive.


Trophic transfer of carbon nanofibers among eisenia fetida, danio rerio and oreochromis niloticus and their toxicity at upper trophic level.
Although the toxicity of carbon-based nanomaterials has already been demonstrated in several studies, their transfer in the food chain and impact on the upper trophic level remain unexplored. Thus, based on the experimental food chain "Eisenia fetida → Danio rerio → Oreochromis niloticus", the current study tested the hypothesis that carbon nanofibers (CNFs) accumulated in animals are transferred to the upper trophic level and cause mutagenic and cytotoxic changes. E. fetida individuals were exposed to CNFs and offered to D. rerio, which were later used to feed O. niloticus. The quantification of total organic carbon provided evidence of CNFs accumulation at all evaluated trophic levels. Such accumulation was associated with higher frequency of erythrocyte nuclear abnormalities such as constricted erythrocyte nuclei, vacuole, blebbed, kidney-shaped and micronucleated erythrocytes in Nile tilapia exposed to CNFs via food chain. The cytotoxic effect was inferred based on the smaller size of the erythrocyte nuclei and on the lower "nuclear/cytoplasmic" area ratio in tilapia exposed to CNFs via food chain. Our study provided pioneering evidence about CNFs accumulation at trophic levels of the experimental chain, as well as about the mutagenic and cytotoxic effect of these materials on O. niloticus.