High glucose induces alteration of gap junction permeability and phosphorylation of connexin-43 in cultured aortic smooth muscle cells.
Gap junction is thought to have a crucial role in maintaining tissue homeostasis. We examined the effect of a high glucose level on gap junctional intercellular communication (GJIC) activity in cultured vascular smooth muscle cells (VSMCs) using the fluorescent dye transfer method. After a 48-h incubation with 22 mmol/l glucose (high glucose level), GJIC activity of VSMCs was significantly reduced compared with incubation with 5.5 mmol/l glucose (normal glucose level) (P < 0.05). Treatment of the cells with 12-O-tetradecanoylphorbol-13-acetate (TPA; 5 x 10(-8) mol/l), a protein kinase C (PKC) activator, for 1 h also reduced GJIC activity (P < 0.01). In addition, treatment of the cells with calphostin C, a specific PKC inhibitor, for 3 h completely restored the GJIC activity inhibited by the high glucose level. Western blot analysis showed that connexin 43 (Cx43), which is the major functional protein of gap junction, is present in multiphosphorylated forms: a nonphosphorylated form (P0) and phosphorylated forms (P1, P2, and P3). Incubation of VSMCs with a high glucose level significantly increased the density ratio of P3/P0 compared with a normal glucose level (P < 0.05). Similarly, treatment of the cells with TPA significantly increased the P3/P0 ratio compared with controls (P < 0.01). In addition, the increase in the P3/P0 density ratio induced by a high glucose level was restored to the control level by both staurosporine and calphostin C. These results suggest that the high glucose level induced the inhibition of GJIC activity in cultured VSMCs through excessive phosphorylation of Cx43, mediated by PKC activation. This may contribute to the development of the macroangiopathy associated with diabetes.


Synthesis and self-assembly of temperature-responsive copolymers based on N-vinylpyrrolidone and triethylene glycol methacrylate.
Polyvinylpyrrolidone (PVP) is a biocompatible, water-soluble polymer with unique physicochemical properties and attractive biological features that has found widespread use in several industries. Owing to advances in controlled polymerisation techniques, PVP can be easily synthesised with robust control over its architecture. However, the synthesis of PVP copolymers, which can allow tailoring of its properties and expand the scope of this polymeric material, is challenging and rarely reported. Here, we demonstrate the synthesis of well-defined, temperature-responsive polyvinylpyrrolidone-co-poly(triethylene glycol methacrylate) (PVP-co-pTEGMA) block copolymers via successive Reversible Addition-Fragmentation chain Transfer (RAFT) and Activators ReGenerated by Electron Transfer Atom Transfer Radical Polymerisation (ARGET-ATRP) techniques. We show that PVP-co-pTEGMA block copolymers display temperature-responsive behaviour and self-assemble above their cloud point temperature (Tcp) to form spherical nanostructures of 100-200 nm in diameter. Finally, we demonstrate stabilisation of these assemblies below their Tcp by cross-linking through the PVP block.


Digital micromirror device as a spatial illuminator for fluorescence lifetime and hyperspectral imaging.
Time-domain fluorescence lifetime imaging (FLIM) and hyper-spectral imaging (HSI) are two advanced microscopy techniques widely used in biological studies. Typically both FLIM and HSI are performed with either a whole-field or raster-scanning approach, which often prove to be technically complex and expensive, requiring the user to accept a compromise among precision, speed, and spatial resolution. We propose the use of a digital micromirror device (DMD) as a spatial illuminator for time-domain FLIM and HSI with a laser diode excitation source. The rather unique features of the DMD allow both random and parallel access to regions of interest (ROIs) on the sample, in a very rapid and repeatable fashion. As a consequence both spectral and lifetime images can be acquired with a precision normally associated with single-point systems but with a high degree of flexibility in their spatial construction. In addition, the DMD system offers a very efficient way of implementing a global analysis approach for FLIM, where average fluorescence decay parameters are first acquired for a ROI and then used as initial estimates in determining their spatial distribution within the ROI. Experimental results obtained on phantoms employing fluorescent dyes clearly show how the DMD method supports both spectral and temporal separation for target identification in HSI and FLIM, respectively.


Residual biliary tract stones: nonoperative treatment of 570 patients.
A nonperative method has been employed over a period of 11 years in the treatment of 570 patients with residual biliary duct lithiasis. A successful instrumental extraction was accomplished in 543 of these patients (95.3%). These extractions were performed via the transcholedochal and the transcholecystic routes, the latter being used both for choledochal and gallbladder stones. The high percentage of successes, the extremely few complications, and the good long-term results turn this method into a definitely useful adjunct to the conventional therapy for biliary pathology.


Influence of epoietinum therapy on the oxidative stress in haemodialysis patients.
The causes of oxidative stress in haemodialysis (HD) patients are still controversial. Beside the uraemic state and dialysis-related factors, adjuvant drug therapies such as epoietinum (rHuEpo) and intravenous iron were involved. Several parameters related to oxidative stress were assessed by spectrophotometry in stable HD patients, treated for at least 2 months with epoietinum (n = 14; mean dose = 97.7 +/- 19.1 U/kg/week) or not (n = 15), none of them on iron therapy, and in 13 controls. Plasma thiobarbituric acid-reactive substances (TBARS) were used as markers of reactive species generation. Erythrocyte and plasma antioxidant systems, reflected by non-protein erythrocyte thiols, and erythrocyte enzyme activities -- superoxide dismutase (SOD), glutathione peroxidase, catalase and plasma total thiols, respectively -- were also investigated. There were no differences between HD subgroups regarding haemoglobin levels. Plasma TBARS was increased in all HD patients as opposed to controls, irrespective of rHuEpo therapy. In addition, no change in antioxidant status parameters between rHuEpo-treated and -untreated patients was observed. Except for SOD, the other antioxidant indices were higher in all HD patients versus controls. These results suggest that (1) chronic HD patients appear to have simultaneously enhanced reactive species generation and antioxidative systems efficiency, and (2) epoietinum therapy did not change their oxidative status, at least in the absence of concomitant iron supplementation and at similar haemoglobin levels.


Syphilis-Associated Proteinuria and Hepatitis in the Setting of Human Immunodeficiency Virus (HIV) Co-Infection.
Syphilis has long been known as "the great imitator", mimicking a wide variety of diseases, and often its diagnosis is delayed or missed. It remains an important public health issue that continues to occur at high rates among patients with HIV. We report a case of a 52-year-old man who presented with a constellation of unusual symptoms highlighting that syphilis should be included in the differential diagnosis in patients with HIV presenting with abnormal liver enzymes, rash, proteinuria, conjunctivitis, and/or sexual risk factors.


Effect of para-chlorophenylalanine on male rats: histopathological and biochemical changes in the testes.
Spermatogenically active testes of rat challenged by 100 mg/kg body weight of p- Chlorophenylalanine for 45 days displayed marked and drastic changes in the seminiferous epithelium. Degenerative changes followed by immense necrosis of germ cells lead to complete breakdown of seminiferous tubules. Leydig cells, however, remained unaffected histologically in the treated animals. Among the accessory sex organs, epididymis alone showed a marked decrease in its weight. A biochemical study in the drug treated rats revealed a significant accumulation of glycogen in the testes accompanied by increase in the activities of enzymes like the succinic dehydrogenase, glucose-6-phosphatase, ATP-ase and acid phosphatases. However, a marked decrease was noticed in the activities of enzymes like alkaline phosphatase, phosphohexose isomerase and lactate dehydrogenase. No significant change was found in the protein, DNA and RNA concentrations in the drug treated testes. The histological and biochemical changes induced in the testes by p-CPA suggest the deleterious effect of the drug on the seminiferous tubules of the testes.


An effective and safe supplement for stem cells expansion ex vivo: cord blood serum.
Mesenchymal stem cells (MSCs) are potential and optimal stem cells in clinical cell therapy, and fetal bovine serum (FBS) is widely used for expansion of MSCs, despite the risks of infectious disease transmission and immunological reaction of the xenogenic origin. This study was designed to compare human four blood group cord blood serum (CBS) with FBS in culturing human placenta-derived mesenchymal stem cells (hPDMSCs), which were derived from four blood group donors. The expansion medium included: 10% FBS, 10% A-CBS, 10% B-CBS, 10% O-CBS, and 10% AB-CBS. Cumulative population doubling, generation time, fold expansion rates and differentiation capacity, cell cycle, and immunophenotype were also assessed. The results showed that fold expansion rate and cumulative population doubling of hPDMSCs significantly increased during long-term MSC expansion in CBS medium, but the generation time decreased compared with FBS. CBS might be an effective supplement for stem cells expand rapidly ex vivo. Cell cycle and differentiation assays showed that most of the hPDMSCs expanding in the presence of CBS were in stationary phase, which was the characteristic of stem cells, and they retained their ability to differentiate into chondrogenic and endothelial cells. By comparing these four blood groups of CBS, we found that there was no significant difference among different blood groups in culturing hPDMSCs, which were isolated from different blood group donors. So CBS may be an optimal replacement to avoid the risks of FBS application in expansion of stem cell for clinical cell therapy and tissue engineering.


Scapulothoracic Dissociation: Evaluation and Management.
Scapulothoracic dissociation is a rare, potentially limb- and life-threatening injury of the shoulder girdle. The injury is characterized by lateral displacement of the scapula resulting from traumatic disruption of the scapulothoracic articulation. The typical physical examination findings consist of substantial swelling of the shoulder girdle, along with weakness, numbness, and pulselessness in the ipsilateral upper extremity. Radiographic evaluation includes measurement of the scapular index on a nonrotated chest radiograph and assessment for either a distracted clavicle fracture or a disrupted acromioclavicular or sternoclavicular joint. Although vascular injury occurs in most patients, emergent surgery is performed only in patients with either limb-threatening ischemia or active arterial hemorrhage. Management of neurologic injury can be delayed if necessary. The location and severity of neurologic injury determine whether observation, nerve grafting, nerve transfer, or above-elbow amputation is performed. Skeletal stabilization procedures include plate fixation of clavicle fractures and reduction of distracted acromioclavicular or sternoclavicular joints. The extent of neurologic injury determines clinical outcomes. Medical Outcomes Study 36-Item Short Form scores are significantly lower in patients with complete brachial plexus avulsion injury than in patients with postganglionic injury.


Early home-based intervention in the Netherlands for children at familial risk of dyslexia.
Dutch children at higher familial risk of reading disability received a home-based intervention programme before formal reading instruction started to investigate whether this would reduce the risk of dyslexia. The experimental group (n=23) received a specific training in phoneme awareness and letter knowledge. A control group (n=25) received a non-specific training in morphology, syntax, and vocabulary. Both interventions were designed to take 10 min a day, 5 days a week for 10 weeks. Most parents were sufficiently able to work with the programme properly. At post-test the experimental group had gained more on phoneme awareness than the control group. The control group gained more on one of the morphology measures. On average, these specific training results did not lead to significant group differences in first-grade reading and spelling measures. However, fewer experimental children scored below 10th percentile on word recognition.


The occurrence of home and personal care products in the Haihe River catchment and estimation of human exposure.
A sub-catchment of the Haihe River basin goes through the Beijing-Tianjin region with a population of 26 million, therefore, the use and release of home and personal care product ingredients (HPCPs) to the river catchment could be potentially substantial. Many HPCPs have been shown to be toxic to human and animals. So, it is essential to know the exposure level of HPCPs in the river basin. The average concentrations of five preservatives, three disinfectants and an antioxidant were found to be 398, 352 and 77.7 ng L-1, respectively, in the dry season. The chemical concentrations in the effluents of wastewater treatment plants (WWTPs) and untreated wastewater discharge were respectively ca. 1.3-2.2 and 1.6-7.5 times higher than those in river water. The mass flux of ∑HPCPs has been estimated to be 8.7 g/h at the outfall of the Shahe Reservoir and 181 g/h and 214 g/h at the estuary of the Haihe River and Yongdingxin River to Bohai Bay, respectively. The attenuation of ∑HPCPs was over 79% along the Wenyu River. By using the backward method, the estimated average loadings to WWTPs ranged from 0.51 to 2.0 mg/day/cap for the various individual compounds. They were 1-3 orders of magnitude higher than the estimation from the forward calculation for parabens. This indicates the possible underestimation of chemical usage and human exposure levels by the current published studies or the probably additional industrial release to the target catchment. Such a study provides useful information for the development of chemical management approaches and indicates that further research is needed to improve the estimation of HPCPs usage and emissions to aquatic environment.


MIDAS: Regionally linear multivariate discriminative statistical mapping.
Statistical parametric maps formed via voxel-wise mass-univariate tests, such as the general linear model, are commonly used to test hypotheses about regionally specific effects in neuroimaging cross-sectional studies where each subject is represented by a single image. Despite being informative, these techniques remain limited as they ignore multivariate relationships in the data. Most importantly, the commonly employed local Gaussian smoothing, which is important for accounting for registration errors and making the data follow Gaussian distributions, is usually chosen in an ad hoc fashion. Thus, it is often suboptimal for the task of detecting group differences and correlations with non-imaging variables. Information mapping techniques, such as searchlight, which use pattern classifiers to exploit multivariate information and obtain more powerful statistical maps, have become increasingly popular in recent years. However, existing methods may lead to important interpretation errors in practice (i.e., misidentifying a cluster as informative, or failing to detect truly informative voxels), while often being computationally expensive. To address these issues, we introduce a novel efficient multivariate statistical framework for cross-sectional studies, termed MIDAS, seeking highly sensitive and specific voxel-wise brain maps, while leveraging the power of regional discriminant analysis. In MIDAS, locally linear discriminative learning is applied to estimate the pattern that best discriminates between two groups, or predicts a variable of interest. This pattern is equivalent to local filtering by an optimal kernel whose coefficients are the weights of the linear discriminant. By composing information from all neighborhoods that contain a given voxel, MIDAS produces a statistic that collectively reflects the contribution of the voxel to the regional classifiers as well as the discriminative power of the classifiers. Critically, MIDAS efficiently assesses the statistical significance of the derived statistic by analytically approximating its null distribution without the need for computationally expensive permutation tests. The proposed framework was extensively validated using simulated atrophy in structural magnetic resonance imaging (MRI) and further tested using data from a task-based functional MRI study as well as a structural MRI study of cognitive performance. The performance of the proposed framework was evaluated against standard voxel-wise general linear models and other information mapping methods. The experimental results showed that MIDAS achieves relatively higher sensitivity and specificity in detecting group differences. Together, our results demonstrate the potential of the proposed approach to efficiently map effects of interest in both structural and functional data.


Adhesion and the cell cycle in cultured L929 and CHO cells.
The adhesiveness of L929 and CHO-K1 cells was monitored throughout their respective cell cycles. The cell cycle stages were identified by a combination of measuring, histochemical and autoradiographic techniques. Adhesiveness was shown to be maximal during the S and late G2 phases, and minimal at mitosis. S cells adhered selectively to other S cells, while late G2 phase cells adhered selectively to other late G2 cells. Mixing of the two cell lines, L929 and CHO-K1, resulted in the S phase cells adhering to each other irrespective of the cell line to which they belonged. The phases of increased adhesiveness and selectivity coincided with a decrease in cell surface negative charge, corresponding to well documented changes in the morphology of the cells.


[Review of psychopharmacological treatments in adolescents and adults with autistic disorders].
Autism is an early developmental disorder. It leads to severe and durable disturbances. Given this problem, no treatment can be excluded a priori. Thus, many approaches are used to deal with autistic disorders. In France, pharmacological treatments are, for instance, largely and mostly used in adults. In the USA, these treatments concern 50% of persons with autism of any age. Nevertheless, they are rarely based on controlled studies. At the present, however, prescriptions and expected effects appear to be hard to localize. Furthermore, only few controlled studies validate their use. Aim - We offer a review of studies about medical treatments used in adolescents and adults with autism. They are classified in 3 categories: the first (category I) includes drugs used for their neurochemical effects focusing on autistic signs. The second (category II) covers drugs used for treatment of behavioural disorders frequently associated with autism. The third (category III) corresponds to a wide range of drugs or vitamins for wich only few case studies exist reporting irregular positive effects. The main hypothesis of this review is that autism involves a dysfunction of the neuromediation systems. This hypothesis opens new perspectives in the research of medical treatments in autism by focusing on molecules, which are supposed to have an effect on neuromediation systems. Method - Our review is based on studies, which have been published during the past twenty years. For many studies, data are limited to adolescents and adults. So we expanded our review to data available in children. The data bases that we have used are medline and psyclit. Keywords have been chosen according to: pharmacological considerations (psychotropic, psychoactive drugs, psychopharmacology) and clinical symptoms (autism, automutilations, aggressive behavior, and hyperactivity). Hypothesis of a dysfunction in the neuromediation systems in autism - Many studies exist about biochemical abnormalities in autism. As in schizophrenia and mental retardation, dysfunctions of the neuromediation systems are considered to be etiological factors. In 30% of people with autism the most regular dysfunction is the increase of serotonine. This led to the serotoninergic hypothesis in autism and to the use of active drugs in the serotonine system. However, the presence of other neurometabolic abnormalities also motivates the use of drugs, supposed to be active in other neuromediation systems. Pharmacological treatments in autism - Category I section sign 1 Active drugs in the dopamine system. Haloperidol (Dopamine antagonist): The effects of this molecule have been broadly studied in autism. Results indicate high efficiency in some symptoms of autism (lack in social behaviour, stereotypical behaviour) and in behavioural impairments that may be associated with autism (aggressive behaviour, hyperactivity). Its side effects, particulary the risk of late dyskinesy, make atypical antipsychotics preferable because of their lower risks. Risperidone (Dopamine and serotonine antagonist): Among several studies only few have been controlled. They indicate that Risperidone has positive effects on the behaviour and is quite well tolerated. section sign 2 Active drugs in the serotonine system. Clomipramine: after promising results, the medium-term efficiency has decreased and severe side effects have limited its use. Fluvoxamine, Fluoxétine, Sertraline (Specific serotonine drugs): Their efficiency has been mainly tested through open studies and their results are contrasted. In some cases, social behaviours have improved and aggressiveness and stereotyped behaviours have decreased. Fenfluramine: At present, this drug is removed from the market. Yet, some studies have suggested that it improves behavioural disturbances as well as performances in autism. section sign 3 Active drugs in the opiate system. Naltrexone: Several controlled studies have indicated an improvement in social and aggressive behaviours. Nevertheless, these studies have used small size sample and have not been replicated. Category II. This category correspond to drugs supposed to be active on neurochemical disturbances found in autism but their target symptoms are not autism specific signs as defined by the ICD 10. Buspirone: This serotonine agonist may have a good impact on emotional disorders and sleeping confusions. Methylphenidate: Most of the current studies about this noradrenergic drug concern children. The results are variable. Paradoxical effects may exist in children with severe mental retardation. Propanolol: Some isolated studies habe reported its efficiency on behavioural disturbances. Clonidine: This adrenergic drug treats efficiently some cases of aggressive behaviour and hyperactivity. Category III. This category contains a wide range of drugs, vitamins or method used in autism after sporadic observations of their positive effects. Secretine: An important improvement has been reported in isolated cases. However, controlled studies in children do not confirm these results. Vitamines B6, B12 and Magnesium: An improvement in socialization and in behavioural disorders have been reported in some cases, but these results are not yet confirmed. Lithium, Carbamazépine, Valproate: Results of some case studies have found it to be efficient in cyclic disorders. Gluten and casein free diet: An improvement of social behaviour have been reported by some parents after these diets. No controlled study has validated this observation. Conclusion - There is no consensus on the use of psychopharmacological treatments in autism. Although there exist many clinical observations, only few controlled studies have validated the efficiency and safety of these treatments. At the present time and until having sufficient studies, drugs are generally limited to severe disorders, for which usual psycho-educational approaches are insufficient.


Consolidating and Upscaling Molecular Research Capacity in Nigeria: On Who's Account?
Molecular research and researchers engage in studies that seek to understand the structures, functions, and interactions of biomolecules as the basis for cellular and systemic effects in living organisms. This research approach was made possible by considerable technological advancements that equip researchers with tools to view biomolecules. Although molecular research holds great promises for improving lives and living, the technological requirements and equipment to undertake molecular research are quite expensive, often requiring a heavy start-up capital or investment. In developing countries such as Nigeria, where the majority of the population lives below the poverty line and research funding is abysmally low, such heavy investments into research that do not provide immediate solutions to societal problems are difficult. This is mostly due to limited resources available to tackle many urgent and pressing needs, and limited perspective and understanding of policymakers, leading to infrastructural and skilled personnel deficit to support molecular research. Despite all these, the field of molecular research continues to grow exponentially globally, hence, funding and investments into this critical life science research area have become imperative. With the rich biodiversity of humans, animals, and plants in Nigeria, and the huge burden of infectious diseases in the country or region, global advances in genomics and proteomics studies will be incomplete without adequate contribution from Nigeria and sub-Saharan Africa region. This paper examines the progression and challenges of undertaking molecular research in Nigeria, and how Nigerian molecular research scientists are tackling these issues, with recommendations for improved molecular research capacity and output in the country or region.


[The value of serum ferritin, serum iron and iron-binding capacity in the differential diagnosis of microcytic hypochromic anemia].
Ferritin, iron, total iron binding capacity and transferrin saturation were measured in the serum of 247 patients with microcytic hypochromic anemia. Differentiation into various categories of microcytic hypochromic anemia was based on clinical criteria and on the response to iron treatment. This produced 147 patients with iron deficiency anemia, 35 patients with anemia secondary to infection, 27 patients with anemia due to tumor and 38 patients with thalassemia. Analysis of the iron parameters revealed the reliability of both serum ferritin and transferrin saturation in distinguishing between the various forms of anemia. However, measurement of serum ferritin is slightly more reliable and much less expensive than determination of transferrin saturation.


Defocused mode in depressed mood and its changes in time-frequency attention-related beta.
Depressed mood has been proposed to possibly possess a unique mode of defocused attention. However, this argument needs to be supported by experimental evidence based on attentional performance. The present study used a perceptual load paradigm, combining factors of perceptual load, distractor-target compatibility, and eccentricity, to investigate the degree of attentional distraction in depressed mood. In addition, the mode of attentional distraction associated with depressed mood was explored with the time-frequency features of electroencephalography (EEG). The behavioral results showed that the high depressed mood (HD) group had significantly higher attentional distraction than the low depressed mood (LD) group. EEG results showed that 1) the beta power (especially beta-2, 18-30 Hz) of the two groups differed in the medio-late part of the attentional distraction, with significantly lower power in the HD group than in the LD group; 2) the results of the correlation between beta-2 power and depression scores revealed a significant negative correlation. These results imply that beta-2 is a potential marker that may be sensitive to depressed mood during attentional processing, which was further supported by the classification results of the support vector machine (SVM) with 80.65% accuracy between the HD and LD groups.


Ezetimibe Lipid-Lowering Trial on Prevention of Atherosclerotic Cardiovascular Disease in 75 or Older (EWTOPIA 75): A Randomized, Controlled Trial.
Evidence regarding the primary prevention of coronary artery disease events by low-density lipoprotein cholesterol (LDL-C) lowering therapy in older individuals, aged ≥75 years, is insufficient. This trial tested whether LDL-C-lowering therapy with ezetimibe is useful for the primary prevention of cardiovascular events in older patients. This multicenter, prospective, randomized, open-label, blinded end-point evaluation conducted at 363 medical institutions in Japan examined the preventive efficacy of ezetimibe for patients aged ≥75 years, with elevated LDL-C without history of coronary artery disease. Patients, who all received dietary counseling, were randomly assigned (1:1) to receive ezetimibe (10 mg once daily) versus usual care with randomization stratified by site, age, sex, and baseline LDL-C. The primary outcome was a composite of sudden cardiac death, myocardial infarction, coronary revascularization, or stroke. Overall, 3796 patients were enrolled between May 2009 and December 2014, and 1898 each were randomly assigned to ezetimibe versus control. Median follow-up was 4.1 years. After exclusion of 182 ezetimibe patients and 203 control patients because of lack of appropriate informed consent and other protocol violations, 1716 (90.4%) and 1695 (89.3%) patients were included in the primary analysis, respectively. Ezetimibe reduced the incidence of the primary outcome (hazard ratio [HR], 0.66; 95% CI, 0.50-0.86; P=0.002). Regarding the secondary outcomes, the incidences of composite cardiac events (HR, 0.60; 95% CI, 0.37-0.98; P=0.039) and coronary revascularization (HR, 0.38; 95% CI, 0.18-0.79; P=0.007) were lower in the ezetimibe group than in the control group; however, there was no difference in the incidence of stroke, all-cause mortality, or adverse events between trial groups. LDL-C-lowering therapy with ezetimibe prevented cardiovascular events, suggesting the importance of LDL-C lowering for primary prevention in individuals aged ≥75 years with elevated LDL-C. Given the open-label nature of the trial, its premature termination and issues with follow-up, the magnitude of benefit observed should be interpreted with caution. Clinical Registration: URL: https://www.umin.ac.jp. Unique identifier: UMIN000001988.


[Oncogenes RET/PTC and mechanisms of their involvement in thyroid cancerogenesis].
Papillary thyroid carcinomas are the most common type of thyroid oncopathology, and are rather often associated with the expression of RET/PTC oncogens. The first oncogen RET/PTC1 was isolated more than 20 years ago. Now 13 different forms of RET/PTC are known, and 12 different partner-genes are described, that could be involved in formation of RET/PTC oncogenes. The most common of them are RET/PTC1 and RET/PTC3 forms. The great majority of oncogens RET/PTC, except for two--ELKS-RET and HOOK3-RET, have been founded in radioaction-induced thyroid tumors. There is an opinion that the key role in development of papillary thyroid carcinomas belongs to RET/PTC oncogens. The data about different types of RET/PTC oncogens, factors, that lead to their formation have been described in the present review. Also different mechanisms of activation of transduction pathways and gene's expression in thyroid cells after RET/PTC induction have been presented.


Hyperbaric oxygen is still needed in the management and prevention of mandibular necrosis: a response to Mr. Richard Clarke.
Mr. Richard Clarke presents in this Journal his arguments against continued application of hyperbaric oxygen (HBO2) therapy to the pre-extraction neoadjuvant treatment or the treatment of frank mandibular ORN. In the same article he advocates a promising renewed interest in HBO2 as a radiosensitizer. Arguments against HBO2 prior to extractions are based on several papers which consistently include low-risk patients. The just-released HOPON trial reports a negative pre-extraction outcome for HBO2, but patients were enrolled with radiation doses as low as 50Gy. For advanced mandibular necrosis (Marx Stage III) requiring resection, fibular free flap reconstruction is advocated. A high complication rate with free flaps is acknowledged but the magnitude of these complications is not discussed. A cost savings for this procedure is suggested, but no mention is made of the typical cost of the procedure ($90,000) or the requirement of a typical one-week hospital stay, including an initial one or two days in the ICU. Nor is mention made of the very low rate of subsequent dental rehabilitation. The success reported by Delainian, et al. employing pentoxifylline, Vitamin E and sometimes a bisphosphonate is equated to the four decades of HBO2 success with the Marx protocol for Stage I and II ORN. In the phase II trial by Delainian (not randomized) six of her 54 patients died secondary to sepsis, and she graded patients as complete responders if 5mm or less bone was exposed. Even at entry patients had an average of only 1.7 cm exposed bone and treatment was prolonged (16 + or -9 months). Any cost comparison studies will have to account for the indirect expenses of this prolonged treatment including lost productivity.


The Depth-Depended Fungal Diversity and Non-depth-Depended Aroma Profiles of Pit Mud for Strong-Flavor Baijiu.
Microorganisms in pit mud are the essential factor determining the style of strong flavor Baijiu. The spatial distribution characteristics of fungal communities and aroma in the pit mud for strong flavor Baijiu from Xinjiang, China, were investigated using Illumina MiSeq high-throughput sequencing and electronic nose technology. A total of 138 fungal genera affiliated with 10 fungal phyla were identified from 27 pit mud samples; of these, Saccharomycopsis, Aspergillus, and Apiotrichum were the core fungal communities, and Aspergillus and Apiotrichum were the hubs that maintain the structural stability of fungal communities in pit mud. The fungal richness and diversity, as well as aroma of pit mud, showed no significant spatial heterogeneity, but divergences in pit mud at different depths were mainly in pH, total acid, and high abundance fungi. Moisture, NH4 +, and lactate were the main physicochemical factors involved in the maintenance of fungal stability and quality in pit mud, whereas pH had only a weak effect on fungi in pit mud. In addition, the fungal communities of pit mud were not significantly associated with the aroma. The results of this study provide a foundation for exploring the functional microorganisms and dissecting the brewing mechanism of strong flavor Baijiu in Xinjiang, and also contributes to the improvement of pit mud quality by bioaugmentation and controlling environmental physicochemical factors.


Dose-Response Associations of Physical Activity and Sitting Time with All-Cause Mortality in Older Japanese Adults.
Although examining the dose-response curves of physical activity (PA) and sitting time with health-related outcomes is an important research agenda, the results for older Japanese adults are extremely limited. We examined the dose-response associations of PA and sitting time with all-cause mortality among older Japanese. Initially, 8,069 non-disabled residents (4,073 men; 3,996 women) aged 65-84 years of Ota City, Japan, were recruited. Moderate-to-vigorous PA (MVPA) and sitting time were evaluated using the International Physical Activity Questionnaire-Short Form. Multivariate-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) of MVPA and sitting time for all-cause mortality were calculated, and the dose-response curves were examined using restricted cubic splines (RCS). During 4.1 years of follow-up, 458 participants (5.7%; 331 men and 127 women) died. Compared with the low MVPA (<600 metabolic equivalents [METs]·minutes/week) group, HR for mortality gradually reduced in moderate (600-3000 METs·minutes/week) and high (>3000 METs·minutes/week) MVPA groups (moderate: HR 0.66; 95% CI, 0.54-0.82; high: HR 0.58; 95% CI, 0.45-0.75; P <0.001 for trend). RCS showed that the HR for mortality reduced linearly up to approximately 2000 METs·minutes/week of MVPA, and maximal risk reduction was seen at approximately 3000-4500 METs·minutes/week of MVPA. No significant dose-response association of sitting time with mortality was observed. Higher MVPA levels reduced all-cause mortality risk, in a significant inverse non-linear dose-response manner. Sitting time was not significantly associated with all-cause mortality. It is important to disseminate the significance of even a slight increase in the MVPA.


The Genetic Evolution of Metastasis.
Cancer is an evolutionary process that is characterized by the emergence of multiple genetically distinct populations or clones within the primary tumor. Intratumor heterogeneity provides a substrate for the selection of adaptive clones, such as those that lead to metastasis. Comparative molecular studies of primary tumors and metastases have identified distinct genomic features associated with the development of metastases. In this review, we discuss how these insights could inform clinical decision-making and uncover rational antimetastasis treatment strategies.


[Fluoride treatment: a good choice in osteoporosis].
Fluoride changes the composition of the hydroxyapatite crystal, reducing the solubility of the bone crystals. Fluoride also stimulates the production of DNA and new osteoblasts, which results in formation of new bone. The therapeutical range of fluoride is narrow. With monofluorophosphate in a dose of 76 mg twice daily positive effect on bone mass is obtained and measurement of serum fluoride concentration is unnecessary. For the treatment of osteoporosis the best result could be obtained with a combined treatment with monofluorophosphate, stimulating osteoblasts and production of new bone, and a bisphosphonate, which simultaneously inhibits osteoclasts and bone resorption. Using this combined treatment only few side effects and no lower extremity pain syndrome were observed. Patients > 65 years of age and younger patients showed similar positive effects on bone mass.


Membrana shrapnelli of maturing rats. The occurrence of mast cells, and histamine content in relation to otitis media with effusion.
The membrana shrapnelli or pars flaccida of 12 day old rats is thick (84 micron) and almost completely fills the lateral attic compartment. Histologically it is characterized by a thickened layer of stratified squamous epithelium and a lamina propria whereas an inner epithelial lining is difficult to discern. Mast cells are numerous and distributed throughout the lamina propria. At day 21 and day 30 the thickness of the pars flaccida is reduced and histologically it resembles that of the adult. Mast cells are mainly located in the vicinity of blood vessels and in a layer just beneath the squamous epithelium. The number of mast cells per unit area of pars flaccida is considerably higher in young rats as compared to adults. At day 12 their number is roughly 2.1 times that of the adult and at day 21 and 30 about 1.5 times. Except for a slightly raised level of histamine at day 30 the concentration of histamine in the pars flaccida of young rats did not differ from values obtained in the adult. Previous experimental studies have indicated that mast cells in the pars flaccida may play an important role in the pathogenesis of OME. It is interesting to note that the pars flaccida of young rats contains more mast cells than that of the adult. Should this also be the case in the pars flaccida of young humans it is tempting to suggest that mast cells may play a role in the pathogenesis of OME in children too.


CXCR4 expression in the bone marrow microenvironment is required for hematopoietic stem and progenitor cell maintenance and early hematopoietic regeneration after myeloablation.
The bone marrow (BM) microenvironment/niche plays a key role in regulating hematopoietic stem and progenitor cell (HSPC) activities; however, mechanisms regulating niche cell function are not well understood. In this study, we show that niche intrinsic expression of the CXCR4 chemokine receptor critically regulates HSPC maintenance during steady state, and promotes early hematopoietic regeneration after myeloablative irradiation. At steady state, chimeric mice with wild-type (WT) HSPC and marrow stroma that lack CXCR4 show decreased HSPC quiescence, and their repopulation capacity was markedly reduced. Mesenchymal stromal cells (MSC) were significantly reduced in the BM of CXCR4 deficient mice, which was accompanied by decreased levels of the HSPC supporting factors stromal cell-derived factor-1 (SDF-1) and stem cell factor (SCF). CXCR4 also plays a crucial role in survival and restoration of BM stromal cells after myeloablative irradiation, where the loss of BM stromal cells was more severe in CXCR4-deficient mice compared to WT mice. In addition, transplantation of WT donor HSPC into CXCR4-deficient recipient mice demonstrated reduced HSPC homing and early hematopoietic reconstitution. We found that CXCR4 signaling attenuates irradiation-induced BM stromal cell loss by upregulating the expression of the antiapoptotic protein Survivin via the PI3K pathway. Our study suggests that SDF-1-CXCR4 signaling in the stromal microenvironment cells plays a crucial role in maintenance of HSPCs during homeostasis, and promotes niche regeneration and early hematopoietic reconstitution after transplantation. Modulation of CXCR4 signaling in the HSPC microenvironment could be a means to enhance hematopoietic recovery after clinical hematopoietic cell transplantation.


Incidence of pin track infections in children's fractures treated with Kirschner wire fixation.
Smooth Kirschner wires (K-wires) are frequently used in pediatric orthopaedics; however, the infection rate associated with these devices is not known. A retrospective study on infection rates in pediatric patients who had fractures treated with percutaneous smooth K-wire fixation was performed. A total of 202 patients with an average age of 9.7 years were included in the study. Upper extremity fractures made up 93% of the cases. The overall infection rate was 7.9%. The classification system of Green was used to describe major and minor infections. There were 16 infections noted; 12 (5.9%) of these were minor, and 4 (2%) were major. Infection rates did not correlate with length of fixation. There was also no correlation of increased infection rates with open fractures or fractures that required open reduction. The infection rates associated with the use of smooth K-wires in pediatric fracture patients is low. The major infection rate is only 2% and does not correlate to length of fixation or type of fracture.


Differential responses of sensory neurones innervating glycolytic and oxidative muscle to protons and capsaicin.
Activation of thin fibre muscle afferent nerves by metabolic by-products plays a critical role in the initiation and maintenance of the autonomic response to exercise and the metabolic profile of active muscle can influence the response. The purpose of this report was to determine the responsiveness of sensory neurones innervating muscles comprising predominantly glycolytic and oxidative fibres to protons and capsaicin using whole-cell patch clamp methods. Dorsal root ganglion (DRG) neurones from 4- to 6-week-old rats were labelled by injecting the fluorescence tracer DiI into the muscle 3-5 days prior to the recording experiments. The percentage of the DRG neurones innervating glycolytic and oxidative muscle was similar in response to both protons and capsaicin. However, the neurones innervating glycolytic muscle had greater inward current amplitude responses to protons and capsaicin as compared with oxidative muscle. The peak current amplitudes to pH 6.0 were 0.84 +/- 0.06 nA (oxidative muscle) versus 1.36 +/- 0.07 nA (glycolytic muscle, P < 0.05). The capsaicin-induced current amplitudes were 2.3 +/- 0.15 nA (oxidative muscle) versus 3.1 +/- 0.21 nA (glycolytic muscle, P < 0.05). Of neurones that responded to pH 6.0 with a sustained current, 88% also responded to capsaicin. Capsaicin exposure enhanced the proton responsiveness in the neurones innervating the muscle; and protons also increased the capsaicin response. These data suggest that (1) receptors mediating protons and capsaicin responses coexist in the DRG neurones innervating muscle; (2) the responsiveness of acidosis and capsaicin can be sensitized by each other; and (3) DRG neurones with nerve endings in a glycolytic muscle developed greater inward current responses to protons and capsaicin than did those with nerve endings in an oxidative muscle.


Differences in the mechanism of collagen lattice contraction by myofibroblasts and smooth muscle cells.
Both rat derived vascular smooth muscle cells (SMC) and human myofibroblasts contain α smooth muscle actin (SMA), but they utilize different mechanisms to contract populated collagen lattices (PCLs). The difference is in how the cells generate the force that contracts the lattices. Human dermal fibroblasts transform into myofibroblasts, expressing α-SMA within stress fibers, when cultured in lattices that remain attached to the surface of a tissue culture dish. When attached lattices are populated with rat derived vascular SMC, the cells retain their vascular SMC phenotype. Comparing the contraction of attached PCLs when they are released from the culture dish on day 4 shows that lattices populated with rat vascular SMC contract less than those populated with human myofibroblast. PCL contraction was evaluated in the presence of vanadate and genistein, which modify protein tyrosine phosphorylation, and ML-7 and Y-27632, which modify myosin ATPase activity. Genistein and ML-7 had no affect upon either myofibroblast or vascular SMC-PCL contraction, demonstrating that neither protein tyrosine kinase nor myosin light chain kinase was involved. Vanadate inhibited myofibroblast-PCL contraction, consistent with a role for protein tyrosine phosphatase activity with myofibroblast-generated forces. Y-27632 inhibited both SMC and myofibroblast PCL contraction, consistent with a central role of myosin light chain phosphatase.


Proteinuria precedes podocyte abnormalities inLamb2-/- mice, implicating the glomerular basement membrane as an albumin barrier.
Primary defects in either podocytes or the glomerular basement membrane (GBM) cause proteinuria, a fact that complicates defining the barrier to albumin. Laminin beta2 (LAMB2) is a GBM component required for proper functioning of the glomerular filtration barrier. To investigate the GBM's role in glomerular filtration, we characterized GBM and overlying podocyte architecture in relation to development and progression of proteinuria in Lamb2-/- mice, which model Pierson syndrome, a rare congenital nephrotic syndrome. We found ectopic deposition of several laminins and mislocalization of anionic sites in the GBM, which together suggest that the Lamb2-/- GBM is severely disorganized, although it is ultrastructurally intact. Importantly, albuminuria was detectable shortly after birth and preceded podocyte foot process effacement and loss of slit diaphragms by at least 7 days. Expression and localization of slit diaphragm and foot process-associated proteins appeared normal at early stages. GBM permeability to the electron-dense tracer ferritin was dramatically elevated in Lamb2-/- mice, even before widespread foot process effacement. Increased ferritin permeability was not observed in nephrotic CD2-associated protein-null (Cd2ap-/-) mice, which have a primary podocyte defect. Together these data show that the GBM serves as a barrier to protein in vivo and that the glomerular slit diaphragm alone is not sufficient to prevent the passage of albumin into the urinary space.


Preclinical characteristics of the hepatitis C virus NS3/4A protease inhibitor ITMN-191 (R7227).
Future treatments for chronic hepatitis C virus (HCV) infection are likely to include agents that target viral components directly. Here, the preclinical characteristics of ITMN-191, a peptidomimetic inhibitor of the NS3/4A protease of HCV, are described. ITMN-191 inhibited a reference genotype 1 NS3/4A protein in a time-dependent fashion, a hallmark of an inhibitor with a two-step binding mechanism and a low dissociation rate. Under preequilibrium conditions, 290 pM ITMN-191 half-maximally inhibited the reference NS3/4A protease, but a 35,000-fold-higher concentration did not appreciably inhibit a panel of 79 proteases, ion channels, transporters, and cell surface receptors. Subnanomolar biochemical potency was maintained against NS3/4A derived from HCV genotypes 4, 5, and 6, while single-digit nanomolar potency was observed against NS3/4A from genotypes 2b and 3a. Dilution of a preformed enzyme inhibitor complex indicated ITMN-191 remained bound to and inhibited NS3/4A for more than 5 h after its initial association. In cell-based potency assays, half-maximal reduction of genotype 1b HCV replicon RNA was afforded by 1.8 nM; 45 nM eliminated the HCV replicon from cells. Peginterferon alfa-2a displayed a significant degree of antiviral synergy with ITMN-191 and reduced the concentration of ITMN-191 required for HCV replicon elimination. A 30-mg/kg of body weight oral dose administered to rats or monkeys yielded liver concentrations 12 h after dosing that exceeded the ITMN-191 concentration required to eliminate replicon RNA from cells. These preclinical characteristics compare favorably to those of other inhibitors of NS3/4A in clinical development and therefore support the clinical investigation of ITMN-191 for the treatment of chronic hepatitis C.


Contributions of Johann jacob Huber to the surface anatomy of the spinal cord and meninges.
From prehistoric times, man has been aware that injury to the spine may result in paralysis of the limbs; this is reflected in bas-relief figures found at Nineweh in ancient Mesopotamia, in a hunting scene that depicts a lioness wounded by King Ashurbanipal. The Edwin Smith papyrus gives many case illustrations of spinal cord injury resulting in paralysis, yet early physicians were unaware of the anatomy of the spinal cord. Galen performed prospective studies in animals by sectioning the spinal cord at varying levels and observing the commensurate paralysis and sensory loss. Real advances in the understanding of spinal cord anatomy did not occur until human cadaveric dissections were undertaken; even then, the knowledge of the anatomy of the spinal cord lagged behind that of other body structures. Johann Jacob Huber appears to be the first anatomist to focus on the spinal cord almost exclusively. His descriptions, and especially his illustrations that depict spinal cord surface anatomy, are impressive with regard to their accuracy and their sense of photorealism. Indeed, his illustrations seem to compare well with the anatomic drawings in contemporary anatomic texts. Yet, we were unable to find a single article in the entire English-language literature depicting his illustrations. We conclude that the description and anatomic illustrations by Johann Jacob Huber remain a hidden gem in the history of human spinal anatomy.


Hypoglycemia in non-critically ill, hospitalized patients with diabetes: evaluation, prevention, and management.
Hypoglycemia among hospitalized patients with diabetes is a common problem. Of the > 8 million patients admitted to US hospitals annually with a diagnosis of diabetes, up to 25% may have a low blood glucose level during hospitalization. As a widely recognized cause of acute, potentially fatal events, hypoglycemia remains a significant barrier to optimal inpatient glycemic control. Although iatrogenic hypoglycemia is associated with adverse outcomes, it may be a marker for illness rather than causal in itself. Several factors, such as administration of exogenous insulin, mismatch of insulin administration with nutrition, and the loss of normal counterregulatory responses, place patients with diabetes at higher risk for hypoglycemia than patients without diabetes. Causes and predictors of hypoglycemia in hospitalized patients with diabetes are discussed. Careful attention to contributing factors, responsiveness to changes in clinical status, and specific institutional protocols and policies can reduce the risk of hypoglycemia. Use of subcutaneous basal-bolus insulin dosing consistent with national guidelines and correction rather than sliding-scale insulin may minimize both hyper- and hypoglycemia. A majority of the literature on inpatient hypoglycemia has been limited to the critical-care setting. This review therefore focuses on hypoglycemia among non-critically ill inpatients with diabetes.


Upconversion nanoparticles in biological labeling, imaging, and therapy.
Upconversion refers to non-linear optical processes that convert two or more low-energy pump photons to a higher-energy output photon. After being recognized in the mid-1960s, upconversion has attracted significant research interest for its applications in optical devices such as infrared quantum counter detectors and compact solid-state lasers. Over the past decade, upconversion has become more prominent in biological sciences as the preparation of high-quality lanthanide-doped nanoparticles has become increasingly routine. Owing to their small physical dimensions and biocompatibility, upconversion nanoparticles can be easily coupled to proteins or other biological macromolecular systems and used in a variety of assay formats ranging from bio-detection to cancer therapy. In addition, intense visible emission from these nanoparticles under near-infrared excitation, which is less harmful to biological samples and has greater sample penetration depths than conventional ultraviolet excitation, enhances their prospects as luminescent stains in bio-imaging. In this article, we review recent developments in optical biolabeling and bio-imaging involving upconversion nanoparticles, simultaneously bringing to the forefront the desirable characteristics, strengths and weaknesses of these luminescent nanomaterials.


LncRNA LINK-A regulates ROCK1 expression in early-stage pancreatic adenocarcinoma.
Long intergenic non-coding RNA for kinase activation (LINK-A) has been characterized as an oncogenic long non-coding (lnc)RNA in triple-negative breast cancer and ovarian carcinoma, but its involvement in other malignancies remains elusive. In the present study, it was determined that the plasma levels of LINK-A lncRNA and Rho-associated protein kinase 1 (ROCK1) were significantly increased in patients with pancreatic adenocarcinoma compared with those in healthy controls. The plasma levels of LINK-A lncRNA were positively correlated with the plasma levels of ROCK1 in pancreatic adenocarcinoma patients, but not in healthy controls. Silencing of LINK-A led to inhibition of pancreatic adenocarcinoma cell proliferation, migration and invasion. Simultaneous overexpression of ROCK1 attenuated the inhibitory effect of LINK-A silencing on cancer cell proliferation, migration and invasion. Overexpression of LINK-A lncRNA led to upregulation of ROCK1 expression, while overexpression of ROCK1 had no significant effect on LINK-A lncRNA expression. It may therefore be concluded that LINK-A lncRNA may have a role in pancreatic adenocarcinoma, at least in part, by promoting ROCK1 expression.


Biological assessment of a calcium silicate incorporated hydroxyapatite-gelatin nanocomposite: a comparison to decellularized bone matrix.
Our laboratory utilized biomimicry to develop a synthetic bone scaffold based on hydroxyapatite-gelatin-calcium silicate (HGCS). Here, we evaluated the potential of HGCS scaffold in bone formation in vivo using the rat calvarial critical-sized defect (CSD). Twelve Sprague-Dawley rats were randomized to four groups: control (defect only), decellularized bone matrix (DECBM), and HGCS with and without multipotent adult progenitor cells (MAPCs). DECBM was prepared by removing all the cells using SDS and NH4OH. After 12 weeks, the CSD specimens were harvested to evaluate radiographical, histological, and histomorphometrical outcomes. The in vitro osteogenic effects of the materials were studied by focal adhesion, MTS, and alizarin red. Micro-CT analysis indicated that the DECBM and the HGCS scaffold groups developed greater radiopaque areas than the other groups. Bone regeneration, assessed using histological analysis and fluorochrome labeling, was the highest in the HGCS scaffold seeded with MAPCs. The DECBM group showed limited osteoinductivity, causing a gap between the implant and host tissue. The group grafted with HGCS+MAPCs resulting in twice as much new bone formation seems to indicate a role for effective bone regeneration. In conclusion, the novel HGCS scaffold could improve bone regeneration and is a promising carrier for stem cell-mediated bone regeneration.


Inhibition of adjuvant-induced arthritis by nasal administration of novel synthetic peptides from heat shock protein 65.
Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease mediated by T cells. The aim of the present study was to investigate the therapeutic efficacy of synthetic peptides (HP-R1, HP-R2 and HP-R3), derived from the sequence of 65-kD mycobacterial heat shock protein (HSP), in the treatment of RA using adjuvant-induced arthritis (AA) animal model. AA was induced by a single intradermal injection Freund's complete adjuvant in male Lewis rats. At the first clinical sign of disease, rats were administered nasally by micropipette of peptides or phosphate buffer saline (PBS). Disease progression was monitored by measurement of body weight, arthritis score and paw swelling. The changes of histopathology were assessed by hematoxylin eosin staining. The serum levels of tumor necrosis factor (TNF) - alpha and interleukin (IL)-4 were measured by enzyme-linked immunosorbent assay (ELISA). The peptides efficiently inhibited the footpad swelling and arthritic symptoms in AA rats. The synthetic peptides displayed significantly less inflammatory cellular infiltration and synovium hyperplasia than model controls. This effect was associated with a suppression of pro-inflammatory cytokine TNF-alpha production and an increase of anti-inflammatory cytokine IL-4 production after peptides treatment. These results suggest that the synthetic peptides derived from HSP65 induce highly effective protection against AA, which is mediated in part by down-regulation of inflammatory cytokines, and support the view that the synthetic peptides is a potential therapy for RA that may help to diminish both joint inflammation and destruction.


Adverse events and safety issues associated with physical activity and exercise for adults with osteoporosis and osteopenia: A systematic review of observational studies and an updated review of interventional studies.
We conducted a systematic review to identify adverse effects of physical activity and/or exercise for adults with osteoporosis/osteopenia. We synthesised evidence from observational studies, and updated three previously published systematic reviews. We searched MEDLINE, EMBASE, CINAHL, Web of Science, grey literature and reference lists of relevant studies. Selection criteria were: (1) observational studies in patients with osteoporosis/osteopenia; and (2) in accordance with the criteria used in the previous reviews. A narrative synthesis was conducted for the observational data. Random effects meta-analysis was undertaken for the review updates. For the observational synthesis 14 studies were included. The majority of studies reported no adverse events, reduced incidence/improvement, or no significant change after physical activity or exercise. Activities that involved spinal flexion (certain yoga moves and sit-ups) were associated with a greater risk of vertebral fractures but these events were rare. For the update of reviews, 57 additional studies were identified. Exercise was generally associated with a greater number of minor adverse events including mild muscle/joint pain. Serious adverse events were rare and could not be attributed to the intervention. Patients with osteoporosis/osteopenia can safely participate in structured exercise programmes, whether at home or in supervised facilities. Systematic review registration for observational studies: PROSPERO 2017: CRD42017070551.


Complement system on the attack in autoimmunity.
The antiphospholipid syndrome is characterized clinically by fetal loss and thrombosis and serologically by the presence of autoantibodies to lipid-binding proteins. In a model of this procoagulant condition in which these antibodies are injected into pregnant mice, fetal loss was prevented by blocking of complement activation. Specifically, interaction of complement component 5a (C5a) with its receptor is necessary for thrombosis of placental vasculature. Inhibition of complement activation may have a therapeutic role in this disease.


Echinacoside rescues the SHSY5Y neuronal cells from TNFalpha-induced apoptosis.
We investigated the neuroprotective effect of echinacoside, one of the phenylethanoids isolated from the stems of Cistanches salsa, a Chinese herbal medicine, on tumor necrosis factor-alpha (TNFalpha)-induced apoptosis in human neuroblastoma (SHSY5Y) cells. Treatment of cultured SHSY5Y cells with TNFalpha 100 ng ml(-1) for 36 h stimulated apoptosis, as demonstrated by typical morphological changes, cell viability, DNA laddering, annexin-V binding, intracellular reactive oxygen species, mitochondrial membrane potential and caspase-3 activity. However, simultaneous treatment with echinacoside (1, 10 or 100 microg ml(-1)) attenuated the TNFalpha-mediated apoptosis. The antiapoptotic action of echinacoside was partially dependent on antioxidative stress effects, maintenance of mitochondria function, inhibition of caspase-3 activity and was also associated with increasing the expression of the antiapoptotic protein Bcl2. Thus, echinacoside has the neuroprotective capacity to antagonize TNFalpha-induced apoptosis in SHSY5Y cells and may be useful in treating some neurodegenerative diseases.


Cardiac PET-CT.
Integrated positron emission tomography computed tomography (PET/CT) scanners allow a true integration of the structure and function of the heart. Myocardial perfusion PET provides a high sensitivity (91%) and specificity (89%) for the diagnosis of obstructive coronary artery disease (CAD). But, as with single photon emission CT, relative perfusion PET often uncovers only the territory subtended by the most severe coronary stenosis, leading to underestimation of the extent of CAD. In contrast, quantitative PET provides a noninvasive assessment of myocardial blood flow and coronary flow reserve and improves detection of preclinical and multivessel coronary atherosclerosis. Similarly, CT coronary angiography is an accurate means to image the entire continuum of anatomic coronary atherosclerosis from nonobstructive to obstructive CAD. However, not all coronary stenoses are hemodynamically significant and <50% of the patients with obstructive CAD on CT angiography demonstrate stress induced perfusion defects. Stress PET data complement the anatomic information on the CT angiogram by providing instant readings about the ischemic burden of coronary stenoses. Thus, combined PET/CT may be potentially superior to CT angiography alone for the guiding revascularization decisions. Further, fusion of the PET and CT angiogram images allows identification of the culprit stenosis in patients presenting with chest pain. Finally, the advances in molecular imaging and image fusion may soon make noninvasive detection of vulnerable coronary plaques a clinical reality. In summary, integrated PET/CT is a powerful new noninvasive modality that offers the potential for refined diagnosis and management of the entire spectrum of coronary atherosclerosis.


Acute disease episodes in a Wuchereria bancrofti-endemic area of Papua New Guinea.
Acute disease episodes of Bancroftian filariasis were monitored prospectively in a rural area of Papua New Guinea. The frequency and duration of episodes were recorded for the leg, arm, scrotum, and breast. A very high incidence of acute disease was observed; 0.31 episodes per person-year in the leg alone. Incidence generally increased with age, except in the breast, where episodes were concentrated in the reproductive age range. Males had slightly higher incidence than females in the leg and arm. Chronic disease was strongly associated with acute disease incidence in all locations. Microfilaremia had a statistically significant association with acute disease in the leg, arm, and breast, but not the scrotum. This study again demonstrates the high burden of acute manifestations of lymphatic filariasis, and provides new information on risk factors, which may lead to better understanding of etiology and control prospects.


ULK1 Inhibition as a Targeted Therapeutic Strategy for Psoriasis by Regulating Keratinocytes and Their Crosstalk With Neutrophils.
Psoriasis is a common inflammatory skin disease resulting from an interplay of keratinocytes and immune cells. Previous studies have identified an essential role of autophagy in the maintenance of epidermal homeostasis including proliferation and differentiation. However, much less is known about the role of autophagy-related proteins in the cutaneous immune response. Herein, we showed that ULK1, the key autophagic initiator, and its phosphorylation at Ser556 were distinctively decreased in the epidermis from lesional skin of psoriasis patients. Topical application of SBI0206965, a selective ULK1 inhibitor, significantly attenuated epidermal hyperplasia, infiltration of neutrophils, and transcripts of the psoriasis-related markers in imiquimod (IMQ)-induced psoriasiform dermatitis (PsD). In vitro, ULK1 impairment by siRNA and SBI0206965 arrested cell proliferation and promoted apoptosis of keratinocytes but had a marginal effect on the expression of proinflammatory mediators under steady status. Surprisingly, SBI0206965 blocked the production of chemokines and cytokines in keratinocytes stimulated by neutrophils. Of interest, the pro-apoptotic and anti-inflammatory effects of ULK1 inhibition cannot be fully replicated by autophagic inhibitors. Our findings suggest a self-regulatory process by downregulating ULK1 to maintain the immune homeostasis of psoriatic skin via regulating keratinocytes and their crosstalk with neutrophils, possibly through both autophagy-dependent and independent mechanisms. ULK1 might be a potential target for preventing or treating psoriasis.


Deep brain stimulation versus anterior capsulotomy for obsessive-compulsive disorder: a review of the literature.
Obsessive-compulsive disorder (OCD) is a chronic and debilitating psychiatric condition. Traditionally, anterior capsulotomy (AC) was an established procedure for treatment of patients with refractory OCD. Over recent decades, deep brain stimulation (DBS) has gained popularity. In this paper the authors review the published literature and compare the outcome of AC and DBS targeting of the area of the ventral capsule/ventral striatum (VC/VS) and nucleus accumbens (NAcc). Patients in published cases were grouped according to whether they received AC or DBS and according to their preoperative scores on the Yale-Brown Obsessive-Compulsive Scale (YBOCS), and then separated according to outcome measures: remission (YBOCS score < 8); response (≥ 35% improvement in YBOCS score); nonresponse (< 35% improvement in YBOCS score); and unfavorable (i.e., worsening of the baseline YBOCS score). Twenty studies were identified reporting on 170 patients; 62 patients underwent DBS of the VC/VS or the NAcc (mean age 38 years, follow-up 19 months, baseline YBOCS score of 33), and 108 patients underwent AC (mean age 36 years, follow-up 61 months, baseline YBOCS score of 30). In patients treated with DBS there was a 40% decrease in YBOCS score, compared with a 51% decrease for those who underwent AC (p = 0.004). Patients who underwent AC were 9% more likely to go into remission than patients treated with DBS (p = 0.02). No difference in complication rates was noted. Anterior capsulotomy is an efficient procedure for refractory OCD. Deep brain stimulation in the VC/VS and NAcc area is an emerging and promising therapy. The current popularity of DBS over ablative surgery for OCD is not due to nonefficacy of AC, but possibly because DBS is perceived as more acceptable by clinicians and patients.


A human monoclonal antibody against varicella-zoster virus glycoprotein III.
Hybridomas producing human monoclonal antibodies (HMAbs) against varicella-zoster virus (VZV) were generated by fusing murine myeloma cells with human lymphocytes immunized in vitro. An assay system was developed to select anti-glycoprotein (gp)III HMAbs from the pool of anti-VZV HMAbs. A murine anti-gpIII MAb, 4B7, did not react with a VZV-infected cell homogenate, but did react with a VZV-infected cell monolayer, whereas anti-gpI and anti-gpII MAbs reacted with both antigens. Hybridomas were screened to obtain HMAbs having a reaction profile similar to that of 4B7 and one such clone, V3, stably produces human IgG1 (kappa). HMAb V3 immunoprecipitated a VZV antigen of 115K to 120K, which was not immunoabsorbed by an anti-gpII HMAb, implying that V3 recognizes gpIII. V3 neutralized VZV independently of complement, unlike anti-gpI and anti-gpII HMAbs. All five strains of VZV tested were completely neutralized by V3, and the dose of V3 required to reduce the number of virus plaques by 50% ranged from 0.027 to 0.15 micrograms/ml. V3 was also able to inhibit the spread of virus infection from infected to uninfected cells, whereas anti-gpI and anti-gpII HMAbs could not. In addition, V3 mediated antibody-dependent cellular cytotoxicity but not complement-dependent cytotoxicity of VZV-infected cells. The results suggest that an anti-gpIII HMAb may provide a new means of passive immunoprophylaxis and also help to identify an antigenic epitope appropriate for a subunit vaccine.


Endoglin promotes angiogenesis in cell- and animal-based models of retinal neovascularization.
We investigated endoglin expression in hypoxic microvascular endothelial cells and retinal endoglin expression in rats that develop experimental oxygen-induced retinopathy (OIR). We also tested neutralizing antibodies (Abs) against endoglin (anti-CD105 Ab) and VEGF (anti-VEGF Ab) either alone or in combination for efficacy against serum-induced retinal microvascular endothelial cell proliferation and retinal neovascularization (NV) in OIR rats. To our knowledge, this marks the first time that a biologic agent has been used to target retinal endoglin and modulate retinal neovascularization. Induction of endoglin by hypoxia was measured by immunohistochemical analysis and ELISA. Proliferation was quantified using a colorimetric 5-bromo-2-deoxyuridine ELISA. Western blots were used to measure endoglin levels in retinas of OIR rats. Immunohistochemical staining was also preformed in OIR rats using anti-CD105 and fluorescein isothiocyanate-conjugated isolectin B4 antibodies. Anti-CD105 Ab and Anti-VEGF Ab, administered alone or in combination, reduced serum-induced retinal microvascular endothelial cell proliferation. Additionally, in a rat model of oxygen-induced retinopathy, retinal endoglin was significantly increased at 14(2), 14(3), 14(4) and 14(6) compared with retinal levels in control rats. At 14(2), immunohistochemical analysis demonstrated that endoglin was elevated in newly developed vessels at the peripheral extent of major veins, precisely where NV is expected to develop in OIR rats. Neutralizing anti-CD105 reduced retinal NV in OIR rats. Our data support other studies showing that reduction of endoglin expression inhibits retinal NV. Our findings demonstrate that retinal endoglin immunolocalization overlaps with nascent neovascular structures in OIR rats. Therefore, endoglin may serve as a useful predictor of incipient neovascular disease.


Single chain antibody vaccination in mice against human ovarian cancer enhanced by microspheres and cytokines.
Experimental studies using monoclonal antibody have demonstrated that anti-idiotypic immunity can be enhanced by microspheres or cytokines. The underlying physiological principles behind these strategies involve sustained release of antigen and arousal of the immune system. In this study, a murine model was used to investigate the combination effect of microspheres and cytokines on anti-idiotypic response. A single chain antibody scFv-pDL1.0, which recognizes the human ovarian cancer antigen CA125 was formulated in microspheres and injected to mice alone or in the presence of colony stimulating factor (GM-CSF) or tumor necrosis factor (TNF-alpha). The immunization of mice with formulated single chain antibody and cytokines resulted in enhanced production of anti-idiotypic antibody, and which subsequently induced the production of anti-anti-idiotypic antibody. These results raise the possibility of cancer immunotherapy by administration of single chain antibody encapsulated in microspheres with GM-CSF or TNF-alpha.


Antiviral activity of polyoxomolybdoeuropate PM-104 against human immunodeficiency virus type 1.
A polyoxomolybdoeuropate PM-104 (NH4)12H2[Eu4(MoO4)(H2O)16(Mo7O24)4].13H2O was found to be a potent inhibitor of the growth of human immunodeficiency virus type 1 (HIV-1), a causative agent of acquired immunodeficiency syndrome (AIDS). On the basis of TI50 [median cytotoxic concentration (CC50)/median effective concentration (EC50)], the in vitro anti-HIV-1 activity of PM-104 is favorably comparable to that of a heteropolyoxotungstate PM-19 K7[PTi2W10O40].6H2O, which is one of the most potent HIV-1 inhibitors among the polyoxometalates so far tested. The heteropolyoxomolybdate with a potent anti-HIV-1 activity is introduced for the first time in this communication.


A New Microfluidic Device to Facilitate Functional Precision Medicine Assays.
Functional precision medicine (FPM) has emerged as a new approach to improve cancer treatment. Despite its potential, FPM assays present important limitations such as the number of cells and trained personnel required. To overcome these impediments, here we describe a novel microfluidic platform that can be used to perform FPM assays, optimizing the use of primary cancer cells and simplifying the process by using microfluidics to automatize the process.


NSD1 mutations deregulate transcription and DNA methylation of bivalent developmental genes in Sotos syndrome.
Sotos syndrome (SS), the most common overgrowth with intellectual disability (OGID) disorder, is caused by inactivating germline mutations of NSD1, which encodes a histone H3 lysine 36 methyltransferase. To understand how NSD1 inactivation deregulates transcription and DNA methylation (DNAm), and to explore how these abnormalities affect human development, we profiled transcription and DNAm in SS patients and healthy control individuals. We identified a transcriptional signature that distinguishes individuals with SS from controls and was also deregulated in NSD1-mutated cancers. Most abnormally expressed genes displayed reduced expression in SS; these downregulated genes consisted mostly of bivalent genes and were enriched for regulators of development and neural synapse function. DNA hypomethylation was strongly enriched within promoters of transcriptionally deregulated genes: overexpressed genes displayed hypomethylation at their transcription start sites while underexpressed genes featured hypomethylation at polycomb binding sites within their promoter CpG island shores. SS patients featured accelerated molecular aging at the levels of both transcription and DNAm. Overall, these findings indicate that NSD1-deposited H3K36 methylation regulates transcription by directing promoter DNA methylation, partially by repressing polycomb repressive complex 2 (PRC2) activity. These findings could explain the phenotypic similarity of SS to OGID disorders that are caused by mutations in PRC2 complex-encoding genes.


Bi-criteria Pareto optimization to balance irradiation time and dosimetric objectives in proton arc therapy.
Objective. Proton arc therapy (PAT) is a new delivery technique that exploits the continuous rotation of the gantry to distribute the therapeutic dose over many angular windows instead of using a few static fields, as in conventional (intensity-modulated) proton therapy. Although coming along with many potential clinical and dosimetric benefits, PAT has also raised a new optimization challenge. In addition to the dosimetric goals, the beam delivery time (BDT) needs to be considered in the objective function. Considering this bi-objective formulation, the task of finding a good compromise with appropriate weighting factors can turn out to be cumbersome.Approach. We have computed Pareto-optimal plans for three disease sites: a brain, a lung, and a liver, following a method of iteratively choosing weight vectors to approximate the Pareto front with few points. Mixed-integer programming (MIP) was selected to state the bi-criteria PAT problem and to find Pareto optimal points with a suited solver.Main results. The trade-offs between plan quality and beam irradiation time (staticBDT) are investigated by inspecting three plans from the Pareto front. The latter are carefully picked to demonstrate significant differences in dose distribution and delivery time depending on their location on the frontier. The results were benchmarked against IMPT and SPArc plans showing the strength of degrees of freedom coming along with MIP optimization.Significance. This paper presents for the first time the application of bi-criteria optimization to the PAT problem, which eventually permits the planners to select the best treatment strategy according to the patient conditions and clinical resources available.


Pilot trial of a group cognitive behavioural therapy program for comorbid depression and obesity.
Depression and obesity are significant global health concerns that commonly occur together. An integrated group cognitive behavioural therapy program was therefore developed to simultaneously address comorbid depression and obesity. Twenty-four participants (63% women, mean age 46 years) who screened positively for depression with a body mass index ≥25 were recruited from a self-referred general population sample. The group therapy program (10 two-hour weekly sessions) was examined in a single-arm, before-after pilot trial, conducted in a behavioural health clinic in Adelaide, Australia. Primary outcomes included survey and assessment-based analyses of depression, anxiety, body image, self-esteem, and weight (kg), assessed at four time-points: baseline, post-intervention, three-months and 12-months post program. Eighteen participants (75%) completed the program and all assessments. Significant improvements in depression, anxiety, self-esteem and body shape concern scores, several quality of life domains, eating behaviours and total physical activity (among others) - but not weight - were observed over the course of the trial. Results from this pilot trial suggest that combining interventions for depression and obesity may be useful. Further development of the program, particularly regarding the potential for physical health benefits, and a randomised controlled trial, are warranted. Trial registration: ANZCTR, ACTRN12617001079336, 13 July 2017. Retrospectively registered after date of the first consent (6 July 2017), but before the date of the first intervention session (20 July 2017).


Essential role of non-canonical Wnt signalling in neural crest migration.
Migration of neural crest cells is an elaborate process that requires the delamination of cells from an epithelium and cell movement into an extracellular matrix. In this work, it is shown for the first time that the non-canonical Wnt signalling [planar cell polarity (PCP) or Wnt-Ca2+] pathway controls migration of neural crest cells. By using specific Dsh mutants, we show that the canonical Wnt signalling pathway is needed for neural crest induction, while the non-canonical Wnt pathway is required for neural crest migration. Grafts of neural crest tissue expressing non-canonical Dsh mutants, as well as neural crest cultured in vitro, indicate that the PCP pathway works in a cell-autonomous manner to control neural crest migration. Expression analysis of non-canonical Wnt ligands and their putative receptors show that Wnt11 is expressed in tissue adjacent to neural crest cells expressing the Wnt receptor Frizzled7 (Fz7). Furthermore, loss- and gain-of-function experiments reveal that Wnt11 plays an essential role in neural crest migration. Inhibition of neural crest migration by blocking Wnt11 activity can be rescued by intracellular activation of the non-canonical Wnt pathway. When Wnt11 is expressed opposite its normal site of expression, neural crest migration is blocked. Finally, time-lapse analysis of cell movement and cell protrusion in neural crest cultured in vitro shows that the PCP or Wnt-Ca2+ pathway directs the formation of lamellipodia and filopodia in the neural crest cells that are required for their delamination and/or migration.


Ethical issues in public health surveillance: a systematic qualitative review.
Public health surveillance is not ethically neutral and yet, ethics guidance and training for surveillance programmes is sparse. Development of ethics guidance should be based on comprehensive and transparently derived overviews of ethical issues and arguments. However, existing overviews on surveillance ethics are limited in scope and in how transparently they derived their results. Our objective was accordingly to provide an overview of ethical issues in public health surveillance; in addition, to list the arguments put forward with regards to arguably the most contested issue in surveillance, that is whether to obtain informed consent. Ethical issues were defined based on principlism. We assumed an ethical issue to arise in surveillance when a relevant normative principle is not adequately considered or two principles come into conflict. We searched Pubmed and Google Books for relevant publications. We analysed and synthesized the data using qualitative content analysis. Our search strategy retrieved 525 references of which 83 were included in the analysis. We identified 86 distinct ethical issues arising in the different phases of the surveillance life-cycle. We further identified 20 distinct conditions that make it more or less justifiable to forego informed consent procedures. This is the first systematic qualitative review of ethical issues in public health surveillance resulting in a comprehensive ethics matrix that can inform guidelines, reports, strategy papers, and educational material and raise awareness among practitioners.


Endoplasmic Reticulum (ER) Stress Enhances Tip60 (A Histone Acetyltransferase) Binding to the Concanavalin A.
Herein, we report that the concanavalin A binding of Tip60 (a target of the human immunodeficiency virus type 1-encoded transactivator Tat interacting protein 60 KD; a histone acetyltransferase; HAT) is enhanced as the result of endoplasmic reticulum (ER) stress. The cell expression of Tip60 combined with site-directed mutagenesis analysis was used to identify the glutamine 324 residue as the lecithin binding (Concanavalin A; Con A) site. The Tip60 N324A mutant strain, which seems to be the Con A binding-deficient, was attenuated the protein-protein interactions with FE65 and its protein stability, but its ability of G0-G1 cell cycle arrest was not interrupted. Interestingly, both HAT activity and the nuclear localization of Tip60 N324A mutant were enhanced than those of Tip60 WT. Thus, our results indicate that the Con A binding deficient of Tip60 seems to be one of the most pivotal posttranslational modifications (such as N-glycosylation) for its functional regulation signal, which is generated in response to ER stress.


Identification of an acetylation site of Chlamydomonas alpha-tubulin.
An acetylation site of Chlamydomonas axonemal alpha-tubulins was identified near, or within, the binding site of 6-11B-1, a monoclonal antibody specific for posttranslationally acetylated alpha-tubulins. In a first approach, axonemal proteins were hydrolyzed by formic acid, cyanogen bromide, or chymotrypsin and analyzed with immunoblots. The smallest alpha-tubulin peptide retained on nitrocellulose and containing antibody-binding site(s) was found to span amino acids 37-138 (alpha 37-138). A smaller antibody-binding peptide, identified as alpha 25-50, was obtained by complete digestion of alpha-tubulin with chymotrypsin. This fragment was purified by reversed-phase HPLC and assayed by its ability to bind 6-11B-1 in solution. Determination of the amino acid sequences of alpha 37-138 and alpha 25-50 showed that residue 40 in axonemal alpha-tubulin is epsilon N-acetyllysine. A sequence very similar to Chlamydomonas alpha 25-50 is found in the majority of alpha-tubulins analyzed so far. However, the corresponding region is markedly divergent in some alpha-tubulin isoforms from chicken, Drosophila, and yeast.


Discriminant Subgraph Learning from Functional Brain Sensory Data.
The human brain is a complex system with many functional units interacting with each other. This interacting relationship, known as the functional connectivity network (FCN), is critical for brain functions. To learn the FCN, machine learning algorithms can be built based on brain signals captured by sensing technologies such as EEG and fMRI. In neurological diseases, past research has revealed that the FCN is altered. Also, focusing on a specific disease, some part of the FCN, i.e., a sub-network, can be more susceptible than other parts. However, the current knowledge about disease-specific sub-networks is limited. We propose a novel Discriminant Subgraph Learner (DSL) to identify a functional sub-network that best differentiates patients with a specific disease from healthy controls based on brain sensory data. We develop an integrated optimization framework for DSL to simultaneously learn the FCN of each class and identify the discriminant sub-network. Further, we develop tractable and converging algorithms to solve the optimization. We apply DSL to identify a functional sub-network that best differentiates patients with episodic migraine (EM) from healthy controls based on a fMRI dataset. DSL achieved the best accuracy compared to five state-of-the-art competing algorithms.


Morphologic variations in mandibular premolars: a report of three cases.
To report cases with morphologic variations in mandibular premolars. Mandibular premolars have earned the reputation for having aberrant anatomy. The occurrence of three canals with three separate (type V, Vertucci) foramina in mandibular premolars is very rare. In this report, three cases are presented involving the root canal treatment of mandibular premolars with three different root and canal configurations. The present report also emphasizes the use of the dental operating microscope to enhance visualization. Correlation between radiographic findings, use of dental operating microscope and knowledge of aberrant anatomy helps to identify, locate and treat the aberrant anatomy of the tooth. The treatment was successful as all canals were identified and filled and also as evidenced by relief of symptoms. Success is largely dependent on the use of magnification, which aided in identifying the location of the three individual root canal orifices; thus emphasizing the need to familiarize ourselves with dental microscopy, to obtain maximal anatomic information in endodontic practice.


Single base-pair mutations in centromere element III cause aberrant chromosome segregation in Saccharomyces cerevisiae.
In this paper we show that a 211-base pair segment of CEN3 DNA is sufficient to confer wild-type centromere function in the yeast Saccharomyces cerevisiae. We used site-directed mutagenesis of the 211-base pair fragment to examine the sequence-specific functional requirements of a conserved 11-base pair segment of centromere DNA, element III (5'-TGATTTATCCGAA-3'). Element III is the most highly conserved of the centromeric DNA sequences, differing by only a single adenine X thymine base pair among the four centromere DNAs sequenced thus far. All of the element III sequences contain specific cytosine X guanine base pairs, including a 5'-CCG-3' arrangement, which we targeted for single cytosine-to-thymine mutations by using sodium bisulfite. The effects of element III mutations on plasmid and chromosome segregation were determined by mitotic stability assays. Conversion of CCG to CTG completely abolished centromere function both in plasmids and in chromosome III, whereas conversion of CCG to TCG decreased plasmid and chromosome stability moderately. The other two guanine X cytosine base pairs in element III could be independently converted to adenine X thymine base pairs without affecting plasmid or chromosome stability. We concluded that while some specific nucleotides within the conserved element III sequence are essential for proper centromere function, other conserved nucleotides can be changed.


Primary hyperparathyroidism in Saudi Arabia: a review of 46 cases.
Records of 46 patients who were treated for primary hyperparathyroidism at King Khalid University Hospital, Riyadh Saudi Arabia from 1st July 2000 to 30th June 2006 were reviewed. Mean age at diagnosis was 44 years (range 13 - 70 years) and average duration of symptoms was 39 months (1 month to 11 years). There were 35 females and 11 males with a ratio of 3.2:1. Bone pains were the major symptoms at presentation in 45.7% followed by no symptom in 23.9%, renal stones in 15.2%, polyuria in 6.5%, while 4.3% each presented with depression, and constipation. Males had significantly more severe disease with higher serum calcium, higher urinary calcium excretion, and higher serum creatinine. Ninety six percent of patients had successful surgery and 4% (two patients) each had recurrence and hungry bone syndrome. It is concluded that PHPT in Saudi Arabia continues to be a symptomatic disorder with skeletal and renal manifestations occurring at a younger age and males having more severe disease. Further prospective studies are needed to verify our findings.


Experience with virtual reality-based technology in teaching restorative dental procedures.
This article reports on extensive experience with advanced simulation at the University of Pennsylvania, School of Dental Medicine (UPSDM). Virtual reality-based technology (VRBT) or advanced simulation is currently available for the instruction of dental students in preclinical restorative procedures. UPSDM was one of the first schools in the world to have extensive experience with VRBT technology using an advanced simulation unit (DentSim) from DenX, Ltd. UPSDM's experience consists of several years of research using control and experimental groups, employing students to participate in an investigative project, and using the units for remediation and a supplement to the preclinical laboratory. Currently, all first-year students (Class of 2007 and Class of 2008) are receiving most of their preparative operative training on the VRBT units. UPSDM started with one (beta) version unit in 1998, which was later updated and expanded first to four units and recently to fifteen units. This communication is presenting the studies that were of fundamental importance in making the decision to acquire fifteen units in 2003. The areas of main interest to the SDM concerning this technology were its use in teaching, refreshing, and remediating students in restorative procedures and its effectiveness as a teaching methodology in relation to time, efficiency, and faculty. Several studies with varying parameters were performed at various time points. The limited statistical analysis conducted was not conclusive for all measures, and in some cases the data only suggest areas of possible significance. This is due to the low number of students who could access the limited number of available units and the change of protocols in response to student and faculty input. Overall, the results do suggest, however, that students learn faster, arrive at the same level of performance, accomplish more practice procedures per hour, and request more evaluations per procedure or per hour than in our traditional laboratories. Students' attitudes, as measured by surveys, group interviews, and private interviews, are mixed. Our overall evaluation of our experience with this technology has been positive and led to the purchase of additional units, its full incorporation into our curriculum, and curriculum revision to maximize its potential. Our conclusion is that this technology offers significant potential in the field of dental education and that further use and investigation are both desired and justified.


Hepatitis B virus in patients undergoing hemodialysis: transmission risks and psychosocial reactions.
Thirteen patients undergoing hemodialysis who were carriers of hepatitis B virus (HBV) were screened for inappropriate reactions to HBV transmission risks and for emotional reactions to their renal and viral diseases. Four patients underreacted to HBV transmission risks through denial or misunderstanding. The nine remaining patients reported that their HBV, as compared with their renal disease, resulted in substantially greater restrictions in interpersonal relations and significantly greater feelings of not being accepted by others. In contrast, the nine patients reported that their renal disease, as compared with their HBV, resulted in substantially greater restrictions in leisure and work activities. Thirteen control patients undergoing hemodialysis but not having HBV did not differ from the patients with HBV in psychosocial reactions to renal disease. As shown in a one-month follow-up, counseling was effective in enhancing awareness of HBV transmission risks and in improving emotional adjustments to the renal and viral diseases.


Telemedicine: a solution to disparities in human immunodeficiency virus prevention and pre-exposure prophylaxis uptake, and a framework to scalability and equity.
As the group with the highest risk and the lowest pre-exposure prophylaxis (PrEP) utilization, black and Hispanic young men who have sex with men (MSM) face the brunt of the current human immunodeficiency virus (HIV) epidemic within the United States. In this narrative review, we summarized recent interventions that utilize technology (telemedicine) to improve HIV PrEP uptake. We examined the general model of those telemedicine programs and evaluated their success and limitations in addressing PrEP accessibility for at-risk individuals. We also identified the current bottlenecks in the scalability of telemedicine programs, offered suggestions to overcome these barriers, and advocated for a centralized source of funding to increase the accessibility of telemedicine programs for PrEP.


Endocannabinoids exert CB1 receptor-mediated neuroprotective effects in models of neuronal damage induced by HIV-1 Tat protein.
In the era of combined antiretroviral therapy (cART), human immunodeficiency virus type 1 (HIV-1) is considered a chronic disease that specifically targets the brain and causes HIV-1-associated neurocognitive disorders (HAND). Endocannabinoids (eCBs) elicit neuroprotective and anti-inflammatory actions in several central nervous system (CNS) disease models, but their effects in HAND remain unknown. HIV-1 does not infect neurons, but produces viral toxins, such as transactivator of transcription (Tat), that disrupt neuronal calcium equilibrium and give rise to synaptodendritic injuries and cell death, the former being highly correlated with HAND. Consequently, we tested whether the eCBs N-arachidonoylethanolamine (anandamide/AEA) and 2-arachidonoyl-glycerol (2-AG) offer neuroprotective actions in a neuronal culture model. Specifically, we examined the neuroprotective actions of these eCBs on Tat excitotoxicity in primary cultures of prefrontal cortex neurons (PFC), and whether cannabinoid receptors mediate this neuroprotection. Tat-induced excitotoxicity was reflected by increased intracellular calcium levels, synaptodendritic damage, neuronal excitability, and neuronal death. Further, upregulation of cannabinoid 1 receptor (CB1R) protein levels was noted in the presence of HIV-1 Tat. The direct application of AEA and 2-AG reduced excitotoxic levels of intracellular calcium and promoted neuronal survival following Tat exposure, which was prevented by the CB1R antagonist rimonabant, but not by the CB2R antagonist AM630. Overall, our findings indicate that eCBs protect PFC neurons from Tat excitotoxicity in vitro via a CB1R-related mechanism. Thus, the eCB system possesses promising targets for treatment of neurodegenerative disorders associated with HIV-1 infection.