Detection of human papillomavirus DNA in anogenital condylomata in men using in situ DNA hybridisation applied to paraffin sections.
An in situ DNA hybridisation method was used to detect human papillomavirus (HPV) DNA (HPV types 6, 11, 16, and 18), and an immunoperoxidase (IP-PAP) method to detect HPV structural protein expression in paraffin sections of biopsy specimens from 133 men treated for penile (in 114 cases) and anal (in 19 cases) warts. The anatomical distribution on the penis of classic condyloma acuminatum and of papular and flat condylomata was practically identical. The gross appearance of the warts did not correlate with their morphology on light microscopy. The detection rate of dysplasia was very different in the three types of lesions (25% in flat, 50% in acuminatum, and 75% in papular warts). Of 133 lesions, 59 (44.4%) contained HPV antigens, their expression being inversely related to the grade of dysplasia; only 17% of HPV 16 lesions had detectable HPV antigen compared with 50% to 67% in lesions of the other three HPV types. HPV 16 and HPV 18 DNA were most commonly (11%) detectable in Bowenoid lesions; however, most of the HPV 16 and 18 positive cases were found among the flat and acuminatum type of lesions. Though the overall detection rate of HPV DNA (76%) did not correlate with the grade of dysplasia, a clear cut association of HPV 16 and HPV 18 with dysplastic lesions was found, none of the HPV 16 and 25% of the HPV 18 positive cases being devoid of concomitant dysplasia. The corresponding figures for HPV 6 and HPV 11 were 59.2% and 68.8%, respectively. The implications of these findings are discussed in terms of epidemiologically established connections between penile and cervical cancer, with special emphasis of the high risk HPV types 16 and 18. The applicability of in situ DNA hybridisation as a powerful tool in the analysis of specific HPV DNA sequences in routinely processed biopsy specimens from these lesions is emphasised.


Discrepancies between Perceived and Measured Cognition in Kidney Transplant Recipients: Implications for Clinical Management.
Cognitive impairment is common in kidney transplant (KT) recipients and affects quality of life, graft survival, morbidity, and mortality. Failure to identify patients with cognitive impairment can withhold appropriate and timely intervention. This study determines whether measured cognition with standard screening tools offers any advantage over perceived cognition in screening transplant patients for cognitive impairment. Cognition was assessed in 157 KT recipients using the Montreal Cognitive Assessment (MoCA; measured cognition). In addition, transplant physicians and nurse coordinators were asked to rate transplant recipients' level of cognition after routine clinical interactions (perceived cognition). Physicians and nurses were blind to MoCA scores. Perceived cognition scores were compared to MoCA scores. Perceived cognition scores fairly correlated with MOCA scores (γ = 0.24, p = 0.001 for physicians and γ = 0.33, p < 0.0001 for nurses). Physician scores moderately correlated with nurses scores (κ = 0.44, p < 0.0001). Clinical perception had a low accuracy for identifying patients with cognitive impairment (sensitivity 66% for physicians, 65% for nurses), and those without cognitive impairment (specificity 67% for physicians, 76% for nurses). Clinical perception is inaccurate at detecting cognitive impairment in KT recipients. Objective tests should be considered to screen KT recipients for cognitive impairment.


Antarctica: The Microbiology of an Unfrozen Saline Pond.
A saline pond in a region in Antarctia where other lakes and ponds are frozen remains unfrozen at the prevailing low temperatures. The ecology of the pond is unique. A distinctive aerobic microbial population, though restricted to this natural habitat, adapts to growth in artificial culture. The growth habit of these organisms, as seen in nature and in laboratory culture, indicates a possible relationship between growth at high salt concentration, at low temperatures, and in media of low organic content.


Incidence and baseline clinical characteristics of treated neovascular age-related macular degeneration in a well-defined region of the UK.
To analyse the incidence and baseline clinical characteristics of patients with neovascular age-related macular degeneration (nAMD) treated with intravitreal anti-vascular endothelial growth factor (VEGF) injections in a defined UK region. A standardised dataset was collected prospectively using an electronic medical record (EMR) system from 1 January 2008 to 21 June 2012 for all patients living in Gloucestershire who received intravitreal anti-VEGF injections for nAMD. Over the study period, 1207 eyes from 1033 patients began intravitreal anti-VEGF injections for nAMD. The annual incidence in the years after National Institute for Health and Care Excellence (NICE) technology appraisal 155 implementation was stable at 120 (95% CI 110 to 138) eyes or 100 (89 to 115) people per 100 000 population. The most common indication was occult choroidal neovascularisation (51%). Median baseline visual acuity (VA) was significantly higher for second treated than first treated eyes (66 and 56 letters, respectively; p<0.0001). Median baseline VA of fellow eyes increased from 47 (2008) to 67 letters (2012; p<0.005). The proportion of patients with baseline VA in the better eye ≥70 letters increased from 27.6% (2008) to 51.4% (2012; p<0.0001), while the proportion eligible at baseline for full or partial certificate of visual impairment decreased from 13.8% (2008) to 7.1% (2012; p<0.05). The incidence of patients undergoing anti-VEGF therapy for nAMD increased substantially following NICE approval of ranibizumab (August 2008), and has been stable since 2009. This equates to an annual UK incidence of 26 850 (21 320 to 32 440) eyes, similar to NICE estimates. Patients eligible for blindness certification before treatment decreased by half from 2008-2012. Prospective data collection using an EMR system is invaluable for efficient monitoring of real-world clinical care.


Nouns and verbs in Chintang: children's usage and surrounding adult speech.
Analyzing the development of the noun-to-verb ratio in a longitudinal corpus of four Chintang (Sino-Tibetan) children, we find that up to about age four, children have a significantly higher ratio than adults. Previous cross-linguistic research rules out an explanation of this in terms of a universal noun bias; instead, a likely cause is that Chintang verb morphology is polysynthetic and difficult to learn. This hypothesis is supported by the fact that the development of Chintang children's noun-to-verb ratio correlates significantly with the extent to which they show a similar flexibility with verbal morphology to that of the surrounding adults, as measured by morphological paradigm entropy. While this development levels off around age three, children continue to have a higher overall noun-to-verb ratio than adults. A likely explanation lies in the kinds of activities that children are engaged in and that are almost completely separate from adults' activities in this culture.


Hepatic hydrothorax. Cause and management.
Significant pleural effusions are infrequently noted in patients with cirrhosis of the liver. A large effusion (hepatic hydrothorax) occasionally appears during the course of the disease. The fluid in the pleural space is believed to be derived from ascitic fluid that may accompany hepatic cirrhosis. Although the exact mechanism is somewhat controversial, it appears that the ascitic fluid is transported directly into the pleural space. A therapeutic thoracentesis, usually accompanied by a paracentesis, may be necessary to relieve acute symptoms. Long-term management, however, centers around eliminating or reducing the formation of ascites. When this is not successful, tube thoracostomy followed by chemical pleurodesis, primary repair of diaphragmatic defects with pleural sclerosis, or peritoneovenous shunting in conjunction with chemical pleurodesis may be attempted. These interventions may or may not be successful. Management of hepatic hydrothorax remains a clinical challenge.


Aripiprazole Induced Acute Transient Bilateral Myopia: A Case Report.
Aripiprazole is an atypical antipsychotic drug. Acute transient myopia is a rare response to systemic medication. Unspecified ocular side effects of aripiprazole have rarely been reported. To the best of our knowledge, only 3 cases of aripiprazole induced myopia and diplopia have been reported in the literature. The aim of this article is to present a case of aripiprazole-induced acute transient myopia. A 30 year-old woman underwent treatment for 5 days with aripiprazole and presented with bilateral painless blurred vision. The patient's uncorrected visual acuity was 2/10 in both eyes and her best corrected visual acuity was 10/10 in both eyes with a refractive error of -3.00 diopters using a standard Snellen chart. Adding 2 mgs of biperiden a day to her treatment regimen decreased her blurred vision symptoms. After discontinuation of aripiprazole treatment and cross-switching to Paliperidon palmitate (75 mg/month) her blurred vision completely resolved. The same side effect did not re-occur when checked on her 6-month follow up. Ophthalmologists as well as psychiatrists must be aware of this myopic shift and should also ask these patients about medicine usage, especially aripiprazole. Ophthalmologists should consult the prescribing psychiatrist and stop the drug immediately to reverse this temporary condition.


The evolving role of chemotherapy in androgen-independent (hormone-refractory) prostate cancer.
Over the last decade, much progress has been made in the treatment of advanced hormone-refractory prostate cancer (HRPC). The mitoxantrone-prednisone drug combination was approved in the United States based on palliative end points and became the standard of care despite the lack of survival benefit. Docetaxel demonstrated clinical activity in HRPC on weekly and every-3-weeks schedules as evidenced by decreases in prostate-specific antigen (PSA) values that were higher than those traditionally seen with other chemotherapy regimens. The TAX 327 phase 3 trial was designed to further define the role of docetaxel in HRPC. A total of 1006 patients were randomized to receive docetaxel 75 mg/m2 every 3 weeks, docetaxel 30 mg/m2 weekly for 5 of 6 weeks, or mitoxantrone 12 mg/m2 every 3 weeks. All patients received daily prednisone. The primary study objective was survival, with secondary objectives of pain response, PSA decrease, measurable response, and quality of life (QOL). Compared with mitoxantrone, a survival benefit was demonstrated for docetaxel every 3 weeks (P = 0.009) and for the combined docetaxel arms (P = 0.03, respectively). Decreases in PSA values were superior with docetaxel (45% for the every-3-weeks regimen, P = 0.0005; 47% for the weekly regimen, P < 0.0001). Pain improvement was superior in the every-3-weeks docetaxel arm (P = 0.0107), and QOL was significantly better with both docetaxel regimens (every 3 weeks, P = 0.009; weekly, P = 0.005). Serious toxicity was rare in all arms. Given these compelling results, it can be concluded that docetaxel represents a new standard of care for the treatment of HRPC.


Human cutaneous protothecosis: report of a case and literature review.
The Prototheca species is achlorophyllic algae and rarely causes human infection. Human protothecosis presents clinically as a cutaneous infection, olecranon bursitis, and disseminated systemic disease. We report a case of human cutaneous protothecosis involving the left wrist. A 68-year-old man presented with an ill-defined erythematous lesion with crust at the dorsal aspect of his left wrist. A punch biopsy was performed to reveal the histologic features of granulomatous inflammation with necrosis at the upper dermis, containing Prototheca organisms, of which, the characteristic features were highlighted by special staining. Through a molecular study, the Prototheca zopfii species was identified.


Haplotype analysis improves molecular diagnostics of autosomal recessive polycystic kidney disease.
Autosomal recessive polycystic kidney disease (ARPKD) is characterized by wide phenotypic variability, ranging from in utero detection with enlarged, echogenic kidneys to an adult presentation with congenital hepatic fibrosis. The ARPKD gene, PKHD1 , covers about 470 kb of DNA (67 exons), and mutation studies have found marked allelic heterogeneity with a high level of novel missense changes and neutral polymorphisms. To improve the prospects for molecular diagnostics and to study the origin of some relatively common mutations, the authors have developed a strategy for improved ARPKD haplotyping. A protocol of multiplex PCR and fluorescence genotyping in a single capillary has been developed to assay 7 highly informative simple sequence repeat (SSR) markers that are intragenic or closely flanking PKHD1. Examples in which haplotype analysis, used in combination with mutation screening, improved the utility of molecular diagnostics, especially in families in which just a single PKHD1 mutation has been identified, are illustrated. The new markers also allow screening for larger DNA deletions, detecting unknown consanguinity and exploring the disease mechanism. Analysis of 8 recurring mutations has shown likely common haplotypes for each, and the divergence from the ancestral haplotype, by recombination, can be used to trace the history of the mutation. The common mutation, T36M, was found to have a single European origin, about 1,225 years ago. Improved haplotype analysis of ARPKD complements mutation-based diagnostics and helps trace the history of common PKHD1 mutations.


Genetic dissection of the vav2-rac1 signaling axis in vascular smooth muscle cells.
Vascular smooth muscle cells (vSMCs) are key in the regulation of blood pressure and the engagement of vascular pathologies, such as hypertension, arterial remodeling, and neointima formation. The role of the Rac1 GTPase in these cells remains poorly characterized. To clarify this issue, we have utilized genetically engineered mice to manipulate the signaling output of Rac1 in these cells at will using inducible, Cre-loxP-mediated DNA recombination techniques. Here, we show that the expression of an active version of the Rac1 activator Vav2 exclusively in vSMCs leads to hypotension as well as the elimination of the hypertension induced by the systemic loss of wild-type Vav2. Conversely, the specific depletion of Rac1 in vSMCs causes defective nitric oxide vasodilation responses and hypertension. Rac1, but not Vav2, also is important for neointima formation but not for hypertension-driven vascular remodeling. These animals also have allowed us to dismiss etiological connections between hypertension and metabolic disease and, most importantly, identify pathophysiological programs that cooperate in the development and consolidation of hypertensive states caused by local vascular tone dysfunctions. Finally, our results suggest that the therapeutic inhibition of Rac1 will be associated with extensive cardiovascular system-related side effects and identify pharmacological avenues to circumvent them.


Differences in neuronal firing rates in pallidal and cerebellar receiving areas of thalamus in patients with Parkinson's disease, essential tremor, and pain.
The motor symptoms of Parkinson's disease (PD) are thought to result from increased inhibitory outflow from the basal ganglia to the pallidal receiving areas of thalamus (ventral oral anterior and posterior-Voa,Vop). To test this hypothesis, we examined the firing rates of neurons in pallidal and cerebellar receiving areas of thalamus in five PD patients and compared them to those of neurons in comparable regions of motor thalamus in two other patient groups where hyperactivity of GPi is not believed to occur [essential tremor (ET), pain]. Neuronal recordings were made during microelectrode-guided functional stereotactic neurosurgery. The mean spontaneous firing rate (MSFR) of neurons classified as voluntary neurons and presumed to be in pallidal receiving areas of thalamus in PD patients [7.4 +/- 1.0 (SE) Hz] was significantly lower (P < 0.01) than in the ET (18.1 +/- 3.0 Hz) and pain (19.0 +/- 1.9Hz) groups. In contrast, the MSFR of neurons classified as kinesthetic and presumed to be primarily in the cerebellar receiving area of thalamus (ventral intermediate-Vim), although some are probably in the deep shell region of the ventrocaudal nucleus (VPLa), was significantly greater in ET patients (25.8 +/- 3.5 Hz) than in the PD (14.3 +/- 1.6 Hz; P < 0.01) and pain (16.1 +/- 1.5 Hz; P < 0.05) groups. Similar findings were obtained when the neurons were grouped according to their estimated locations in Voa/Vop and Vim of motor thalamus. These data provide support for the prediction of the classical pathophysiological model of PD and moreover suggest that pathophysiology in the cerebello-thalamo-cortical pathway may be a possible cause of tremor in ET patients.


Role of mini-invasive surgery in the treatment of enteric duplications in paediatric age: a survey of 15 years.
Gastrointestinal tract duplications (ATD) are rare malformations that occur with an incidence of 1 in 4000-5000 live births, with heterogeneous clinical pictures in relation to the different location and size. The purpose of this study was to analyze the role of minimally invasive surgery in the management of ATDs, through a critical analysys of 15 years of experience of the Department of Paediatric Surgery, University of Bologna. The medical records of 22 children diagnosed with ATDs were retrospectively reviewed (January 1995-August 2010). The study analyses: clinical presentation, preoperative diagnosis, site, anatomic type, treatment, and outcome. Children were 16 males and 6 females, with age ranged from 1 day to 10 years. 20 ATDs (91%) were cystic type, while 2 cases (9%) were tubular one. During the first period of our experience, 10 (45.5%) cases were approached with an open surgery. Then subsequent 8 (36.4%) cases were treated with a diagnostic laparoscopy. This approach permitted also to perform a minilaparotomy, close to the site of the malformation, with a short length of the scar. In 2 cases (9%), we realized an ileal resection with end-to-end anastomosis with a trans-umbilical video-assisted procedure. In 2 cases (9%), we performed a complete removal of the lesions, after complete ligation of the vascular pedicle through a laparoscopic approach. 10 cases (45.5%) were located in the ileum, 6 cases (27.3%) were esophageal duplication, 3 cases (13.6%) were gastric duplications, 2 cases (9%) were located in the colon-rectum. The postoperative course was uneventful in all the cases. this study shows how, in the management of intestinal duplications, a mininvasive approach, is increasingly taking the field, along with increasing the "learning curve" with laparoscopy. In experienced hands, the laparoscopic approach allows an accurate definition of the exact site of duplication and a miniinvasive treatment with similar principles of open techniques.


Chitosan and chitosan chlorhydrate based various approaches for enhancement of dissolution rate of carvedilol.
Carvedilol nonselective β-adrenoreceptor blocker, chemically (±)-1-(Carbazol-4-yloxy)-3-[[2-(o-methoxypHenoxy) ethyl] amino]-2-propanol, slightly soluble in ethyl ether; and practically insoluble in water, gastric fluid (simulated, TS, pH 1.1), and intestinal fluid (simulated, TS without pancreatin, pH 7.5) Compounds with aqueous solubility less than 1% W/V often represents dissolution rate limited absorption. There is need to enhance the dissolution rate of carvedilol. The objective of our present investigation was to compare chitosan and chitosan chlorhydrate based various approaches for enhancement of dissolution rate of carvedilol. The different formulations were prepared by different methods like solvent change approach to prepare hydrosols, solvent evaporation technique to form solid dispersions and cogrind mixtures. The prepared formulations were characterized in terms of saturation solubility, drug content, infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD), electron microscopy, in vitro dissolution studies and stability studies. The practical yield in case of hydrosols was ranged from 59.76 to 92.32%. The drug content was found to uniform among the different batches of hydrosols, cogrind mixture and solid dispersions ranged from 98.24 to 99.89%. There was significant improvement in dissolution rate of carvedilol with chitosan chlorhdyrate as compare to chitosan and explanation to this behavior was found in the differences in the wetting, solubilities and swelling capacity of the chitosan and chitosan salts, chitosan chlorhydrate rapidly wet and dissolve upon its incorporation into the dissolution medium, whereas the chitosan base, less water soluble, would take more time to dissolve. This technique is scalable and valuable in manufacturing process in future for enhancement of dissolution of poorly water soluble drugs.


[The effects of L-arginine and NG-nitro-L-arginine methylester on lipopolysaccharide and lipoteichoic acid induced liver injury in the rat].
Whether nitric oxide (NO) protects or impairs the liver function and structure during the early phase of sepsis is still controversial. The aim of this study is to evaluate the effect of NO on lipopolysaccharide (LPS) or lipoteichoic acid (LTA) induced liver injury in rats. One hundred twenty-six Wistar rats were assigned randomly and equally to LTA and LPS groups. Each group was divided into three subgroups which received saline, L-Arginine (L-Arg) or NG-nitro-L-arginine methylester (L-NAME) consecutively, one hour prior to either 5 mg/kg LPS or 3 mg/kg LTA injection. Plasma nitrite plus nitrate [NOx] were measured. Liver injury was assessed by measuring the rises in circulating liver enzymes and by scoring the extent of liver necrosis. Administration of L-NAME+LPS not only reduced [NOx] production but also enhanced liver damage. LNAME+ LTA caused an increase in the plasma levels of [NOx] (p=0.0006), and produced sinusoidal enlargement in liver. L-Arg protected the hepatocytes against LPS injury, whereas, enhanced the liver damage in the LTA group. Our data indicated that overproduction of NO exerts detrimental effect on LTA-treated rats while providing a protective function in LPS group.


Integrating Non-monotonic Logical Reasoning and Inductive Learning With Deep Learning for Explainable Visual Question Answering.
State of the art algorithms for many pattern recognition problems rely on data-driven deep network models. Training these models requires a large labeled dataset and considerable computational resources. Also, it is difficult to understand the working of these learned models, limiting their use in some critical applications. Toward addressing these limitations, our architecture draws inspiration from research in cognitive systems, and integrates the principles of commonsense logical reasoning, inductive learning, and deep learning. As a motivating example of a task that requires explainable reasoning and learning, we consider Visual Question Answering in which, given an image of a scene, the objective is to answer explanatory questions about objects in the scene, their relationships, or the outcome of executing actions on these objects. In this context, our architecture uses deep networks for extracting features from images and for generating answers to queries. Between these deep networks, it embeds components for non-monotonic logical reasoning with incomplete commonsense domain knowledge, and for decision tree induction. It also incrementally learns and reasons with previously unknown constraints governing the domain's states. We evaluated the architecture in the context of datasets of simulated and real-world images, and a simulated robot computing, executing, and providing explanatory descriptions of plans and experiences during plan execution. Experimental results indicate that in comparison with an "end to end" architecture of deep networks, our architecture provides better accuracy on classification problems when the training dataset is small, comparable accuracy with larger datasets, and more accurate answers to explanatory questions. Furthermore, incremental acquisition of previously unknown constraints improves the ability to answer explanatory questions, and extending non-monotonic logical reasoning to support planning and diagnostics improves the reliability and efficiency of computing and executing plans on a simulated robot.


Water interactions with hydrophobic groups: assessment and recalibration of semiempirical molecular orbital methods.
In this work, we present a study of the ability of different semiempirical methods to describe intermolecular interactions in water solution. In particular, we focus on methods based on the Neglect of Diatomic Differential Overlap approximation. Significant improvements of these methods have been reported in the literature in the past years regarding the description of non-covalent interactions. In particular, a broad range of methodologies has been developed to deal with the properties of hydrogen-bonded systems, with varying degrees of success. In contrast, the interactions between water and a molecule containing hydrophobic groups have been little analyzed. Indeed, by considering the potential energy surfaces obtained using different semiempirical Hamiltonians for the intermolecular interactions of model systems, we found that none of the available methods provides an entirely satisfactory description of both hydrophobic and hydrophilic interactions in water. In addition, a vibrational analysis carried out in a model system for these interactions, a methane clathrate cluster, showed that some recent methods cannot be used to carry out studies of vibrational properties. Following a procedure established in our group [M. I. Bernal-Uruchurtu, M. T. C. Martins-Costa, C. Millot, and M. F. Ruiz-López, J. Comput. Chem. 21, 572 (2000); W. Harb, M. I. Bernal-Uruchurtu, and M. F. Ruiz-López, Theor. Chem. Acc. 112, 204 (2004)], we developed new parameters for the core-core interaction terms based on fitting potential energy curves obtained at the MP2 level for our model system. We investigated the transferability of the new parameters to describe a system, having both hydrophilic and hydrophobic groups, interacting with water. We found that only by introducing two different sets of parameters for hydrophilic and hydrophobic hydrogen atom types we are able to match the features of the ab initio calculated properties. Once this assumption is made, a good agreement with the MP2 reference is achieved. The results reported in this work provide therefore a direction for future developments of semiempirical approaches that are still required to investigate chemical processes in biomolecules and in large disordered systems.


Impact of hypoglycemic agents on myocardial ischemic preconditioning.
Murry et al in 1986 discovered the intrinsic mechanism of profound protection called ischemic preconditioning. The complex cellular signaling cascades underlying this phenomenon remain controversial and are only partially understood. However, evidence suggests that adenosine, released during the initial ischemic insult, activates a variety of G protein-coupled agonists, such as opioids, bradykinin, and catecholamines, resulting in the activation of protein kinases, especially protein kinase C (PKC). This leads to the translocation of PKC from the cytoplasm to the sarcolemma, where it stimulates the opening of the ATP-sensitive K(+) channel, which confers resistance to ischemia. It is known that a range of different hypoglycemic agents that activate the same signaling cascades at various cellular levels can interfere with protection from ischemic preconditioning. This review examines the effects of several hypoglycemic agents on myocardial ischemic preconditioning in animal studies and clinical trials.


[Transplantation in pulmonary hypertension].
Pulmonary hypertension is one of the indications for lung transplantation. Recent advances on medical management of the disease have dramatically decreased the number of patients who required lung transplantation. In reported series, single or double lung transplantation have been successfully undertaken by many transplant centers. In patients, whose cardiac functions are irreversibly damaged heart-lung transplantation remains as an only option for long term survival. Transplantation should only be considered in those patients who are having optimal medical support and relatively good condition for a major operation.


His bundle electrogram in patients with acute myocardial infarction complicated by atrioventricular or intraventricular conduction disturbances.
Seventy-two patients with acute myocardial infarction complicated by atrioventricular or bundle-branch block or a combination of both had His bundle electrogram studies performed during their stay in the coronary care unit. In 19 of the 72 patients a repeat His bundle electrogram was performed before discharge from hospital. These studies demonstrated that 30 of the 32 patients with atrioventricular block and narrow QRS complexes had a block above the origin othe His spike (proximal block). Eleven patients in this group had repeat His bundle electrograms performed before discharge and in 3 patients there was evidence of residual atrioventricular nodal dysfunction. Both the hospital and follow-up mortality in this group was low and there was no evidence to suggest that permanent pacing would benefit these patients. Of the 18 patients with bundle-branch block and a normal PR interval, 9 had prolongation of the HV interval, but there was no difference in mortality in patients with normal or prolonged HV intervals. Twenty-two patients with bundle-branch block also developed atrioventricular block. In 5 of these patients the site of the AV block was proximal and in 14 it was distal, while 3 patients had both proximal and distal block. The hospital mortality in those patients who progressed to second- or third-degree atrioventricular block was considerably higher than in those patients who remained in first-degree atrioventricular block.


Cadmium toxicity: A role in bone cell function and teeth development.
Cadmium (Cd) is a widespread environmental contaminant that causes severe bone metabolism disease, such as osteoporosis, osteoarthritis, and osteomalacia. The present review aimed to explore the molecular mechanisms of Cd-induced bone injury starting from bone cell function and teeth development. Cd inhibits the differentiation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts, and directly causes BMSC apoptosis. In the case of osteoporosis, Cd mainly affects the activation of osteoclasts and promotes bone resorption. Cd-induces osteoblast injury and oxidative stress, which causes DNA damage, mitochondrial dysfunction, and endoplasmic reticulum stress, resulting in apoptosis. In addition, the development of osteoarthritis (OA) might be related to Cd-induced chondrocyte damage. The high expression of metallothionein (MT) might reduce Cd toxicity toward osteocytes. The toxicity of Cd toward teeth mainly focuses on enamel development and dental caries. Understanding the effect of Cd on bone cell function and teeth development could contribute to revealing the mechanisms of Cd-induced bone damage. This review explores Cd-induced bone disease from cellular and molecular levels, and provides new directions for removing this heavy metal from the environment.


Evidence-based development of liver allocation: a review.
Liver transplantation has undergone a rapid evolution from a high-risk experimental procedure to a mainstream therapy for thousands of patients with a wide range of hepatic diseases. Its increasing success has been accompanied by progressive imbalance between organ donor supply and the patients who might benefit. Where demand outstrips supply in transplantation, a system of organ allocation is inevitably required to make the wisest use of the available, but scarce, organs. Early attempts to rationally allocate donor livers were particularly hampered by lack of available and suitable data, leading to imperfect solutions that created or exacerbated inequities in the system. The advent and maturation of evidence-based predictors of waiting list mortality risk led to more objective criteria for liver allocation, aided by the increasing availability of data on large numbers of patients. Until now, the vast majority of allocation systems for liver transplantation have relied on estimation of waiting list mortality. Evidence-based allocation systems that incorporate measures of post-transplant outcomes are conceptually attractive and these transplant benefit-based allocation systems have been developed, modeled, and subjected to computer simulation. Future implementations of benefit-based liver allocation await continued refinement and additional debate in the transplant community.


Determination of Penicillium griseofulvum-oriented pyripyropene A, a selective inhibitor of acyl-coenzyme A:cholesterol acyltransferase 2, in mouse plasma using liquid chromatography-tandem mass spectrometry and its application to pharmacokinetic studies.
In this study, we developed a method for the determination of Penicillium griseofulvum-oriented pyripyropene A (PPPA), a selective inhibitor of acyl-coenzyme A:cholesterol acyltransferase 2, in mouse and human plasma and validated it using liquid chromatography-tandem mass spectrometry. Pyripyropene A (PPPA) and an internal standard, carbamazepine, were separated using a Xterra MS C18 column with a mixture of acetonitrile and 0.1% formic acid as the mobile phase. The ion transitions monitored in positive-ion mode [M + H]+ of multiple-reaction monitoring (MRM) were m/z 148.0 from m/z 584.0 for PPPA and m/z 194.0 from m/z 237.0 for the internal standard. The detector response was specific and linear for PPPA at concentrations within the range from 1 to 5,000 ng/mL. The intra-/inter-day precision and accuracy of the method was acceptable by the criteria for assay validation. The matrix effects of PPPA ranged from 97.6 to 104.2% and from 93.3 to 105.3% in post-preparative mouse and human plasma samples, respectively. PPPA was also stable under various processing and/or handling conditions. Finally, PPPA concentrations in the mouse plasma samples could be measured after intravenous, intraperitoneal, or oral administration of PPPA, suggesting that the assay is useful for pharmacokinetic studies on mice and applicable to human studies.


Immunomodulatory effects of dietary protein during Nippostrongylus brasiliensis re-infection in lactating rats.
Periparturient relaxation of immunity (PPRI) to secondary infection with nematodes is believed to have a nutritional basis due to differential partitioning of scarce nutrient resources, particularly protein, to reproductive rather than immune functions. At times of protein scarcity, an increase in protein supply has been reported to assuage this phenomenon. The Nippostrongylus brasiliensis reinfected lactating rat model is now being utilized to investigate the immune reactions underlying the modifying role of dietary protein on PPRI. Herein, we demonstrate that lactating rats reinfected with N. brasiliensis under high protein (HP) dietary conditions exhibit decreased worm burdens and reduced colon egg counts compared to their low protein (LP) counterparts. These reductions correlated with increased mastocytosis and greater goblet cell hyperplasia. Additionally, the local antibody profile revealed that HP reinfected lactating rats developed a stronger antigen specific IgG2b response earlier in infection in comparison with their LP counterparts. Our study provides evidence that increased dietary protein content reduces the PPRI to N. brasiliensis re-infection in the lactating rat through improved mucosal immune responses.


TFAP2 paralogs regulate midfacial development in part through a conserved ALX genetic pathway.
Cranial neural crest development is governed by positional gene regulatory networks (GRNs). Fine-tuning of the GRN components underly facial shape variation, yet how those in the midface are connected and activated remain poorly understood. Here, we show that concerted inactivation of Tfap2a and Tfap2b in the murine neural crest, even during the late migratory phase, results in a midfacial cleft and skeletal abnormalities. Bulk and single-cell RNA-seq profiling reveal that loss of both Tfap2 family members dysregulates numerous midface GRN components involved in midface morphogenesis, patterning, and differentiation. Notably, Alx1/3/4 (Alx) transcript levels are reduced, while ChIP-seq analyses suggest TFAP2 directly and positively regulates Alx gene expression. TFAP2 and ALX co-expression in midfacial neural crest cells of both mouse and zebrafish implies conservation of this regulatory axis across vertebrates. Consistent with this notion, tfap2a zebrafish mutants present abnormal alx3 expression patterns, Tfap2a binds alx loci, and tfap2a-alx3 genetic interactions are observed. Together, these data demonstrate TFAP2 paralogs regulate vertebrate midfacial development in part by activating expression of ALX transcription factor genes.


[The action of a new taurine derivative in different variants of hypoxic states].
A new N-phenylalkyl derivative of taurin was tested. The results show that this compound exhibits a pronounced general antihypoxant activity and is capable of increasing the stability of heart with respect to hypoxia and anoxia, protecting heart against the ischemic damage, and facilitating the adaptation capacity of myocardium.


Neurocognitive improvements after best-practice intervention for chronic fatigue syndrome: Preliminary evidence of divergence between objective indices and subjective perceptions.
Neurocognitive difficulties are commonly reported by patients suffering from chronic fatigue syndrome (CFS). Moderate improvements from 'best practice' therapy are promising, but to date reported efficacy is based entirely on subjective measures. This is problematic, given the well-documented divergence between subjective perceptions and actual neurocognitive performance, including in this patient group. Subjective and objective measures of neurocognitive performance were obtained from 25 patients with well-characterized CFS before and after the completion of a 12-week graded-activity program incorporating a cognitive training component. Additionally, self-reported symptoms, cardiac autonomic activity (a relevant biomarker of stress responsivity), and their relation to neurocognitive improvements were examined. Substantive post-intervention improvements in subjective (p=0.006) and objective (including faster responses speeds and greater accuracy, p's<0.001) neurocognitive performance were documented. Participants also demonstrated reduced autonomic reactivity to the cognitive challenge at follow-up (p's≤0.01). These improvements were accompanied by improvements in symptom ratings (p's≤0.01). However, subjective ratings of neurocognitive difficulties, and CFS-related symptoms were not linked to objective performance improvements. These initial data provide the first evidence of objective neurocognitive performance improvements accompanied by a significant reduction in responsiveness in stress-related neural pathways consequent to cognitive-behavioral/graded exercise therapy programs. These findings provide support for the effectiveness of such programs in remediating clinical status. These promising findings warrant further investigation, including replication in a larger sample utilizing more controlled study designs.


Voluntary exercise alters GABA(A) receptor subunit and glutamic acid decarboxylase-67 gene expression in the rat forebrain.
Voluntary exercise improves stress coping and lowers anxiety. Because of the role of GABA in these processes, we investigated changes in the central GABAergic system in rats with free access to a running wheel for 4 weeks. The control animals had no access to a running wheel. Using insitu hybridisation histochemistry, we studied changes in gene expression of various GABA(A) receptor subunits as well as the GABA-synthesising enzyme glutamic acid decarboxylase-67 (GAD67) in the forebrain. There were region-specific decreases in alpha2, beta3 and gamma2 subunit mRNA expression and region-specific increases in beta1 subunit expression. The alpha5 and delta subunits, in the forebrain specifically associated with extrasynaptic GABA(A) receptors in the hippocampus, showed differential increases in expression levels. Expression of GAD67 mRNA was increased in many forebrain regions including all hippocampal cell layers, peri-paraventricular nucleus, bed nucleus stria terminalis, nucleus accumbens core and motor cortex, suggesting that long-term voluntary exercise enhances forebrain GABA synthesis capacity but in a region-specific manner. Thus, regular performance of exercise results in extensive changes in the forebrain GABAergic system that may be implicated in the changes in stress sensitivity and emotionality observed in exercising subjects.


Relationship Between Ginsenoside Rg3 and Metabolic Syndrome.
Metabolic syndrome is an important public health issue and is associated with a more affluent lifestyle. Many studies of metabolic syndrome have been reported, but its pathogenesis remains unclear and there is no effective treatment. The ability of natural compounds to ameliorate metabolic syndrome is currently under investigation. Unlike synthetic chemicals, such natural products have proven utility in various fields. Recently, ginsenoside extracted from ginseng and ginseng root are representative examples. For example, ginseng is used in dietary supplements and cosmetics. In addition, various studies have reported the effects of ginsenoside on metabolic syndromes such as obesity, diabetes, and hypertension. In this review, we describe the potential of ginsenoside Rg3, a component of ginseng, in the treatment of metabolic syndrome.


PARALYTIC EFFECTS OF "PARALYTIC SHELLFISH POISON" ON FROG NERVE AND MUSCLE.
A purified extract of toxic lamellibranchs, Saxidomus giganteus (Deshayes), containing "paralytic shellfish poison," has been tested for its effects on conduction and contraction in frog nerve and muscle. The poison was very toxic and concentrations within the range 0.025 to 0.1 mug/ml. paralysed isolated muscle preparations, with abolition of the muscle action potential. The poison did not readily penetrate the perineurium, but in desheathed sciatic nerves the conduction of nerve impulses was rapidly blocked by concentrations of 0.05 to 0.1 mug/ml. There was no evidence that the poison had any specific curarizing action at the neuromuscular junction, and the paralysis was not accompanied by any appreciable depolarization of the muscle membrane.


Degradation of Carbonyl Hydroperoxides in the Atmosphere and in Combustion.
Oxygenates with carbonyl and hydroperoxy functional groups are important intermediates that are generated during the autoxidation of organic compounds in the atmosphere and during the autoignition of transport fuels. In the troposphere, the degradation of carbonyl hydroperoxides leads to low-vapor-pressure polyfunctional species that may precipitate in clouds and fog droplets or to the formation of secondary organic aerosols (SOAs). In combustion, the fate of carbonyl hydroperoxides is important for the performance of advanced combustion engines, especially for autoignition. A key fate of the carbonyl hydroperoxides is reaction with OH radicals, for which kinetics data are experimentally unavailable. Here, we study 4-hydroperoxy-2-pentanone (CH3C(═O)CH2CH(OOH)CH3) as a model compound to clarify the kinetics of OH reactions with carbonyl hydroperoxides, in particular H atom abstraction and OH addition reactions. With a combination of electronic structure calculations, we determine previously missing thermochemical data, and with multipath variational transition state theory (MP-VTST), a multidimensional tunneling (MT) approximation, multiple-structure anharmonicity, and torsional potential anharmonicity, we obtained much more accurate rate constants than the ones that can computed by conventional single-structure harmonic transition state theory (TST) and than the empirically estimated rate constants that are currently used in atmospheric and combustion modeling. The roles of various factors in determining the rates are elucidated. The pressure-dependent rate constants for the addition reaction are computed using system-specific quantum RRK theory. The calculated temperature range is 298-2400 K, and the pressure range is 0.01-100 atm. The accurate thermodynamic and kinetics data determined in this work are indispensable in the global modeling of SOAs in atmospheric science and in the detailed understanding and prediction of ignition properties of hydrocarbons and alternative fuels.


Serum interleukin 10 and interleukin 11 in patients with acute pancreatitis.
Proinflammatory and anti-inflammatory cytokines are involved in the pathogenesis of acute pancreatitis. To measure the serial serum levels of interleukin 10 and interleukin 11 in patients with acute pancreatitis and analyse the relation of these anti-inflammatory cytokines to disease severity. In 50 patients with acute pancreatitis, the serum concentrations of interleukin 10 and interleukin 11 were determined on days one, two, three, four, and seven after admission. Serum C reactive protein levels were evaluated on days one and two. Severity of pancreatitis was determined according to the Atlanta criteria. Serum concentrations of interleukin 10 on days one to seven were significantly higher in patients with severe pancreatitis than in those with mild pancreatitis. Patients with severe attacks had significantly elevated serum interleukin 11 concentrations on days two to four compared with those with mild attacks, but not on days one and seven. With cut off levels of 30 pg/ml for interleukin 10, 10.5 pg/ml for interleukin 11, and 115 mg/l for C reactive protein, the accuracy rates for detecting severe pancreatitis were 84%, 64%, and 78% respectively on day one and 82%, 74%, and 84% respectively on day two. Serum interleukin 10 and interleukin 11 concentrations reflect the severity of acute pancreatitis. Interleukin 10 is a useful variable for early prediction of the prognosis of acute pancreatitis.


Brain-Gut Axis: Invasive and Noninvasive Vagus Nerve Stimulation, Limitations, and Potential Therapeutic Approaches.
Inflammatory bowel disease (IBD) is a chronic relapsing condition with no known etiology and is characterized by disrupted gut homeostasis, chronic inflammation, and ulcerative lesions. Although current treatments can reduce disease activity, IBD frequently recurs once treatments are discontinued, indicating that treatments are ineffective in providing long-term remission. The lack of responsiveness and reluctance of some affected persons to take medications because of potential adverse effects has enhanced the need for novel therapeutic approaches. The vagus nerve (VN) is likely important in the pathogenesis of IBD, considering the decreased activity of the parasympathetic nervous system, especially the VN, and the impaired interaction between the enteric nervous system and central nervous system in patients with IBD. Vagus nerve stimulation (VNS) has demonstrated anti-inflammatory effects in various inflammatory disorders, including IBD, by inhibiting the production of inflammatory cytokines by immune cells. It has been suggested that stimulating the vagus nerve to induce its anti-inflammatory effects may be a potential therapeutic approach for IBD. Noninvasive techniques for VNS have been developed. Considering the importance of VN function in the brain-gut axis, VNS is a promising treatment option for IBD. This review discusses the potential therapeutic advantages and drawbacks of VNS, particularly the use of noninvasive transcutaneous auricular vagus nerve stimulation.


New prospectives in the delivery of galantamine for elderly patients using the IntelliDrug intraoral device: in vivo animal studies.
The transbuccal delivery of drugs could assist several categories of chronic, especially elderly, patients in adhering to a correct dosage regimen. In particular, patients suffering from dementia have several difficulties in following the prescribed dosage, in addition to problems associated with swallowing tablets. Galantamine is currently used for treating patients with mild to moderate Alzheimer's-type dementia. The transbuccal delivery of this drug could be an interesting non- invasive and safe administration route. Several studies have been performed in vitro and ex vivo within the framework of a European Commission funded Project (IntelliDrug-FP6), aimed at developing a device which would be fitted for controlled delivery of drugs by an electronic and software-driven system. The primary objective of this study was to evaluate the efficacy of a prototype of the IntelliDrug device in vivo on 6 pigs, following a single Galantamine dose to be delivered through the buccal mucosae, as compared to intravenous drug injection. The secondary objectives were: a) to verify Galantamine bioavailability through buccal delivery; b) to evaluate the permeation enhancement effect of iontophoresis; and, finally, c) to assess any histomorphological changes in the buccal mucosae after transbuccal delivery. The results suggested that transbuccal delivery has the potential to cause long-lasting and controlled blood levels of Galantamine. The latter crosses the entire buccal mucosae, reaching systemic circulation after about 30 minutes and its plasmatic peak approximately 120 minutes after administration. The histological analysis of the buccal mucosae did not reveal any evidence of flogosis or tissue injury. Our results have clearly confirmed that the buccal delivery of Galantamine is a reliable tool with which to overcome the drawbacks associated with the conventional administration route. In general, transbuccal drug delivery has been shown to be an interesting, non-invasive and safe administration route for delivering systemically-acting drugs.


Development and characterization of a novel antiacne niosomal gel of rosmarinic acid.
The antibacterial and anti-inflammatory potential of rosemarinic acid (ROA), a naturally occurring ester of caffeic acid has been well reported. Antibacterial effect of ROA is attributed to nucleoid damage with an increase in spatial division and condensation of genetic material. ROA has been found dynamic against many human pathogenic bacterial strains but its inhibitory prospective has never been established against skin inflammations caused by Propionibacterium acne. The skin surface in acne prone areas is colonized with Staphylococcus aureus and Propionibacterium acnes which contribute to inflammation and acne. Resistance to current antimicrobial therapies suggested the need to explore new antimicrobial agents against acne. Present work included the preparation of ROA-loaded niosomes and their in vitro antimicrobial evaluation against P. acne and S. aureus. This work also included the development of niosomal gel of rosmarinic acid for sustained delivery to bacteria infected cells. Niosomes of rosmarinic acid were formulated by reverse phase evaporation method using different ratio of span 85 and cholesterol. The prepared formulations were evaluated for its vesicle size, entrapment efficiency, in vitro release study and antibacterial activity. In vivo study of developed formulation was conducted on Swiss albino mice in comparison with solution of plain drug and a marketed formulation of benzoyl peroxide. It was evident that niosomes are novel carrier for delivery of naturally occurring antimicrobial agents, in deeper tissues of skin. The results showed that drug-loaded niosomes dispersed in the gelling agent are an effective delivery system for treatment of acne vulgaris.


Soluble factors and the development of rod photoreceptors.
Photoreceptors are the most abundant cell type in the vertebrate neural retina. Like the other retinal neurons and the Müller glia, they arise from a population of precursor cells that are multipotent and intrinsic to the retina. Approximately 10 years ago, several studies demonstrated that retinal precursor cells (RPCs) are competent to respond to environmental factors that promote cell type determination and differentiation. Since those studies, significant effort has been directed at identifying the molecular nature of these environmental signals and understanding the precise mechanisms they employ to drive RPCs towards the different retinal fates. In this review, we describe the recent progress toward understanding how environmental factors influence the development of vertebrate rod photoreceptors.


A computationally efficient approach to the estimation of two- and three-dimensional hidden Markov models.
Statistical modeling methods are becoming indispensable in today's large-scale image analysis. In this paper, we explore a computationally efficient parameter estimation algorithm for two-dimensional (2-D) and three-dimensional (3-D) hidden Markov models (HMMs) and show applications to satellite image segmentation. The proposed parameter estimation algorithm is compared with the first proposed algorithm for 2-D HMMs based on variable state Viterbi. We also propose a 3-D HMM for volume image modeling and apply it to volume image segmentation using a large number of synthetic images with ground truth. Experiments have demonstrated the computational efficiency of the proposed parameter estimation technique for 2-D HMMs and a potential of 3-D HMM as a stochastic modeling tool for volume images.


DNMT3L is a regulator of X chromosome compaction and post-meiotic gene transcription.
Previous studies on the epigenetic regulator DNA methyltransferase 3-Like (DNMT3L), have demonstrated it is an essential regulator of paternal imprinting and early male meiosis. Dnmt3L is also a paternal effect gene, i.e., wild type offspring of heterozygous mutant sires display abnormal phenotypes suggesting the inheritance of aberrant epigenetic marks on the paternal chromosomes. In order to reveal the mechanisms underlying these paternal effects, we have assessed X chromosome meiotic compaction, XY chromosome aneuploidy rates and global transcription in meiotic and haploid germ cells from male mice heterozygous for Dnmt3L. XY bodies from Dnmt3L heterozygous males were significantly longer than those from wild types, and were associated with a three-fold increase in XY bearing sperm. Loss of a Dnmt3L allele resulted in deregulated expression of a large number of both X-linked and autosomal genes within meiotic cells, but more prominently in haploid germ cells. Data demonstrate that similar to embryonic stem cells, DNMT3L is involved in an auto-regulatory loop in germ cells wherein the loss of a Dnmt3L allele resulted in increased transcription from the remaining wild type allele. In contrast, however, within round spermatids, this auto-regulatory loop incorporated the alternative non-coding alternative transcripts. Consistent with the mRNA data, we have localized DNMT3L within spermatids and sperm and shown that the loss of a Dnmt3L allele results in a decreased DNMT3L content within sperm. These data demonstrate previously unrecognised roles for DNMT3L in late meiosis and in the transcriptional regulation of meiotic and post-meiotic germ cells. These data provide a potential mechanism for some cases of human Klinefelter's and Turner's syndromes.


Identifying the essential components of cultural competence in a Chinese nursing context: A qualitative study.
This qualitative study using semi-structured interviews was conducted to identify the essential components of cultural competence from the perspective of Chinese nurses. A purposive sample of 20 nurse experts, including senior clinical nurses, nurse administrators, and educators in transcultural nursing, was recruited. Using thematic analysis, four themes: awareness, attitudes, knowledge, and skills, with two subthemes for each, were identified. Notably, culture in China was understood in a broad way. The participants' responses focused upon demographic attributes, individuality, and efforts to facilitate quality care rather than on the cultural differences of ethnicity and race and developing the capacity to change discrimination or health disparities. A greater understanding of cultural competence in the Chinese nursing context, in which a dominant cultural group exists, is essential to facilitate the provision of culturally competent care to diverse populations.


Ventral striatal network connectivity reflects reward learning and behavior in patients with Parkinson's disease.
A subgroup of Parkinson's disease (PD) patients treated with dopaminergic therapy develop compulsive reward-driven behaviors, which can result in life-altering morbidity. The mesocorticolimbic dopamine network guides reward-motivated behavior; however, its role in this treatment-related behavioral phenotype is incompletely understood. Here, mesocorticolimbic network function in PD patients who develop impulsive and compulsive behaviors (ICB) in response to dopamine agonists was assessed using BOLD fMRI. The tested hypothesis was that network connectivity between the ventral striatum and the limbic cortex is elevated in patients with ICB and that reward-learning proficiency reflects the extent of mesocorticolimbic network connectivity. To evaluate this hypothesis, 3.0T BOLD-fMRI was applied to measure baseline functional connectivity on and off dopamine agonist therapy in age and sex-matched PD patients with (n = 19) or without (n = 18) ICB. An incentive-based task was administered to a subset of patients (n = 20) to quantify positively or negatively reinforced learning. Whole-brain voxelwise analyses and region-of-interest-based mixed linear effects modeling were performed. Elevated ventral striatal connectivity to the anterior cingulate gyrus (P = 0.013), orbitofrontal cortex (P = 0.034), insula (P = 0.044), putamen (P = 0.014), globus pallidus (P < 0.01), and thalamus (P < 0.01) was observed in patients with ICB. A strong trend for elevated amygdala-to-midbrain connectivity was found in ICB patients on dopamine agonist. Ventral striatum-to-subgenual cingulate connectivity correlated with reward learning (P < 0.01), but not with punishment-avoidance learning. These data indicate that PD-ICB patients have elevated network connectivity in the mesocorticolimbic network. Behaviorally, proficient reward-based learning is related to this enhanced limbic and ventral striatal connectivity. Hum Brain Mapp 39:509-521, 2018. © 2017 Wiley Periodicals, Inc.


Overexpression of Acyl-CoA-Binding Protein 1 (ChACBP1) From Saline-Alkali-Tolerant Chlorella sp. Enhances Stress Tolerance in Arabidopsis.
A large proportion of the world's arable land is saline-alkali land, and this is becoming an urgent environmental problem for agriculture. One approach to address this problem is to develop new varieties of stress-resistant plants through genetic engineering. The algae (Chlorella sp.) JB6, which was previously isolated from saline-alkali land, was found to exhibit strong NaHCO3 tolerance. Here, we explored saline-alkali-tolerance genes in this alga that might be useful for producing abiotic stress-resistant transgenic plants. We identified a gene encoding acyl-CoA-binding protein 1 (ACBP1) from JB6 by screening a full-length cDNA library in yeast under NaHCO3 stress. Northern blot analyses showed that the ChACBP1 mRNA levels were significantly up-regulated under abiotic stresses such as salinity, oxidation, heavy metals, and low temperature stresses. The recombinant ChACBP1 protein was found to bind phosphatidylcholine in vitro. Green fluorescent protein-labeled ChACBP1 was localized to the cytosol. Overexpression of ChACBP1 in yeast and Arabidopsis increased their resistance to high salinity, oxidation, heavy metals, and low temperature stresses. These results suggested that ChACBP1 may mediate plant abiotic stress adaptation through phospholipid metabolism. Thus, ChACBP1 may be useful to genetically improve the tolerance of plants to saline-alkali soil.


Squamous cell carcinoma of the maxillary sinus.
Eighty-five patients with squamous cell cancer of the maxillary sinus received all of their treatment at The University of Texas M.D. Anderson Cancer Center between the years 1971 and 1986. Their records were evaluated according to stage, disease at presentation, symptoms and signs at presentation, treatment, and outcome. There were no differences in locoregional control or survival between groups treated with surgery alone vs surgery plus radiotherapy. Careful analysis of the data indicates that there was almost certainly some selection bias for the patients undergoing combination therapy, as most of this group had historically adverse prognostic factors identified. Those patients who underwent radiotherapy alone or chemotherapy presented with either metastatic or locally advanced disease and were treated with palliative intent; therefore, comparison between this group and standard therapy groups was impossible in this retrospective review. Although it is tempting to speculate that combination therapy improved locoregional control and survival in patients with more advanced disease, none of the data presented in this review reach statistical significance. Furthermore, there is no difference in survival in this population compared with a study at this institution 20 years ago. Squamous cell cancer of the maxillary sinus continues to be a challenging neoplasm. Radiotherapy may improve locoregional control and survival in a group of patients with more advanced disease and may have its greatest utility in earlier-stage disease. A multi-institutional prospective trial is needed to find ways to improve outcome in this patient population.


Effects of cotransplanted fetal liver on fetal pancreas isografts.
Ricordi et al. described a hepatotrophic effect mediated by pancreatic islets on cotransplanted hepatocytes. We found a reciprocal salutary effect of fetal liver (FL) on fetal pancreas (FP) in the intramural small bowel (ISB) site. To further investigate this intriguing finding, composite FP/FL isografts were transplanted to the conventional renal subcapsular (RSC) site and the accessible but historically inhospitable intramuscular site in streptozotocin-diabetic Lewis rats. A comparison of recipients of FP/FL and FP alone found the proportion rendered normoglycemic was site dependent. All recipients of either composite FP/FL grafts or FP alone transplanted in the ISB site became normoglycemic. The proportion of normoglycemic recipients was lower in the RSC site (71% FP and 40% FP/FL) and the i.m. site (14% FP and 67% FP/FL). Importantly, regardless of site, normoglycemia was established with an accelerated time course in recipients of FP/FL versus FP alone (24 +/- 8 vs. 67 +/- 43 days; P = 0.001). Normal (or more rapid) glucose clearance after challenge was achieved in all normoglycemic recipients except those transplanted in the RSC site. On histological examination of excised FP/FL grafts, hepatocytes were present in association with islets. Cyclosporine-induced islet toxicity could not be overcome in 6 recipients of FP alone, but 6 of 8 recipients of FP/FL became normoglycemic (P less than 0.01). To assess the effect of FP on hepatocytes, allografts (Wistar donors) of FP or FP/FL were cotransplanted in the ISB of enzyme-deficient jaundiced Gunn rats. Immunosuppression consisted of rapamycin (0.8 mg/kg/day) infused intravenously for 4 weeks. In the FP/FL group (n = 4), the mean serum bilirubin level decreased from 8.6 to 4.9 mg/dl at 6 weeks after transplantation. This was a significant difference as compared with the increased mean serum bilirubin from 6.9 to 7.8 mg/dl (P less than 0.05; paired Student's t test) in recipients of FL alone (n = 4). In conclusion, we found a mutual paracrine effect on islets and hepatocytes transplanted as a composite FP/FL graft. FL hastened the establishment of normoglycemia following transplantation of FP in diabetic rats, and FP enhanced FL transplant function in Gunn rats.


Variation of single particle mid-infrared emission spectrum with particle size.
The emission spectra of single particles of inorganic solids as a function of particle size have been recorded from 6 micro to 11.8 micro. For small optically thin particles, an emission maximum is produced in the reststrahlen region. The emission behavior is dominated by scattering and can be adequately described in terms of Mie absorption efficiency factors. As the particle size is increased, the emission band reverses its polarity, and the spectrum approaches that of a polished plate. The data provide source functions necessary for determining the emission behavior of particulate samples in which temperature gradients exist, such as on the lunar surface. The data are of particular interest for interpreting the spectral behavior of circumstellar silicate particles.


Temporal lobe abnormalities in a patient with schizophrenia who has word-finding difficulty: use of high-resolution magnetic resonance imaging and auditory P300 event-related potentials.
Postmortem, magnetic resonance, and event-related potential studies suggest the presence of temporal lobe abnormalities in schizophrenia. Analyses using convergent measurements of brain structure and function, however, have rarely been done in the same patients. We recently developed a protocol using high-spatial-resolution magnetic resonance scans, auditory P300 event-related potentials, and thought disorder scales to examine temporal lobe structure and function in the same patients. We report a case of schizophrenia that showed left-lateralized volume reduction in the superior temporal gyrus, hippocampus, and parahippocampal gyrus (also on right), with associated P300 amplitude reduction and thought disorder marked by word-finding difficulties and perseverations.


Malignant fibrous histiocytoma in a baboon.
A recurrent, invasive, soft tissue tumor was observed in the subcutaneous tissue of the back of an adult baboon (Papio cynocephalus). The neoplasm was diagnosed as a malignant fibrous histiocytoma.


The first reported cases of human cryptosporidiosis caused by Cryptosporidium hominis in Slovak Republic.
Cryptosporidiosis belongs to the important parasitic infections with zoonotic potential and the occurrence in European countries is rare. The first cases of cryptosporidiosis caused by Cryptosporidium hominis detected in the Slovak republic were described here. Collection of examined humans consisted of five family members. Faecal specimens were examined by formalin sedimentation, by the Sheather's sugar flotation and by immunochromatography and visualised by the Ziehl-Neelsen acid fast stain. A fragment of the Cryptosporidium small subunit ribosomal RNA gene was amplified by nested polymerase chain reaction and species was determined by restriction fragment length polymorphism analysis with the endonucleases SspI and VspI. C. hominis was found in faeces of two immunocompetent siblings (a 7-year-old boy and a 2-year-old girl). The symptoms occurred only in the boy as gastrointestinal disorders lasting 5 days, and manifested by abdominal pain, an elevated body temperature (37.2 °C), mild diarrhoea, accompanied by lassitude, depression and anorexia. Ultrasonic scan revealed enlarged spleen and mezenteric lymph nodes. Microscopic examination of the stool sample revealed numerous Cryptosporidium oocysts. The DNA typing identified C. hominis subtype IbA10G2. Cryptosporidium was also detected in the boy's sister without any complications and symptoms. Their father, mother and grandmother were parasitologically negative. The source of infection remained unknown. Human cases in present study reflect necessity of systematic attention on intestinal parasites diagnostic inclusive of cryptosporidia.


In vitro infection with Theileria parva is associated with IL10 expression in all bovine lymphocyte lineages.
The protozoan parasite Theileria parva infects and transforms bovine lymphocytes, giving rise to a fatal lymphoproliferative condition known as East Coast fever. Although immune cattle mount strong cytolytic T lymphocyte responses to the parasite, naive animals appear unable to respond and develop severe immunopathological lesions. We have investigated the patterns of cytokine mRNA expressed by 19 bulk and cloned parasite-infected lymphoblast cell lines using a multiplex PCR system. Considerable variation was observed in the cytokine profiles of these lines and only IL10 was universally expressed. Investigation of cloned lines representing the major bovine lymphocyte populations failed to reveal a lineage-specific pattern of cytokine mRNA expression that could be associated with infection. Nonetheless, analysis of a CD4+ T cell clone before and after transformation with the parasite indicated that infection does alter the pattern of cytokine expression, with apparent upregulation of IL10. These observations raise the possibility that IL10 derived from infected cells may influence the immune responses of naive cattle to challenge.


The antigenic and molecular alterations of C3 in the fluid phase during an immune reaction in normal human serum. Demonstration of a new conversion product, C3x.
During the reaction of an immune precipitate with fresh human serum, C3 undergoes a number of molecular alterations with the formation of conversion products differing from those obtained when purified components react. Those products which remain in the fluid phase, the subject of the present paper, have been identified by their reaction with monospecific antisera to the three antigenic determinants of C3, A, B, and D, after electrophoresis in agar or polyacrylamide gel. When purified C3 reacts with EAC1,4,2, C3i is found in the fluid phase. C3i, a loose complex of C3a and C3b, is in a conformational state whereby only the A and D antigens, present on its C3b portion, will consume antibody. The B antigen, present on the C3a portion of C3i, is unavailable for combination with antibody until C3i dissociates. In the fluid phase of the reaction of an immune precipitate with whole serum, C3i, C3a, and C3b, formed when purified components react, cannot be found. Instead the end products of the reaction appear to be C3c, which contains the A antigen, and C3d, which contains the D antigen. C3c and C3d are similar to the beta1A and alpha2D produced by the aging of serum but differ in their mobilities in acrylamide gel and in agar. The C3c and C3d generated by an immune precipitate also differ slightly from the C3c and C3d produced by the reaction of trypsin with C3 in whole human serum. As human serum reacts with an immune complex, native C3 appears to undergo a primary alteration before conversion. This alteration results in a molecular species of C3 which is labile at 56 degrees C for 30 min, fails to expose additional A and D antigenic sites upon aging, and which forms beta1A and C3d rather than beta1A and alpha2D during aging. In addition to this altered form of native C3, a new conversion product, C3x, is formed as whole serum reacts with an immune complex. C3x is not found in systems utilizing pure complement components. C3x is like C3 in that it bears all three antigenic determinants but differs in that it has a slightly faster mobility in polyacrylamide gel than does native C3. C3x is not only found in the fluid phase but is also bound to the immune precipitate. Finally, the fluid-phase kinetics of each of the antigens of C3 have been determined as normal human serum reacts with an immune precipitate. These illustrate that nearly the entire population of native C3 molecules undergoes conversion rapidly as manifested by the disappearance of the B antigen from the fluid phase. Moreover, the kinetics of the fluid-phase A and D antigens reflect that the conversion of C3 in serum is quantitatively not the same as when purified C3 reacts with C4,2.


Playgroups offering health and well-being support for families: a systematic review.
This systematic review explores the health and well-being outcomes that families experience as a result of their participation in playgroups that provide health and well-being services. A systematic review of peer-reviewed literature resulted in 12 articles for inclusion. A meta-synthesis method, using reciprocal translation, was implemented to synthesize findings. Findings suggest that programs support children's social development, transition to school, and overall health. For parents, programs provide targeted social and health support, and knowledge sharing and learning opportunities. Future research in this area will benefit from participatory research approaches and exploring how universal approaches to program provision benefit families.


Distribution of High-Volume Ankle Replacement Surgeons in United States Metropolitan Areas.
Ankle osteoarthritis is increasing, thus creating greater demand for high-volume total ankle arthroplasty (TAA) surgeons. The Medicare Provider Utilization and Payment Data Public Use File (MPUPD-PUF) provides volume and reimbursement data for procedures performed by physicians participating in Medicare. This study analyzes surgeon prevalence, surgeon distribution, and factors affecting surgeon prevalence in metropolitan areas. The MPUPD-PUF was reviewed from 2012 to 2015, and data were extracted for physicians performing ≥11 TAA procedures. Physicians in metropolitan areas (population >1 million) were grouped together, and reimbursement, number of high-volume surgeons, and procedures were calculated. Presence of an American Orthopaedic Foot and Ankle Society (AOFAS) fellowship program was analyzed for associations with high-volume TAA surgeons. Fifty-three surgeons performed ≥11 TAA procedures (1,960 total) covered by Medicare. Of these surgeons, 66% practice in metropolitan areas with a population >1 million. Fifty-one percent of US major metropolitan areas contained no surgeon who submitted >10 traditional Medicare claims for TAA. Areas with an AOFAS fellowship had nominally more TAA claims submitted. The distribution of high-volume TAA surgeons among major metropolitan areas in the United States is highly unequal. Analyzing the data with this method aids in targeting TAA surgeons to currently underserved areas. Level IV: Retrospective-comparative study.


[Comparison of ATP-pyrophosphate exchange reactions, catalyzed by two different "one-site" forms of tryptophanyl-tRNA-synthetase and by native enzyme].
Kinetic parameters of the reaction of ATP-PPi exchange, catalysed by "one-site" form of beef pancreas tryptophanyl-tRNA-synthetase are determined. The one-site form was obtained by affinity labelling of one active centre of the enzyme with alkylated analog of tryptophan. The comparison of kinetic parameters of this one-site form with the other one produced earlier by affinity labelling with analog of tryptophanyl-tRNA and the native enzyme was carried out. It was shown that the presence of tRNA improves the discrimination properties of the enzyme and diminishes the concentrations of "dead-end" complexes. Negative cooperativity in binding of two molecules of tryptophan is observed, which disappears in the presence of ATP, leading to an apparent synergistic effect. The molecule of tryptophan, which is present in the active centre, diminishes affinity of pyrophosphate.


Opposing peripheral fates of tissue-restricted self antigen-specific conventional and regulatory CD4+ T cells.
The development of self antigen-specific T cells is influenced by how the self antigen is expressed. Here, we created a mouse in which a model self antigen is conditionally expressed in different tissue environments. Using peptide:MHCII tetramer-based cell enrichment methods, we examined the development of corresponding endogenous self antigen-specific CD4+ T cell populations. While ubiquitous self antigen expression resulted in efficient deletion of self antigen-specific T cells in the thymus, some tissue-restricted expression patterns resulted in partial deletion of the population in peripheral lymphoid organs. Deletion specifically affected Foxp3- conventional T cells (Tconv) with a bias towards high avidity TCR expressing cells in the case of thymic, but not peripheral deletion. In contrast, Foxp3+ Treg exhibited elevated frequencies with increased TCR avidity. T cells surviving deletion were functionally impaired, with Tconv cells exhibiting more impairment than Tregs. Collectively, our results illustrate how postthymic recognition of tissue-restricted self antigens results in opposing developmental fates for Tconv and Treg cell subsets.


Beam sampling: taking samples at the micro-scale.
When using a beam to make a measurement in situ, irrespective of scale, the process implicitly includes the taking of a sample. Therefore, the uncertainty of the measurement result needs to include the uncertainty generated by the sampling process, which is usually dominated by the heterogeneity of the analyte at that scale. Reliable estimates of the uncertainty of beam measurements are essential to judge their fitness-for-purpose (FFP) and hence to enable their rigorous interpretation. This approach can be applied to a wide range of techniques for the analytical assessments of materials, from handheld portable X-ray Fluorescence (pXRF) at the millimeter scale, to Secondary Ion Mass Spectrometry (SIMS) at the micron scale.


Tea consumption enhances endothelial-dependent vasodilation; a meta-analysis.
Tea consumption is associated with a lower risk of cardiovascular disease including stroke. Direct effects of tea components on the vasculature, particularly the endothelium, may partly explain this association. We performed a meta-analysis of controlled human intervention studies on the effect of tea on flow-mediated dilation (FMD) of the brachial artery, a measurement of endothelial function, which is suggested to be associated with cardiovascular risk. Human intervention studies were identified by systematic search of the databases Medline, Embase, Chemical Abstracts and Biosis through March 2009 and by hand-searching related articles. Studies were selected based on predefined criteria: intervention with tea as the sole experimental variable, placebo-controlled design, and no missing data on FMD outcome or its variability. A random effects model was used to calculate the pooled overall effect on FMD due to the intake of tea. The impact of various subject and treatment characteristics was investigated in the presence of heterogeneity. In total, 9 studies from different research groups were included with 15 relevant study arms. The overall absolute increase in FMD of tea vs. placebo was 2.6% of the arterial diameter (95% CI: 1.8-3.3%; P-value <0.001) for a median daily dose of 500 mL of tea (2-3 cups). This is a relative increase of approximately 40% compared to the average FMD of 6.3% measured under placebo or baseline conditions. There was significant heterogeneity between studies (P-value <0.001) that might partly be explained by the cuff position either distal or proximal to the area of FMD measurement. No indication for publication bias was found. Moderate consumption of tea substantially enhances endothelial-dependent vasodilation. This may provide a mechanistic explanation for the reduced risk of cardiovascular events and stroke observed among tea drinkers.


A new deep-reef scorpionfish (Teleostei, Scorpaenidae, Scorpaenodes) from the southern Caribbean with comments on depth distributions and relationships of western Atlantic members of the genus.
A new species of scorpionfish, Scorpaenodes barrybrowni Pitassy & Baldwin, sp. n. which is described, was collected during submersible diving in the southern Caribbean as part of the Smithsonian's Deep Reef Observation Project (DROP). It differs from the other two western Atlantic species of the genus, Scorpaenodes caribbaeus and Scorpaenodes tredecimspinosus, in various features, including its color pattern, having an incomplete lateral line comprising 8-10 pored scales, tending to be more elongate, usually having the 11(th)-12(th) pectoral-fin rays elongate, and by 20-23% divergence in the cytochrome c oxidase I (COI) DNA barcode sequences. It further differs from one or the other of those species in head spination and in numbers of soft dorsal-fin rays, pectoral-fin rays, and precaudal + caudal vertebrae. Inhabiting depths of 95-160 m, the new species is the deepest western Atlantic member of the genus (Scorpaenodes caribbaeus occurs at depths < 35 m and Scorpaenodes tredecimspinosus from 7 to 82 m). DNA barcode data do not rigorously resolve relationships among the ten species of the genus for which those data are available.


[Effect of adaptation to cold and moderate cooling on respiration and oxidative phosphorylation in the muscle mitochondria of albino rats].
In rats, exposed to cooling at 2 degrees within 2 hrs, total gas exchange and the respiratory activity of sceletal muscles mitochondria were studied. The adapted rats, which were kept at 5-7 degrees within 13-17 days, were compared with unadapted animals (kept at 18 degrees). In the group of adapted rats cooling caused the increase in the total O2 utilization by 105% with increased free oxidation in mitochondria and the distinct decrease in P/O ratio. Cooling of unadapted rats was accompanied by the increase in O2 utilization by 85% without any marked alteration in P/O ratio in sceletal muscles mitochondria. The body temperature was not altered.


Successes and failures of built environment interventions: Using concept mapping to assess stakeholder perspectives in four Canadian cities.
Healthy communities can be supported through built environment interventions that redesign cities for improved health outcomes. Understanding the context of these interventions is critical for assessing how an intervention impacts population health; such context is often poorly documented. This study uses concept mapping to capture stakeholders' perspectives on the factors that influence the success and failure of built environment interventions across cities. The INTErventions, Research, and Action in Cities Team (INTERACT) research program undertook concept mapping exercises related to interventions in four Canadian cities: public bike share program (Vancouver); a cycling network (Victoria); a bus rapid transit program (Saskatoon); and interventions related to the Montreal Sustainability Plan (Montreal). Concept mapping synthesised stakeholder perceptions and Go zones were used to prioritise factors based on importance and feasibility. Resulting themes were integrated into implementation science frameworks. Across four cities, 95 stakeholders participated. An average of 38 factors were identified in each city, resulting in 5 emerging clusters in Victoria and Montreal and 6 clusters in Vancouver and Saskatoon. Clusters covered domains of economic and political context, intervention planning, equity considerations, user experience, and stakeholder engagement. Common across all cities was the importance of stakeholder engagement. Concerns for citizen safety were prominent in Victoria, Vancouver, and Saskatoon. Interventions in Saskatoon and Vancouver were related to programs, and reliability of service and ease of use emerged. Go zones highlighted 2-5 items in each city, which can inform priority actions. Our study provides stakeholders' collective thinking on the contextual factors that influence the success and failure of built environment interventions. Organising context within an implementation science framework can provide a common language to synthesise stakeholder perspectives across settings. Go zone items can inform city-specific priority actions to support a common vision around implementing built environment interventions in pursuit of designing equitable and healthy cities.


Bystander Ethics and Good Samaritanism: A Paradox for Learning Health Organizations.
In 2012, a U.S. Institute of Medicine report called for a different approach to health care: "Left unchanged, health care will continue to underperform; cause unnecessary harm; and strain national, state, and family budgets." The answer, they suggested, would be a "continuously learning" health system. Ethicists and researchers urged the creation of "learning health organizations" that would integrate knowledge from patient-care data to continuously improve the quality of care. Our experience with an ongoing research study on atrial fibrillation-a trial known as IMPACT-AFib-gave us some insight into one of the challenges that will have to be dealt with in creating these organizations. Although the proposed educational intervention study placed no restrictions on what providers and health plans could do, the oversight team argued that the ethical principle of beneficence did not allow the researchers to be "bystanders" in relation to a control group receiving suboptimal care. In response, the researchers designed a "workaround" that allowed the project to go forward. We believe the experience suggests that what we call "bystander ethics" will create challenges for the kinds of quality improvement research that LHOs are designed to do.


A Study of Parameters Affecting Atom Probe Tomography Specimen Survivability.
Specimen survivability is a primary concern to those who utilize atom probe tomography (APT) for materials analysis. The state-of-the-art in understanding survivability might best be described as common-sense application of basic physics principles to describe failure mechanisms. For example, APT samples are placed under near-failure mechanical-stress conditions, so reduction in the force required to initiate field evaporation must provide for higher survivability-a common sense explanation of survivability. However, the interplay of various analytical conditions (or instrumentation) and how they influence survivability (e.g., decreasing the applied evaporation field improves survivability), and which factors have more impact than others has not been studied. In this paper, we report on the systematic analysis of a material composed of a silicon-dioxide layer surrounded on two sides by silicon. In total, 261 specimens were fabricated and analyzed under a variety of conditions to correlate statistically significant survivability trends with analysis conditions and other specimen characteristics. The primary result suggests that, while applied field/force plays an obvious role in survivability for this material, the applied field alone does not predict survivability trends for silicon/silicon-dioxide interfaces. The rate at which ions are extracted from the specimen (both in terms of ions-per-pulse and pulse-frequency) has similar importance.


Measles virus nucleotide sequences: detection by hybridization in situ.
A tritium-labeled probe that detects measles virus nucleotide sequences was hybridized in situ to cells infected with measles virus and to sections of brain tissue from patients with subacute sclerosing panencephalitis and from patients with multiple sclerosis. The measles virus genome was detected in many cells in subacute sclerosing panencephalitis where this virus would have been missed by methods such as immunofluorescence. Measles virus sequences were also found in two foci in one of four cases of multiple sclerosis. This refined method of hybridization in situ, which can be useful in the search for covert virus infections of man, provides evidence that viruses may be involved in multiple sclerosis.


Isolation and Characterization of Diesel-Degrading Bacteria from Hydrocarbon-Contaminated Sites, Flower Farms, and Soda Lakes.
Hydrocarbon-derived pollutants are becoming one of the most concerning ecological issues. Thus, there is a need to investigate and develop innovative, low-cost, eco-friendly, and fast techniques to reduce and/or eliminate pollutants using biological agents. The study was conducted to isolate, characterize, and identify potential diesel-degrading bacteria. Samples were collected from flower farms, lakeshores, old aged garages, asphalt, and bitumen soils and spread on selective medium (Bushnell Haas mineral salt agar) containing diesel as the growth substrate. The isolates were characterized based on their growth patterns using optical density measurement, biochemical tests, and gravimetric analysis and identified using the Biolog database and 16S rRNA gene sequencing techniques. Subsequently, six diesel degraders were identified and belong to Pseudomonas, Providencia, Roseomonas, Stenotrophomonas, Achromobacter, and Bacillus. Among these, based on gravimetric analysis, the three potent isolates AAUW23, AAUG11, and AAUG36 achieved 84%, 83.4%, and 83% diesel degradation efficiency, respectively, in 15 days. Consequently, the partial 16S rRNA gene sequences revealed that the two most potent bacterial strains (AAUW23 and AAUG11) were Pseudomonas aeruginosa, while AAUG36 was Bacillus subtilis. This study demonstrated that bacterial species isolated from hydrocarbon-contaminated and/or uncontaminated environments could be optimized to be used as potential bioremediation agents for diesel removal.


Dietary reference intake (DRI) value for dietary polyphenols: are we heading in the right direction?
Dietary Reference Intake (DRI) values exist for vitamins and minerals, and provide a guideline on the optimal dose range to avoid deficiency and prevent toxicity. Polyphenols are widely distributed in plant foods, and have been linked to improved human health through reduced risk of chronic diseases, especially cardiovascular. Although they do not cause classical deficiencies, recently they have been discussed as 'lifespan essentials because they are needed to achieve a full lifespan by reducing the risk of a range of chronic diseases. A recent meta analysis shows promising actions of polyphenols from cocoa, soya and tea on flow mediated dilation, blood pressure and LDL cholesterol. Many epidemiological studies support the action of polyphenols or polyphenol-rich foods on health, but there are still many gaps in our knowledge. More adequately powered, randomised, placebo controlled human studies are needed on polyphenols. There is a large number of structurally different polyphenols which are relevant for health, and obtaining enough information to set a DRI for each of these will not be feasible in the foreseeable future. A new approach is needed, and a new way of thinking, which would apply not only to polyphenols but also to other phytochemicals. Today, a target intake value of polyphenols as 'lifespan essentials' needs to be based on the amount of polyphenols in '5-a-day'. We are heading in the right direction towards a DRI, but bioavailability and dose-effects, including toxic levels, need to be established before DRIs can be considered.


Tamoxifen Sensitizes Acute Lymphoblastic Leukemia Cells to Cannabidiol by Targeting Cyclophilin-D and Altering Mitochondrial Ca2+ Homeostasis.
Cytotoxic effects of cannabidiol (CBD) and tamoxifen (TAM) have been observed in several cancer types. We have recently shown that CBD primarily targets mitochondria, inducing a stable mitochondrial permeability transition pore (mPTP) and, consequently, the death of acute lymphoblastic leukemia (T-ALL) cells. Mitochondria have also been documented among cellular targets for the TAM action. In the present study we have demonstrated a synergistic cytotoxic effect of TAM and CBD against T-ALL cells. By measuring the mitochondrial membrane potential (ΔΨm), mitochondrial calcium ([Ca2+]m) and protein-ligand docking analysis we determined that TAM targets cyclophilin D (CypD) to inhibit mPTP formation. This results in a sustained [Ca2+]m overload upon the consequent CBD administration. Thus, TAM acting on CypD sensitizes T-ALL to mitocans such as CBD by altering the mitochondrial Ca2+ homeostasis.


The influence of extracellular matrix composition on the differentiation of neuronal subtypes in tissue engineered innervated intestinal smooth muscle sheets.
Differentiation of enteric neural stem cells into several appropriate neural phenotypes is crucial while considering transplantation as a cellular therapy to treat enteric neuropathies. We describe the formation of tissue engineered innervated sheets, where intestinal smooth muscle and enteric neuronal progenitor cells are brought into close association in extracellular matrix (ECM) based microenvironments. Uniaxial alignment of constituent smooth muscle cells was achieved by substrate microtopography. The smooth muscle component of the tissue engineered sheets maintained a contractile phenotype irrespective of the ECM composition, and generated equivalent contractions in response to potassium chloride stimulation, similar to native intestinal tissue. We provided enteric neuronal progenitor cells with permissive ECM-based compositional and viscoelastic cues to generate excitatory and inhibitory neuronal subtypes. In the presence of the smooth muscle cells, the enteric neuronal progenitor cells differentiated to functionally innervate the smooth muscle. The differentiation of specific neuronal subtypes was influenced by the ECM microenvironment, namely combinations of collagen I, collagen IV, laminin and/or heparan sulfate. The physiology of differentiated neurons within tissue engineered sheets was evaluated. Sheets with composite collagen and laminin had the most similar patterns of Acetylcholine-induced contraction to native intestinal tissue, corresponding to an increased protein expression of choline acetyltransferase. An enriched nitrergic neuronal population, evidenced by an increased expression of neuronal nitric oxide synthase, was obtained in tissue engineered sheets that included collagen IV. These sheets had a significantly increased magnitude of electrical field stimulated relaxation, sensitive maximally to nitric oxide synthase inhibition. Tissue engineered sheets containing laminin and/or heparan sulfate had a balanced expression of contractile and relaxant motor neurons. Our studies demonstrated that neuronal subtype was modulated by varying ECM composition. This observation could be utilized to derive enriched populations of specific enteric neurons in vitro prior to transplantation.


Occlusion of 22Na+ and 86Rb+ in membrane-bound and soluble protomeric alpha beta-units of Na,K-ATPase.
In this work, we examined occlusion of 22Na+ and 86Rb+ in membranous and detergent-solubilized Na,K-ATPase from outer renal medulla. Optimum conditions for occlusion of 22Na+ were provided by formation of the phosphorylated complex from the beta,gamma-bidentate complex of chromium (III) with ATP (CrATP). Release of occluded cations occurred at equally slow rates in soluble and membrane-bound Na,K-ATPase. Values of 22Na+ occlusion as high as 11 nmol/mg of protein were measured, corresponding to 1.8-2.7 mol of Na+/mol of phosphorylated Na,K-ATPase as determined by 32P incorporation from [gamma-32P]CrATP. Maximum capacity for phosphorylation from [gamma-32P]CrATP was 6 nmol/mg of protein and equal to capacities for binding of [48V]vanadate and [3H]ouabain. The stoichiometry for occlusion of Rb+ was close to 2 Rb+ ions/phosphorylation site. In an analytical ultracentrifuge, the soluble Na+- or Rb+-occluded complexes showed sedimentation velocities (S20,w = 6.8-7.4) consistent with monomeric alpha beta-units. The data show that soluble monomeric alpha beta-units of Na,K-ATPase can occlude Rb+ or Na+ with the same stoichiometry as the membrane-bound enzyme. The structural basis for occlusion of cations in Na,K-ATPase is suggested to be the formation of a cavity inside a monomeric alpha beta-unit constituting the minimum protein unit required for active Na,K-transport.


Immune complex-driven neutrophil activation and BAFF release: a link to B cell responses in SLE.
The role of neutrophils in driving pathogenic B cell responses in SLE is not fully understood. In this study, we explored the link between immune complex (IC)-driven neutrophil activation, the release of B cell pro-survival factor BAFF and B cell activation using SLE clinical samples. BAFF levels were analysed in serum samples from patients with SLE (n=60) and healthy controls (HCs, n=20) by ELISA and correlated with markers of neutrophil activation and circulating IC levels. Neutrophils were stimulated with RNP/IgG ICs and neutrophil activation, the release of BAFF, and neutrophil-mediated B cell responses were studied in vitro. Levels of BAFF in patients with SLE were associated with markers of disease activity, including anti-dsDNA antibody titres (r=0.33, p<0.05), serum C3 levels (r=-0.57, p<0.001) and levels of circulating ICs (r=0.39, p<0.05). Stimulation of neutrophils from healthy individuals with RNP-ICs in vitro induced the release of BAFF (p<0.05), concomitant with formation of neutrophil extracellular traps (NETs) (p<0.05). In culture, neutrophils promoted B cell survival (p<0.05), proliferation (p<0.05) and CD27hiCD38hi plasmablast differentiation. Our results support a new mechanism by which ICs, on NET formation, induce the release of B cell pro-survival factor BAFF by neutrophils. Furthermore, neutrophils directly promoted B cell activation and cell differentiation. Targeting neutrophil-B cell interactions can be further explored as an approach for inhibiting pathogenic B cell responses in SLE.


Re-programming immunosurveillance in persistent non-infectious ocular inflammation.
Ocular function depends on a high level of anatomical integrity. This is threatened by inflammation, which alters the local tissue over short and long time-scales. Uveitis due to autoimmune disease, especially when it involves the retina, leads to persistent changes in how the eye interacts with the immune system. The normal pattern of immune surveillance, which for immune privileged tissues is limited, is re-programmed. Many cell types, that are not usually present in the eye, become detectable. There are changes in the tissue homeostasis and integrity. In both human disease and mouse models, in the most extreme cases, immunopathological findings consistent with development of ectopic lymphoid-like structures and disrupted angiogenesis accompany severely impaired eye function. Understanding how the ocular environment is shaped by persistent inflammation is crucial to developing novel approaches to treatment.