Detection and quantitation of octopine in normal plant tissue and in crown gall tumors.
Octopine has been detected in normal tobacco leaf and stem tissue, normal sunflower stem tissue, pinto bean leaves, and normal tobacco callus tissue in culture. Octopine was identified in extracts by means of electrophoresis and chromatography in several solvent systems. Tobacco and sunflower tumor lines induced by various strains of Agrobacterium tumefaciens were found to contain from 1 to 240 times as much octopine as the normal plant tissues examined. Several strains of A. tumefaciens produce undifferentiated tobacco tumors containing high levels of octopine, but produce undifferentiated sunflower tumors containing normal levels of octopine and high levels of arginine. Further, strain CGIC of A. tumefaciens produces in tobacco an undifferentiated tumor which contains high levels of octopine and a teratoma which contains normal levels of octopine. This evidence shows that there is no consistent relationship between the causative strain of A. tumefaciens and the octopine content of the resulting crown gall tumor.


Effects of U-71,184 and several other CC-1065 analogues on cell survival and cell cycle of Chinese hamster ovary cells.
CC-1065 is a very potent antitumor antibiotic which selectively binds in the minor groove of DNA with alkylation at N-3 of adenine. Since therapeutic doses of CC-1065 caused delayed deaths in mice, analogues were synthesized, some of which had significant antitumor activity. The effects of several of these analogues on inhibition of CHO cell survival, cell progression, and their phase-specific toxicity are reported. CC-1065, U-66,664, U-66,819, U-66,694, and U-71,184 all have a left hand segment with an intact cyclopropyl group but have different tail segments. Lethality of these compounds after 2 h drug exposure was in the following order (50% lethal dose in nM in parentheses): CC-1065 (0.06) greater than U-71,184 (1.3) greater than U-66,694 (3.2) greater than U-68,819 (171) greater than U-66,664 (greater than 1200). In general, these compounds did not inhibit progression from G1 to S but slowed progression through S and blocked cells in G2-M. The phase-specific toxicity of U-71,184 and U-66,694 was different from that of CC-1065. CC-1065 was most cytotoxic to cells in M and early G1 and toxicity decreased as cells entered late G1 and S. In contrast, U-66,694 and U-71,184 were most toxic to cells in late G1. The biochemical and cellular effects of U-71,184 were then studied in detail since it was the most active among these analogues. After a 2-h exposure to 3 ng/ml U-71,184, 90% cell kill or growth inhibition was observed whereas 100 ng/ml was needed for similar inhibition of DNA and RNA synthesis. This discrepancy between the doses suggested that inhibition of nucleic acid synthesis may not be causally related to lethality. Further studies showed that when drug was removed after 2 h exposure, DNA synthesis continued to be inhibited whereas RNA and protein synthesis reached levels higher than the control. Therefore, it is likely that at cytotoxic doses the low level of inhibition of DNA synthesis combined with the stimulation of RNA and protein synthesis leads to unbalanced growth and cell death.


A Proposal for a Consolidated Structural Model of the CagY Protein of Helicobacter pylori.
CagY is the largest and most complex protein from Helicobacter pylori's (Hp) type IV secretion system (T4SS), playing a critical role in the modulation of gastric inflammation and risk for gastric cancer. CagY spans from the inner to the outer membrane, forming a channel through which Hp molecules are injected into human gastric cells. Yet, a tridimensional structure has been reported for only short segments of the protein. This intricate protein was modeled using different approaches, including homology modeling, ab initio, and deep learning techniques. The challengingly long middle repeat region (MRR) was modeled using deep learning and optimized using equilibrium molecular dynamics. The previously modeled segments were assembled into a 1595 aa chain and a 14-chain CagY multimer structure was assembled by structural alignment. The final structure correlated with published structures and allowed to show how the multimer may form the T4SS channel through which CagA and other molecules are translocated to gastric cells. The model confirmed that MRR, the most polymorphic and complex region of CagY, presents numerous cysteine residues forming disulfide bonds that stabilize the protein and suggest this domain may function as a contractile region playing an essential role in the modulating activity of CagY on tissue inflammation.


Using indocyanine green angiography to achieve complete engraftment of pectoralis major myocutaneous flaps.
Although the pectoralis major myocutaneous (PMMC) flap is among the useful reconstructive materials following oral cancer ablation, this flap has an unstable blood circulation that could result in partial necrosis of the skin paddle. This report describes the usefulness of indocyanine green angiography (ICGA) to achieve complete engraftment of the PMMC flap. Five patients with oral cancer underwent reconstruction with a PMMC flap after cancer ablation. During the skin paddle design and flap elevation, the blood supply to the flap was assessed by ICGA. Areas of the skin paddle that showed no ICG fluorescence were excised. Consequently, prior to transfer to the recipient site, the blood supply to all flaps was confirmed with indocyanine green visible at the edge of the skin paddle, and complete engraftment was achieved without partial necrosis. Based on the results observed, ICGA would make a useful contribution to complete engraftment of the PMMC flap.


Novel therapies for myelofibrosis.
Myelofibrosis (MF), including primary, post-essential thrombocythemia and post-polycythemia vera MF, associates with a reduced quality of life and shortened life expectancy. Dysregulation of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway is prominent, even in the absence of the JAK2(V617F) mutation. Therefore, all symptomatic MF patients may potentially derive benefit from JAK inhibitors. Despite the efficacy of JAK inhibitors in controlling signs and symptoms of MF, they do not eradicate the disease. Therefore, JAK inhibitors are currently being tested in combination with other novel therapies, a strategy which may be more effective in reducing disease burden, either by overcoming JAK inhibitor resistance or targeting additional mechanisms of pathogenesis. Additional targets include modulators of epigenetic regulation, pathways that work downstream from JAK/STAT (i.e. mammalian target of rapamycin/AKT/phosphoinositide 3-kinase) heat shock protein 90, hedgehog signaling, pro-fibrotic factors, abnormal megakaryocytes and telomerase. In this review, we discuss novel MF therapeutic strategies.


Nosocomial outbreak of parvovirus B19 infection in a renal transplant unit.
Parvovirus B19 (B19) infection is known to cause chronic infection leading to anemia in immunocompromised patients. Although nosocomial B19 infections in immunocompetent patients have been documented, no outbreaks in immunocompromised patients have been previously reported. Whether transmission can occur from a patient with chronic infection is also unknown. An outbreak of B19 infection in a renal transplant unit was investigated by molecular analysis of the virus strains and a case-control study. Three patients had genetically identical virus strains suggesting the occurrence of nosocomial transmission. The index case transmitted infection many weeks after the onset of her clinical symptoms. Other patients at risk of acquiring infection were those most intensively immunosuppressed. Viral load in the serum correlated with the hematological response. A rebound in the viral load was associated with clinical relapse and the failure of i.v. immunoglobulin therapy. Nosocomial transmission of B19 can occur from immunocompromised patients even when they are in the chronic stage of the infection. The clinical and virological response to i.v. immunoglobulin therapy is variable and depends on the overall level of immunosuppression of the patient.


Direct and Indirect Costs of Infectious Conjunctivitis in a Commercially Insured Population in the United States.
To assess the direct and indirect costs of infectious conjunctivitis and quantify medical costs due to conjunctivitis transmission in families. In this retrospective claims analysis from the OptumHealth Care Solutions, Inc. database (1998-2016), beneficiaries with or without at least one diagnosis of infectious conjunctivitis were identified. Direct and indirect costs (in 2016 US$) during the 60 days post conjunctivitis diagnosis (or imputed date for controls) were compared using cost differences in linear regressions. For transmission cost analysis, the total cost of each conjunctivitis episode was the sum of the primary episode (seed patient) and the secondary episode (infected family members) costs. A generalized estimating equation model adjusted for seed patient characteristics was used to assess the impact of number and rate of transmissions on episode cost. Health care resource utilization and direct costs were significantly higher for patients with conjunctivitis (n=1,002,188) versus controls (n=4,877,210): 1.67 all-cause visits per person per month (PPPM) versus 0.79 visits PPPM, respectively; total mean direct cost of $396.04 PPPM versus $289.63 PPPM, respectively. The cost of medically related absenteeism was $105.42 (95% confidence interval [CI], $104.18-$106.75) higher for patients with conjunctivitis than for controls. Episode cost, without transmission due to seed patient, was $669.43 (95% CI, $654.67-$684.85); it increased with each additional infected family member and with increased infection transmission time between family members. Conjunctivitis was associated with a notable economic burden in terms of direct medical costs and medically related absenteeism. Family health care costs increased with transmission time and with each family member infected with conjunctivitis.


Cell coupling and impulse propagation in the failing heart.
Cell coupling and impulse propagation were investigated in the ventricle of cardiomyopathic hamsters at an advanced stage of heart failure. An appreciable decline in junctional conductance was found, a phenomenon in part related to activation of the plasma and cardiac renin-angiotensin systems. Decreased expression of connexin43 or an alteration of junctional proteins also might be implicated in the decreased cell coupling. Morphologic abnormalities such as fibrosis, necrosis, and rupture of cell contacts contribute to the decline of conduction velocity or to the blockade of impulse propagation in some areas of the ventricle, creating the conditions for anisotropic conduction and cardiac arrhythmias. The decrease in membrane potential found in myopathic cells is related in part to depression of Na-KATPase activity, and the lack of action of beta-adrenergic agonists on junctional conductance is explained by down-regulation of beta receptors and an abnormality of adenyl cyclase.


D-antigen determination in polio vaccine production: comparison of gel diffusion and ELISA methods.
Up to now gel diffusion is widely used for in vitro determination of polio D-antigen. This method has several disadvantages, its relatively low sensitivity being the main one. Generally samples have to be concentrated, which requires fairly large amounts of material and often results in some loss of antigen due to adsorption. Two ELISA methods have been developed for determination of polio D-antigen. These tests have a high sensitivity, approximately 1 DU/ml, and require very small amounts of sample and antisera. Using the indirect method only one HRPO conjugated antiserum is needed for quantitation of type 1, 2 and 3 D-antigen. Comparison of D-antigen values obtained by ELISA and gel diffusion for a number of vaccines revealed a good correlation between the different methods. The indirect ELISA method has also been applied successfully for D-antigen quantitation of A1PO4 adsorbed DT-and DPT-polio vaccines.


A Hierarchical Image Matting Model for Blood Vessel Segmentation in Fundus Images.
In this paper, a hierarchical image matting model is proposed to extract blood vessels from fundus images. More specifically, a hierarchical strategy is integrated into the image matting model for blood vessel segmentation. Normally the matting models require a user specified trimap, which separates the input image into three regions: the foreground, background and unknown regions. However, creating a user specified trimap is laborious for vessel segmentation tasks. In this paper, we propose a method that first generates trimap automatically by utilizing region features of blood vessels, then applies a hierarchical image matting model to extract the vessel pixels from the unknown regions. The proposed method has low calculation time and outperforms many other state-of-art supervised and unsupervised methods. It achieves a vessel segmentation accuracy of 96.0%, 95.7% and 95.1% in an average time of 10.72s, 15.74s and 50.71s on images from three publicly available fundus image datasets DRIVE, STARE, and CHASE DB1, respectively.


The effects of adolescence sports and exercise on adulthood leisure-time physical activity in educational groups.
Physical inactivity has become a major public health problem and clear educational differences in physical activity have been reported across Europe and USA. The origins of adulthood physical activity are suggested to be in childhood and adolescence physical activity. Hardly any studies have, however, examined if the educational differences in physical activity might also be due to educational differences in early experiences in physical activity. Thus, our aim was to examine how competitive sports in youth, and exercise in late adolescence, and opinions on physical education (PE) in childhood determined adulthood leisure-time physical activity (LTPA) in different educational groups. We used cross-sectional population-based National FINRISK 2002 data for 1918 men and 2490 women aged 25 to 64 years. Competitive sports in youth, exercise in late adolescence, and opinions on PE in childhood were assessed retrospectively via self-reports. Adulthood LTPA was collected with 12-month recall. In 2008, we calculated structural equation models including latent variables among the low- (<12 years) and high- (>/=12 years) educated. Men more often than women reported that their experience of PE was interesting and pleasant as well as having learned useful skills during PE classes. Men, compared to women, had also been more active in the three selected competitive sports in youth and exercised in late adolescence. Participation in competitive sports in youth among the low-educated and exercise in late adolescence among the high-educated had a direct effect on adulthood LTPA. Among the low-educated, opinions on PE in childhood had an indirect effect on adulthood LTPA through participation in competitive sports in youth whereas among the high-educated, the indirect effect went through exercise in late adolescence. The effects were mainly similar between genders. Our study answers to a strong need to assess the determinants of leisure-time physical activity to promote physical activity in low-educated individuals. The pathways of physical activity from childhood to adulthood LTPA may be different depending on the pursued educational career. Further prospective studies are needed to confirm our results.


Mechanochemical model of microtubule structure and self-assembly kinetics.
Microtubule self-assembly is largely governed by the chemical kinetics and thermodynamics of tubulin-tubulin interactions. An important aspect of microtubule assembly is that hydrolysis of the beta-tubulin-associated GTP promotes protofilament curling. Protofilament curling presumably drives the transition from tip structures associated with growth (sheetlike projections and blunt ends) to those associated with shortening (rams' horns and frayed ends), and transitions between these structures have been proposed to be important for growth-shortening transitions. However, previous models for microtubule dynamic instability have not considered such structures or mechanics explicitly. Here we present a three-dimensional model that explicitly incorporates mechanical stress and strain within the microtubule lattice. First, we found that the model recapitulates three-dimensional tip structures and rates of assembly and disassembly for microtubules grown under standard conditions, and we propose that taxol may stabilize microtubule growth by reducing flexural rigidity. Second, in contrast to recent suggestions, it was determined that sheetlike tips are more likely to undergo catastrophe than blunt tips. Third, partial uncapping of the tubulin-GTP cap provides a possible mechanism for microtubule pause events. Finally, simulations of the binding and structural effects of XMAP215 produced the experimentally observed growth and shortening rates, and tip structure.


Implementing Effective Educational Practices at Scales of Social Importance.
Implementing evidence-based practices is becoming both a goal and standard across medicine, psychology, and education. Initial successes, however, are now leading to questions about how successful demonstrations may be expanded to scales of social importance. In this paper, we review lessons learned about scaling up evidence-based practices gleaned from our experience implementing school-wide positive behavioral interventions and supports (PBIS) across more than 23,000 schools in the USA. We draw heavily from the work of Flay et al. (Prev Sci 6:151-175, 2005. doi: 10.1007/s11121-005-5553-y ) related to defining evidence-based practices, the significant contributions from the emerging "implementation science" movement (Fixsen et al. in Implementation research: a synthesis of the literature, University of South Florida, Louis de la Parte Florida Mental Health Institute, The National Implementation Research Network (FMHI Publication #231), Tampa 2005), and guidance we have received from teachers, family members, students, and administrators who have adopted PBIS.


The effect of daily protein supplementation, with or without resistance training for 1 year, on muscle size, strength, and function in healthy older adults: A randomized controlled trial.
Protein supplementation alone or combined with resistance training has been proposed to be effective in counteracting age-related losses of muscle mass and strength. To investigate the effect of protein supplementation alone or combined with light-intensity or heavy-load resistance exercise on muscle size, strength, and function in older adults. In a 1-y randomized controlled trial, 208 healthy older adults (>65 y) were randomly assigned to 1 of 5 interventions: 1) carbohydrate supplementation (CARB); 2) collagen protein supplementation (COLL); 3) whey protein supplementation (WHEY); 4) light-intensity resistance training 3-5 times/wk with whey protein supplementation (LITW); and 5) heavy resistance training 3 times weekly with whey protein supplementation (HRTW). Protein supplements contained 20 g protein + 10 g carbohydrate, whereas CARB contained 30 g of carbohydrates. All intervention groups received the supplement twice daily. The primary outcome was change in the quadriceps cross-sectional area (qCSA). Secondary outcomes included measures of lower extremity strength and power, functional capabilities, and body composition. There were 184 participants who completed the study. COLL and WHEY did not affect any measured parameter compared to CARB. Compared to WHEY, HRTW improved the qCSA size (between-group difference, +1.68 cm2; 95% CI, +0.41 to +2.95 cm2; P = 0.03), as well as dynamic (+18.4 Nm; 95% CI, +10.1 to +26.6 Nm; P < 10-4) and isometric knee extensor strength (+23.9 Nm; 95% CI, +14.2 to +33.6 Nm; P < 10-5). LITW did not improve the qCSA size, but increased dynamic knee extensor strength compared to WHEY (+13.7 Nm; 95% CI, +5.3 and +22.1 Nm; P = 0.01). Recommending protein supplementation as a stand-alone intervention for healthy older individuals seems ineffective in improving muscle mass and strength. Only HRTW was effective in both preserving muscle mass and increasing strength. Thus, we recommend that future studies investigate strategies to increase long-term compliance to heavy resistance exercise in healthy older adults. This trial was registered at clinicaltrials.gov as NCT02034760.


Chloroquine prophylaxis against vivax malaria in pregnancy: a randomized, double-blind, placebo-controlled trial.
To assess the safety of chloroquine (CQ) as prophylaxis against Plasmodium vivax infection during pregnancy. One thousand pregnant Karen women were enrolled in a randomized, double-blind, placebo-controlled trial of chemoprophylaxis with chloroquine (500 mg phosphate (or 300 mg base) weekly). Women received a median (range) chloroquine phosphate total dose of 9500 (1500-17 500) mg. The mothers were actively followed from inclusion to delivery and their infants until 12 months of age. Chloroquine prophylaxis completely prevented P. vivax episodes; 10.1% (95%CI: 7.3-14.5) of women in the placebo group experienced at least one episode of vivax malaria but no episode occurred in women in the CQ group. By contrast, the numbers of P. falciparum episodes were similar in each group: 7.4% (95%CI: 3.7-11.1) and 5.6% (95%CI: 3.3-7.9) in the placebo and CQ groups respectively (P = 0.56). Chloroquine prophylaxis was well tolerated and there was no difference in the proportions of reported side effects between CQ treated and placebo groups except for the duration of palpitations and sleeping disorders which were more frequent in those who had received CQ. Chloroquine prophylaxis had no impact on maternal anaemia, birth weight, gestational age, development of newborns or on growth, neurological development or visual acuity in infants at 1 year of age. Chloroquine is safe and effective as prophylaxis against P. vivax during pregnancy in this population.


Melatonin Regulates Angiogenic and Inflammatory Proteins in MDA-MB-231 Cell Line and in Co-culture with Cancer-associated Fibroblasts.
Cancer-associated fibroblast (CAFs) are the most abundant cells in the tumor microenvironment, able to secrete growth factors and act on tumor progression. Melatonin is associated with several mechanisms of action with oncostatics and oncoprotectors effects, and also participate in the reduction of synthesis of surrounding fibroblasts and endothelial cells in breast cancer. The objectives of this study were to determine the effectiveness of melatonin in cell viability and expression of proteins involved in angiogenesis and inflammation in triplenegative mammary tumor cell line (MDA-MB-231) and in co-culture with CAFs. Cell viability was measured by MTT assay and the protein expression was evaluated by Membrane Antibody Array after melatonin treatment. Melatonin treatment (1 mM) for 48 hours reduced the cell viability of MDA-MB-231, CAFs and co-culture (p < 0.05). The semi-quantitative protein analysis showed that when monoculture of tumor cells were compared with co-culture of CAFs, there was a regulation of angiogenic and inflammatory proteins (p < 0.05). Melatonin treatment also leads a differential expression of angiogenic and inflammatory proteins in both monoculture and co-culture of tumor cells and CAFs (p < 0.05). The influence of CAFs under the tumor microenvironment was confirmed, increasing the malignancy of the tumor. In addition, melatonin is effective in both monoculture and co-culture, regulating angiogenic and inflammatory proteins that contribute to tumor progression. This study show an overview of melatonin ability in regulating angiogenic and inflammatory proteins, and opens the way for exploration of each individual protein in further studies.


Host-virus evolutionary dynamics with specialist and generalist infection strategies: Bifurcations, bistability, and chaos.
In this work, we have investigated the evolutionary dynamics of a generalist pathogen, e.g., a virus population, that evolves toward specialization in an environment with multiple host types. We have particularly explored under which conditions generalist viral strains may rise in frequency and coexist with specialist strains or even dominate the population. By means of a nonlinear mathematical model and bifurcation analysis, we have determined the theoretical conditions for stability of nine identified equilibria and provided biological interpretation in terms of the infection rates for the viral specialist and generalist strains. By means of a stability diagram, we identified stable fixed points and stable periodic orbits, as well as regions of bistability. For arbitrary biologically feasible initial population sizes, the probability of evolving toward stable solutions is obtained for each point of the analyzed parameter space. This probability map shows combinations of infection rates of the generalist and specialist strains that might lead to equal chances for each type becoming the dominant strategy. Furthermore, we have identified infection rates for which the model predicts the onset of chaotic dynamics. Several degenerate Bogdanov-Takens and zero-Hopf bifurcations are detected along with generalized Hopf and zero-Hopf bifurcations. This manuscript provides additional insights into the dynamical complexity of host-pathogen evolution toward different infection strategies.


An Analysis of Sex-Based Outcomes Following Transcarotid Artery Revascularization.
Several studies suggest that females have higher perioperative adverse events and decreased benefit from carotid artery revascularization with transfemoral carotid artery stenting and carotid endarterectomy (CEA) compared to males. However, there are limited data of sex-based outcomes for transcarotid artery revascularization (TCAR). A retrospective review of prospectively maintained system-wide TCAR databases was performed between December 2015-January 2022. Patients who underwent TCAR were stratified based on sex. Relevant demographics, medical conditions, anatomical characteristics, intra- and postoperative courses, and adverse events were captured. 729 patients underwent TCAR, 486 (66.6%) male and 243 (33.3%) female. Males were more likely to be diagnosed with coronary artery disease (56.9% vs 47.7%, P<.01) and were active smokers (30.4% vs 21.4%, P < .01). Age, symptomatic status, BMI, hypertension, hyperlipidemia, diabetes mellitus, arrhythmia, chronic obstructive pulmonary disease, history of myocardial infarction, heart failure with reduced ejection fraction <30%, end-stage renal disease and Charlson Comorbidity Index were similar. In the perioperative period, there was no significant difference in reintervention rates (1.6% vs 1.2%, P = .75), cranial nerve palsy (.6% vs .4%, P > .99), ipsilateral stroke (1.9% vs 3.3%, P = .29), stent thrombosis (.4% vs .8%, P > .99), myocardial infarction (0% vs 0%, P > .99) and death (1.2% vs 1.2%, P > .99). In follow-up, no significant difference was found in reintervention, ipsilateral stroke, contralateral stroke, myocardial infarction, in-stent restenosis >50%, stent thrombosis, and death. Males and females did not have a statistically significant difference in outcomes when comparing ipsilateral stroke, in-stent thrombosis, conversion to CEA, and death after TCAR. However, our cohort comprised predominantly male patients and may conceal statistical significance as the females in our cohort did have a higher tendency toward developing complications. Future studies with a larger female cohort should be conducted to determine whether there is a true disparity of outcomes between the males and females undergoing TCAR.


Therapeutic effects of chitin from Pleurotus eryngii on high-fat diet induced obesity in rats.
Recent shifts in lifestyles and diets have caused the incidence of obesity to increase rapidly, resulting in a serious threat to modern human health. There is a growing interest the use of plant or fungi derived supplements as a safe and effective means to treat obesity. In recent times, edible-medicinal fungi have garnered attention as therapeutics owing to their biocompatibility and effectiveness. Attempts to determine the therapeutic effects of these fungi have become a prime focus in drug discovery programs. Therefore, this study aimed to determine the anti-obesity effects of P. eryngii chitin in rats with obesity induced by administration of a high fat diet. To investigate the therapeutic effects of chitin from Pleurotus eryngii on high fat diet-induce obesity, we treated obese rats with different concentrations of chitin from P. eryngii for 4 weeks, using Lipitor as positive control. The living condition, food intake, body weight, perirenal adipose tissue, periepididymal adipose tissue, adipose tissue coefficient, serum lipid levels, including total cholesterol (TC), total glyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), were measured, and levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), themalonaldehyde (MDA), and superoxide dismutase (SOD) activity in liver were determined. The rats were also monitored for pathological changes in the liver and aorta. These studies indicated that administration of chitin from P. eryngii could significantly decrease obese rat food utilization rates and accumulation of adipose tissue in the body, thus preventing development of increased body weight. The treatment also significantly reduced serum lipid levels, including levels of TC, TG and LDL-C. Treatment with P. eryngii-derived chitin also enhanced ALT and AST enzymatic activity, enhanced SOD enzymatic activity, and reduced the MDA content of the liver, as well as significantly reducing the liver index and alleviating liver steatosis and aortic atherosclerosis resulting from obesity. In conclusion, chitin from P. eryngii had therapeutic effects on hyperlipidemia, fatty liver, and aortic atherosclerosis resulting from obesity in rats.


The psychometric properties of the Iranian version of revised prenatal coping inventory (Nu PCI) in healthy pregnant women: a confirmatory factor analysis.
The Revised Prenatal Coping Inventory (Nu PCI) is a self-report instrument that focuses on the coping styles of pregnant women during the prenatal period. The present study aimed to translate the scale and evaluate the psychometric properties of the Iranian version of the Nu PCI in healthy pregnant women. Beaton's recommendations have been used for the translation and cultural adaptation of this scale. Reliability, content, and face validity were performed to assess the psychometric properties of the scale. The results indicated that the Nu PCI was successfully translated into the Persian language, with both the total scale and subscales exhibiting both content validity and internal consistency. Confirmatory factor analyses revealed that the conceptual three-factor structure of the measure had good fit indices. Reliability was evaluated using internal consistency. Cronbach Alpha coefficient was 0.92 and intra-class correlation coefficient (ICC) was 0.91. Therefore, the Nu PCI is a reliable instrument to measure prenatal coping patterns in primigravida healthy pregnant women.


MicroRNA-21 is Required for Hematopoietic Cell Viability After Radiation Exposure.
Radiation therapy is an essential intervention used in the treatment of more than half of cancer patients. With the increasing use of hypofractionated radiation regimens, concurrent use of radiation and chemotherapy, targeted agents and immunotherapy, the risk of radiation-induced toxicities is increased. However, much remains unknown about the molecular underpinnings responsible for radiation-induced toxicity. MicroRNA (miRNA) are small, non-coding RNA involved in post-transcriptional regulation of gene expression. miR-21 is an oncomiR that is dysregulated in a significant fraction of human malignancies, and its overexpression is linked to poor overall survival, chemoresistance, and radioresistance in several human cancers. However, the contribution of miR-21 in governing radiation sensitivity in normal, untransformed cells, and the impact of silencing this miRNA in normal tissues remains largely unexplored. miR-21 levels were evaluated in tissues by qRT-PCR without and after total body irradiation (TBI). Mice lacking miR-21 were genetically engineered, subjected to TBI, and monitored for survival. Hematopoietic stem and progenitor cell (HSPC) numbers and function were assessed using flow cytometry, histology, complete blood cell counts, and bone marrow transplantation. miR-21 expression was increased in radiosensitive tissues, but not in radioinsensitive tissues following TBI in wild-type mice, suggesting it may have a critical function in the normal tissue response to irradiation. Compared to wild-type mice, mice lacking one or both alleles of miR-21 showed reduced numbers of HSPCs and increased sensitivity to an LD50/30 dose of TBI with evidence of bone marrow failure. Transplantation of wild-type bone marrow into irradiated miR-21-deficient mice rescued the mice from death. Our data identify miR-21 as a critical component of HSPC viability and essential for bone marrow recovery following irradiation. Further investigation is warranted to determine whether miR-21 can be used to stratify patients at risk for hematopoietic toxicity following irradiation.


Intravenous nitroglycerin in acute coronary artery disease.
Intravenous nitroglycerin (IVNG) was used in 12 instances (11 patients) to relieve pain, following myocardial infarction or unstable angina. The strength of IVNG infusion could be easily controlled and relief of pain was prompt and satisfactory. Excessive tachycardia or severe headache was not encountered and marked hypotension was seen in one patient only. This patient had three vessel coronary artery disease, extensive myocardial damage and vein graft closure. Cardiovascular stability was provided by IVNG, prior to and during cardiac catheterisation and cineangiography, transportation to another hospital and vein graft surgery. IVNG appears to be a safe and effective drug in the management of patients with acute coronary artery disease (unstable angina and myocardial infarction). In those with both right and left ventricular wall infarction, however, the usefulness of IVNG needs to be established.


Prediction of response to pegylated interferon plus ribavirin in HIV/hepatitis C virus (HCV)-coinfected patients using HCV genotype, IL28B variations, and HCV-RNA load.
This study aimed at developing a predictive algorithm based on interleukin 28B (IL28B) genotype, hepatitis C virus (HCV) genotype, and plasma HCV-RNA load, which could accurately allow us to define the probability of response to pegylated interferon (Peg-IFN) plus ribavirin (RBV) therapy in HIV/HCV-coinfected patients. Five hundred and twenty-one treatment-naive HIV-infected patients, who initiated HCV therapy with Peg-IFN/RBV, were analysed in an on-treatment basis. Patients were categorized as unlikely responders, uncertain responders, and anticipated responders (<20%, 20-60%, and >60% probability to achieve SVR, respectively). HCV genotype, baseline HCV-RNA load, and IL28B genotype were confirmed as independent predictors of SVR in a logistic regression analysis. A stepwise algorithm based on these three variables was created based on 321 patients and evaluated in the remaining 200 patients. Unlikely responders included patients with genotype 1 or 4, HCV-RNA load ≥600,000IU/ml, and rs12979860 non-CC (rate of SVR: 17.3%). Anticipated responders were those with HCV genotype 2-3, patients harboring HCV genotype 4 and IL28B CC, as well as those who simultaneously bore HCV genotype 1, HCV-RNA load <600,000IU/ml, and IL28B CC (rate of SVR 74.1%, 77.8%, and 64.4%, respectively). The area under the receiver operating characteristic curve of the model was 0.77 (0.733-0.814). The combined use of IL28B genotype, HCV genotype, and HCV-RNA load enables to easily identify patients with a high and very low likelihood of SVR. HCV therapy could be deferred in the latter patients, until more effective options are available, at least if they do not show advanced liver fibrosis.


Defective ureagenesis in mice carrying a liver-specific disruption of hepatocyte nuclear factor 4alpha (HNF4alpha ). HNF4alpha regulates ornithine transcarbamylase in vivo.
Hepatocyte nuclear factor 4alpha (HNF4alpha) regulates the expression of many genes preferentially expressed in liver. HNF4alpha-null mice die during embryogenesis precluding the analysis of its function in the adult. To circumvent this problem, liver-specific HNF4alpha-null mice were produced. Mice lacking hepatic HNF4alpha expression exhibited increased serum ammonia and reduced serum urea. This disruption in ureagenesis may be explained by a marked decrease in expression and activity of hepatic ornithine transcarbamylase (OTC). To determine the molecular mechanisms involved in transcriptional regulation of the mouse OTC gene, the OTC promoter region was analyzed. Sequence analysis revealed the presence of two putative HNF4alpha-binding sites in the mouse OTC promoter region. By using transient transfection analysis, it was established that high levels of promoter activity were dependent on both HNF4alpha-binding sites and the expression of HNF4alpha. Furthermore, the proximal HNF4alpha-binding site was found to be more important than the distal one for transactivating OTC promoter. These data demonstrate that HNF4alpha is critical for urea homeostasis by direct regulation of the OTC gene in vivo.


Diagnostic and prognostic value of genetics in undifferentiated peripheral inflammatory arthritis: a systematic review.
To evaluate the diagnostic and prognostic utility of genetic testing in undifferentiated peripheral inflammatory arthritis (UPIA). A systematic literature search was performed in Medline, Embase, the Cochrane Library, and abstracts presented at the 2007 and 2008 meetings of the American College of Rheumatology and the European League Against Rheumatism. The target studies were those evaluating diagnostic or prognostic value of genetic markers specifically in UPIA. Two reviewers independently screened titles and abstracts and reviewed included articles in detail. All data were collected using ad hoc standard forms, permitting the calculation of positive and negative likelihood ratios of each genetic marker for diagnoses of different rheumatic diseases and for the development of relevant outcomes. Of the 3109 articles retrieved, 26 original studies fulfilled criteria of the systematic review. The most frequent diagnosis tested was rheumatoid arthritis, followed by inflammatory polyarthritis, and spondyloarthropathies. The main prognostic outcome evaluated was development of erosions, followed by median Larsen score, remission, Health Assessment Questionnaire (HAQ) score, and persistent synovitis. In total, 122 genetic markers were tested. No genetic marker had a high likelihood ratio for the diagnosis of a specific rheumatic disease. The shared epitope was associated with poor prognosis (erosions, HAQ > 1, mortality, and persistent synovitis). Other genes did not predict outcome in undifferentiated arthritis. Other outcomes for persistent disease or disability were not studied in depth. In isolation, no studied genetic marker is very informative of a future diagnosis in patients with UPIA. The shared epitope has a slight association with poor prognosis of UPIA.


Supplementation of corn-soy-based diets with an Eschericia coli-derived phytase: effects on broiler chick performance and the digestibility of amino acids and metabolizability of minerals and energy.
The effect of the supplementation of diets containing low available P concentrations with low and supra-activities of an Eschericia coli 6-phytase was assessed using growing broiler chicks. A total of 384 female Ross broiler chicks were weighed at d 1 of life and assigned to 1 of 8 experimental treatments. There were 12 replicate cages with 4 chicks per cage, and the diets were fed from d 1 of life for a period of 16 d. A positive control diet (5 g/kg of available P) and a negative control diet (3 g/kg of available P) were used, and 6 more diets were manufactured by supplementing the negative control diet with 150, 300, 600, 1,200, 2,400, and 24,000 U/kg of exogenous phytase. Body weight gain and feed conversion ratios were determined, as were nutrient digestibility coefficients and toe ash values. Birds fed the negative control diet had lower (P < 0.05) weight gains than those fed the positive control diet. The addition of exogenous phytase above 150 U/kg improved (P < 0.05) weight gain, toe ash percentage, and nutrient utilization of the birds fed the negative control diet. Furthermore, the 24,000 U/kg of diet improved (P < 0.05) toe ash percentage and the utilization of several nutrients beyond that of the lower doses of phytase. It can be concluded that the supplementation of diets containing 3 g/kg of available P with exogenous phytase can improve the performance of chicks to that of birds fed a diet containing 5 g/kg of available P. In addition, the use of high doses of phytase (> 1,000 U/kg of diet) can improve nutrient availability in poultry diets beyond that of diets containing lower (< 1,000 U/kg) phytase activities. These results may be mediated partially by reduced endogenous loss as well as an increase in the availability of dietary nutrients as indicated by improvements in digestibility coefficients.


Altered inflammasome machinery as a key player in the perpetuation of Rett syndrome oxinflammation.
Rett syndrome (RTT) is a progressive neurodevelopmental disorder mainly caused by mutations in the X-linked MECP2 gene. RTT patients show multisystem disturbances associated with an oxinflammatory status. Inflammasomes are multi-protein complexes, responsible for host immune responses against pathogen infections and redox-related cellular stress. Assembly of NLRP3/ASC inflammasome triggers pro-caspase-1 activation, thus, resulting in IL-1β and IL-18 maturation. However, an aberrant activation of inflammasome system has been implicated in several human diseases. Our aim was to investigate the possible role of inflammasome in the chronic subclinical inflammatory condition typical of RTT, by analyzing this complex in basal and lipopolysaccharide (LPS)+ATP-stimulated primary fibroblasts, as well as in serum from RTT patients and healthy volunteers. RTT cells showed increased levels of nuclear p65 and ASC proteins, pro-IL-1β mRNA, and NLRP3/ASC interaction in basal condition, without any further response upon the LPS + ATP stimuli. Moreover, augmented levels of circulating ASC and IL-18 proteins were found in serum of RTT patients, which are likely able to amplify the inflammatory response. Taken together, our findings suggest that RTT patients exhibited a challenged inflammasome machinery at cellular and systemic level, which may contribute to the subclinical inflammatory state feedback observed in this pathology.


Renal transplantation in patients above 60 years of age in the modern era: a single center experience with a review of the literature.
A retrospective study was conducted of 797 patients receiving renal transplants from January 1985 to March 1997. Patient and graft survival was compared for patients above and below the age of 60. Sixty-nine patients < or =60 years old received 73 kidneys. Race: 73% Caucasian, 26% Black, 1% Other. Sex: 68% M. Hypertension (19) and PCKD (15) were the most common diagnoses. Mean peak panel reactive antibody (PRA) was 37.7%. Donor age was 2 to 66 years. Mean Cold ischemic time was 28.1 hours. Follow-up was until death or until 8/30/97. Patients <60 years included: 62% Caucasian, 34% Black, 4% Other; 60% male, Mean PRA 39.3. Of the 69 study patients, 27 died: 19 with a functioning graft, 8 within one year of transplantation. Cardiovascular causes (19 patients, 72%) and infection (7 patients, 24%) were most common. Common causes of graft loss were death with a functioning graft (19) and chronic rejection (15); other causes were acute rejection and primary non-function. Univariate analysis of 18 risk factors showed CHF and past history of vascular surgery significantly (p < 0.05) affected time of return to dialysis. Multi variate analysis did not show these independent variables to be significant. Abnormal ejection fraction and presence of q waves on EKG significantly affected time to death (p < 0.05) on uni- and multi-variate analysis. After censoring patients that died with functioning grafts, difference in graft survival between > or =60 and <59 years was not significant (p > 0.2). In this study, 68% of older patients had allografts functioning at 1 year. The fact that older patients succumb over time from natural causes should not keep patients from transplantation. Immunosuppressive agents need to be limited to reduce the incidence of infection. Criteria need to be refined to define those who are at prohibitive risk, who may not be candidates for transplantation.


Phenylalkylamine stimulants, hallucinogens, and designer drugs.
Phenylalkylamine derivatives produce several types of behavioral effects including central stimulation and hallucinogenic activity. SAR are being formulated and already (a) it has been demonstrated that each of these types of activities is associated with a distinct SAR, and (b) it is now possible to use these SAR to make predictions as to whether the stimulus effects of certain PAAs are primarily AMPH-like or DOM-like. The AMPH-like nature of PAAs seems to involve a dopaminergic mechanism whereas DOM-like activity involves a serotonergic (in particular a 5-HT2) mechanism. It is apparent, however, that there is an additional type of activity emerging from studies with some PAAs that is neither solely AMPH-like nor DOM-like. MDA seems to produce both types of actions and may even produce this third type of effect. MDMA produces AMPH-like and MDA-like effects, but does not produce DOM-like effects. Other agents, such as MDE and PMMA, produce neither AMPH-like nor DOM-like effects but clearly produce MDMA-like stimulus effects. Thus, there is a third type of SAR that may be formulated. In all likelihood, however, few PAAs will be shown to produce a single "pure" activity and because there are some similarities in the different SARs (even though there are some very clear differences) it is not unreasonable to assume that many PAAs will produce more than one type of effect or will display vestiges of one or more different components of action. Therefore, although a PAA may be classified as primarily producing one type of effect, it should be understood that the other types of effects are not necessarily absent.(ABSTRACT TRUNCATED AT 250 WORDS)


Pharmaco-thermodynamics of deuterium-induced oedema in living rat brain via 1H2O MRI: implications for boron neutron capture therapy of malignant brain tumours.
In addition to its common usage as a tracer in metabolic and physiological studies, deuterium possesses anti-tumoural activity and confers protection against gamma-irradiation. A more recent interest in deuterium emanates from the search for alternatives capable of improving neutron penetrance whilst reducing healthy tissue radiation dose deposition in boron neutron capture therapy of malignant brain tumours. Despite this potential clinical application, deuterium induces brain oedema, which is detrimental to neutron capture therapy. In this study, five adult male rats were titrated with deuterated drinking water while brain oedema was monitored via water proton magnetic resonance imaging. This report concludes that deuterium, as well as deuterium-induced brain oedema, possesses a uniform brain bio-distribution. At a steady-state blood fluid deuteration value of 16%, when the deuterium isotope fraction in drinking water was 25%, a mean oedematous volume change of 9 +/- 2% (p-value <0.001) was observed in the rat brain-this may account for neurological and behavioural abnormalities found in mammals drinking highly deuterated water. In addition to characterizing the pharmaco-thermodynamics of deuterium-induced oedema, this report also estimates the impact of oedema on thermal neutron enhancement and effective dose reduction factors using simple linear transport calculations. While body fluid deuteration enhances thermal neutron flux penetrance and reduces dose deposition, oedema has the opposite effect because it increases the volume of interest, e.g., the brain volume. Thermal neutron enhancement and effective dose reduction factors could be reduced by as much as approximately 10% in the presence of a 9% water volume increase (oedema).


Service-line management/bottom-line management.
To survive in the sturm und drang of health care administration, hospitals and health care systems will have to restructure themselves in ways that emphasize their specific clinical strengths, control their costs, and manage the delivery and outcomes of care. Structuring the organization along clinical lines of service (e.g., oncology, cardiology, rehabilitation) cedes total bottom-line authority for all aspects of that service to the service, or product, line manager. This article discusses the qualifications, compensation, and responsibilities of service-line managers in well-integrated health care systems and describes how they and managed care organizations view each other. It also suggests which organizations will, and will not, benefit from restructuring along service lines.


A transgenic zebrafish model of a human cardiac sodium channel mutation exhibits bradycardia, conduction-system abnormalities and early death.
The recent exponential increase in human genetic studies due to the advances of next generation sequencing has generated unprecedented numbers of new gene variants. Determining which of these are causative of human disease is a major challenge. In-vitro studies and murine models have been used to study inherited cardiac arrhythmias but have several limitations. Zebrafish models provide an attractive alternative for modeling human heart disease due to similarities in cardiac electrophysiology and contraction, together with ease of genetic manipulation, external development and optical transparency. Although zebrafish cardiac mutants and morphants have been widely used to study loss and knockdown of zebrafish gene function, the phenotypic effects of human dominant-negative gene mutations expressed in transgenic zebrafish have not been evaluated. The aim of this study was to generate and characterize a transgenic zebrafish arrhythmia model harboring the pathogenic human cardiac sodium channel mutation SCN5A-D1275N, that has been robustly associated with a range of cardiac phenotypes, including conduction disease, sinus node dysfunction, atrial and ventricular arrhythmias, and dilated cardiomyopathy in humans and in mice. Stable transgenic fish with cardiac expression of human SCN5A were generated using Tol2-mediated transgenesis and cardiac phenotypes were analyzed using video microscopy and ECG. Here we show that transgenic zebrafish expressing the SCN5A-D1275N mutation, but not wild-type SCN5A, exhibit bradycardia, conduction-system abnormalities and premature death. We furthermore show that SCN5A-WT, and to a lesser degree SCN5A-D1275N, are able to compensate the loss of endogenous zebrafish cardiac sodium channels, indicating that the basic pathways, through which SCN5A acts, are conserved in teleosts. This proof-of-principle study suggests that zebrafish may be highly useful in vivo models to differentiate functional from benign human genetic variants in cardiac ion channel genes in a time- and cost-efficient manner. This article is part of a Special Issue entitled "Na(+) Regulation in Cardiac Myocytes".


[Effect of tongmal recipe on atherosclerosis in patients with early lower extremity arteriopathy caused by type 2 diabetes mellitus].
To observe the effect of Tongmai Recipe (TMR) on atherosclerosis (AS) in patients with early lower extremity arteriopathy disease (LEAD) caused by type 2 diabetes mellitus (T2DM). Patients were randomly assigned to two groups. The 23 patients in the treatment group were treated by TMR and the 22 in the control group were given Cilostazo, all for 3 months. Before and after treatment, lower extremity vascular color Doppler image, blood levels of glucose, insulin, serum matrix metalloproteinase-9 (MMP-9) and interleukin-6 (IL-6) were measured. The thickness of AS plaque and that of arterial intima-media obviously reduced (P < 0.01 and P < 0.05), insulin sensitivity improved noticeably (P < 0.01), and serum levels of MMP-9 and IL-6 lowered obviously (P < 0.05) in the treatment group after treatment, but these indexes were unchanged in the control group statistically. TMR has remarkable action in reducing the thickness of AS plaque and intima-media of artery, its mechanism might be related with improving of insulin sensitivity and anti-inflammatory reaction, and reducing of serum MMP-9 level as well in patients.


[Surgical choice of posterior osteotomy way for senile osteoporotic thoracolumbar fracture with kyphosis].
To investigate the surgical choice of posterior osteotomy way by the observation of clinical outcome of Smith-Petersen osteotomy (SPO), pedicle subtraction osteotomy (PSO) and vertebral column re-section (VCR) for senile osteoporotic thoracolumbar fracture with kyphosis. From June 2015 to August 2017, an amount of 8 elderly patients with thoracolumbar kyphosis caused by osteoporotic vertebral fracture underwent osteotomy approach for posterior osteotomy. All patients were old osteoporotic vertebral fracture more than 6 months and received invalid conservative treatment for 3 months including nonsteroidal anti-inflammatory and analgesic drugs, anti-osteoporosis drugs and acupuncture, etc. There were 3 males and 5 females, with an average age of 73.4 years (66 to 83 years), with an average course of the disease of 34.6 months (8 to 60 months). Eight patients had a total of 8 vertebral fractures, and fracture segment was in T10 of 1 case, T11 of 1 case, T12 of 3 cases, L1 of 2 cases, L2 of 1 case. Eight patients showed kyphosis caused by wedge deformation of single segmental vertebral fractures. The thoracolumbar kyphosis and symptoms were progressively developing into central sagittal imbalance. SPO osteotomy was performed in 3 cases, PSO osteotomy in 3 cases, and VCR osteotomy in 2 cases. Orthopaedic effects were analyzed by imaging measurements, including pre- and post-operative kyphosis Cobb angle, localized kyphosis (LK), thoracic kyphosis (TK), lumbar lordosis (LL), sacral tilt angle (ST) and sagittal vertical axis (SVA). Visual analogue scale (VAS) and Oswestry Disability Index (ODI) were used to evaluate the pain and lumbar function. All the eight patients were followed up from 8 to 24 months with an average of 13.5 months and all the symptoms of low back pain have significantly reduced or disappeared. The VAS score decreased from 5-8 points (mean 6.5 points) before surgery to 1-4 points (mean 1.88 points) at the final follow-up, and the score was significantly improved. The ODI score decreased from 36-78 points (mean 60.25 points) before surgery to 10-32 points (mean 20.38 points) at the final follow-up, and the functional score improved significantly. During the follow-up period, X-ray examination showed that some patients had a slight decrease in the height of the intervertebral fusion, and the bone graft was healed. There was no obvious corrected degree loss and internal fixation loosening, and the thoracolumbar kyphosis was significantly improved. The mean Cobb angle of T10-L2 was reduced from 25.3° to 2.8° with corrected rate of 89.3% ; LK was reduced from 43.4° to 7.1° with corrected rate of 86.2% ; TK was reduced from 49.9° to 30.6°, LL was reduced from 43.6° to 30.8°, and ST was changed from 24.0° to 32.1°, SVA was changed from 6.23 cm to 2.40 cm. For the different pathological features and deformities of senile osteoporotic thoracolumbar fracture combined with kyphosis, SPO, PSO or VCR can achieve good orthopedic effect and clinical efficacy.


[Carbohydrate metabolic disturbances and obstructive sleep apnea syndrome].
The paper analyzes an association between obstructive sleep apnea syndrome (OSAS) and carbohydrate metabolic disturbances. It presents the data of clinical trials that consider OSAS as a risk factor of type 2 diabetes mellitus (T2DM). The most important factors influencing the progression of carbohydrate metabolic disturbances in patients with OSAS are analyzed. Whether OSAS can be corrected as a modifiable risk factor in patients with T2DM is considered.


An unusual variety of the extensor digiti muscles: report with notes on repetition strain injuries.
In over 500 human cadaveric dissections of arms and forearms, performed to the present date, we find frequent anatomical variations, corresponding to classic descriptions. Last year, we found a singular anatomic variation of the extensor muscles of the forearm, which seems previously undescribed. It is our strong belief that gross anatomy studies, and gross dissection should be updated and reintroduced in modern anatomical studies, for teaching, research, or surgical training purposes. We detected a peculiar anatomical variant of the Superficial Extensor Digiti Muscles in the forearm of a human 73 year old male Caucasian cadaver. We clearly identified a thick bundle of muscular fibres, connecting the main muscular shafts of the Extensor Digiti Minimi, and the Extensor Digitorum Communis Muscles, in a perfectly defined muscular expansion, bridging obliquely downwards and outwards, between the two main muscular shafts. In our series, this is the first occurrence of such anatomical disposition. Anatomical variations of the extensor tendons to the fingers are frequently detected in the wrist, hand and fingers compartments. The careful analysis of the variants of muscular shafts in the forearm compartment, as commonly reported in the earliest anatomical descriptions will bring renewed light to the functional assessment of the extensor mechanism of the human fingers. In this sense, we reviewed the oldest anatomical descriptions, from the 16th century to the present date.


Advanced carcinoma of the pancreas: phase II study of combined chemotherapy, beta-interferon, and retinoids.
Because of the poor response of pancreatic cancer to conventional therapy, the authors performed a phase II pilot study to evaluate whether beta-interferon and retinoids, added to active chemotherapeutic agents, could increase response rate and survival in a group of patients who had metastatic disease. Twenty-three chemotherapy-naive patients were treated as follows: epirubicin, 60 mg/m2, and mitomycin C, 10 mg/m2, intravenously on day 1; folinic acid, 200 mg/m2, and 5-fluorouracil (5-FU), 370 mg/m2, intravenously for 5 consecutive days. beta-Interferon, 1 x 10(6) IU/m2, subcutaneously three times a week, and retinol palmitate, 50,000 IU orally twice a day, were given between chemotherapy cycles. Patients having responses and disease stabilization were maintained with the same dose of beta-interferon and retinol palmitate. Treatment was given every 4 weeks for four courses or until onset of progression. A median of three courses of chemotherapy was delivered to each patient. All patients were evaluable. Eight patients responded (35%) and 8 (35%) had stable disease. Median time to progression and survival for all patients were, respectively, 6.1 months and 11 months. Toxicity was severe: 60% of patients had hematologic toxicity, 40% had gastrointestinal toxicity, 13% had cardiac toxicity, and 1 patient had a hemolitic-uremic syndrome. The combination of chemotherapy, beta-interferon, and retinoids shows activity in metastatic pancreatic carcinoma. Toxicity was high but patients who had responses and disease stabilization had prolonged symptom palliation.


Effects of food abundance and early clutch predation on reproductive timing in a high Arctic shorebird exposed to advancements in arthropod abundance.
Climate change may influence the phenology of organisms unequally across trophic levels and thus lead to phenological mismatches between predators and prey. In cases where prey availability peaks before reproducing predators reach maximal prey demand, any negative fitness consequences would selectively favor resynchronization by earlier starts of the reproductive activities of the predators. At a study site in northeast Greenland, over a period of 17 years, the median emergence of the invertebrate prey of Sanderling Calidris alba advanced with 1.27 days per year. Yet, over the same period Sanderling did not advance hatching date. Thus, Sanderlings increasingly hatched after their prey was maximally abundant. Surprisingly, the phenological mismatches did not affect chick growth, but the interaction of the annual width and height of the peak in food abundance did. Chicks grew especially better in years when the food peak was broad. Sanderling clutches were most likely to be depredated early in the season, which should delay reproduction. We propose that high early clutch predation may favor a later reproductive timing. Additionally, our data suggest that in most years food was still abundant after the median date of emergence, which may explain why Sanderlings did not advance breeding along with the advances in arthropod phenology.


Synergistic anti-proliferative and apoptotic effect of NVP-BEZ235 and curcumin on human SH-SY5Y neuroblastoma cells.
Neuroblastoma, a tumor of the sympathetic nervous system, is one of the most common tumors found in children. Most patients develop resistance to therapy and show poor prognosis, thus there is a need of novel therapeutic agents for the treatment of neuroblastoma. NVP-BEZ235 is a dual Phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) kinase inhibitor that induces apoptosis and suppresses the growth of cancer. Curcumin acts as an anticancer agent in certain cancers. This study investigated the synergetic effect of NVP-BEZ235 and curcumin against neuroblastoma SH-SY5Y cell line. In the current study, the synergic effect of NVP-BEZ235 and curcumin in SH-SY5Y was examined in terms of the cell growth by cell viability and colony forming assay, cell cycle and apoptotic cell death by flow cytometry and mRNA expression levels by quantitative Real Time Polymerase Chain Reaction (qRT-PCR). Curcumin, NVP-BEZ235 or a combination of both, showed cytotoxicity in a dose and time dependent manner in SH-SY5Y cells. 10 µM curcumin and 200 nM NVP-BEZ235 were chosen as combination therapy, as the combination index showed synergism. Colony forming assay showed decrease in cell growth in combination group. The cell cycle distribution for combination group demonstrated a decrease in G0/G1 phase at 48 h. Annexin V showed an anticancer effect in combination group when compared to control group. Moreover, qRT-PCR results showed a significant increase in caspase 3, caspase 7, Bax and p53 genes, while a decrease in Bcl-2 gene expression levels. These findings suggest that combination therapy of NVP-BEZ235 and curcumin may be a promising therapeutic candidate for treatment of neuroblastoma.


Determinants of sensitivity and resistance to rapamycin-chemotherapy drug combinations in vivo.
The phosphatidylinositol-3-OH kinase [PI(3)K] pathway is frequently activated in human cancers and represents a rational target for therapeutic intervention. We have previously shown that enforced expression of Akt, which is a downstream effector of PI(3)K, could promote tumorigenesis and drug resistance in the Emu-myc mouse lymphoma model, and that these tumors were particularly sensitive to inhibition of mammalian target of rapamycin (mTOR) with rapamycin when combined with conventional chemotherapy. We now show that reduced dosage of PTEN, a negative regulator of PI(3)K signaling, is sufficient to activate Akt, but has only a modest effect on lymphomagenesis in the same model. Nonetheless, loss of even one PTEN allele resulted in lymphomas that were resistant to conventional chemotherapy yet sensitive to rapamycin/chemotherapy combinations. These effects could be recapitulated by using RNA interference to suppress PTEN expression in lymphomas, which were previously established in the absence of PI(3)K lesions. Finally, the introduction of lesions that act downstream of mTOR (eIF4E) or disable apoptosis (Bcl-2 and loss of p53) into PTEN+/- lymphomas promoted resistance to rapamycin/chemotherapy combinations. Thus, whether activation of the PI(3)K pathway confers sensitivity or resistance to therapy depends on the therapy used as well as secondary genetic events. Understanding these genotype-response relationships in human tumors will be important for the effective use of rapamycin or other compounds targeting the PI(3)K pathway in the clinic.


Oxidative phosphorylation by a cell-free particulate system from unfertilized arbacia eggs.
1. A cell-free particulate system capable of effecting oxidative phosphorylation has been prepared from unfertilized eggs of Arbacia punctulata. A substantial increase in phosphorylation can be produced by addition of alpha-ketoglutarate, oxalacetate, or succinate, the magnitude of the increase being greatest with alpha-ketoglutarate. The activity of the phosphorylating system is sharply dependent on maintenance of a comparatively narrow pH range during both the preparation of the particulate system and its subsequent incubation with oxidizable substrate. 2. The maximum oxygen consumption of the cell-free particulate system derived from a given weight of unfertilized eggs is about three times that of the same weight of intact unfertilized eggs and approximately the same as that of an equal weight of fertilized eggs. 3. The data indicate that generation of high-energy phosphate bonds in the Arbacia egg is coupled, as it is in mammalian liver or kidney, with the functioning of the tricarboxylic acid cycle.


Protective functions of Lycium barbarum polysaccharides in H2O2-injured vascular endothelial cells through anti-oxidation and anti-apoptosis effects.
Cell injury in the cardiovascular endothelia caused by oxidative stress is among the major inducers of endothelium dysfunction and serves an important role in initiating cardiovascular diseases (CVDs). Therefore, protecting and improving the normal function of endothelial cells are considered key measures against CVDs. As a traditional Chinese medicinal component, Lycium barbarum is regarded to have high medicinal value. The present study aimed to investigate the potential anti-apoptosis and anti-oxidation effects of Lycium barbarum polysaccharides (LBPs) on injured rat artery endothelial cells, to demonstrate the experimental and medicinal values of LBPs. In the present study, the aortic endothelial cells of rats were cultivated and randomly divided into five groups: A control group, H2O2-injured group (H2O2 group), H2O2+LBPs (110 µg/ml) group (low-dose group, LT), H2O2+LBPs (220 µg/ml) group (medium-dose group, MT) and H2O2+LBPs (440 µg/ml) group (high-dose group, HT). Among these, the activity of superoxide dismutase (SOD), and the levels of malondialdehyde (MDA) and nitric oxide (NO) were detected by colorimetry. Additionally, the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) were detected by western blotting. It was observed that SOD activity and NO content decreased while MDA content increased significantly in the H2O2 group (P<0.05 vs. control); that SOD activity in the MT and HT group, and NO content in all three LBP groups were increased, while MDA content in the three LBP groups was decreased, compared with the H2O2 group (all P<0.05); that Bcl-2 expression decreased significantly in the H2O2 group while the expression of Bax increased significantly compared with the control group (both P<0.05); and that Bcl-2 expression in all three LBP groups increased, while Bax expression in the MT and HT groups decreased compared with the H2O2 group (all P<0.05), with these altered Bax levels being statistically similar to those in the control group (P>0.05). On light microscopy, the cells in the control group exhibited spindle-shaped morphology, consistent sizes, defined boundaries, and distinct nuclei of equivalent sizes with round or oval morphology. Additionally, the chromatin in the nuclei was evenly distributed, and all cells were adhered in a paving-stone arrangement. Notably, only few cells died. Conversely, the cells in the H2O2 group exhibited signs of damage and enlarged gaps, and focal cells died. In the HT group, the cells once again appeared adherent and exhibited similar morphological status to the normal cells. Overall, these results indicate that LBPs serve a protective role in oxidative-injured vascular endothelial cells through anti-apoptosis and anti-oxidation effects.


Self-replicating and self-repairing multicellular automata.
Biological organisms are among the most intricate structures known to man, exhibiting highly complex behavior through the massively parallel cooperation of numerous relatively simple elements, the cells. As the development of computing systems approaches levels of complexity such that their synthesis begins to push the limits of human intelligence, engineers are starting to seek inspiration in nature for the design of computing systems, both at the software and at hardware levels. We present one such endeavor, notably an attempt to draw inspiration from biology in the design of a novel digital circuit: a field-programmable gate array (FPGA). This reconfigurable logic circuit will be endowed with two features motivated and guided by the behavior of biological systems: self-replication and self-repair.


Tunable multi-channel inverse optomechanically induced transparency and its applications.
In contrast to the optomechanically induced transparency (OMIT) defined conventionally, the inverse OMIT behaves as coherent absorption of the input lights in the optomechanical systems. We characterize a feasible inverse OMIT in a multi-channel fashion with a double-sided optomechanical cavity system coupled to a nearby charged nanomechanical resonator via Coulomb interaction, where two counter-propagating probe lights can be absorbed via one of the channels or even via three channels simultaneously with the assistance of a strong pump light. Under realistic conditions, we demonstrate the experimental feasibility of our model by considering two slightly different nanomechanical resonators and the possibility of detecting the energy dissipation of the system. In particular, we find that our model turns to be a unilateral inverse OMIT once the two probe lights are different with a relative phase, and in this case the relative phase can be detected precisely.


Opinion versus evidence for the need to move away from animal testing.
Science is based on facts and their discourse. Willingly or unwillingly, facts are mixed with opinion, i.e., views or judgments formed, not necessarily based on fact or knowledge. This is often necessary, where we have controversial facts or no definitive evidence yet, because we need to take decisions or have to prioritize. Evidence-based approaches aim at identifying the facts and their quality objectively and transparently; they are now increasingly embraced in toxicology, especially by employing systematic reviews, meta-analyses, quality scoring, risk-of-bias tools, etc. These are core to Evidence-based Toxicology. Such approaches aim at minimizing opinion, the "eminence-based" part of science. Animal experiments are the basis of a lot of our textbook knowledge in the life sciences, have helped to develop desperately needed therapies, and have made this world a safer place. However, they represent only one of the many possible approaches to accomplish all these things. Like all approaches, they come with shortcomings, and their true contribution is often overrated. This article aims to summarize their limitations and challenges beside the ethical and economical concerns (i.e., costs and duration as well as costs following wrong decisions in product development): they include reproducibility, inadequate reporting, statistical under-powering, lack of inter-species predictivity, lack of reflection of human diversity and of real-life exposure. Each and every one of these increasingly discussed aspects of animal experiments can be amended, but this would require enormous additional resources. Together, they prompt a need to engineer a new paradigm to ensure the safety of patients and consumers, new products and therapies.


Hyperbranched CdTe nanostructures via a self-assembly route: optical properties.
In this work, we report a luminescent nanobundle structure formed by a hierarchical self-assembly process of thioglycolic acid (TGA)-capped CdTe quantum dots (QDs). The luminescence intensity of CdTe nanostructures is high enough to get a clear one-photon excitation confocal image. High contrast two-photon excitation confocal images suggest that the nonlinear properties of pristine QDs are well inherited by the formed CdTe nanostructures. The controllability of the structures and inheritance of the optical properties of the building units make the self-assembled nanostructures new generation materials.


Frequency of recurrent urinary tract infection in patients with pelvic organ prolapse.
The aim of the study was to investigate the existence of a relationship between pelvic organ prolapse (POP) and recurrent urinary tract infection (UTI). The hospital database was searched for women diagnosed with pelvic floor disorders and all medical records were reviewed for recurrent UTI, diagnosed by two or more positive urine cultures taken within 12 months of each other. The control group was created using one-to-one matching for age and menopausal status. The prevalence of recurrent UTI in these patients were compared. The mean age of the 210 participants was 54.64±5.15 years. We found no association between POP and recurrent UTI. In the prolapse group, 22 women (21%) had recurrent UTI compared with 19 women (18%) in the control group (P=0.316). Post-void residual (PVR) volumes >50 mL were associated with increased prevalence of recurrent UTI. POP is not a risk factor for recurrent UTI, but women with POP are more likely to have high PVR volumes. High PVR volumes increase the risk of recurrent UTI. Clinical examination and ultrasound assessment of PVR should be performed in all women presenting with prolapse and UTI. Elevated PVR is the most significant risk factor, linking POP with recurrent UTI.


Phosphatases involved in modulation of gap junctional intercellular communication and dephosphorylation of connexin43 in hamster fibroblasts: 2B or not 2B?
12-O-Tetradecanoylphorbol-13-acetate (TPA) caused strong suppression of gap junctional intercellular communication, altered phosphorylation status of the gap junction protein, connexin43, and disappearance of immunorecognizible connexin43-containing gap junction plaques in V79 fibroblasts. When TPA was removed, all parameters normalized during a 3- to 4-h period. The normalizations were independent of protein synthesis, suggesting the possible involvement of phosphatases. None of the phosphatase inhibitors okadaic acid, calyculin A, cyclosporin A, or FK506 affected intercellular communication or connexin43 phosphorylation status on their own. In sequential exposures to TPA and phosphatase inhibitors, only the protein-phosphatase 2B (PP2B) inhibitors cyclosporin A and FK506 delayed the recovery of the studied parameters. Rapamycin binds to the same set of proteins as does FK506, but without inhibiting PP2B. Rapamycin did not affect the recovery of intercellular communication, but it delayed the normalization of connexin43 band pattern and immunorecognition of gap junction plaques. Dephosphorylation of immunoprecipitated connexin43 was studied using PP1, 2A, 2B, and 2C. PP2A was the most efficient (by 100-fold on a molar basis). Connexin43 immunoprecipitated from TPA-exposed cells was a poor substrate for PP1, 2B, and 2C. Thus, PP2B appeared to play a role in normalization of intercellular communication, but not necessarily in direct dephosphorylation of connexin43. Peptidyl-prolyl isomerase activity of cyclosporin/FK506/rapamycin-binding proteins may promote the dephosphorylation of connexin43 in cells.


Chromosome 9p deletions identify an aggressive phenotype of clear cell renal cell carcinoma.
The authors investigated whether deletion of chromosome 9p in clear cell renal cell carcinoma (ccRCC) predicted worse disease-specific survival (DSS) and recurrence-free survival (RFS) and whether it was associated with more aggressive behavior in small renal masses. In total, 703 ccRCC tumors were analyzed using fluorescence in situ hybridization (316 tumors) and cytogenetics (388 tumors). Tumor grade, classification, and size; 9p status; Eastern Cooperative Oncology Group performance status (ECOG PS); lymph node involvement; and the presence of metastasis were recorded. Outcomes were stratified by 9p status, and a Cox proportional hazards models was constructed using TNM staging, ECOG PS, tumor size, tumor grade, and 9p status. Deletions of 9p were detected in 97 tumors (13.8%). At presentation, 9p-deleted tumors were larger and were more likely to be high grade (grade 3 or 4), to have a high tumor (T) classification (T3-T4), and to have lymph node or distant metastases (P < .01). The median DSS for patients with and without 9p deletions was 37 months and 82 months, respectively (P < .01). In patients with localized disease, the median RFS in those who had 9p deletions was 53 months and was not reached in those without 9p deletions (P < .01). In patients who had localized lesions that measured ≤4 cm in greatest dimension, 9p-deleted tumors were more likely to recur (19% vs 2%; P = .01). Deletion of chromosome 9p in ccRCC occurred in 14% of patients and was associated with higher grade and T classification, and the presence of lymph node and distant metastases. In addition, 9p deletion independently conferred a worse prognosis for patients with localized ccRCC, and most noteworthy, in patients with localized, small renal masses. Preoperatively identifying patients with 9p deletions will improve risk stratification and will help to select appropriate patients for surveillance protocols or aggressive treatment.


Silicone Particles in Capsules Around Breast Implants: An Investigation Into Currently Available Implants in North America.
Breast implants have always been composed of a silicone elastomer envelope filled with either silicone gel or saline. Breast Implant Illness (BII) is a set of symptoms which has previously been linked to the leakage of silicone particles from the implants into the body. Our research aimed to quantify the number of silicone particles present in the capsules of breast implants available in North American. Thirty-five peri-prosthetic capsules were sampled and analyzed, and silicone particles were counted and measured. The capsule surface area was then measured and used to calculate particle density and total number of silicone particles. Eighty-five percent of capsules analyzed from silicone gel implants contained silicone, with an average of 62 particles per mm3 of capsular tissue. These patients had approximately 1 million silicone particles per capsule. In contrast, none of the saline implant capsules contained silicone. Capsules from macro-textured tissue expanders contained fewer and larger silicone particles. Silicone gel implants presented silicone particle bleeding into the peri-prosthetic capsule, totaling on average 1 million silicone particles per capsule. On the other hand, no silicone particle bleeding was observed with saline breast implants. This data suggests that particle bleeding comes from the inner silicone gel, and not from the smooth outer silicone shell. Previous studies have demonstrated the presence of breast-implant illness (BII) in patients with both silicone and saline filled implants. Therefore, our data suggests that silicone migration alone does not seem to be the sole cause of BII.


A brief history of total ankle replacement and a review of the current status.
Total ankle replacement (TAR) was first attempted in the early 1970s, but poor early results lead to it being abandoned in favour of arthrodesis. Arthrodesis is not totally satisfactory, often causing further hindfoot arthritis and this has lead to a resurgence of interest in joint replacement. New designs which more closely approximated the natural anatomy of the ankle and associated biomechanics have produced more encouraging results and led to renewed interest in total ankle replacement. Three prostheses dominate the market: Agility, Buechel-Pappas and STAR, and improving clinical results with these devices have led to more designs appearing on the market. Modern designs of prosthetic ankles almost exclusively consist of three part prostheses with a mobile bearing component, similar to the Buechel-Pappas and STAR. Clinical results of these newer designs are limited and short-term and have often been carried out by the designers of the implants. This paper presents a brief history of the development of total ankle replacement and a review of the current status.


Reinnervation of glabrous skin in baboons: properties of cutaneous mechanoreceptors subsequent to nerve transection.
1. A total of 758 fibers were isolated from previously transected and repaired ulnar nerves of five baboons. These fibers were compared to fibers from normal and previously crushed nerves studied in an earlier experiment. 2. The conduction velocities of the proximal portion of the injured axons dropped below normal, and this reduction persisted until reinnervation appeared nearly complete. 3. The receptive-field organization and response characteristics of 79 cutaneous afferent fibers serving the glabrous skin were studied in detail and compared to cutaneous afferent fibers of normal and previously crushed nerves studied earlier. 4. Initially, receptive fields were small and irregular, and often one fiber served several distinct skin regions. Ten months later, most of these abnormalities were no longer apparent. 5. Thresholds for single impulses elicited by von Frey hairs remained elevated for up to 4 mo after the receptive field reappeared, but then dropped abruptly to a near-normal range. 6. After reinnervation, rapidly adapting fibers displayed tuning curves characteristic of their submodality, but thresholds were elevated and only began to approach the normal range 6 mo after reinnervation. 7. After reinnervation, slowly adapting fibers displayed stimulus-response curves with elevated thresholds and they tended to saturate at lower stimulus intensities than normal fibers. 8. When compared to the return of function following a crushing injury, axons that had been transected displayed a slower time course for the return to normal values of conduction velocity and threshold. Receptive-field organization also remained abnormal for a longer time period. 9. These data support the hypotheses that a) breaking the continuity of the Schwann cells and extracellular matrix that occurs during transection but not during crush is a major factor leading to errors of axonal regeneration in the distal stump, b) submodality specificity is a property of the regenerating axon, and c) regenerating axons are influenced by an internal or external cue, causing them to form and maintain a single relatively homogeneous receptive field.


Investigating the formation and growth of alpha-particle radiation-induced foci of altered hepatocytes: a model-based approach.
The effect of alpha-particle radiation on the formation and increase in volume of preneoplastic liver lesions was investigated in an animal experiment. Mice were divided into four groups; two groups received different doses of the alpha-particle-labeled antibody (213)Bi-anti CD19 ((213)Bi-CD19), Thorotrast was administered to one group, and one group was left untreated. Hematoxylin and eosin-stained liver sections were evaluated for preneoplastic foci of altered hepatocytes 6, 12 and 17 months after treatment. The density and size distribution of focal transections were described by a mechanistic model for the formation and growth of foci of altered hepatocytes. The negative control and the (213)Bi-CD19 groups were combined to investigate the dose-response relationship for model parameters describing the formation and growth of foci of altered hepatocytes. Although (213)Bi-CD19 was given by single injection, the effect on formation of foci of altered hepatocytes lasted for the entire experiment. Likelihood-ratio tests comparing nested models showed that (213)Bi-CD19 increases the rates of both the formation and growth of foci of altered hepatocytes. Comparing the effects of Thorotrast with those of (213)Bi-CD19 revealed that Thorotrast had an effect similar to that of a low dose of (213)Bi-CD19, but the effect on focus formation was slightly smaller whereas the effect on focus growth was slightly higher for Thorotrast, in contrast to a low dose of (213)Bi-CD19.


Current management of massive lower gastrointestinal bleeding.
Massive lower gastrointestinal hemorrhage originates in the colon in 70% of cases. In half the remaining cases it originates in the small bowel, and in the other half in the duodenum. Currently available diagnostic techniques are: mesenteric angiography, barium contrast meal studies, colonoscopy, and laparotomy. Causes of massive lower gastrointestinal bleeding are: diverticulosis, angiodysplasia, ischemic colitis, inflammatory bowel disease and intestinal varices. Accurate localization of the bleeding site is essential if surgical treatment is to be successful.


Influence of behavior and mating success on brood-specific contribution to fish recruitment in ponds.
One source of uncertainty in predicting the response of populations to exploitation is individual differences within a population in both vulnerability to capture and contribution to population renewal. For species with parental care, individuals engaged in nesting behavior are often targeted for exploitation, but predicting outcomes of this nonrandom vulnerability will depend in part on an understanding of how parental traits are related to potential for brood contribution to the population. Variation in brood-specific contribution to recruitment of largemouth bass (Micropterus salmoides), a fish species with extended parental care, was quantified to determine if differences in mating success, parental care behaviors, and timing of reproduction influenced offspring recruitment. Dependence of these relationships on brood predation was tested in communities that differed in the presence of bluegill, Lepomis macrochirus, an important nest predator. Daily snorkel surveys were conducted in experimental ponds during spring to monitor male spawning and parental care behaviors in populations of largemouth bass. Tissue samples collected from larvae in nests were used to develop brood-specific DNA fingerprints for determining nest origins of fall recruits. Largemouth bass spawning period in bluegill ponds was longer and more variable in duration, with lower, more variable mating success, than in ponds without bluegill. In all populations, only one or two broods provided the majority of recruits, and these were broods produced during the earliest days of spawning by the oldest, largest males. In bluegill ponds, brood contribution from earliest nests also increased with brood size. Earliest nesters were the oldest males, and recruits from these nests were often above average in body size. Offspring needed to be guarded to at least swim-up larval stage to contribute any recruits. Termination of parental protection before offspring were free swimming mainly occurred with broods guarded by smaller males in ponds with brood predators. These age- and size-specific differences in timing of spawning and duration of parental care are consistent with influences of residual reproductive value and energetic constraints on reproductive behavior. Furthermore, these patterns of individual contribution to recruitment imply that fisheries that selectively target either nesting individuals or larger, older males could potentially decrease recruitment at the population scale.


Value of pilocarpine test in early diagnosis of leprosy.
Modified Pilocarpine Test was carried out in 48 patients under investigation for Leprosy. 37 cases showed deficient sweating of varying degree. Many of these patients did not reveal confirmatory histopathological changes and were bacteriologically negative. In the absence of other causes of neuropathy this finding of deficient sweating by modified pilocarpine test, was significant. Same test was carried out in 8 confirmed cases of Tuberculoid leprosy. All cases showed positive results.


An overview of slow channel blocking drugs: pharmacological basis for therapeutic applications.
The use of voltage clamps in cardiac muscle has permitted the separation of two inward depolarizing membrane currents. The first is the fast sodium current, with rapid kinetics of activation and inactivation, which is blocked by local anesthetics. The second is the slow current carried largely by calcium ions, having slow kinetics with a long time constant of inactivation. Agents which selectively block this current are called Ca antagonists. Such agents are heterogeneous structurally; they also block calcium currents in smooth muscle and therefore produce coronary as well as peripheral vasodilatation. The most significant agents are verapamil, gallopamil, nifedipine, diltiazem and tiapamil. They have a varying spectrum of pharmacological effects with respect to changes they produce on heart rate, afterload, preload, contractility, coronary flow and on the AV node. A similar spectrum of therapeutic effects is apparent in different clinical disorders, in particular vasospastic angina, stable and unstable angina, cardiac arrhythmias, hypertension, hypertrophic cardiomyopathy and possibly other disorders. Indeed, the full range of clinical disorders which may respond to calcium antagonists is still not fully delineated and may well be at least as extensive as that for beta-adrenoceptor blocking drugs. The advent of calcium antagonists in the last decade represents one of the most significant advances in cardiovascular therapeutics.


Selection and Characterization of a DNA Aptamer Specifically Targeting Human HECT Ubiquitin Ligase WWP1.
Nucleic acid aptamers hold promise as therapeutic tools for specific, tailored inhibition of protein targets with several advantages when compared to small molecules or antibodies. Nuclear WW domain containing E3 ubiquitin ligase 1 (WWP1) ubiquitin ligase poly-ubiquitinates Runt-related transcription factor 2 (Runx2), a key transcription factor associated with osteoblast differentiation. Since WWP1 and an adapter known as Schnurri-3 are negative regulators of osteoblast function, the disruption of this complex has the potential to increase bone deposition for osteoporosis therapy. Here, we develop new DNA aptamers that bind and inhibit WWP1 then investigate efficacy in an osteoblastic cell culture. DNA aptamers were selected against three different truncations of the HECT domain of WWP1. Aptamers which bind specifically to a C-lobe HECT domain truncation were observed to enrich during the selection procedure. One particular DNA aptamer termed C3A was further evaluated for its ability to bind WWP1 and inhibit its ubiquitination activity. C3A showed a low µM binding affinity to WWP1 and was observed to be a non-competitive inhibitor of WWP1 HECT ubiquitin ligase activity. When SaOS-2 osteoblastic cells were treated with C3A, partial localization to the nucleus was observed. The C3A aptamer was also demonstrated to specifically promote extracellular mineralization in cell culture experiments. The C3A aptamer has potential for further development as a novel osteoporosis therapeutic strategy. Our results demonstrate that aptamer-mediated inhibition of protein ubiquitination can be a novel therapeutic strategy.


Expression of human milk beta-casein in Escherichia coli: comparison of recombinant protein with native isoforms.
Studies on physiological function and on structure-function relationships of human milk beta-casein have been limited. In this study, we have introduced the human beta-casein cDNA into vectors designed for expression in Escherichia coli. The inducible T7-based expression system resulted in high-level expression of recombinant beta-casein. The recombinant beta-casein, localized intracellularly in E. coli, was purified to homogeneity and compared with purified native beta-casein, in particular with respect to phosphorylation. The E. coli-produced beta-casein was found to comigrate with the full-length, nonphosphorylated native human beta-casein isoform on SDS-PAGE. An N-terminal peptide containing all tentative phosphorylation sites was isolated from the recombinant protein and analyzed by mass spectrometry. The molecular mass as well as the migration of this peptide on reversed-phase chromatography confirmed that it was unphosphorylated.


Consensus-degenerate hybrid oligonucleotide primers (CODEHOP) for the detection of novel papillomaviruses and their application to esophageal and tonsillar carcinomas.
Human papillomaviruses (HPVs) have been identified in 99% of cervical tumors. Considerable speculation exists about the role of HPV in cancer at other sites, such as skin, esophagus, and lung. One barrier to the study of HPV in extragenital tumors may be the inability to detect novel HPV types. Our objective was to design broad-range primers for detecting papillomaviruses from any group in the HPV Sequence Database. Complete L1 protein sequences were aligned and 11 blocks of conserved protein sequences were identified. Primers were designed in the corresponding nucleotide regions of five blocks. One pair of primers, ME and MH, derived from blocks E and H, appeared promising when compared with established consensus primers. To improve the sensitivity for detection of multiple papillomavirus types, ME/MH primers were split into several primers with the 5' regions matched to individual papillomavirus supergroups. Compared with the commonly used MY09/11 primer set, our primer sets were broader in range and more sensitive against most papillomavirus types tested. Application of these primers to esophageal and tonsillar carcinomas identified no novel HPV types but confirmed a high level of sensitivity for detecting HPV from these clinical samples. These primers should facilitate the search for novel papillomaviruses.


A New Oxadiazole-Based Topsentin Derivative Modulates Cyclin-Dependent Kinase 1 Expression and Exerts Cytotoxic Effects on Pancreatic Cancer Cells.
Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal form of cancer characterized by drug resistance, urging new therapeutic strategies. In recent years, protein kinases have emerged as promising pharmacological targets for the treatment of several solid and hematological tumors. Interestingly, cyclin-dependent kinase 1 (CDK1) is overexpressed in PDAC tissues and has been correlated to the aggressive nature of these tumors because of its key role in cell cycle progression and resistance to the induction of apoptosis. For these reasons, CDK1 is one of the main causes of chemoresistance, representing a promising pharmacological target. In this study, we report the synthesis of new 1,2,4-oxadiazole compounds and evaluate their ability to inhibit the cell growth of PATU-T, Hs766T, and HPAF-II cell lines and a primary PDAC cell culture (PDAC3). Compound 6b was the most active compound, with IC50 values ranging from 5.7 to 10.7 µM. Molecular docking of 6b into the active site of CDK1 showed the ability of the compound to interact effectively with the adenosine triphosphate binding pocket. Therefore, we assessed its ability to induce apoptosis (which increased 1.5- and 2-fold in PATU-T and PDAC3 cells, respectively) and to inhibit CDK1 expression, which was reduced to 45% in Hs766T. Lastly, compound 6b passed the ADME prediction, showing good pharmacokinetic parameters. These data demonstrate that 6b displays cytotoxic activity, induces apoptosis, and targets CDK1, supporting further studies for the development of similar compounds against PDAC.


A validation of 11 body-condition indices in a giant snake species that exhibits positive allometry.
Body condition is a gauge of the energy stores of an animal, and though it has important implications for fitness, survival, competition, and disease, it is difficult to measure directly. Instead, body condition is frequently estimated as a body condition index (BCI) using length and mass measurements. A desirable BCI should accurately reflect true body condition and be unbiased with respect to size (i.e., mean BCI estimates should not change across different length or mass ranges), and choosing the most-appropriate BCI is not straightforward. We evaluated 11 different BCIs in 248 Burmese pythons (Python bivittatus), organisms that, like other snakes, exhibit simple body plans well characterized by length and mass. We found that the length-mass relationship in Burmese pythons is positively allometric, where mass increases rapidly with respect to length, and this allowed us to explore the effects of allometry on BCI verification. We employed three alternative measures of 'true' body condition: percent fat, scaled fat, and residual fat. The latter two measures mostly accommodated allometry in true body condition, but percent fat did not. Our inferences of the best-performing BCIs depended heavily on our measure of true body condition, with most BCIs falling into one of two groups. The first group contained most BCIs based on ratios, and these were associated with percent fat and body length (i.e., were biased). The second group contained the scaled mass index and most of the BCIs based on linear regressions, and these were associated with both scaled and residual fat but not body length (i.e., were unbiased). Our results show that potential differences in measures of true body condition should be explored in BCI verification studies, particularly in organisms undergoing allometric growth. Furthermore, the caveats of each BCI and similarities to other BCIs are important to consider when determining which BCI is appropriate for any particular taxon.