Immunohistochemical distribution of cell cycle proteins p27, p57, cyclin D3, PCNA and Ki67 in normal and diabetic human placentas.
The placenta is a regulator organ for many metabolic activities between mother and fetus. Therefore, fetal growth is directly related to the placental development. Placental development is a series of events that depend on the coordinated action of trophoblasts' proliferation, differentiation and invasion. Studies on cell cycle related proteins which control these events are fairly limited. How placental tissue proliferation is affected by diabetes is not exactly known yet. Therefore in this study, the immunohistochemical localizations of cell cycle related proteins like PCNA, Ki67, cyclin D3, p27 and p57 in the differentiation, proliferation and apoptosis mechanisms of normal and diabetic placentas were investigated. Information on cell cycle related proteins that control these events is limited and how they are affected in diabetes mellitus is not fully understood yet. Therefore, in this study, to understand the role of cell cycle regulators in diabetic placentas we aimed to determine the spatio-temporal immunolocalizations of cell cycle regulators in diabetic and normal human term placentas. Term placentas were obtained from diabetic women and from normal pregnancies with informed consent following caesarean deliveries. Placental samples were stained via immunohistochemistry with PCNA, Ki67, cyclin D3, p27 and p57 antibodies and were examined by light microscopy. When compared to control placentas, PCNA, Ki67 and cyclin D3 staining intensities significantly increased in villous parts of diabetes group. Moreover, Ki67 and cyclin D3 stainings also significantly increased in basal plates and chorionic plate respectively. In chorionic plates, p27 and p57 staining intensities significantly decreased in diabetic group. p57 staining also significantly decreased in villous parts of diabetic placentas. Placental abnormalities seen in diabetic placentas could be associated with proliferation and cell cycle arrest mechanisms' alterations occurred in diabetes mellitus.


Catheter Ablation for Atrial Fibrillation in Patients ≤30 Years of Age.
Pulmonary vein (PV) automaticity is an established trigger for paroxysmal atrial fibrillation (PAF), making PV isolation (PVI) the cornerstone of catheter ablation. However, data on triggers for atrial fibrillation (AF) and catheter ablation strategy in very young patients aged <30 years are sparse. A total of 51 young patients (mean age 24.0 ± 4.2 years, 78.4% men) with drug-refractory PAF underwent electrophysiology (EP) study and ablation at 5 EP centers. None of the patients had structural heart disease or family history of AF. EP study induced supraventricular tachycardia (SVT) in 12 patients (n = 12, 23.5%): concealed accessory pathway mediated orthodromic atrioventricular reentrant tachycardia in 3 patients, typical atrioventricular nodal reentrant tachycardia in 6 patients, left superior PV tachycardia in 1 patient, left atrial appendage tachycardia in 1 patient, and typical atrial flutter in 1 patient. In patients with induced SVTs, SVT ablation without PVI was performed as an index procedure, except for the patient with atrial flutter who received cavotricuspid isthmus ablation in addition to PVI. Remaining patients underwent radiofrequency (n = 15, 29.4%) or second-generation cryoballoon-based PVI (n = 24, 47%). There were no major complications related to ablation procedures. Follow-up was based on outpatient visits including 24-hour Holter-electrocardiogram at 3, 6, and 12 months after ablation, or additional Holter-electrocardiogram was ordered in case of symptoms suggesting recurrence. Recurrence was defined as any atrial tachyarrhythmia (ATA) episode >30 seconds after a 3-month blanking period. A total of 2 patients with atrioventricular nodal reentrant tachycardia, 1 with left atrial appendage tachycardia, experienced AF recurrence within the first 3 months and received PVI. After the 3-month blanking period, during a median follow-up of 17.0 ± 10.1 months, 44 of 51 patients (86.2%) were free of ATA recurrence. In the PVI group, 33 of 39 patients (84.6%) experienced no ATA recurrence. In conclusion, SVT substrate is identified in around a quarter of young adult patients with history of AF, and targeted ablation without PVI may be sufficient in the majority of these patients. PVI is needed in the majority and is safe and effective in this population.


Quantum-Mechanical/Molecular-Mechanical Studies of CYP11A1-Catalyzed Biosynthesis of Pregnenolone from Cholesterol Reveal a C-C Bond Cleavage Reaction That Occurs by a Compound I-Mediated Electron Transfer.
We explore here a long-standing mechanistic question by using quantum-mechanical/molecular-mechanical (QM/MM) methodology. The question concerns the mechanism of steroid hormone biosynthesis, whereby the P450 enzyme, CYP11A1, catalyzes the C20-C22 bond-cleavage in the 20,22-hydroxylated cholesterol, 20R,22R-DiOHCH, leading to pregnenolone, which is critical for the subsequent production of all steroid hormones. This is an unusual feat whereby the P450 enzyme breaks two O-H bonds and one C-C bond, while making two C═O bonds. How does the enzyme perform such a complex and highly energy-demanding reaction? Our computational results rule out the previously proposed Compound I (Cpd I) electrophilic attack mechanism via the formation of a peroxide intermediate as well as the H-abstraction-mediated C-C cleavage mechanism. Notably, oxygen-rebound cannot transpire, in spite of the fact that the classical active species, Cpd I, participates in the catalytic process. Our findings reveal a mechanism whereby C-C bond cleavage is mediated by an electron transfer from the C22-O--deprotonated substrate to Cpd I. As such, our QM/MM calculations demonstrate that Cpd I acts as an electron sink that facilitates the C-C bond cleavage.


Applications of contrast-enhanced ultrasound in the pediatric abdomen.
Contrast-enhanced ultrasound (CEUS) is a radiation-free, safe, and in specific clinical settings, highly sensitive imaging modality. Over the recent decades, there is cumulating experience and a large volume of published safety and efficacy data on pediatric CEUS applications. Many of these applications have been directly translated from adults, while others are unique to the pediatric population. The most frequently reported intravenous abdominal applications of CEUS in children are the characterization of focal liver lesions, monitoring of solid abdominal tumor response to treatment, and the evaluation of intra-abdominal parenchymal injuries in selected cases of blunt abdominal trauma. The intravesical CEUS application, namely contrast-enhanced voiding urosonography (ceVUS), is a well-established, pediatric-specific imaging technique entailing the intravesical administration of ultrasound contrast agents for detection and grading of vesicoureteral reflux. In Europe, all pediatric CEUS applications remain off-label. In 2016, the United States Food and Drug Administration (FDA) approved the most commonly used worldwide second-generation ultrasound contrast SonoVue®/Lumason® for pediatric liver and intravesical applications, giving new impetus to pediatric CEUS worldwide.


Hepatic amebic abscess masquerading as acute cholecystitis.
Hepatobiliary scintigraphy has disclosed a host of intrahepatic and extrahepatic lesions which mimic gallbladder disease. Two cases of hepatic amebic abscess are reported in which the presentations masqueraded as acute cholecystitis. Correlative, noninvasive imaging rapidly diagnosed both cases and in 1 case identified a serious complication.


Pyridoxal 5'-phosphate, a fluorescent probe in the active site of aspartate transcarbamylase.
Pyridoxal-P reacts specifically with a single lysine residue at the active site of Escherichia coli aspartate transcarbamylase (Greenwell, P., Jewett, S. L., and Stark, G. R. (1973) J. Biol. Chem. 248, 5994-6001). Reduction of the Schiff base with sodium borohydride, succinylation of the remaining lysine residues, and digestion with trypsin result in formation of a single pyridoxyl peptide, which was purified to homogeneity after chromatography on DEAE-cellulose, treatment with alkaline phosphatase, and rechromatography. Amino acid composition and the results of limited sequential degradation showed that this peptide corresponds to residues 62 to 98 in the sequence of Konigsberg and co-workers, and contains 2 residues of lysine (Henderson, L., Roy, D., Martin, D., and Konigsberg, W., personal communication). By similar isolation, a second peptide was obtained from unsuccinylated catalytic subunit, containing only the pyridoxylated lysine, which corresponds to Lys-80. Derivatives of catalytic subunit containing an average of either one, two, or three pyridoxamine-P moieties per trimer have been prepared by reduction. These species, which retain catalytic activity in proportion to their unmodified active sites, were recombined with regulatory subunit to prepare partially modified derivatives of native aspartate transcarbamylase. At pH 8, fluorescence emission bands were observed at 340 nm, due to aromatic amino acids in the protein, and at 395 nm, due to the pyridoxamine-P moiety. Upon excitation at 280 nm energy transfer from protein to pyridoxamine-P was approximately 15%. The properties of the probe were used to study changes accompanying the binding of substrates and inhibitors. The effects of CTP and ATP were small. With the transition state analog N-(phosphonacetyl)-L-aspartate (PALA) or the substrate carbamyl-P, two types of response were observed. Derivatives of catalytic subunit and native enzyme which contain some unmodified sites and hence retain partial catalytic activity gave large increases in fluorescence at 395 nm. However, fully modified inactive derivatives gave much smaller increases. A derivative of native enzyme containing one triply modified and one unmodified catalytic subunit behaved like the other partially modified species. These results indicate that there is communication among the active sites of different catalytic trimers in modified native enzyme, as well as among active sites within the same modified catalytic trimer. The increases in fluorescence result from a red shift of the absorption maximum of the pyridoxamine-P moiety from 315 to 325 nm, which increases the absorbance at the excitation wavelength for fluorescence. At pH 7, the absorption spectrum is already shifted and, consequently, the binding of PALA and carbamyl-P has little effect on the fluorescence. Therefore, the binding of these compounds at pH 8.0 must cause a structural change in the protein, which in turn causes protonation of a group in the modified active sites, altering the spectral properties.


Convulsive therapy in delusional disorders.
Delusions are psychopathologic expressions associated with schizophrenia, affective disorders, and systemic and toxic illnesses. In each condition, the efficacy of ECT is well defined. For some illnesses, such as major depression with psychosis, manic delirium, and catatonia, ECT is effective in relieving the delusional states. This activity of ECT as an antidelusional agent compels us to consider the mode of action of ECT as broader than as an antidepressant treatment alone. The experience also compels greater consideration of the unitary classification of severe mental disorders.


Modelling foot-and-mouth disease: a comparison between the UK and Denmark.
Whilst the UK 2001 FMD (foot-and-mouth disease) outbreak provides an extremely rich source of spatio-temporal epidemic data, it is not clear how the models and parameters from the UK can be translated to other scenarios. Here we consider how the model framework used to capture the UK epidemic can be applied to a hypothetical FMD outbreak in Denmark. Whilst pigs played a relatively minor role in the UK epidemic (being the infected animal on just 18 farms), they dominate the Danish livestock landscape. In addition, it is not clear whether transmission parameters from the UK will transfer to Denmark where farming practices may be significantly different. We therefore explore a large volume of high-dimensional parameter space, but seek to relate final epidemic size, risk of spread to Danish islands and potential success of control measures, to early indicators of epidemic dynamics. The results of this extensive modelling exercise therefore allow us to provide timely advice on control options based on the observed behaviours of the first few generations.


Oxygen uptake by isolated perfused fish hearts with differing myoglobin concentrations under hypoxic conditions.
Hearts from three species of fish with varying myoglobin content were perfused with stepwise changes in input perfusate PO2 from approximately 160 to 10 mmHg. Flow through the heart, rate of contraction, and afterload were kept constant. This standardized stroke volume and bulk flow of perfusate to the myocytes since these hearts are nourished by the fluid in the ventricular lumen. In some cases NaNO2 was added to the perfusion medium to decrease existing levels of functional myoglobin. Myoglobin-rich hearts were able to extract a constant amount of oxygen until perfusate PO2 had fallen below 80 mmHg. At this point oxygen uptake began to decline. These hearts consumed oxygen until input PO2 was 10 mmHg or less. When normoxic conditions were restored the myoglobin-rich hearts showed complete recovery. Performance was maintained at a constant level over the entire range of input PO2. Myoglobin-poor hearts and nitrite-treated hearts were unable to sustain constant levels of oxygen consumption in the face of a declining perfusate PO2. These hearts were unable to extract oxygen from the medium and failed at perfusate PO2's of 40 mmHg for naturally myoglobin-poor hearts and 30 mmHg for nitrite-treated hearts. Half-maximal oxygen consumptions were attained by myoglobin-rich hearts at lower input PO2's than either myoglobin-poor or nitrite-treated hearts. The impact of myoglobin in intact heart is apparent at relatively high extracellular PO2's (40-80 mmHg) in this model system.


Crystalline Nanoparticles of Water-Soluble Covalent Basket Cages (CBCs) for Encapsulation of Anticancer Drugs.
We herein describe the preparation, assembly, recognition characteristics, and biocompatibility of novel covalent basket cage CBC-11, composed of four molecular baskets linked to four trivalent aromatic amines through amide groups. The cage is tetrahedral in shape and similar in size to small proteins (Mw =8637 g/mol) with a spacious nonpolar interior for accommodating multiple guests. While 24 carboxylates at the outer surface of CBC-11 render it soluble in aqueous phosphate buffer (PBS) at pH=7.0, the amphiphilic nature prompts its assembly into nanoparticles (d=250 nm, DLS). Cryo-TEM examination of nanoparticles revealed their crystalline nature with wafer-like shapes and hexagonally arranged cages. Nanoparticulate CBC-11 traps anticancer drugs irinotecan and doxorubicin, with each cage binding up to four drug molecules in a non-cooperative manner. The inclusion complexation resulted in nanoparticles growing in size and precipitating. In media containing mammalian cells (HCT 116, human colon carcinoma), the IC50 value of CBC-11 was above 100 μM. While this work presents the first example of a large covalent organic cage operating in water at the physiological pH and forming crystalline nanoparticles, it also demonstrates its biocompatibility and potential to act as a polyvalent binder of drugs for their sequestration or delivery.


Predictive motor activation: Modulated by expectancy or predictability?
Predicting actions is a fundamental ability that helps us to comprehend what is happening in our environment and to interact with others. The motor system was previously identified as source of action predictions. Yet, which aspect of the statistical likelihood of upcoming actions the motor system is sensitive to remains an open question. This EEG study investigated how regularities in observed actions are reflected in the motor system and utilized to predict upcoming actions. Prior to measuring EEG, participants watched videos of action sequences with different transitional probabilities. After training, participants' brain activity over motor areas was measured using EEG while watching videos of action sequences with the same statistical structure. Focusing on the mu and beta frequency bands we tested whether activity of the motor system reflects the statistical likelihood of upcoming actions. We also explored two distinct aspects of the statistical structure that capture different prediction processes, expectancy and predictability. Expectancy describes participants' expectation of the most likely action, whereas predictability represents all possible actions and their relative probabilities. Results revealed that mu and beta oscillations play different roles during action prediction. While the mu rhythm reflected anticipatory activity without any link to the statistical structure, the beta rhythm was related to the expectancy of an action. Our findings support theories proposing that the motor system underlies action prediction, and they extend such theories by showing that multiple forms of statistical information are extracted when observing action sequences. This information is integrated in the prediction generated by the neural motor system of which action is going to happen next.


Shaping the spectrum - From autoinflammation to autoimmunity.
Historically, autoimmune-inflammatory disorders were subdivided into autoinflammatory vs. autoimmune diseases. About a decade ago, an immunological continuum was proposed, placing "classical" autoinflammatory disorders, characterized by systemic inflammation in the absence of high-titer autoantibodies or autoreactive T lymphocytes, at the one end, and autoimmune disorders at the other end. We provide an overview of recent developments and observations, filling in some of the gaps and showing strong interconnections between innate and adaptive immune mechanisms, indicating that disorders from both ends of the immunological spectrum indeed share key pathomechanisms. We focus on three exemplary disorders: i) systemic juvenile idiopathic arthritis representing "classical" autoinflammatory disorders; ii) psoriasis, a mixed pattern disease; and iii) systemic lupus erythematosus, a prototypical autoimmune disease. We summarize scientific observations suggesting that, depending on disease stages and/or duration, individualized treatment targeting innate or adaptive immune mechanisms in disorders from either end of the immunological spectrum may control disease activity.


Gene expression profiling in glioblastoma and immunohistochemical evaluation of IGFBP-2 and CDC20.
Thirty-nine glial tumours (28 glioblastomas (GB) and 11 low-grade gliomas) were investigated with DNA microarrays to reveal a possible specific gene expression profile. Unsupervised classification through hierarchical cluster analysis identified two groups of tumours, the first composed of low-grade gliomas and the second mainly composed of GB. Nine genes were identified as most informative: seven were over-expressed in low-grade gliomas and under-expressed in GB; on the contrary, two genes, insulin-like growth factor binding protein 2 (IGFBP-2) and cell division cycle 20 homologue (CDC20), were over-expressed in GB and under-expressed in low-grade tumours. This same genetic profile was confirmed by reverse transcriptase polymerase chain reaction. Immunohistochemistry for IGFBP-2 was positive in 88.8% of the cases of GB and in only one low-grade glioma, whilst CDC20 immunostained 74.1% of the cases of GB and none low-grade glioma. This was confirmed in an additional series of cases studied with immunohistochemistry only. In conclusion, over-expression of mRNA levels of IGFBP-2 and CDC20 is highly related to GB, IGFBP-2 and CDC-20 gene and protein expressions are strongly correlated, and IGFBP-2 and CDC20 immunopositivity can be useful for the identification of GB in small biopsies.


Electron microscopy of flagella, primary cilia, and intraflagellar transport in flat-embedded cells.
Intraflagellar transport (IFT) is an evolutionarily highly conserved, microtubule-based, bidirectional transport system found in eukaryotic cilia/flagella and is indispensable for their assembly, maintenance, and sensory functions. Powered by two different motor complexes, linear arrays of protein particles, called IFT trains, are transported from the base to the tip of the cilium/flagellum and back, carrying axonemal precursors to the tip for assembly and turnover products back to the cell body for recycling. The dynamics of IFT can be visualized using various types of live-cell microscopy techniques, but for analyzing the ultrastructure of IFT trains, transmission electron microscopy is indispensable. The focus of this chapter is to describe the application of the flat embedding technique to Chlamydomonas reinhardtii and monolayers of mammalian culture cells. Such flat embeddings are well suited for the analysis of the ultrastructure of the IFT system by standard electron microscopy and electron tomography.


Functional properties of selected starter cultures for sour maize bread.
This paper focuses on the functional properties of maize sour-dough microflora selected and tested for their use as starter cultures for sour maize bread. Lactic acid bacteria and yeasts isolated from spontaneously fermented maize dough were selected based on dominance during fermentation and presence at the end of fermentation. Functional properties examined included acidification, leavening and production of some antimicrobial compounds in the fermenting matrix. The organisms previously identified as Lactobacillus plantarum, Lb. brevis, Lb. fermentum, Lb. acidophilus, Pediococcus acidilactici, Leuconostoc mesenteroides and Leuconostoc dextranicum and Saccharomyces cerevisiae were used singly and as mixed cultures in the fermentation (fermentation time: 12h at 28+/-2 degrees C) of maize meal (particle size >0.2mm). The pH fell from an initial value of 5.62-3.05 in maize meals fermented with Lb. plantarum; 4.37 in L. dextranicum+S. cerevisiae compared with the value for the control (no starter) of 4.54. Significant differences (P <or =0.05) were observed in values obtained for the functional properties tested when starters were inoculated compared with the control (no starter) except for leavening. Bivariate correlations at 0.01 levels (two-tailed) showed that significant correlations existed among pH and production of antimicrobial compounds in the fermenting meals, the highest correlation being between production of diacetyl and acid (0.694), a positive correlation indicating that production of both antimicrobial compounds increase together with time. Antimicrobial activities of the fermented maize dough were confirmed by their abilities to inhibit the growth of Salmonella typhi, Escherichia coli, Staphylococcus aureus and Aspergillus flavus from an initial inoculum concentration of 7 log cfu ml(-1)) for test bacteria and zone of inhibition of up to 1.33 cm for aflatoxigenic A. flavus. The findings of this study form a database for further studies on the development of starter cultures for sour maize bread production as an alternative bread specialty.


Cutaneous manifestations in patients with systemic lupus erythematosus: Forms at the beginning of the disease and during follow-up. Retrospective analysis.
To describe skin involvement (SI) in patients with systemic lupus erythematosus (SLE) at onset and during follow-up of the disease and to determine factors associated with SI at lupus diagnosis. Retrospective, observational, and descriptive study, from a single centre in patients diagnosed with SLE (ACR 1982-97 or SLICC 2012 criteria). The modified Gilliam classification for SI was used. Descriptive statistics and bivariate and multivariate analysis were performed to evaluate the factors associated with SI at diagnosis of the disease. 149 patients were included, 91.3% women with a median age at diagnosis of 33 years. SI at onset of the disease occurred in 125 patients (83.9%), followed by joint involvement in 120 cases (80.5%). Non-specific skin lesions were more frequent than specific lesions, 92.8% versus 66.4%, respectively. In the bivariate analysis, a longer delay to diagnosis, the presence of joint involvement, a lower presence of thrombocytopenia, and a higher SLEDAI-2K score were associated with the presence of SI at onset of the disease. In the multivariate analysis, the variable that remained independently associated was joint involvement (OR 2.8%-95% CI 1.1-7.5, p: .04). During follow-up, 4/24 patients who had not presented SI at diagnosis and 51/125 patients who had, had at least one new skin flare (range: 1-5 outbreaks). Our study demonstrates the high frequency of skin involvement in SLE, both diagnostically and evolutionarily, and confirms previously reported data regarding the existence of a skin-articular phenotype.


Dysregulated synthesis of intracellular type 1 and type 2 cytokines by T cells of patients with cutaneous T-cell lymphoma.
Mycosis fungoides (MF) and Sezary syndrome (SS) are the two main clinical entities of cutaneous T-cell lymphoma (CTCL). As the disease progresses from MF to SS, a switch from a type 1 (interleukin [IL]-2 and gamma interferon [IFN-gamma]) to a type 2 (IL-4) cytokine production profile occurs. Although roles for type 1 and type 2 cytokines in the pathogenesis of CTCL have been proposed, the cellular origins of these cytokines are unclear. Using flow cytometry to identify individual T-cell subsets, we studied cytokine synthesis by the T cells of 13 patients with SS and 12 with MF and 9 hematologically healthy donors. Upon activation with phorbol 12-myristate 13-acetate (PMA), the numbers of T cells synthesizing IL-2 were similar for all study groups. Whereas the predominant T-cell producing IL-2 in healthy donors and in those with MF was CD7(+), in patients with SS, it was CD7(-). Although the number of IL-4(+) CD4(+) T cells was low for all study groups, there was a significantly higher number of IL-4(+) CD8(+) T cells in patients with MF than in those with SS or healthy donors. There was a decline in the number of IFN-gamma-producing T cells in CTCL donors compared to that in healthy donors. More importantly, there was a significant decrease in the number of IFN-gamma-producing T cells with disease progression from MF to SS. The inability of these T cells to synthesize IFN-gamma may have prognostic value in CTCL, since it may be responsible for the progression of the disease from MF to SS.


Investigating the regulatory function of the ANO1-AS2 on the ANO1 gene in infertile men with asthenozoospermia and terato-asthenozoospermia.
Long non-coding RNAs (lncRNAs) have a particular expression in the testicular tissue and exhibit a regulatory function on the reproduction system. ANO1-AS2 (linc02584), as an lncRNA is located near the anoctamin1 (ANO1) gene. ANO1 is an important component of the transmembrane system exhibiting expression modifications in the idiopathic infertile men. Therefore, the present study was conducted to investigate the relationship between ANO1-AS2 and ANO1 gene expression with sperm motility and morphology in the patients with asthenozoospermia (AZ) and terato- asthenozoospermia (TAZ). The study population included 32 patients with AZ, 35 patients with TAZ, and 34 people with normozoospermia (NZ, control). The expression levels of ANO1 gene and ANO1-AS2 in the spermatozoa were measured by the quantitative real-time polymerase chain reaction (PCR). Docking analysis was performed to investigate the interactions of the ANO1 gene promoter and intermediate elements with ANO1-AS2. ANO1 gene expression was significantly (P < 0.05) downregulated in the patients however; ANO1-AS2 expression was significantly upregulated (P < 0.05). The subsequent analysis confirmed the inverse correlation between ANO1 and ANO1-AS2. ANO1 gene expression level was significantly positively correlated with sperm motility and morphology (P < 0.05). Moreover, ANO1-AS2 expression showed an inverse correlation with sperm motility and morphology (P < 0.05). Docking analysis confirmed that ANO1-AS2 could stably interact with ANO1 gene promoter. In conclusion, ANO1-AS2 is likely to downregulate the ANO1 gene by interacting with ANO1 gene promoter, which can influence the sperm motility and morphology.


The crystal structure of JNK from Drosophila melanogaster reveals an evolutionarily conserved topology with that of mammalian JNK proteins.
The c-Jun N-terminal kinases (JNKs), members of the mitogen-activated protein kinase (MAPK) family, engage in diverse cellular responses to signals produced under normal development and stress conditions. In Drosophila, only one JNK member is present, whereas ten isoforms from three JNK genes (JNK1, 2, and 3) are present in mammalian cells. To date, several mammalian JNK structures have been determined, however, there has been no report of any insect JNK structure. We report the first structure of JNK from Drosophila melanogaster (DJNK). The crystal structure of the unphosphorylated form of DJNK complexed with adenylyl imidodiphosphate (AMP-PNP) has been solved at 1.79 Å resolution. The fold and topology of DJNK are similar to those of mammalian JNK isoforms, demonstrating their evolutionarily conserved structures and functions. Structural comparisons of DJNK and the closely related mammalian JNKs also allow identification of putative catalytic residues, substrate-binding sites and conformational alterations upon docking interaction with Drosophila scaffold proteins. The DJNK structure reveals common features with those of the mammalian JNK isoforms, thereby allowing the mapping of putative catalytic and substrate binding sites. Additionally, structural changes upon peptide binding could be predicted based on the comparison with the closely-related JNK3 structure in complex with pepJIP1. This is the first structure of insect JNK reported to date, and will provide a platform for future mutational studies in Drosophila to ascertain the functional role of insect JNK.


Functional recreation of age-related CD8 T cells in young mice identifies drivers of aging- and human-specific tissue pathology.
Mitigating effects of aging on human health remains elusive because aging impacts multiple systems simultaneously, and because experimental animals exhibit critical aging differences relative to humans. Separation of aging into discrete processes may identify targetable drivers of pathology, particularly when applied to human-specific features. Gradual homeostatic expansion of CD8 T cells dominantly alters their function in aging humans but not in mice. Injecting T cells into athymic mice induces rapid homeostatic expansion, but its relevance to aging remains uncertain. We hypothesized that homeostatic expansion of T cells injected into T-deficient hosts models physiologically relevant CD8 T cell aging in young mice, and aimed to analyze age-related T cell phenotype and tissue pathology in such animals. Indeed, we found that such injection conferred uniform age-related phenotype, genotype, and function to mouse CD8 T cells, heightened age-associated tissue pathology in young athymic hosts, and humanized amyloidosis after brain injury in secondary wild-type recipients. This validates a model conferring a human-specific aging feature to mice that identifies targetable drivers of tissue pathology. Similar examination of independent aging features should promote systematic understanding of aging and identify additional targets to mitigate its effects on human health.


Efficacy and safety of natalizumab in Crohn's disease patients treated at 6 Boston academic hospitals.
Despite trials demonstrating its efficacy, many physicians harbor concerns regarding the use of natalizumab in the treatment of patients with refractory Crohn's disease (CD). The purpose of this study was to perform a descriptive analysis of a series of CD patients not currently enrolled in a clinical trial. A retrospective case review of patients treated with natalizumab at 6 sites in Massachusetts: Boston Medical Center, Beth Israel Deaconess Medical Center, Brigham & Women's Hospital, Lahey Clinic, Massachusetts General Hospital, and UMass Medical Center. Data on 69 CD patients on natalizumab were collected. At the start of treatment, patients' disease duration was 12 years. A high proportion of patients were women (68%), presented with perianal disease (65%) and upper gastrointestinal tract involvement (14%). Prior nonbiologic therapies were steroids (96%), thiopurines (94%), antibiotics (74%), methotrexate (58%), and at least two anti-tumor necrosis factor agent failures (81%). Sixty-nine percent (44 of 64 patients) with available medical evaluation had a partial or complete clinical response. Loss of response was 13% after an average of 1 year of treatment. Adverse events were infusion reactions, headaches, fever, and infections. No case of progressive multifocal leukoencephalopathy was observed. In our clinical experience outside the context of a clinical trial, natalizumab is largely reserved for CD patients with extensive ileocolonic disease who have failed conventional immunosuppressants and of at least 2 anti-tumor necrosis factor agents. This drug is, however, well tolerated and offers significant clinical improvement for more than a year in one-third of these difficult-to-treat CD patients.


Intracellular localization of lipoplexed siRNA in vascular endothelial cells of different mouse tissues.
Liposomally formulated siRNA can be used for RNAi applications in vivo. Intravenous bolus administration of lipoplexed siRNA has been shown to reduce gene expression in the vascular endothelium. Here, we applied immunofluorescence staining for different endothelial markers (PECAM-1, CD34, laminin) on paraffin sections to compare the respective expression pattern with the intracellular localization of intravenously administered, fluorescently labeled siRNA (siRNA-Cy3-lipoplex). By confocal microscopy, lipoplexed siRNA-Cy3 was detected inside vascular endothelial cells in vivo, which where identified with co-staining of endothelial markers. Consequently, the finding of intracellular siRNA uptake by vascular endothelial cells correlated with RNAi based specific protein reduction in situ as revealed by PECAM-1 specific immunofluorescence staining in lung tissue sections. Therefore, by using a cell biological approach these in situ data emphasize the functional uptake of liposomal siRNA molecules in vascular endothelial cells of different mouse tissues as indicated in our previous molecular study.


Asymmetric dimethylarginine (ADMA) in human blood: effects of extended haemodialysis in the critically ill patient with acute kidney injury, protein binding to human serum albumin and proteolysis by thermolysin.
Free, non-protein bound asymmetrically guanidine-dimethylated arginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthesis. Human erythrocytic membrane comprises considerable amounts of large (>50 kDa) ADMA-containing proteins. Location in the erythrocyte membrane and identity and physiological functions of ADMA-containing proteins are unrevealed. In healthy subjects, the concentration of free ADMA in heparinised plasma is almost identical to that of serum. We hypothesised that the robustness of free ADMA concentration in human blood is due to a remarkable resistance of erythrocytic ADMA-containing proteins against proteases. In vivo, we investigated the course of the concentration of ADMA in serum and EDTA plasma of a critically ill patient with acute kidney injury during extended haemodialysis. In vitro, we studied the effects of thermolysin, a useful experimental proteolytic enzyme of erythrocyte membrane proteins, on erythrocytic ADMA. The protein binding (PB) of ADMA to human serum albumin (HSA) was also determined. In these studies, ADMA was measured by a previously reported, fully validated GC-MS/MS method. We measured almost identical ADMA concentrations in plasma and serum samples of the patient. During dialysis, the circulating ADMA concentration decreased slowly and moderately indicating removal of this substance, which was however much less than expected from its low molecular weight (202 Da) and high water solubility. After dialysis, circulating ADMA concentration increased again, a phenomenon called rebound, and ADMA reached higher levels compared to the baseline. The PB value of ADMA to HSA was about 30 %. This surprisingly high PB value of ADMA to HSA may be an explanation for the rather poor dialysance of ADMA. Washed human erythrocytes suspended in phosphate-buffered physiological saline were found not to release appreciable amounts of free and ADMA-containing proteins. The lack of effect of coagulation or anticoagulation on the concentration of circulating free ADMA in humans is likely to be due to a remarkable resistance of ADMA-containing proteins in the erythrocyte membrane against proteases in vivo in humans. Our study suggests that free ADMA is released in the circulating blood at relatively high rates. The considerable PB of ADMA to HSA is likely to add to the apparently poor dialysability of ADMA. Other contributing factors could be redistribution of free ADMA between plasma and erythrocytes in favour of plasma ADMA and parallel formation of free ADMA from erythrocytic ADMA-containing proteins during haemodialysis.


VEGF inhibits PDGF-stimulated calcium signaling independent of phospholipase C and protein kinase C.
Despite advances in both open and endovascular techniques for treatment of arterial occlusive disease, restenosis because of neointimal hyperplasia continues to be a major cause of graft failure and restenosis. This phenomenon has been attributed to vascular smooth muscle cell (VSMC) activation by several potent mitogens including platelet derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) released at the site of injury. PDGF is known to stimulate calcium influx in VSMC that has been shown to be critical for VSMC migration and proliferation. We have previously shown that VEGF inhibits PDGF-stimulated VSMC proliferation. The objective of this set of experiments was to investigate whether VEGF modulated PDGF-stimulated Ca2+ influx in VSMC. Primary cultured human aortic SMC were grown to subconfluency and assigned to the following groups: no stimulation, stimulation with PDGF-BB (20 ng/ml), stimulation with VEGF165 (40 ng/ml), or a combination of PDGF-BB + VEGF165. Ca2+ influx was measured using a Fura-2 fluorescence assay. The intracellular Ca2+ fraction was assayed with the Fura-2 assay by using Ca2+-free media. Phospholipase Cgamma1 (PLCgamma1), protein kinase C (PKC), and Akt phosphorylation was assessed with standard immunoblotting techniques at 1, 5, and 10 min time points. Ca2+-calmodulin kinase II (CaMKII) activity was extrapolated from the phosphorylation of Phospholamban B (PLB), a well-known protein substrate, at 1, 5, and 10 min time points. PDGF stimulation resulted in a 328 +/- 9 nm total calcium influx in VSMC. The combination of VEGF + PDGF resulted in a 273 +/- 21 nm total calcium influx, an amount significantly less than with PDGF alone (P < 0.04). PDGF stimulation resulted in a 72 +/- 35 nm intracellular calcium release. The addition of VEGF to PDGF resulted in an intracellular calcium release of only 15 +/- 11 nm, a significant decrease compared to PDGF alone (P < 0.01). The phosphorylation of PLCgamma1, PKC, and Akt was equivalent at 1, 5, and 10 min between the PDGF and the PDGF + VEGF treatment groups. There was an increase in CaMKII activity at 1 and 5 min time points in both the PDGF and PDGF + VEGF treatment groups suggesting that extracellular calcium influx is sufficient for CaMKII activation. VEGF inhibits PDGF-stimulated total calcium influx and, in particular, PDGF-stimulated intracellular calcium release in VSMC. The equivalent phosphorylation of PLCgamma1, PKC, and Akt suggests that the inhibitory mechanism by VEGF on calcium influx occurs downstream of these proximal mediators. The inhibition of intracellular calcium release did not inhibit CaMKII activity. VEGF may play an important role in modulating PDGF induced VSMC proliferation by specifically inhibiting intracellular calcium release in response to PDGF.


A Cell-Cell Fusion Assay to Assess Arenavirus Envelope Glycoprotein Membrane-Fusion Activity.
For many viruses that enter their target cells through pH-dependent fusion of the viral and endosomal membranes, cell-cell fusion assays can provide an experimental platform for investigating the structure-function relationships that promote envelope glycoprotein membrane-fusion activity. Typically, these assays employ effector cells expressing the recombinant envelope glycoprotein on the cell surface and target cells engineered to quantitatively report fusion with the effector cell. In the protocol described here, Vero cells are transfected with a plasmid encoding the arenavirus envelope glycoprotein complex GPC and infected with the vTF7-3 vaccinia virus expressing the bacteriophage T7 RNA polymerase. These effector cells are mixed with target cells infected with the vCB21R-lacZ vaccinia virus encoding a β-galactosidase reporter under the control of the T7 promoter. Cell-cell fusion is induced upon exposure to low-pH medium (pH 5.0), and the resultant expression of the β-galactosidase reporter is quantitated using a chemiluminescent substrate. We have utilized this robust microplate cell-cell fusion assay extensively to study arenavirus entry and its inhibition by small-molecule fusion inhibitors.


Functional analysis of jasmonic acid-responsive secondary metabolite transporters.
Jasmonic acid (JA) is a plant hormone that mediates a wide variety of plant developmental processes and defense responses. One of the major roles of JA is the versatile enhancement of the production of secondary metabolites that function as second messengers in plant defense responses. Recently, several genes have been identified as coding for JA-responsive transporters involved in the membrane transport of various secondary metabolites. Although in the literature such transport activities have been explored by a number of methods, only a few studies systematically provide a detailed technical basis of the transport assay. Here, we describe the established method to functionally analyze secondary metabolite transporters by means of a yeast cellular transport system. Moreover, the advantages and disadvantages of the method are summarized and the relevant technical points are noted.


Therapeutic improvement of glucoregulation in newly diagnosed type 2 diabetes patients is associated with a reduction of IL-17 levels.
We explored the effect of therapeutic glucoregulation on the blood levels of proinflammatory T helper (Th)17 cytokines interleukin (IL)-17 and IL-23, and Th1 cytokines interferon (IFN)-γ and IL-12 in newly diagnosed type 2 diabetes patients. The investigated group consisted of 23 subjects (17 men and 6 women, age 26-64). The cytokine serum levels, glycated hemoglobin (HbA1c) as a marker of glucoregulation, homeostasis model assessment index as a measure of insulin resistance (HOMA-IR), and body mass index (BMI) were determined before and after 12 weeks of therapy consisting of standard lifestyle modification and metformin (1000 mg b.i.d.). The levels of Th17 and Th1 cytokines before treatment did not correlate with age, BMI or HOMA-IR. The patients with poor glucoregulation (HbA1c>7%, n=12), compared to those with good glucoregulation (HbA1c≤7%, n=11), had higher serum levels of Th17 and Th1 cytokines, but only the differences in IL-17 (median 21.2 pg/ml vs. 4.8 pg/ml) and IFN-γ 5 (0.6 pg/ml vs. 27.7 pg/ml) reached statistical significance (p=0.003 and p=0.012, respectively). The reduction of HbA1c values (from 8.6 to 5.9%, p=0.000) observed upon treatment in patients with poor glucoregulation was associated with a significant decrease in the concentration of IL-17 (from 21.2 to 12.9 pg/ml, p=0.020), but not IFN-γ (50.6 vs. 52.3, p=0.349). These data indicate that therapeutic improvement of glucoregulation might contribute to a reduction of IL-17 levels in newly diagnosed type 2 diabetes patients.


Detection, characterization, and antimicrobial susceptibility of Yersinia enterocolitica in different types of wastewater in the Czech Republic.
The purpose of the study was to evaluate the occurrence of Yersinia enterocolitica in different types of wastewater and to characterize the isolates by biotyping, serotyping, and antimicrobial susceptibility testing. In addition, cultivation protocols were evaluated. The occurrence of Y. enterocolitica was determined in treated and untreated municipal wastewater, as well as in hospital, slaughterhouse, and cowshed wastewater. Y. enterocolitica was detected in 84.1% of the wastewater samples, while the main sources were untreated municipal and slaughterhouse wastewater. In contrast, the lowest incidence was found in hospital wastewater. An exclusive occurrence of biotype 1A (98.3%) was detected. Pathogenic bio-serotypes 4/O:3 and 3/O:3 were isolated only from slaughterhouse wastewater. The highest resistance rates were observed for ampicillin (92.5%) and amoxicillin-clavulanic acid (36.8%). Y. enterocolitica was commonly detected in wastewater, although the prevalence varied depending on the origin of the wastewater. No single cultivation protocol was able to recover Y. enterocolitica isolates from such a complex matrix as wastewater. Our study provided data that can contribute to the evaluation of wastewater as a source of Y. enterocolitica and to understanding the threat of wastewater isolates to human health.


Multifunctional role of β-1, 3 glucan binding protein purified from the haemocytes of blue swimmer crab Portunus pelagicus and in vitro antibacterial activity of its reaction product.
β-1, 3 glucan binding protein (β-GBP) was isolated from the haemocytes of blue swimmer crab, Portunus pelagicus and purified by laminarin coupled Sephadex G-100 affinity column chromatography. The purified β-GBP has the molecular mass of 100 kDa, confirmed by SDS-PAGE. The X-ray diffraction analysis of purified β-GBP indicates the crystalline nature of the protein and also the presence of single peak confirming the existence of β-glucan molecule. The results of agglutination assay showed that the purified β-GBP had the ability to agglutinate with yeast cell, Saccharomyces cerevisiae and mammalian erythrocytes. β-GBP can agglutinate with yeast cells at the concentration of 50 μg/ml. The phagocytic and encapsulation activity of purified β-GBP from P. pelagicus was determined with yeast cell S. cerevisiae and sepharose bead suspension respectively. This reveals that, β-GBP have the ability to detect the pathogen associated molecular patterns (PAMP) found on the surface of fungi and bacteria. The recognition of invading foreign substances and in the involvement of functional activities induces the activation of prophenoloxidase. This revealed that β-GBP play a major role in the innate immune system of crustaceans by stimulating the prophenoloxidase system. Moreover, it was obvious to note that β-GBP reaction product exhibited antibacterial and antibiofilm activity against Gram positive and Gram negative bacteria. This study concludes the functional aspects of β-GBP purified from P. pelagicus and its vital role in the stimulation of prophenoloxidase cascade during the pathogenic infection.


The number of parallel fibre-Purkinje dendrite synapses. A morphometric evaluation.
Quantitative parameters concerning synapses were studied in the cerebellar molecular layer of 4 cats using ultrastructural morphometric methods. The number of parallel fibre-Purkinje dendrite synapses was estimated to be about 200,000.


Synergistic "Anchor-Capture" Enabled by Amino and Carboxyl for Constructing Robust Interface of Zn Anode.
While the rechargeable aqueous zinc-ion batteries (AZIBs) have been recognized as one of the most viable batteries for scale-up application, the instability on Zn anode-electrolyte interface bottleneck the further development dramatically. Herein, we utilize the amino acid glycine (Gly) as an electrolyte additive to stabilize the Zn anode-electrolyte interface. The unique interfacial chemistry is facilitated by the synergistic "anchor-capture" effect of polar groups in Gly molecule, manifested by simultaneously coupling the amino to anchor on the surface of Zn anode and the carboxyl to capture Zn2+ in the local region. As such, this robust anode-electrolyte interface inhibits the disordered migration of Zn2+, and effectively suppresses both side reactions and dendrite growth. The reversibility of Zn anode achieves a significant improvement with an average Coulombic efficiency of 99.22% at 1 mA cm-2 and 0.5 mAh cm-2 over 500 cycles. Even at a high Zn utilization rate (depth of discharge, DODZn) of 68%, a steady cycle life up to 200 h is obtained for ultrathin Zn foils (20 μm). The superior rate capability and long-term cycle stability of Zn-MnO2 full cells further prove the effectiveness of Gly in stabilizing Zn anode. This work sheds light on additive designing from the specific roles of polar groups for AZIBs.


Molecular identification of vaginal fluid by microbial signature.
The discrimination of body fluids in forensic examinations can play an important role in crime scene reconstruction. Conventional methods rely on the detection of antigens or enzymatic activity, limiting detection sensitivity and specificity, particularly on old forensic samples. Methods based on human RNA analysis are not easily applicable to samples exposed to harsh and degrading environments. An alternative approach based on the identification of prokaryotic genomes was developed. Specific bacterial communities are characteristic typical of different human non-sterile body fluids: the molecular characterization of a microbial signature, and not the typing of single bacterial species, can effectively lead to univocal identification of these fluids. A multiplex real time PCR assay was developed using oligonucleotide mixtures targeting genomes specific for a selected group of bacteria. Microflora DNA (mfDNA) was extracted from vaginal, oral and fecal clinical swabs. In addition forensic samples were processed. Vaginal samples showed a strong specific signal for bacteria of the female genital tract. Oral samples clearly showed signal for bacteria present in saliva, and in fecal samples the main signal was from Enterococcaceae. Vaginal casework samples showed results comparable to freshly collected ones; moreover the DNA extracted was successfully used for STR typing. Also mixtures of body fluids were analyzed, providing a microbiological signature compatible with the presence of microbes of oral, fecal and vaginal origin. The presented method can be useful in identifying biological fluids, and it is based on DNA technologies already available in forensic laboratories and feasible for further high throughput automation.


Sarcomatous proliferation of the vasculature in a subependymoma.
A case of a recurrent posterior fossa subependymoma in a 60-year-old man is reported. In the recurrent tumor, the vasculature had undergone apparent sarcomatous proliferation of the endothelial cells. The bizarre cells expanded and replaced the vessel walls, and stained for Factor VIII-related antigen. The literature on gliosarcomas arising in brain tumors other than high-grade gliomas is reviewed.


The effect of molecular adsorbent recirculating system on pathophysiological parameters in patients with acute liver failure.
To investigate the effect of the molecular adsorbent recirculating system (MARS) on physiological variables in patients with acute liver failure. A prospective, observational study of MARS in addition to standard medical therapy in the management of patients presenting with acute liver failure. A regional liver transplant centre. Ten consecutive patients admitted with acute liver failure with a grade III or IV hepatic encephalopathy. MARS therapy for 8 h on 2 consecutive days. Standard monitoring included the use of a pulmonary artery catheter and an intracranial pressure monitor. During the first MARS treatment there was a significant increase in systemic vascular resistance index (SVRI) from 1114+/-196 to 1432+/-245 dyne s(-1) cm(-5) m(-2) with a reduction in cardiac index from 5.5+/-0.6 to 4.2+/-0.4 l min(-1) m(-2). The changes were maintained between the start of the first and second sessions but not to the end of second. Significant clearance of urea and creatinine was observed. Intracranial pressure did not change during the treatments. Overall mortality was 70%. MARS therapy was well tolerated, with significant increases in vascular tone during the first session. This increase was not sustained over the duration of the study with a return to baseline values by the end of the second session. Based on our experience we cannot recommend the routine use of MARS therapy in acute liver failure outside of a clinical trial.


Surgical considerations in the use of platelet-rich plasma.
Platelet-rich plasma is an autologous source of platelet-derived growth factors that enhance surgical soft- and hard-tissue wound healing. Surgeons should be aware of its mechanisms of action and benefits, as well as the controversy regarding its use.


Dynamic monitoring of green credit risk using a hybrid gray correlation degree-TOPSIS method.
Under the background of the new development pattern, the green credit market has ushered in a huge space for development. The monitoring and control of green credit risk is conducive to solving the negative impact of green credit risk and promoting the healthy development and smooth operation of the green credit market. This study first takes different provinces in China as samples; takes the data of government departments, financial institutions, enterprises, and other stakeholders as the monitoring content; and then uses the hybrid gray correlation degree-TOPSIS method to analyze. Next, this paper compares the green credit risk monitoring values of different regions at the same time and the same regions at different times. And it compares the green credit risk monitoring values of green finance pilot cities before and after the pilot. The research results show that there are obvious differences in eastern, central, and western regions in different time periods, and the green credit risk is also different in green finance pilot cities before and after the pilot. Stakeholders such as government departments, financial institutions, and enterprises need to take active measures to improve the efficiency of green credit risk management and achieve the sustainability of green credit development.


Enhanced human contrast sensitivity with increased stimulation of melanopsin in intrinsically photosensitive retinal ganglion cells.
The intrinsically photosensitive retinal ganglion cells (ipRGCs) are known to serve non-image-forming functions, such as photoentrainment of the circadian rhythm and pupillary light reflex. However, how they affect human spatial vision is largely unknown. The spatial contrast sensitivity function (CSF), which measures contrast sensitivity as a function of spatial frequency, was used in the current study to investigate the function of ipRGCs in pattern vision. To compare the effects of different background lights on the CSF, we utilized the silent substitution technique. We manipulated the stimulation level of melanopsin (i.e., the visual pigment of ipRGCs) from the background light while keeping the cone stimulations constant, or vice versa. We conducted four experiments to measure the CSFs at various spatial frequencies, eccentricities, and levels of background luminance. Results showed that melanopsin stimulation from the background light enhances spatial contrast sensitivity across different eccentricities and luminance levels. Our finding that melanopsin contributes to CSF, combined with the receptive field analysis, suggests a role for the magnocellular pathway and challenges the conventional view that ipRGCs are primarily responsible for non-visual functions.


New roles for insulin-like hormones in neuronal signalling and protection: new hopes for novel treatments of Alzheimer's disease?
Type 2 diabetes has been identified as a risk factor for Alzheimer's disease (AD). This is most likely due to the desensitisation of insulin receptors in the brain. Insulin acts as a growth factor and supports neuronal repair, dendritic sprouting, and differentiation. This review discusses the potential role that insulin-like hormones could play in ameliorating the reduced growth factor signalling in the brains of people with AD. The incretins glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) have very similar properties in protecting neurons from toxic effects, and are capable of reversing the detrimental effects that beta-amyloid fragments have on synaptic plasticity. Therefore, incretins show great promise as a novel treatment for reducing degenerative processes in AD.


Peroxisome proliferator-activated receptor gamma agonists inhibit HIV-1 replication in macrophages by transcriptional and post-transcriptional effects.
Previous studies have demonstrated that cyclopentenone prostaglandins (cyPG) inhibit human immunodeficiency virus type 1 (HIV-1) replication in various cell types. We investigated the role of PG in the replication of HIV-1 in primary macrophages. The cyPG, PGA(1) and PGA(2), inhibited HIV-1 replication in acutely infected human monocyte-derived macrophages (MDM). Because PGA(1) and PGA(2) have previously been shown to be peroxisome proliferator-activated receptor gamma (PPARgamma) agonists, we examined the effect of synthetic PPARgamma agonists on HIV replication. The PPARgamma agonist ciglitazone inhibited HIV-1 replication in a dose-dependent manner in acutely infected human MDM. In addition, cyPG and ciglitazone reduced HIV replication in latently infected and viral entry-independent U1 cells, suggesting an effect at the level of HIV gene expression. Ciglitazone also suppressed HIV-1 mRNA levels as measured by reverse transcriptase PCR, in parallel with the decrease in reverse transcriptase activity. Co-transfection of PPARgamma wild type vectors and treatment with PPARgamma agonists inhibited HIV-1 promoter activity in U937 cells. Activation of PPARgamma also decreased HIV-1 mRNA stability following actinomycin D treatment. In summary, our experimental findings implicate PPARgamma as an important factor in the suppression of HIV-1 gene expression in MDM by cyPG. Thus natural and synthetic PPARgamma agonists may play a role in controlling HIV-1 infection in macrophages.


Differential expression and regulation of anti-hypertrophic genes Npr1 and Npr2 during β-adrenergic receptor activation-induced hypertrophic growth in rats.
We sought to determine the effect of chronic activation of β-adrenergic receptor (β-AR) on the left ventricular (LV) expression profile of Npr1 and Npr2 (coding for NPR-A and NPR-B, respectively) genes, and the functional activity of these receptors in adult Wistar rat hearts. The Npr1 gene expression was markedly reduced (3.5-fold), while the Npr2 gene expression was up regulated (4-fold) in Isoproterenol (ISO)-treated heart as compared with controls. A gradual reduction in NPR-A protein (3-fold), cGMP levels (75%) and a steady increased expression of NPR-B protein (4-fold), were noticed in ISO hearts. Further, in-vitro membranes assay shows that NPR-A dependent guanylyl cyclase (GC) activity was down-regulated (2-fold), whereas NPR-B dependent GC activity was increased (5-fold) in ISO treated hearts. Atenolol treatment normalized the altered expression of Npr1 and Npr2 genes. In conclusion, the chronic β-AR activation differentially regulates Npr1 and Npr2 genes in the heart. Npr1 down regulation is positively associated with the development of left ventricular hypertrophy (LVH) in ISO rats.


Human papillomavirus, Epstein-Barr virus, human herpesvirus 8 and human cytomegalovirus involvement in salivary gland tumours.
Human papillomaviruses (HPV) are involved in the etiology of both benign and malignant epithelial lesions. The occurrence of HPV types 16 and 18 in gynecological squamous cell carcinomas is also well known. Of the herpesviruses, Epstein-Barr virus (EBV) is associated with, for example, undifferentiated nasopharyngeal carcinoma, endemic Burkitt's lymphoma and immunoblastic lymphoma, and human herpesvirus 8 (HHV-8) with Kaposi's sarcoma. As little is known about the etiological factors of salivary gland tumours, the presence of HPV, EBV, HHV-8 and human cytomegalovirus (CMV) in these tumours were examined. Fresh tissue samples obtained from 19 consecutive pleomorphic adenomas and 19 malignant salivary gland tumours were analyzed with polymerase chain reaction. Two samples showed EBV DNA positivity, a lymphoma of the parotid gland and a pleomorphic adenoma arising in the nasal cavity. HPV, HHV-8 and CMV DNA were not detected in any of the tumour samples. The results indicate that HPV, HHV-8 and CMV do not seem to have any role in the etiology of salivary gland neoplasms.


Synthesis, evaluation of anticancer activity and COMPARE analysis of N-bis(trifluoromethyl)alkyl-N'-substituted ureas with pharmacophoric moieties.
A series of new synthesized N-bis(trifluoromethyl)alkyl-N'-substituted ureas have been tested in the National Cancer Institute (NCI, Bethesda, USA) by Program NCI-60 DTP Human Tumor Cell Line Screen at a single high dose (10(-5) M). COMPARE analysis has been carried out for all tested compounds. The tested compounds showed antitumor activity against individual cell lines. The most sensitive cell lines relative to the tested compounds are: 5 g Leukemia RPMI-8226 (GI% 52.7), Non-Small Cell Lung cancer HOP-92 (GI % 88.53), NCI-H522 (GI % 64.41), Melanoma UACC-62 (GI% 53.08), SK-MEL-5 (GI % 74.63), Breast cancer MDA-MB-468 (GI% 51.29), T-47D (GI % 65.1), 5b Leukemia K-562 (GI % 55.55), 7 m Leukemia HL-60(TB) (GI % 51.76).


Don't middle your MIDs: regression to the mean shrinks estimates of minimally important differences.
Minimal important differences (MIDs) for patient-reported outcomes (PROs) are often estimated by selecting a clinical variable to serve as an anchor. Then, differences in the clinical anchor regarded as clinically meaningful or important can be used to estimate the corresponding value of the PRO. Although these MID values are sometimes estimated by regression techniques, we show that this is a biased procedure and should not be used; alternative methods are proposed.


Eugenol inhibits K+ currents in trigeminal ganglion neurons.
Eugenol, a natural capsaicin congener, is widely used in dentistry. Eugenol inhibits voltage-activated Na(+) and Ca(2+) channels in a transient receptor potential vanilloid 1 (TRPV1)-independent manner. We hypothesized that eugenol also inhibits voltage-gated K(+) currents, and investigated this in rat trigeminal ganglion neurons and in a heterologous system using whole-cell patch clamping. Eugenol inhibited voltage-gated K(+) currents, and the inhibitory effects of eugenol were observed in both capsaicin-sensitive and capsaicin-insensitive neurons. Pre-treatment with capsazepine, a well-known antagonist of TRPV1, failed to block the inhibitory effects of eugenol on K(+) currents, suggesting no involvement of TRPV1. Eugenol inhibited human Kv1.5 currents stably expressed in Ltk(-) cells, where TRPV1 is not endogenously expressed. We conclude that eugenol inhibits voltage-gated K(+) currents in a TRPV1-independent manner. The inhibition of voltage-gated K(+) currents is likely to contribute to the irritable action of eugenol. human Kv1.5 channel, hKv1.5; transient receptor potential vanilloid 1, TRPV1.


Opinion on Immune Tolerance Therapeutic Development.
Immune tolerance is defined by an active state of immune system unresponsiveness to foreign and self-antigens. Loss of immune tolerance to self-antigens and the resulting overexpression of autoantibodies can lead to tissue injury and development of various autoimmune diseases. In drug development, the goal of newly emerging immune tolerance therapies is to treat autoimmune disorders by restoring the immunoregulatory capacity of the immune system. Development of immune tolerance targets is initiated with the establishment of pharmacological efficacy in relevant disease animal models, followed by their stepwise translation to humans. This review discusses the major challenges to developing tolerance inducing pharmaceutical drugs, including the selection of appropriate disease models to establish efficacy, adequate, and acceptable in vitro and in vivo safety assessments, relevant biomarkers of human safety and efficacy, and finally, some regulatory guidelines to successfully develop immune tolerance therapeutics. [Box: see text].


Passive transfer studies with type II collagen antibody in B10.D2/old and new line and C57Bl/6 normal and beige (Chediak-Higashi) strains: evidence of important roles for C5 and multiple inflammatory cell types in the development of erosive arthritis.
C5-normal B10.D2/new line mice were susceptible to passively transferred arthritis from purified type II collagen antibody, and in vitro studies demonstrated that, when bound to cartilage, this antibody readily activated complement C5 to C5a. C5-deficient B10.D2/old line mice failed to develop passively transferred arthritis, despite the deposition of antibody and C3 along the cartilage surface. C57Bl/6 mice were susceptible to passively transferred arthritis, which was characterized in histopathologic studies as an erosive synovitis involving multiple inflammatory cell types. In contrast, neither clinical nor histologic evidence of arthritis was observed in C57Bl/6 mice with the beige mutation (Chediak-Higashi syndrome).


Improved sub-areolar breast tissue removal in nipple-sparing mastectomy using hydrodissection.
We report on a new technique of dissection of the nipple-areola-complex (NAC) in nipple-sparing mastectomy (NSM). NACs removed due to the presence of tumor cells beneath them were histologically examined for the presence of normal breast glandular tissue. Cases were divided into cohort 1, where NACs were dissected by sharp isolation, coring the nipple, and cohort 2, where the same procedure was preceded by hydrodissection of the areola. In 20 (17.4%) cases the planned NSM was converted to skin-sparing mastectomy (SSM) because of intraoperative findings of cancer in retro-areolar tissue. Histological examination of 20 NSMs converted to SSM showed the presence of glandular tissue in 12 out of 13 cohort 1 cases (92%) and in 1 out of 7 cohort 2 cases (14%). We conclude that hydrodissection creates a subdermal plane facilitating NAC dissection and permitting a more complete removal of breast tissue in NSM. Such radicality could prove important in the treatment of breast cancer and in BRCA 1-2 mutation carriers because of its potential for reducing the risk of relapse.


The role of imaging in the diagnosis and management of renal stone disease in pregnancy.
The distinction of pain in pregnancy due to urolithiasis from that related to physiological dilation of the renal tract is a common conundrum as renal colic is one of the commonest causes for non-obstetric pain in pregnancy. Ultrasound is the first-line imaging test but although it may demonstrate renal dilation, it may not show the cause. Magnetic resonance imaging (MRI) is able to make the distinction. Physiological dilation will show smooth tapering of the ureter in the middle third as it is compressed between the gravid uterus and the retroperitoneum. Obstruction due to calculi causes renal enlargement and perinephric oedema. When a stone is lodged in the lower ureter, a standing column of dilated ureter will be seen below the physiological constriction. The stone itself may be shown. Computed tomography (CT) is an acceptable alternative if there is a contraindication to MRI, but even low-dose regimes involve some ionising radiation. This paper serves to highlight the role of MRI compared to US and CT in the imaging of renal colic in pregnancy. Multidisciplinary collaboration between obstetricians, urologists, and radiologists is required for effective management.


Antibiotic susceptibility of 206 Haemophilus influenzae isolates collected from children in central Italy.
Susceptibility of 206 H. influenzae isolates was evaluated by disk diffusion method for 11 antimicrobial agents. No isolates were found to be resistant to third-generation cephalosporins, amoxicillin+clavulanic acid, gentamicin and ciprofloxacin. Four untypable isolates (1.9%) were beta-lactamase producing ampicillin-resistant; one of these was also resistant to chloramphenicol. The rate of resistance against rifampin was 0.5 percent.


Billing challenges for residents of skilled nursing facilities.
Consolidated billing is a commonly used but little understood form of reimbursement for medical services provided in skilled nursing facilities.


Effects of the anticancer drug cocktail "UFT" on metastatic potentials of squamous cell carcinoma (O-1N) in a hamster model.
UFT, a mixture of 1-(2-tetrahydrofuryl)-5-fluorouracil (tegafur) and uracil, is one of the most widely used anticancer agents. The tissue localization and the efficacy of UFT in preventing lymph node metastasis were studied in hamsters transplanted with oral squamous cell carcinoma (termed "O-1N") which has high lymph node metastatic potency. After UFT administration for 14 consecutive days, the incidence of lymph node metastasis was significantly decreased (16% vs 58%). When UFT was administered orally for 3 consecutive days to hamsters bearing O-1N, and tissue concentrations of tegafur and 5-FU were measured by gas chromatography, the value of 5-FU was significantly higher in the primary transplant tumour, cervical lymph nodes with or without metastasis, and liver, than in the normal oral tissues. When 14C-UFT(14C-tegafur + uracil, a uracil/FT molar ratio of 4) was administered orally for 3 consecutive days to hamsters bearing O-1N, the 14C-tegafur level of tissue homogenates in the buccal and metastatic tumours was within the range of normal oral tissues, but the localization of silver grains was significantly higher in tumour cells than in the oral normal tissues. The results suggest that UFT would be useful for treatment and prevention of lymph node metastasis of oral carcinomas because of the high accumulation of active metabolites in metastatic and non-metastatic lymph nodes.


Genetic characterization of H5N1 influenza A viruses isolated from zoo tigers in Thailand.
The H5N1 avian influenza virus outbreak among zoo tigers in mid-October 2004, with 45 animals dead, indicated that the avian influenza virus could cause lethal infection in a large mammalian species apart from humans. In this outbreak investigation, six H5N1 isolates were identified and two isolates (A/Tiger/Thailand/CU-T3/04 and A/Tiger/Thailand/CU-T7/04) were selected for whole genome analysis. Phylogenetic analysis of the 8 gene segments showed that the viruses clustered within the lineage of H5N1 avian isolates from Thailand and Vietnam. The hemagglutinin (HA) gene of the viruses displayed polybasic amino acids at the cleavage site, identical to those of the 2004 H5N1 isolates, which by definition are highly pathogenic avian influenza (HPAI). In addition, sequence analyses revealed that the viruses isolated from tigers harbored few genetic changes compared with the viruses having infected chicken, humans, tigers and a leopard isolated from the early 2004 H5N1 outbreaks. Sequence analyses also showed that the tiger H5N1 isolated in October 2004 was more closely related to the chicken H5N1 isolated in July than that from January. Interestingly, all the 6 tiger H5N1 isolates contained a lysine substitution at position 627 of the PB2 protein similar to the human, but distinct from the original avian isolates.


Bacterial colonization, enamel defects and dental caries in 4-6-year-old mono- and dizygotic twins.
Knowledge of the genetic and environmental influences in caries aetiology has relevance for preventive dentistry. This classical twin study compared concordance of mutans streptococci (MS) and lactobacilli (LB) colonization, enamel defects, and caries in a cohort of 4-6-year-old mono- (MZ) and dizygotic (DZ) twin pairs. The twins were examined for prevalence and concordance of enamel opacities and hypoplasia, oral counts of MS and LB, and dental caries. Bacterial counts were assessed using a commercial microbiological kit. Thirty-four MZ and 50 DZ twins (mean gestational age 35.0 ± 2.4 weeks, and birthweight 2.4 ± 0.6 kg) were examined. There were no statistically significant differences between MZ and DZ twins in the prevalence of MS, LB, and enamel hypoplasia. Concordance rates for MS and LB presence and prevalence of enamel defects within MZ and DZ twin pairs were not significantly different. There were more children with caries in DZ compared with MZ twins (18% vs 3%, P = 0.0029), most likely due to increased daily frequency of sugar consumption and less toothbrushing. Concordance data from MZ and DZ twins did not demonstrate any statistically significant difference in susceptibility for enamel defects and colonization of MS and LB.


Restorative materials containing antimicrobial agents: is there evidence for their antimicrobial and anticaries effects? A systematic review.
No study has examined if restorative materials that contain antimicrobials, exert an anticaries effect in vivo.


Ultraviolet-induced DNA-repair synthesis in lymphocytes from patients with chronic lymphatic leukemia.
Ultraviolet-induced DNA-repair synthesis was studied in peripheral lymphocytes from 10 patients with chronic lymphatic leukemia (CLL). An ultraviolet-dose curve was established for each patient and compared with curves for healthy individuals. The same level of DNA-repair synthesis was recorded in CLL and normal lymphocytes. This contrasts with a report from another laboratory. Evidence is presented of the necessity of inhibiting replicative DNA-synthesis with hydroxyurea in order to adequately estimate the DNA-repair synthesis.


5-aminolevulinic acid-photodynamic therapy as a potential approach for kerion.
Kerion is an inflammatory variant of tinea capitis that is caused by ringworm fungi (zoophilic dermatophytes). It often effects prepubertal children. Classically,antifungals are primarily used as therapeutic agents to treat kerion. However, many patients do not respond well to these agents and severe hepatotoxic side reactions may occur with the long-term use of such drugs. New therapeutic approaches are needed. Here, we report a juvenile case of kerion successfully treated by ALA-PDT (5-aminolevulinic acid-photodynamic therapy) after the failure of a 4-week course of itraconazole and terbinafine. We find three published cases of kerion in the literature, all of which were successfully cured by application of ALA-PDT, highlighting a potentially superior therapeutic choice for kerion treatment.


Rainfall characteristics significantly affect the scavenging of water-soluble ions attached to leaves.
Precipitation is considered the most effective way to remove particulate matter from the leaves of plants. Changes in rainfall characteristics can affect the scavenging processes of particulate matter from leaves. In order to better understand the dynamics of PM scavenging during rainfall, especially the water-soluble ions components, leaves from the 11 plant species (trees, shrubs, terrestrial herbs, wetland plants) from the Olympic park were sampled and used in indoor experiments. During the experiments, the rainfall intensity was set at 30 mm/h, 45 mm/h, and 60 mm/h, and the duration was divided into 0-20 min, 20-40 min, and 40-60 min. The sampled plant leaves were set in the experiments at 1 m and 3 m height from the ground. Concentrations and compositions of nine water-soluble ions of rainfall samples were analyzed in this experiment. The results revealed that SO42-, Ca2+, and Na+ were the most abundant ionic species removed from the leaves, and NO3- ranked fourth, followed by Cl-, Mg2+ K+, NH4+, and F-. The ions concentration of rainfall samples decreased when the rain intensity increased from 30 to 45 mm/h and when the rain intensity increased to 60 mm/h. The efficiency of scavenging during different rainfall durations depends on the ionic species. Na+, Mg2+, Ca2+, and SO42- concentrations increased with the increase in rainfall duration, whereas those of NH4+, K+, and Cl- decreased. The effect of leaf height on ions concentration of rainfall samples was also different among the ionic species: Na+, Mg2+, Ca2+, NO3-, and F- concentrations were significantly higher at 1 m compared with 3 m. The principal component analysis of ions in rainfall samples revealed two main sources of particulate matter in our study. One is from vehicle exhaust and industrial and agricultural pollution. The other is agricultural combustion and ground dust sources. The results of the above study can provide a basis and theoretical support for the establishment of urban cleaning systems and the prevention of air pollution.


Prevalence of low birth weight in a scenario of economic depression in Argentina.
Low birth weight (LBW) is considered a general indicator of health because it is related to complications in the life of a newborn infant and is one of the leading causes of infant mortality. It is a multifactorial indicator, and its determinants include socioeconomic factors. To assess the impact of economic inequality on the prevalence of LBW by quantifying its differential effect by maternal age, level of maternal education, and level of care. Epidemiological, cross-sectional study that analyzed all births occurred in Argentina between 2001 and 2013 based on data provided by the National Registry of births, corresponding to the Health Statistics and Information Department. The temporal variation in the prevalence of LBW newborn infants (< 2500 grams) and its relation to demographic and socioeconomic indicators were studied. Its association was assessed using logistic regression models. A total of 9 001 960births were included. The prevalence of LBW newborn infants during the 2001 economic crisis increased -6% in 2002 and 7% in 2003- The impact was heterogeneous and higher on public hospitals (|-1;PR|-3; = 1.03) and adolescent mothers (PR=1.07), but no impact was observed on a low level of maternal education (PR= 0.99). The impact of socioeconomic inequality on the prevalence of LBW was significant and heterogeneous, especially on public hospitals and mothers at the extremes of maternal age.


The use of a bedside assay for plasma B-type natriuretic peptide as a biomarker in the management of patent ductus arteriosus in premature neonates.
To test the utility of the bedside plasma concentration of B-type natriuretic peptide (BNP) assay as a screen for patent ductus arteriosus (PDA) in premature neonates. Newborn infants admitted to the neonatal intensive care unit (NICU) had paired echocardiography and BNP measurements at enrollment and every 4 to 5 days. Twenty neonates (gestational age approximately 28.6 weeks and birth weight approximately 1161 g) had 81 paired echocardiography and BNP determinations. BNP ranged from 5 to 3900 pg/mL. Fifty-six of 81 echocardiograms showed PDA. Significant correlations were found between BNP and ductal size and degree of shunting. Correlation was greater in infants >2 days of age. BNP >300 pg/mL predicted significant PDA, whereas BNP <105 pg/mL predicted absence of significant PDA. Bedside measurement of BNP correlates with magnitude of PDA in premature newborns, particularly beyond day 2, and may be useful in guiding diagnostic and management strategies.


Low-cost sensor system for monitoring the oil mist concentration in a workshop.
Metalworking fluids used in industrial workshops may present a major threat to the health of workers who have been exposed to a high oil mist concentration over a long period of time. Therefore, monitoring the temporal and spatial distribution of particulate matter concentration has great practical significance for the control of oil mist. Traditional particle monitors are generally cumbersome, expensive, and difficult to maintain, which to some extent restricts their extensive use in workshops. Recent years have witnessed tremendous developments in the area of low-cost sensors, which are of great help in obtaining high-density pollution data. In this paper, we evaluate the performance of an inexpensive laser sensor (A4-CG) during long-term oil mist monitoring in a machine shop for the first time. With the use of Lora technology, we developed an online oil mist monitoring network to access real-time concentration, temperature, and humidity information from distributed monitors. According to the results, the sensor data correlated well with measurements by the reference instrument (R2 = 0.96), which means that the distributed sensor network can accurately detect the concentration level of oil mist in the workshop. In fact, most of the sensors demonstrated stable operation for up to half a year according to cluster analysis, while several sensors exhibited serious data drift. Furthermore, the results indicate that the peak oil mist concentration in most areas during production exceeded the value of 0.5 mg m-3 recommended by NIOSH, and it was found that appropriately lowering the relative humidity can make sampling more accurate, while lowering the temperature can reduce the oil mist concentration in the workshop. Thus, measures to control oil mist such as generation and distribution of pollution sources should be on the agenda.


Effects of scopolamine and physostigmine on acquisition of morphine-treated rats in Morris water maze performance.
To investigate effects of morphine on acquisition process of rats a nd interactions of opioid and cholinergic systems by Morris water maze performance. Morris water maze was used to measure the latency of rats with drug s treatment to find the covert platform. Chronic morphine administration (10 mg/kg) impaired the acquisition process of rats in Morris water maze task. Appreciable difference was identified with morphine 10 mg/k g group compared with morphine 3 mg/kg group. Co-administration of morphine (10 mg/kg) and scopolamine (3 mg/kg) aggravated acquisition impairment induced by morphine 1 0 mg/kg or scopolamine alone, though scopolamine itself induced no salient changes in acquisition capabilities of rats. In addition, physostigmine (0.1 mg/kg) could appreciably attenuate morphine-induced acquisition impairment. Morphine 10 mg/kg evidently impaired acquisition process of rats. There was a close relationship between the acquisition capabilities of morphine-treated rats and the functions of cholinergic system.


The effect of pentachlorophenol on DNA adduct formation in p53 wild-type and knockout mice exposed to benzo[a]pyrene.
Previous studies have shown that pentachlorophenol (PCP) has both potentiative and antagonistic effects on the genotoxicity of benzo[a]pyrene (B[a]P). It has been suggested that these effects are due to inhibition and/or induction of enzymes involved in the biotransformation of B[a]P [Carcinogenesis 16 (1995) 2643]. However, B[a]P [J. Biol. Chem. 274 (1999) 35240] and a metabolite of PCP, tetrachlorohydroquinone (TCHQ) [Chem. Biol. Interact. 105 (1997) 1], induce p53 protein synthesis in vitro. To investigate this effect further, C57BL/6Tac trp53+/+ (wild-type, WT) and C57BL/6Tac trp53-/- (knockout, KO) mice were exposed to 55 microg B[a]P/g BW alone or in combination with 25 microg/g PCP. Hepatic and lung DNA were analyzed for the major B[a]P DNA adduct, 7R,8S,9S-trihydroxy-10R-(N2-2'-deoxyguanosyl)-7,8,9,10-tetrahydro-B[a]P (BPDE-N2G) and other minor adducts using the 32P-postlabeling assay. BPDE-N2G adducts were detected in all animals exposed to B[a]P. Similar adduct levels were observed in WT mice exposed to 55 microg/g B[a]P compared with KO mice exposed to B[a]P alone or in combination with PCP. Interestingly, hepatic and lung BPDE-N2G adducts were decreased in WT mice exposed to B[a]P with PCP (P<0.05). Total DNA adducts in the liver (P<0.05) were also decreased in WT mice exposed to B[a]P and PCP. Total DNA adducts in either hepatic or lung DNA isolated from KO mice were not different in mice treated with PCP and B[a]P. These results suggest that the decrease in BPDE-N2G adducts observed in WT mice may be a result of p53 accumulation or induction of repair pathways in response to damage induced by PCP.


A novel model for predicting posthepatectomy liver failure based on liver function and degree of liver resection in patients with hepatocellular carcinoma.
The permissible liver resection rate for preventing posthepatectomy liver failure (PHLF) remains unclear. We aimed to develop a novel PHLF-predicting model and to strategize hepatectomy for hepatocellular carcinoma (HCC). This retrospective study included 335 HCC patients who underwent anatomical hepatectomy at eight institutions between 2013 and 2017. Risk factors, including volume-associated liver-estimating parameters, for PHLF grade B-C were analyzed in a training set (n = 122) via multivariate analysis, and a PHLF prediction model was developed. The utility of the model was evaluated in a validation set (n = 213). Our model was based on the three independent risk factors for PHLF identified in the training set: volume-associated indocyanine green retention rate at 15 min, platelet count, and prothrombin time index (the VIPP score). The areas under the receiver operating characteristic curve of the VIPP scores for severe PHLF in the training and validation sets were 0.864 and 0.794, respectively. In both sets, the VIPP score stratified patients at risk for severe PHLF, with a score of 3 (specificity, 0.92) indicating higher risk. Our model facilitates the selection of the appropriate hepatectomy procedure by providing permissible liver resection rates based on VIPP scores.


Study on PCBs, PCDD/Fs, organochlorine pesticides, heavy metals and arsenic content in freshwater fish species from the River Turia (Spain).
In this study samples of common trout, European eel and barbel have been analysed for the levels of heavy metals such as lead (Pb), cadmium (Cd), zinc (Zn), copper (Cu), and the metalloid arsenic (As). Polychlorinated biphenyls (PCBs), including three non-ortho substituted PCBs, 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and DDT and its two main metabolites were also analysed in some selected samples, in order to evaluate the extent of contamination of the River Turia basin. All samples presented detectable levels of the elements analysed, presenting zinc the highest concentrations. European eel was the species found to highly accumulate these contaminants. Regarding organochlorinated compounds, seven samples exhibited detectable levels of the most toxic PCDD/Fs congener, the 2,3,7,8-TCDD. Non-ortho PCBs were not detected or not quantified in almost all samples and the sum of the individual PCB congeners determined were low compared to other aquatic ecosystems with similar fish species.


The effect of tRNA binding on the structure of 5 S RNA in Escherichia coli. A chemical modification study.
The structure of 5 S RNA within the 70 S ribosome from Escherichia coli was studied using the chemical reagent kethoxal (alpha-keto-beta-ethoxybutyraldehyde) to modify accessible guanosines. The modification pattern of 5 S RNA from free 70 S ribosomes was compared with that of poly(U) programmed ribosomes where tRNA had been bound to both the A- and P-sites. Binding to the ribosomal A-site was achieved enzymatically using the elongation factor Tu and GTP in the presence of deacylated tRNA which blocks the ribosomal P-site. Modified guanosines were identified after partial RNase T1 hydrolysis and separation of the hydrolysis products on sequencing gels. Binding of tRNA to the ribosome leads to a strong protection of 5 S RNA guanosine G-41 and to some degree G-44 from kethoxal modification. The limited RNase T1 hydrolysis pattern provides evidence for the existence of a 5 S RNA conformation different from the known 5 S RNA A- and B-forms which are characterized by their gel electrophoretic mobility. The importance of 5 S RNA for the binding of tRNA to the ribosome is discussed.


Relationship between functional capacity and community responsibility in patients with schizophrenia: differences between independent and assisted living settings.
We examined factors potentially associated with level of community responsibility among middle-aged and older patients with schizophrenia. Participants in residential care facilities engaged in significantly fewer community responsibilities than those residing in the community. However, demographic and clinical characteristics did not explain these differences. Further, greater functional capacity was associated with greater community responsibility among participants residing in the community, but not those in residential care facilities. These results suggest that, despite capacity, patients residing in residential care facilities are not engaging in community responsibilities. Further, among participants residing in the community, functional capacity may predict level of responsibility.


Lack of cross-resistance between certain platinum coordination compounds in mouse leukemia.
Two congeners of cis-platinum diamminodichloride, 1,2-diamminocyclohexylplatinum malonate (NSC 224964) and 1,2-cyclohexyldiamminoplatinum sulfate (NSC 250427), show approximately equal inhibitory activity in vitro against leukemia L1210 and a line of L1210 (L1210/PDD) that has developed resistance to cis-platinum diamminodichloride. These compounds are also active against L1210/PDD in vivo. These observations suggest that they be tried clinically in patients whose disease has become resistant to cis-platinum diamminodichloride.


Contemporary understanding and management of reflux and constipation in the general population and pregnancy: a consensus meeting.
Gastro-oesophageal reflux disease (GERD) and constipation have a major impact on public health; however, the wide variety of treatment options presents difficulties for recommending therapy. Lack of definitive guidelines in pharmacy and general practice medicine further exacerbates the decision dilemma. To address these issues, a panel of experts discussed the principles and practice of treating GERD and constipation in the general population and in pregnancy, with the aim of developing respective treatment guidelines. The panel recommended antacids 'on-demand' as the first-line over-the-counter treatment in reflux, and as rescue medication for immediate relief when reflux breaks through with proton pump inhibitors. Calcium/magnesium-based antacids were recommended as the treatment of choice for pregnant women because of their good safety profile. In constipation, current data do not distinguish a hierarchy between polyethylene glycol (PEG)-based laxatives and other first-line treatments, although limitations are associated with stimulant- and bulk-forming laxatives. Where data are available, PEG is superior to lactulose in terms of efficacy. In pregnancy, PEG-based laxatives meet the criteria for the ideal treatment. The experts developed algorithms that present healthcare professionals with clear treatment options and management strategies for GERD and constipation in pharmacy and general practice medicine.


[Perinatal risk in emergent surgery].
During pregnancy, appendicitis, cholecystitis, choledocholithiasis, intestinal obstruction, and ulcers can lead to complaints and complications which occasion emergent surgery. The disease as such, its complications, the surgical intervention, and the anesthesia represent a risk for both mother and child. Risks arise particularly often due to the delay of diagnosis and therapy. Risk to the child depends on gestational week, fetal lung maturity, and oxygen supply.


Teaching psychiatry in medical school.
Considerable dissatisfaction of students and educators exists in regard to the teaching of psychiatry in medical school. This paper recommends that the liasion model of teaching directed to house and attending staffs be applied to the teaching of medical students during their clerkships throughout the general hospital, with a concomitant de-emphasis of training solely on the psychiatric service.


A soluble factor produced by inoculation of human monocytes with Leishmania donovani promastigotes suppresses IFN-gamma-dependent monocyte activation.
Whereas human cultured monocytes preactivated with IFN-gamma increase their antileishmanial capacity, monocytes inoculated with Leishmania donovani before IFN-gamma treatment fail to respond with an increase of antileishmanial capacity. Cell surface expression of class II MHC products also fails to be increased by IFN-gamma in the infected monocytes, although the accumulation of HLA-DR alpha-chain mRNA is increased to the same extent in infected and noninfected activated monocytes. This inhibition of monocyte activation is caused in a dose-dependent manner by a soluble substance elaborated into the cell supernatant after inoculation of monocytes with L. donovani and is referred to as activation suppressing factor (ASF). ASF activity can be abrogated by dialysis and by treatment with a proteinase, indicating that ASF is a small peptide.


Visual Outcome of Plasma Exchange in Optic Neuritis Associated With Flu Vaccination: A Case Report.
Post-flu-vaccination optic neuritis is an extremely rare condition with an incidence ranging from 0.003 cases to 0.89 per 100 000 population. The exact pathophysiology is not clearly defined. Most of the patients with post-flu-vaccination optic neuritis tend to present with progressive worsening of vision in 2-3 weeks post-vaccine administration. A prompt fundus examination supplemented with MRI imaging of the orbit is required to establish the diagnosis. On diagnosis, early initiation of high-dose oral or IV steroids is recommended to prevent optic atrophy or worsening of vision. Most patients tend to have complete recovery of vision when started on steroids. However, if the patient continues to have worsening symptoms while being treated with a high dose of steroids, plasmapheresis (PLEX) is an effective intervention.


The effects of total sleep deprivation on brain functional organization: mutual information analysis of waking human EEG.
Sleep deprivation can affect the waking electroencephalogram (EEG) that may reflect functional organization of the brain. We examined the effect of total sleep deprivation (TSD) on functional organization between different cortical areas from the waking EEG. Waking EEG data were recorded from 18 healthy male volunteers with eyes closed after 8-h night's sleep and after 24 h of TSD. The averaged cross mutual information (A-CMI) after 24 h of TSD were compared to before TSD. 24 h of TSD yielded the decreased A-CMIs in the inter-hemispheric C3-F4, C3-F8, and C3-C4 pairs: therefore, the electrodes that contribute to pairs with significant decrease of A-CMI were C3, F4, F8, and C4. The decreased A-CMIs between C3 and right frontal and central brain areas after 24 h of TSD may reflect the changes of cortico-cortical functional organization by homeostatic process during TSD. Our results of the frontal-area-related A-CMI decreases may support that the frontal brain regions are related to the homeostatic deterioration of brain function due to TSD.


Rate-dependent electrophysiological effects of OPC-8212: comparison to sotalol.
The rate-dependent effects of OPC-8212 (6 microM), a new positive inotropic drug which lengthens cardiac action potential duration were studied in dog cardiac Purkinje fibers and compared to the effects of sotalol (30 microM), a class III antiarrhythmic using conventional microelectrode technique. Both OPC-8212 and sotalol lengthened action potential duration in a frequency-dependent manner. Both had a greater effect at slower pacing frequencies than at fast. The maximum rising velocity of the action potential upstroke was not affected by either compound at any of the pacing cycle lengths applied (300-2000 ms). The effects of OPC-8212 and sotalol on action potential duration and maximum rising velocity were also compared following abrupt changes in pacing cycle length (i.e. coupling intervals ranging between approximately 250-3000 ms). The effects of both OPC-8212 and sotalol following abrupt changes in pacing cycle length were similar to one another. Neither OPC-8212 nor sotalol altered the time constants for restitution of action potential duration or the kinetics for recovery of the maximum rising velocity of the action potential upstroke. These results indicate that the rate-dependent electrophysiological effects of OPC-8212 closely mimic those of sotalol and suggest that OPC-8212 may be an effective class III antiarrhythmic in addition to being a positive inotropic compound.


Reduced high density lipoprotein in stroke: relationship with elevated triglyceride and hypertension.
Lipids and lipoproteins were analysed in forty-one survivors of stroke, aged less than 65 years, and the same number of age and sex matched controls without vascular disease. The stroke subjects had no evidence of coronary artery or peripheral vascular disease. High density lipoprotein cholesterol was significantly lower (1.19 +/- 0.06 mmol/l) in the stroke subjects than the controls (1.47 +/- 0.07 mmol/l). Triglyceride was also elevated in the stroke subjects, but this was confined to those who were taking antihypertensive treatment which included beta-blockers and/or thiazides. The low levels of high density lipoprotein in stroke were independent of hypertension or its treatment. Thus low levels of high density lipoprotein appear to be associated with cerebrovascular disease, while elevated triglyceride is a complication of anti-hypertensive therapy.


Epitope ratio analysis (ERA): a simple radioimmunological method using monoclonal antibodies for the simultaneous analysis of several antigens.
Monoclonal antibodies specific for human cell surface antigens were used to develop a technique for the simultaneous analysis of several antigenic determinants. The procedure was used to measure the relative expression of specific cell antigens during cultural growth. Epitope ratio analysis (ERA) is applicable to many systems requiring measurement of such differential antigen expression.