Use of multiple logistic analysis in twin zygosity diagnosis.
Stochastic methods were applied to the diagnosis of twin zygosity by mailed questionnaire in a study on adult twins. Multiple logistic discriminant analysis was able to classify twin pairs previously left unclassified (XZ) by a deterministic method, and accuracy of classification was verified by blood testing. The mean intrapair differences of height and weight of XZ pairs were in between dizygotic and monozygotic pairs, but the frequency of personal contact between XZ twins was significantly less than that of MZ or DZ pairs. Both classification methods, when used separately with the same questions, left unclassified 6.5-7.6% of all 11,542 respondent pairs in the Finnish Twin Registry, but deterministic classification followed by logistic analysis misclassified only 1.9% of all twin pairs.


Learning curve takes 65 repetitions of totally extraperitoneal laparoscopy on inguinal hernias for reduction of operating time and complications.
Inguinal hernia repair is one of the most frequently conducted surgical procedures worldwide. Totally extraperitoneal (TEP) hernioplasty has shown many advantages over traditional open surgery. However, because of increased surgical complexity, it requires more practice to achieve optimal results. The aim of this study is to evaluate the learning curve for TEP hernioplasty, analysing parameters related to the mean operating time. The secondary objective is to assess the complications and early discharge rates related to this procedure. A prospective study of 238 consecutive patients undergoing TEP hernioplasty from the same practitioner between May 2009 and May 2014, in a specialised centre for abdominal hernias in Sao Paulo, Brazil, was conducted. All data were obtained through medical records. The study included 137 patients undergoing a total of 157 TEP hernia repairs. Bilaterality and complications can influence the operating time and so were excluded. Patients with unilateral surgery and without complications were included. Patients were chronologically divided into four groups. Groups 1-3 were composed of 35 patients and group 4 of 32. There were no significant variations in clinical characteristics between the groups. The plateau of the learning curve was reached on the 65 th repetition of the surgery, with a mean operating time of 28 min (p < 0,05). Complications were only observed in the first group of 35 patients. 97% of patients were discharged early, defined as patient leaving hospital less than 12 h after surgery. After an initial reduction, the mean operating time stabilised after 65 cases. A reduction in the rate of complications was observed after 35 cases, and a rate of 97% of early discharge was achieved.


Child externalizing behavior problems linked to genetic and non-genetic variation in dental caries.
The association of environmental and genetic variation in caries with child externalizing behavior problems (inattention, hyperactivity, impulsivity, and defiance) was studied in a sample of 239 pairs of 3- to 8-year-old impoverished Brazilian twins. It was hypothesized that externalizing problems would show a stronger positive association with environmental than genetic variation in caries. Univariate twin models were estimated to parse variation in caries into three components: additive genetic (A), shared environment (C) and non-shared environment/error (E). Age-adjusted associations between externalizing problems and each variance component were tested. Contrary to the hypothesis, modest but very consistent negative associations were found between externalizing problems and both genetic and environmental variation in caries. Mutans streptococci and sweetness preference did not explain the negative associations of caries and externalizing problems. Externalizing problems in non-medicated children were associated with less dental decay that could be explained by both genetic and environmental factors.


[Rapid indication of microbiological damage and biodegradation of structural materials in space vehicles].
The study was undertaken to design a kit of indicators for rapid detection and group identification of microbes causing biological damages to the materials. Indication is accomplished by dropwise colorimetric reaction for which modified chromatographic materials serve as substrates. Reactions are made by the indicators whose design allows reagents and samples to be measured on board space vehicles. The designed kit of indicators detects microorganisms, biodegrade products, and identifies them by groups: vegetative or spore-forming bacteria, and lower fungi. The paper considers the characteristics of colorimetric reactions and an algorithm of group identification of microbes by the respective combination of positive and negative results of the reactions. It also presents data on the sensitivity levels of tests provided by the kit, which were obtained from laboratory and field ground tests.


A functional 125-kDa core polypeptide of fission yeast DNA topoisomerase II.
We purified fission yeast DNA topoisomerase II (topo II) to apparent homogeneity. It consists of a single 165-kDa polypeptide in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and, upon treatment with a bifunctional reagent, doubles its molecular weight. Limited proteolysis of intact topo II by papain produces a 125-kDa core, which lacks the N-terminal 75 and the C-terminal approximately 260 amino acids but still contains regions similar to those of bacterial or phage T4 topo II subunits. The core retains relaxing and unknotting activities. Further digestion inactivates the core, cleaving it at the middle of the GyrB-like domain and at the beginning of the GyrA-like domain. Therefore, papain appears to cleave spatially distinct subdomains of topo II. We made top2 mutant genes deleted of the C-terminal 286 or N-terminal 74 amino acids, which can substitute for the wild-type top2+ gene in mitosis and meiosis. However, a mutant containing deletions of both termini cannot rescue the top2 null mutant, despite the fact that the product is enzymatically active. Therefore, the top2 product of the doubly truncated gene may not fulfill all of the in vivo requirements for top2+ function.


Finding words and word structure in artificial speech: the development of infants' sensitivity to morphosyntactic regularities.
To achieve language proficiency, infants must find the building blocks of speech and master the rules governing their legal combinations. However, these problems are linked: words are also built according to rules. Here, we explored early morphosyntactic sensitivity by testing when and how infants could find either words or within-word structure in artificial speech snippets embodying properties of morphological constructions. We show that 12-month-olds use statistical relationships between syllables to extract words from continuous streams, but find word-internal regularities only if the streams are segmented. Seven-month-olds fail both tasks. Thus, 12-month-olds infants possess the resources to analyze the internal composition of words if the speech contains segmentation information. However, 7-month-old infants may not possess them, although they can track several statistical relations. This developmental difference suggests that morphosyntactic sensitivity may require computational resources extending beyond the detection of simple statistics.


Ultrasound-guided pseudoaneurysm compression: efficacy after coronary stenting and angioplasty.
Ultrasound-guided compression of femoral pseudoaneurysms has often obviated the need for open operative repair. Increasing use of percutaneous interventional cardiac procedures has created pseudoaneurysms with a large-caliber arterial defect, often in patients who are placed immediately on anticoagulation therapy. This report describes the prospectively collected information from our vascular laboratory regarding ultrasound-guided compression of these pseudoaneurysms after cardiac procedures, both interventional and diagnostic. Since March 1994 prospective data collection for patients who have undergone pseudoaneurysm compression in our vascular laboratory has recorded information including cardiac procedure, size of catheter or sheath, coagulation parameters, pseudoaneurysm size and location, and time to compression. Forty-one patients underwent attempted ultrasound-guided pseudoaneurysm compression after cardiac procedures: 19 after cardiac catheterization alone, seven after angioplasty, one after atherectomy, two after insertion and subsequent removal of an intraaortic balloon pump, and 12 after coronary stenting. Compression was successful overall in 88% of the patients (36 of 41). Successful compression of the pseudoaneurysm was seen in 95% after catheterization alone, 100% after angioplasty, 100% after atherectomy, and 100% after intraaortic balloon pumping, as compared with 67% after stenting (eight of 12 vs 28 of 29; p = 0.02). A sheath size of 9F or greater was a significant factor in predicting unsuccessful compression (three of eight vs two of 33; p = 0.04). Abnormal coagulation parameters were present in 20 of the 41 patients and was not significantly different in patients who were successfully or unsuccessfully treated (four of five vs 16 of 36; p = 0.40). Pseudoaneurysms after cardiac procedures and interventions can often be successfully compressed with an ultrasound-guided technique. The presence of abnormal coagulation parameters was not identified as a risk factor for compression failure and should not dissuade attempted compression. Stent placement was more likely to result in unsuccessful compression, and this appeared to be a result of the larger size of the arterial defect. Even in this setting, compression achieved obliteration of the pseudoaneurysm in more than half of the patients.


Neuroinflammation as a target for glaucoma therapy.
The pathogenesis of glaucoma is still not fully clarified but a growing body of evidence suggests that neuroinflammation and immune response are part of the sequence of pathological events leading to the optic neuropathy. Indeed, inflammation - involving the activation and proliferation of resident glial cells (astrocytes, Muller cells and microglia) and the release of a plethora of anti- and pro-inflammatory cytokines, chemokines and reactive oxygen species - has been reported as common features in clinical and experimental glaucoma. In the insulted retina, as for other neuronal tissues, pathogenic and reparative aspects coexist in the inflammatory process, with extent and persistency affecting the final outcome. In view of this, therapies aimed at modulating the immune and inflammatory responses may represent a promising approach for limiting the optic nerve damage and the loss of retinal ganglion cells associated with glaucoma.


Exploration of use of SenseCam to support autobiographical memory retrieval within a cognitive-behavioural therapeutic intervention following acquired brain injury.
Delivering effective psychotherapy to address the significant emotional consequences of acquired brain injury (ABI) is challenged by the presence of acquired cognitive impairments, especially retrieval of detailed autobiographical memories of emotional trigger events. Initial studies using a wearable camera (SenseCam) suggest long-term improvements in autobiographical retrieval of recorded events. In this study a single-case experimental design was implemented to explore the use of SenseCam as a memory aid for a man with a specific anxiety disorder and memory and executive difficulties following ABI. We predicted that SenseCam supported rehearsal of memories of events that trigger high levels of anxiety would yield improved retrieval of both factual detail and internal state information (thoughts and feelings) compared with a conventional psychotherapy aid (automatic thought record sheets, ATRs) and no strategy. The findings indicated SenseCam supported retrieval of anxiety trigger events was superior to ATRs or no strategy in terms of both detail and internal state information, with 94% of the information being recalled in the SenseCam condition, compared to 39% for the "no strategy" and 22% for the ATR conditions. It is concluded that SenseCam may be of use as a compensatory aid in psychotherapies relying on retrieval of emotionally salient trigger events.


Time-frequency dynamics of the sum of intra- and extracerebral hemodynamic functional connectivity during resting-state and respiratory challenges assessed by multimodal functional near-infrared spectroscopy.
Monitoring respiratory processes is important for evaluating neuroimaging data, given their influence on time-frequency dynamics of intra- and extracerebral hemodynamics. Here we investigated the time-frequency dynamics of the sum of intra- and extracerebral hemodynamic functional connectivity states during hypo- and hypercapnia by using three different respiratory challenge tasks (i.e., hyperventilation, breath-holding, and rebreathing) compared to resting-state. The sum of intra- and extracerebral hemodynamic responses were assessed using functional near-infrared spectroscopy (fNIRS) within two regions of interest (i.e., the dorsolateral and the medial prefrontal cortex). Time-frequency fNIRS analysis was performed based on wavelet transform coherence to quantify functional connectivity in terms of positive and negative phase-coupling within each region of interest. Physiological measures were assessed in the form of partial end-tidal carbon dioxide, heart rate, arterial tissue oxygen saturation, and respiration rate. We found that the three respiration challenges modulated time-frequency dynamics differently with respect to resting-state: 1) Hyperventilation and breath-holding exhibited inverse patterns of positive and negative phase-coupling. 2) In contrast, rebreathing had no significant effect. 3) Low-frequency oscillations contributed to a greater extent to time-frequency dynamics compared to high-frequency oscillations. The results highlight that there exist distinct differences in time-frequency dynamics of the sum of intra- and extracerebral functional connectivity not only between hypo- (hyperventilation) and hypercapnia but also between different states of hypercapnia (breath-holding versus rebreathing). This suggests that a multimodal assessment of intra-/extracerebral and systemic physiological changes during respiratory challenges compared to resting-state may have potential use in the differentiation between physiological and pathological respiratory behavior accompanied by the psycho-physiological state of a human.


Pemoline abuse.
Addictive use of the stimulant drug pemoline caused a delusional disorder in a middle-aged man. Discontinuance of use of the drug promptly reversed the psychosis, which subsequently recurred when pemoline was again used. Pemoline seems to be an addictive drug and has an additional property that heretofore has been attributed mainly to amphetamines--precipitation of a paranoid psychosis.


Formation of the NO donors glyceryl mononitrate and glyceryl mononitrite from the reaction of peroxynitrite with glycerol.
Peroxynitrite (ONOO-), formed from the rapid reaction of superoxide (O2-.) with NO, is known to generate stable compounds capable of donating NO on reaction with thiols and molecules containing hydroxy groups. Using glycerol as a model compound for the reactions of ONOO- with biomolecules containing hydroxy groups, we separated the products and identified them by HPLC/MS. It was shown that both glyceryl mononitrate and glyceryl mononitrite were formed and released NO on incubation with copper and l-cysteine. The compounds were stable over a period of 4h when shielded from light and kept on ice. Slow spontaneous decomposition occurred in the buffer used for the bioassay, but this was not sufficient to explain the vasorelaxing properties of these NO donors. It is concluded that the stable organic nitrate and nitrite have the capacity to be metabolized by vascular tissues, resulting in vasorelaxation.


A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions.
Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex signaling mechanisms underlying filopodial initiation, elongation and subsequent stabilization or retraction, it is crucial to determine the spatio-temporal protein activity in these dynamic structures. To analyze protein function in filopodia, we recently developed a semi-automated tracking algorithm that adapts to filopodial shape-changes, thus allowing parallel analysis of protrusion dynamics and relative protein concentration along the whole filopodial length. Here, we present a detailed step-by-step protocol for optimized cell handling, image acquisition and software analysis. We further provide instructions for the use of optional features during image analysis and data representation, as well as troubleshooting guidelines for all critical steps along the way. Finally, we also include a comparison of the described image analysis software with other programs available for filopodia quantification. Together, the presented protocol provides a framework for accurate analysis of protein dynamics in filopodial protrusions using image analysis software.


Laser-induced Damage Probability at 1.06 microm and 0.69 microm.
The first directly comparable measurements of the laser-induced surface damage process at both ruby and Nd:YAG laser wavelengths are reported. The most striking feature of the data is that all the materials studied are harder to damage at 0.69 microm than at 1.06 microm. The probabilistic nature of the laser-induced damage process at 1.06 microm was explored further by measuring the distribution of breakdown starting times with an image-converter streak camera. The observed distribution is described by the compound probability that breakdown occurs at a particular time, given that it has not occurred before that time. In addition, several connections between the probabilistic and thresholdlike interpretations of laser-induced damage are discussed. It is shown that these points of view are not totally incompatible.


Dentoskeletal Stability in Conventional Orthognathic Surgery, Presurgical Orthodontic Treatment and Surgery-First Approach in Class-III Patients.
For many years, the conventional approach to orthognathic surgery which was orthodontic treatment prior to orthognathic surgery has been the accepted method of treatment for skeletal class III malocclusion patients. This review compared the dentoskeletal stability of treatment results between conventional orthognathic surgery methods with presurgical orthodontic treatment and surgery-first approach in skeletal class III patients. The study protocol was based on Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement for systematic review and meta-analysis. Electronic and manual searches for literature since 2011 were conducted. PubMed and Medline databases were accessed. Data extraction and analysis were performed by two independent individuals. Seven studies out of hundred-fourteen articles met the inclusion criteria and were selected for qualitative analysis. The included studies were 494 patients with skeletal class-III malocclusion. Stability of treatment was compared between surgery-first approach and conventionally treated patients. The statistical analysis confirmed that surgery-first approach did not show more stability compared with presurgical orthodontics. The surgery-first approach shortened the overall treatment duration. However, more skeletal stability in conventional treatment was assessed. Both surgery-first approach and conventional treatment with presurgical orthodontics resulted in favorable skeletal changes in class-III malocclusion patients. Moreover, these findings should be discussed further due to the variety of study designs, outcomes and biases. Current evidence in this field still needs to be expanded. The authors wish to see more well-designed randomized controlled trials with long-term follow ups to confirm the results.


Findings on MR sleep studies as biomarkers to predict outcome of genioglossus advancement in the treatment of obstructive sleep apnea in children and young adults.
Genioglossus advancement, an operation to pull the tongue anteriorly, is a treatment of obstructive sleep apnea (OSA) secondary to glossoptosis. MRI predictors to identify which children will benefit from genioglossus advancement would be helpful for planning. We reviewed imaging findings on MR sleep studies as biomarkers to predict success or failure of genioglossus advancement in the treatment of OSA in children and young adults. Twenty-eight patients who had undergone genioglossus advancement and preoperative MRI were identified. For each subject, genioglossus advancement, which was performed to treat OSA, was categorized as a success or failure on the basis of polysomnography and clinical criteria. Static and dynamic cine MR sequences were retrospectively evaluated for multiple parameters including measurements of the size of the tongue and of the bony confines of the supraglottic airway, the ratio of tongue size to bony confines size, static size and dynamic changes of the retroglossal airway, tonsil size, and soft palate thickness. Radiologists were blinded to the outcome of genioglossus advancement (i.e., success or failure category). Numeric biomarkers were compared in an analysis-of-covariance model adjusting for patient age. Genioglossus advancement was successful for the treatment of OSA in 17 patients and failed in 11 patients. The relative size of the tongue (tongue-bony confines ratio) was larger in patients with a successful surgical outcome than in those for whom the procedure failed (mean ratio, 0.51 vs 0.46; p = 0.023). Smaller adenoids were associated with a successful outcome (mean size of adenoids, 9.1 vs 12.4 mm; p = 0.049). No other biomarker-including absolute tongue size and airway size or dynamic airway motion-was significant. The relative (not absolute) tongue size and small size of the adenoid tonsils on MRI were predictors of success of genioglossus advancement for the treatment of glossoptosis causing OSA. These findings may be helpful in guiding surgical decision making in children with OSA.


Carbonic anhydrase isozymes IX and XII in gastric tumors.
To systematically study the expression of carbonic anhydrase (CA) isozymes IX and XII in gastric tumors. We analyzed a representative series of specimens from non-neoplastic gastric mucosa and from various dysplastic and neoplastic gastric lesions for the expression of CA IX and XII. Immunohistochemical staining was performed using isozyme-specific antibodies and biotin-streptavidin complex method. CA IX was highly expressed in the normal gastric mucosa and remained positive in many gastric tumors. In adenomas, CA IX expression significantly decreased towards the high grade dysplasia. However, the expression resumed back to the normal level in well differentiated adenocarcinomas, while it again declined in carcinomas with less differentiation. In comparison, CA XII showed no or weak immunoreaction in the normal gastric mucosa and was slightly increased in tumors. These results demonstrate that CA IX expression is sustained in several types of gastric tumors. The variations observed in the CA IX levels support the concept that gastric adenomas and carcinomas are distinct entities and do not represent progressive steps of a single pathway.


Assessment of Resectability of Mediastinal Germ Cell Tumor Using Preoperative Computed Tomography.
Mediastinal germ cell tumor (MGCT) is a relatively rare tumor. Complete resection after chemotherapy is a standard treatment against this disease. However, the risk factors of incomplete resection are unclear. Therefore, we analyzed survival rates and risk factors for incomplete resection based on preoperative imaging. We retrospectively reviewed the medical records of patients (n = 56) with MGCT operated at National Cancer Center Hospital, and analyzed preoperative computed tomography (CT) data in terms of relationship of the tumor and vessels, and investigated survival rate and risk factors for incomplete resection. A total of 56 patients underwent resection of MGCT. The 5-y progression-free survival (PFS) and overall survival (OS) were 79% and 83%. In multivariate analysis, complete resection was the only significant prognostic factor for better PFS (hazard ratio (HR) = 9.083, P= 0.00021) and OS (HR = 5.519, P= 0.0445). The preoperative CT finding of arteries (including the aorta, right brachiocephalic artery, left common carotid artery, and left subclavian artery) surrounded by the tumor was a predictor of incomplete resection (odds ratio = 10.089, P= 0.049). Complete resection is essential for improving the survival of MGCT, and the risk stratification using preoperative CT imaging brings important information to achieve the complete resection.


Protective effect of aqueous extract of the rhizomes of Sansevieria liberica Gérôme and Labroy on carbon tetrachloride induced hepatotoxicity in rats.
The ability of an aqueous extract of the rhizomes of Sansevieria liberica to protect against carbon tetrachloride induced liver injury was investigated in Wistar albino rats. The carbon tetrachloride was prepared 1:5 (v:v) in olive oil, and administered subcutaneously at 1 mL/kg body weight. The extract was administered to both normal and carbon tetrachloride treated rats at 100, 200 and 300 mg/kg. On gas chromatographic analysis of the extract, twenty nine known flavonoids were detected, consisting mainly of 31.94 % apigenin, 20.66 % quercetin, 11.28 % kaempferol, 5.99 % naringenin, 5.83 % (-)-epicatechin, 3.69 % biochanin, 3.58 % (+)-catechin, 2.72 % diadzein, 2.20 % ellagic acid, 2.04 % butein. Compared to test control, the treatment dose dependently produced significantly (P<0.05) lower alkaline phosphatase, aspartate transaminase and alanine transaminase activities. The plasma total bilirubin and total protein levels of the test animals were lower though not significantly. The hepatic histopathological studies showed that carbon tetrachloride caused fatty degeneration of hepatocytes, which was inhibited by pre-treatment with the extract; thus, confirming the results of the biochemical studies. The results of this study indicated that treatment with the plant extracts protects the liver against carbon tetrachloride induced hepatotoxicity. This supports the use of Sansevieria liberica in traditional health care for managing liver problems.


[Neutropenia in HIV infection].
The infection by human immunodeficiency virus (HIV) are commonly associated with haematologic abnormalities (anemia, leucopenia and thrombocytopenia). We review the neutropenia. In patients infected with HIV neutropenia is seen in the 8-50% of them, and also have abnormalities in the neutrophil function. Direct injury of HIV on de bone marrow, anti-neutrophil antibodies, drugs, opportunistic infections of bone marrow, vitamin B12 and folate deficiency, radiations therapy, and hemophagocytic syndrome. These patients have a increased risk of infections, since the neutrophils play an important role in the defense against bacterial and certain fungal infections. It must to treat the causes. When it is not possible, Colony-Stimulating Factor can be use to stimulate the bone marrow granulopoiesis.


An insulin-dependent hypoglycaemia induced by electroacupuncture at the Zhongwan (CV12) acupoint in diabetic rats.
Acupuncture at the Zhongwan acupoint has been widely used in traditional Chinese medicine to relieve symptoms of diabetes mellitus. Our study investigated the effect on plasma glucose of electroacupuncture applied at the Zhongwan acupoint in rat diabetic models. Plasma concentrations of insulin, glucagon and beta-endorphin- were also determined using radioimmunoassay. A decrease in plasma glucose was observed in rats after electroacupuncture (15 Hz, 10 mA) for 30 min at the Zhongwan acupoint. This was observed in normal rats and rat models with Type II (non-insulin-dependent) diabetes mellitus. No significant effect on plasma glucose was observed in rat models with Type I (insulin-dependent) diabetes mellitus: neither the streptozotocin (STZ)-induced diabetic rats nor the genetic (BB/W) rats. Further, the hypoglycaemic action of electroacupuncture stimulation disappeared in rats with insulin-resistance induced by an injection of human long-acting insulin repeated daily to cause the loss of tolbutamide-induced hypoglycaemia. An insulin-related action can thus be hypothesised. This hypothesis is supported by an increase in plasma insulin-like immunoreactivity after electroacupuncture stimulation in normal rats. Participation of glucagon was ruled out because there was no change in plasma glucagon-like immunoreactivity resulting from electroacupuncture stimulation. In addition to an increase in plasma beta-endorphin-like immunoreactivity, the plasma glucose lowering action of electroacupuncture stimulation at Zhongwan acupoint was abolished by naloxone in a sufficient dose to block opioid receptors. Thus we suggest that electroacupuncture stimulation at the Zhongwan acupoint induces secretion of endogenous beta-endorphin which reduces plasma glucose concentration in an insulin-dependent manner.


You Can't Make Me Do It, but I Could Be Persuaded: A Federalism Perspective on the Affordable Care Act.
The Affordable Care Act (ACA) seeks to change fundamentally the US health care system. The responses of states have been diverse and changing. What explains these diverse and dynamic responses? We examine the decision making of states concerning the creation of Pre-existing Condition Insurance Plan programs and insurance marketplaces and the expansion of Medicaid in historical context. This frames our analysis and its implications for future health reform in broader perspective by identifying a number of characteristics of state-federal grants programs: (1) slow and uneven implementation; (2) wide variation across states; (3) accommodation by the federal government; (4) ideological conflict; (5) state response to incentives; (6) incomplete take-up rates of eligible individuals; and (7) programs as stepping-stones and wedges. Assessing the implementation of the three main components of the ACA, we find that partisanship exerts significant influence, yet less so in the case of Medicaid expansion. Moreover, factors specific to the insurance market also play an important role. Finally, we conclude by applying the themes to the ACA and offer an outlook for its continuing implementation. Specifically, we expect a gradual move toward universal state participation in the ACA, especially with respect to Medicaid expansion.


Laser Surgery of Soft Tissue in Orthodontics: Review of the Clinical Trials.
Introduction: Recently, a wide variety of procedures have been done by laser application in orthodontics. Apart from the mentioned range of various treatments, laser has become a tool for many soft tissue surgeries as an alternative to conventional scalpel-based technique during orthodontic treatments in the management of soft tissue. Due to scarce information in the latter subject, this study was designed in order to include clinical trials that included soft tissue ablation by laser in orthodontics. Methods: Literature was searched based on PubMed and Google Scholar databases in 5 years (2010-2015) with English language restriction and clinical trial design. Studies that performed soft tissue application of laser during orthodontic treatment were extracted by the authors. Results: Only eight studies met the inclusion criteria. No significant difference was found between laser ablation and conventional scalpel technique in the matter of treatment outcome. However, few issues remained to clarify the differences in the mentioned procedures. Conclusion: Laser performance can be recommended in case of preceding less bleeding and discomfort during surgical procedure. There are still quandaries among clinical application of scalpel-based surgery in aesthetic region with bracket-bonded teeth. Precaution and knowledge regarding the characteristics of laser beam such as wavelength, frequency, power and timing is extremely needed.


Ultrastructural study of guinea pig tracheal smooth muscle and its innervation.
The ultrastructure of guinea pig tracheal smooth muscle and its innervation was studied by transmission electron microscopy. Smooth muscle cells were arranged into bundles separated by collagen and connective tissue cells. Gap junctions were found only rarely between adjacent smooth muscle cells. Bundles of axons (predominantly unmyelinated) were seen on the periphery of the muscle bundles. Some penetrated deep into the bundles and a few made close neuromuscular contacts (less than 20 nm). Such a spatial arrangement of nerves and smooth muscle suggests a potential for nervous control and limited myogenic activity. Axon profiles containing small agranular vesicles characteristic of cholinergic nerves and small granular vesicles characteristic of adrenergic nerves were seen in close association with airway smooth muscle and with one another. Some large granular vesicles were associated with varicosities of both nerve types. Quantitative studies demonstrated a preponderance of cholinergic over adrenergic nerve profiles in both upper and lower segments. Varicosities, characteristic of adrenergic nerves, were observed more frequently in upper compared with lower segments; however, observed differences were not significant. The morphological findings are consistent with the control functions exerted by cholinergic and adrenergic nerves but provide no basis for control by nonadrenergic inhibitory nerves.


Between Gloom and Transfiguration: Analysis of Petitions conducted by the Ministry of Health of the GDR as a Historical Source for the Description of the Main Problems and the Strategies to handle Problems in the GDR Health Care System
Due to its outstanding political significance the GDR dedicated extraordinary attention to their health care system. By the help of analyses of petitions (complaints and hints) of the citizens conducted by the Ministry of Health of the GDR it is possible to gain a survey about the critiques and expectations in this field that were submitted to the GDR authorities. They form a comprehensive and significant source for the understanding of the GDR health care system, individual and social values as well as institutional reactions and strategies to handle problems. Surprisingly, critiques about the behaviour of the medical staff ranked almost equally to complaints about material deficits. Based on an elaborate system of evaluation these petitions served as an instrument of citizens of "direct democracy" that caused critical reflection of the government and influenced its perception and action. On the other hand the critique concerning the health care system individualized the protests, prevented a collective movement and generally only caused corrections of details.


Toxicological determination and in vitro metabolism of the designer drug methylenedioxypyrovalerone (MDPV) by gas chromatography/mass spectrometry and liquid chromatography/quadrupole time-of-flight mass spectrometry.
A method for the toxicological screening of the new designer drug methylenedioxypyrovalerone (MDPV) is described; with an emphasis on its application for anti-doping analysis. The metabolism of MDPV was evaluated in vitro using human liver microsomes and S9 cellular fractions for CYP450 phase I and uridine 5'-diphosphoglucuronosyltransferase (UGT) and sulfotransferase (SULT) phase II metabolism studies. The resulting metabolites were subsequently liquid/liquid extracted and analyzed using gas chromatography/mass spectrometry (GC/MS) as trimethylsilyl (TMS) derivatives. The structures of the metabolites were further confirmed by accurate mass measurement using a liquid chromatography/quadrupole time-of-flight (LC/QTOF) mass spectrometer. The studies demonstrated that the main metabolites of MDPV are catechol and methyl catechol pyrovalerone, which are in turn sulfated and glucuronated. The method for the determination of MDPV in urine has been fully validated by assessing the limits of detection and quantification, linearity, repeatability, and accuracy. This validation demonstrates the suitability for screening of this stimulant substance for anti-doping and forensic toxicology purposes.


An adaptive large neighborhood search heuristic for Two-Echelon Vehicle Routing Problems arising in city logistics.
In this paper, we propose an adaptive large neighborhood search heuristic for the Two-Echelon Vehicle Routing Problem (2E-VRP) and the Location Routing Problem (LRP). The 2E-VRP arises in two-level transportation systems such as those encountered in the context of city logistics. In such systems, freight arrives at a major terminal and is shipped through intermediate satellite facilities to the final customers. The LRP can be seen as a special case of the 2E-VRP in which vehicle routing is performed only at the second level. We have developed new neighborhood search operators by exploiting the structure of the two problem classes considered and have also adapted existing operators from the literature. The operators are used in a hierarchical scheme reflecting the multi-level nature of the problem. Computational experiments conducted on several sets of instances from the literature show that our algorithm outperforms existing solution methods for the 2E-VRP and achieves excellent results on the LRP.


The role of sea ice for vascular plant dispersal in the Arctic.
Sea ice has been suggested to be an important factor for dispersal of vascular plants in the Arctic. To assess its role for postglacial colonization in the North Atlantic region, we compiled data on the first Late Glacial to Holocene occurrence of vascular plant species in East Greenland, Iceland, the Faroe Islands and Svalbard. For each record, we reconstructed likely past dispersal events using data on species distributions and genetics. We compared these data to sea-ice reconstructions to evaluate the potential role of sea ice in these past colonization events and finally evaluated these results using a compilation of driftwood records as an independent source of evidence that sea ice can disperse biological material. Our results show that sea ice was, in general, more prevalent along the most likely dispersal routes at times of assumed first colonization than along other possible routes. Also, driftwood is frequently dispersed in regions that have sea ice today. Thus, sea ice may act as an important dispersal agent. Melting sea ice may hamper future dispersal of Arctic plants and thereby cause more genetic differentiation. It may also limit the northwards expansion of competing boreal species, and hence favour the persistence of Arctic species.


Cancer incidence and mortality in Newark, N. J. 1970-1974: a national comparison.
New Jersey has acquired the invidious label "Cancer Alley U. S. A." based upon a national cancer mortality analysis. However, a cancer incidence survey conducted in Newark, the largest metropolitan industrial city in New Jersey, showed that age-adjusted Newark rates for all sites were comparable to the Third National Cancer Survey (TNCS) and Surveillance, Epidemiology and End Results (SEER) populations, except for black males who had statistically lower rates compared with the SEER population only. However, Newark did have statistically higher incidence of the following: (a) esophagus cancer among white men, black men, and black women; and (b) cervix, uterus, ovary, and bladder cancers among black women. Age-adjusted Newark cancer mortality for all sites was not statistically different from the SEER experience, except for an excessive cancer mortality among white men for stomach and esophagus; white women for stomach, colon-rectum, and uterus; black men for esophagus and colon-rectum; black women for colon-rectum, cervix, uterus, and ovary. An analysis of Newark mortality/incidence ratios suggests that the excessive cancer burden for the majority of sites studied resulted from poor end results of therapy, probably due to either late diagnosis, poor compliance, and/or suboptimal therapy. The Newark data cast doubt on the validity of the use of mortality data only in referring pejoratively to New Jersey as "Cancer Alley U. S. A."


Response Inhibition and Predicting Response to Pharmacological and Cognitive Behavioral Therapy Treatments for Major Depressive Disorder: A Canadian Biomarker Integration Network for Depression Study.
Major depressive disorder (MDD) is associated with various cognitive impairments, including response inhibition. Deficits in response inhibition may also underlie poor antidepressant treatment response. Recent studies revealed that the neurobiological correlates of response inhibition can predict response to pharmacological treatments. However, the generalizability of this finding to first-line nonpharmacological treatments, particularly cognitive behavioral therapy, remains to be investigated. Data from two independent treatment protocols were combined, one in which 65 patients with MDD underwent treatment with escitalopram, and the other in which 41 patients with MDD underwent a course of cognitive behavioral therapy. A total of 25 healthy control subjects were also recruited. Neural correlates of response inhibition were captured by participants completing a Go/NoGo task during electroencephalography recording. Response inhibition-related measures of interest included the amplitudes of the N2 and P3 event-related potentials. Pretreatment P3 amplitude, which has been linked to both the motor and cognitive aspects of response inhibition, was a significant predictor of change in depressive symptoms following escitalopram and cognitive behavioral therapy treatment. A greater pretreatment P3 amplitude was associated with a greater reduction in depressive severity. In addition, the pretreatment P3 amplitude was found to be significantly greater at baseline in remitters than in nonremitters and healthy control subjects. The integrity of response inhibition may be critical for a successful course of pharmacological or psychological treatment for MDD. Electrophysiological correlates of response inhibition may have utility as a general prognostic marker of treatment response in MDD. Future studies may investigate the benefit of preceding first-line treatments with interventions that improve response inhibition in MDD.


Temperature-induced liquid crystal microdroplet formation in a partially miscible liquid mixture.
Liquid-in-liquid droplets are typically generated by the partitioning of immiscible fluids, e.g. by mechanical shearing with macroscopic homogenisers or microfluidic flow focussing. In contrast, partially miscible liquids with a critical solution temperature display a temperature-dependent mixing behaviour. In this work, we demonstrate how, for a blend of methanol (MeOH) and the thermotropic liquid crystal (LC) 4-Cyano-4'-pentylbiphenyl (5CB), cooling from a miscible to an immiscible state allows the controlled formation of microdroplets. A near-room-temperature-induced phase separation leads to nucleation, growth and coalescence of mesogen-rich droplets. The size and number of the droplets is tunable on the microscopic scale by variation of temperature quench depth and cooling rate. Further cooling induces a phase transition to nematic droplets with radial configuration, well-defined sizes and stability over the course of an hour. This temperature-induced approach offers a scalable and reversible alternative to droplet formation with relevance in diagnostics, optoelectronics, materials templating and extraction processes.


Osmotic water permeability of human red cells.
The osmotic water permeability of human red cells has been reexamined with a stopped-flow device and a new perturbation technique. Small osmotic gradients are used to minimize the systematic error caused by nonlinearities in the relationship between cell volume and light scattering. Corrections are then made for residual systematic error. Our results show that the hydraulic conductivity, Lp, is essentially independent of the direction of water flow and of osmolality in the range 184-365 mosM. the mean value of Lp obtained obtained was 1.8 +/- 0.1 (SEM) X 10-11 cm3 dyne -1 s-1.


Mithramycin suppresses DNA damage repair via targeting androgen receptor in prostate cancer.
The dependency of prostate cancer (PCa) growth on androgen receptor (AR) signaling has been harnessed to develop first-line therapies for high-risk localized and metastatic PCa treatment. However, the occurrence of aberrant expression, mutated or splice variants of AR confers resistance to androgen ablation therapy (ADT), radiotherapy or chemotherapy in AR-positive PCa. Therapeutic strategies that effectively inhibit the expression and/or transcriptional activity of full-length AR, mutated AR and AR splice variants have remained elusive. In this study, we report that mithramycin (MTM), an antineoplastic antibiotic, suppresses cell proliferation and exhibits dual inhibitory effects on expression and transcriptional activity of AR and AR splice variants. MTM blocks AR recruitment to its genomic targets by occupying AR enhancers and causes downregulation of AR target genes, which includes key DNA repair factors in DNA damage repair (DDR). We show that MTM significantly impairs DDR and enhances the effectiveness of ionizing radiation or the radiomimetic agent Bleomycin in PCa. Thus, the combination of MTM treatment with RT or radiomimetic agents, such as bleomycin, may present a novel effective therapeutic strategy for patients with high-risk, clinically localized PCa.


18-month effectiveness of short-course antiretroviral regimens combined with alternatives to breastfeeding to prevent HIV mother-to-child transmission.
We assessed the 18-month effectiveness of short-course (sc) antiretroviral peripartum regimens combined with alternatives to prolonged breastfeeding to prevent mother-to-child transmission (MTCT) of HIV-1 in Abidjan, Côte d'Ivoire. HIV-1 infected pregnant women received from >/=32-36 weeks of gestation scZidovudine (ZDV)+/-Lamivudine (3TC)+single-dose Nevirapine (sdNVP) at delivery within the ANRS 1201/1202 DITRAME-Plus cohort (2001-2003). Neonates received a sdNVP+7-day ZDV prophylaxis. Two infant-feeding interventions were systematically offered free of charge: formula-feeding or exclusive shortened breastfeeding with early cessation from four months. The reference group was the ANRS 049a DITRAME cohort (1994-2000) exposed to scZDV from 36 weeks, then to prolonged breastfeeding. Pediatric HIV infection was defined by a positive plasma HIV-1 RNA at any age, or if aged >/=18 months, a positive HIV-1 serology. Turnbull estimates of cumulative transmission risks (CTR) and effectiveness (HIV-free survival) were compared by exposure group using a Cox model. Among 926 live-born children enrolled, 107 (11.6%) were HIV-infected at 18 months. CTRs were 22.3% (95% confidence interval[CI]:16-30%) in the 238 ZDV long-term breastfed reference group, 15.9% (CI:10-27%) in the 169 ZDV+sdNVP shortened breastfed group; 9.4% (CI:6-14%) in the 195 ZDV+sdNVP formula-fed group; 6.8% (CI:4-11%) in the 198 ZDV+3TC+sdNVP shortened breastfed group, and 5.6% (CI:2-10%) in the 126 ZDV+3TC+sdNVP formula-fed group. Each combination had a significantly higher effectiveness than the ZDV long-term breastfed group except for ZDV+sdNVP shortened breastfed children, ranging from 51% (CI:20-70%) for ZDV+sdNVP formula fed children to 63% (CI:40-80%) for ZDV+3TC+NVPsd shortened breastfed children, after adjustment for maternal eligibility for antiretroviral therapy (ART), home delivery and low birth-weight. Substantial MTCT risk reductions are reachable in Africa, even in short-term breastfed children. The two sc antiretroviral combinations associated to any of the two infant feeding interventions, formula-feeding and shortened breastfeeding, reduce significantly MTCT with long-term benefit until age 18 months and without increasing mortality.


Differences in plasma metabolomics between sows fed DL-methionine and its hydroxy analogue reveal a strong association of milk composition and neonatal growth with maternal methionine nutrition.
The aim of the present study was to determine whether increased consumption of methionine as DL-methionine (DLM) or its hydroxy analogue DL-2-hydroxy-4-methylthiobutanoic acid (HMTBA) could benefit milk synthesis and neonatal growth. For this purpose, eighteen cross-bred (Landrace × Yorkshire) primiparous sows were fed a control (CON), DLM or HMTBA diet (n 6 per diet) from 0 to 14 d post-partum. At postnatal day 14, piglets in the HMTBA group had higher body weight (P= 0·02) than those in the CON group, tended (P= 0·07) to be higher than those in the DLM group, and had higher (P< 0·05) mRNA abundance of jejunal fatty acid-binding protein 2, intestinal than those in the CON and DLM groups. Compared with the CON diet-fed sows, milk protein, non-fat solid, and lysine, histidine and ornithine concentrations decreased in the DLM diet-fed sows (P< 0·05), and milk fat, lactose, and cysteine and taurine concentrations increased in the HMTBA diet-fed sows (P< 0·05). Plasma homocysteine and urea N concentrations that averaged across time were increased (P< 0·05) in sows fed the DLM diet compared with those fed the CON diet. Metabolomic results based on ¹H NMR spectroscopy revealed that consumption of the HMTBA and DLM diets increased (P< 0·05) both sow plasma methionine and valine levels; however, consumption of the DLM diet led to lower (P< 0·05) plasma levels of lysine, tyrosine, glucose and acetate and higher (P< 0·05) plasma levels of citrate, lactate, formate, glycerol, myo-inositol and N-acetyl glycoprotein in sows. Collectively, neonatal growth and milk synthesis were regulated by dietary methionine levels and sources, which resulted in marked alterations in amino acid, lipid and glycogen metabolism.


Calicoblastic neoplasms in Acropora palmata, with a review of reports on anomalies of growth and form in corals.
Colonies of the elkhorn coral (Acropora palmata), possessing raised, whitened, irregularly shaped skeletal protuberances, were discovered at Carysfort Reef and Grecian Rocks, Key Largo National Marine Sanctuary, Key Largo, FL. These lesions exhibited relatively rapid growth and spread along the branches, as polyps were overlaid by coenosteal skeletal material. Histopathological examinations of the soft tissues surrounding and extending into the skeletal masses revealed proliferation of gastrovascular canals and associated calicoblastic epidermis, with loss of normal polyp structures and zooxanthellae. The slightly atypical tumor calicoblasts were cuboidal to columnar, resembling those found in the rapidly growing apical tips. Stable carbon isotope ratios of skeletal samples revealed that the tumor skeleton was isotopically lighter than the skeleton in the normal or apical track regions, indicative of higher tissue metabolic rates and lack of carbon isotope fractionation by zooxanthellae. This condition appears to be a neoplasm of the coral, and it is proposed that it be classified "calicoblastic epithelioma." Several different types of abnormal skeletal deposition, possibly the result of neoplastic processes, have been reported to occur in stony corals and are here reviewed.


Coherent noise reduction in digital holographic phase contrast microscopy by slightly shifting object.
A method to reduce coherent noise in digital holographic phase contrast microscopy is proposed. By slightly shifting the specimen, a series of digital holograms with different coherent noise patterns is recorded. Each hologram is reconstructed individually, while the different phase tilts of the reconstructed complex amplitudes due to the specimen shifts are corrected in the hologram plane by using numerical parametric lens method. Afterward, the lateral displacements of the phase maps from different holograms are compensated in the image plane by using digital image registration method. Thus, all phase images have same distribution, but uncorrelated coherent noise patterns. By a proper averaging procedure, the coherent noise of phase contrast image is reduced significantly. The experimental results are given to confirm the proposed method.


Moraxella catarrhalis activates murine macrophages through multiple toll like receptors and has reduced clearance in lungs from TLR4 mutant mice.
Moraxella catarrhalis is a gram negative bacterium and a leading causative agent of otitis media (OM) in children. Several recent reports have provided strong evidence for an association between toll like receptors and OM. It has been found that both Streptococcus pneumoniae and nontypeable Haemophilus influenzae activate host protective immune responses through toll like receptors (TLRs), however, the precise mechanism by which Moraxella catarrhalis initiates the host immune response is currently unknown. In this report, using murine macrophages generated from a series of knock-out mice, we have demonstrated that M. catarrhalis lipooligosaccharide (LOS) and either heat killed or live bacteria are recognized by one or more TLRs. LOS activates the host immune response through a membrane bound CD14-TLR4 complex, while both heat killed and live M.cat require recognition by multiple toll like receptors such as TLR2, TLR4 and TLR9 without the requirement of CD14. We have also shown that M.cat stimuli are capable of triggering the host innate immune response by both MyD88- and TRIF- dependent signaling pathways. We further showed that M.cat induced activation of mitogen activated protein kinase (MAPK) is essential in order to achieve optimal secretion of pro-inflammatory cytokine TNF-α. We finally showed that TLR4 mutant C3H/HeJ mice produce significantly lower levels of pro-inflammatory cytokines TNF-α and IL-6 in vivo, An increased bacterial loads at 12 and 24 hours (P<0.001) in their lungs upon challenge with live M.cat in an aerosol chamber compared to wild-type (WT) control mice. These data suggest that TLRs are crucial for an effective innate immune response induced by M.cat. The results of these studies contribute to an increased understanding of molecular mechanism and possible novel treatment strategies for diseases caused by M.cat by specifically targeting TLRs and their signaling pathways.


Modulation of Long-Range Connectivity Patterns via Frequency-Specific Stimulation of Human Cortex.
There is increasing interest in how the phase of local oscillatory activity within a brain area determines the long-range functional connectivity of that area. For example, increasing convergent evidence from a range of methodologies suggests that beta (20 Hz) oscillations may play a vital role in the function of the motor system [1-5]. The "communication through coherence" hypothesis posits that the precise phase of coherent oscillations in network nodes is a determinant of successful communication between them [6, 7]. Here we set out to determine whether oscillatory activity in the beta band serves to support this theory within the cortical motor network in vivo. We combined non-invasive transcranial alternating-current stimulation (tACS) [8-12] with resting-state functional MRI (fMRI) [13] to follow both changes in local activity and long-range connectivity, determined by inter-areal blood-oxygen-level-dependent (BOLD) signal correlation, as a proxy for communication in the human cortex. Twelve healthy subjects participated in three fMRI scans with 20 Hz, 5 Hz, or sham tACS applied separately on each scan. Transcranial magnetic stimulation (TMS) at beta frequency has previously been shown to increase local activity in the beta band [14] and to modulate long-range connectivity within the default mode network [15]. We demonstrated that beta-frequency tACS significantly changed the connectivity pattern of the stimulated primary motor cortex (M1), without changing overall local activity or network connectivity. This finding is supported by a simple phase-precession model, which demonstrates the plausibility of the results and provides emergent predictions that are consistent with our empirical findings. These findings therefore inform our understanding of how local oscillatory activity may underpin network connectivity.


The analysis of spinopelvic parameters and stability following long fusions with S1, S2 or iliac fixation.
The purpose of this study was to analyse changes of spinopelvic parameters and stability in the treatment of degenerative lumbar deformity. A retrospective review was carried out on 70 cases of degenerative lumbar deformity treated by long fusion with uni-cortical S1 fixation alone (US1F group, n = 20), bi-cortical S1 fixation alone (BS1F group, n = 20), additional diagonal S2 fixation (DS2F group, n = 14), and additional iliac fixation (ILF group, n = 16) from July 2003 to April 2010. The sagittal vertical axis (SVA), lumbar lordosis (LL), sacral slope (SS), pelvic tilt (PT), pelvic incidence (PI), and stability were used to evaluate radiologic outcomes. The clinical outcome was evaluated using the Oswestry Disability Index (ODI). In all groups, the LL was significantly increased at three months (p < 0.05). The PI and the SS of ILF patients significantly increased at three months (p < 0.05), while none of these values changed over time in non-ILF patients. The PT did not change postoperatively in any groups. The LL, SS, and PI were significantly greater in the ILF group at three months postoperatively and later (p < 0.05). DS2F and ILF had statistically significant stability compared to US1F and BS1F (p < 0.05). The ODI scores were significantly improved after surgery in all groups compared to before surgery (p < 0.05). Patients who had ILF in long fusion for the treatment of degenerative lumbar deformity had significant restoration of spinopelvic parameters compared to the other groups. In addition, DS2F and ILF provided more stability of distal instruments.


Efficacy evaluation of budesonide nebulizer as an adjunctive medication in post-rain asthma acute phase attack.
Bronchospasm attacks occur following syndromic rainfall and are increasing due to air pollution and need effective treatments. In this study, the effect of salbutamol nebulizer in comparison with salbutamol plus budesonide nebulizer in patients referred to the emergency department with dyspnea was investigated. The trial study was conducted on 228 patients with dyspnea after the first rainfall in Ahvaz. Two groups of 114 patients have been randomly allocated. On the course of treatment, the first group received salbutamol plus budesonide nebulizer and the second group received salbutamol alone. In the experimental group, budesonide 0.5 mg with salbutamol was nebulized three times for 20 minutes. In all patients, 20, 40, and 60 minutes after the start of the intervention, forced expiratory volume in 1 second (FEV1) and peak expiratory flow rate (PEFR) and vital signs of size were recorded and analyzed by SPSS and t-test. Data revealed that there were significant differences between PEFR parameters of studied. Groups in minute 40 and 60 after intervention (p = 0.000001). There was better improvement.in PEFR values in minute 40 and 60 in budesonide plus salbutamol study group. There were no significant differences for FEV1 in minute 0, 20, 40, 60 between to studied group. Also there were no significant differences for borg dyspnea scale for minute 0 and 60 between two experimented group. Respiratory rates have significant differences in minutes 20, 40 after intervention and there was better improvement for salbutamol plus budesonide group rather than sulbutamol intervention group alone.(p = 0.001142). Experiment data revealed. that due to the significant difference between PEFR and increased FEV1 in the combination of the two drugs and due to the corticosteroid effects of budesonide in reducing and preventing inflammation and swelling of the lungs, nebulizer salbutamol + budesonide has better effects on moderate breath than in nebulizer salbutamol.


Vaccine Hesitance: Risks, Persuasion, and influence.
In this edition of The Senior Care Pharmacist, we publish a variety of contributions addressing the issue of vaccination. As the Northern Hemisphere enters the autumn/fall period, this matter assumes a renewed significance, with the rollout of immunization as a preventive strategy for the benefit of both individual patients and the wider community. It is, perhaps, ironic that though effective vaccination services are now available for a range of communicable diseases such as influenza, and that these have only become relatively recently available in the course of medical history, health care workers promoting this approach now face a different problem that has sometimes been referred to as "vaccine hesitance."


Rules warp feature encoding in decision-making circuits.
We have the capacity to follow arbitrary stimulus-response rules, meaning simple policies that guide our behavior. Rule identity is broadly encoded across decision-making circuits, but there are less data on how rules shape the computations that lead to choices. One idea is that rules could simplify these computations. When we follow a rule, there is no need to encode or compute information that is irrelevant to the current rule, which could reduce the metabolic or energetic demands of decision-making. However, it is not clear if the brain can actually take advantage of this computational simplicity. To test this idea, we recorded from neurons in 3 regions linked to decision-making, the orbitofrontal cortex (OFC), ventral striatum (VS), and dorsal striatum (DS), while macaques performed a rule-based decision-making task. Rule-based decisions were identified via modeling rules as the latent causes of decisions. This left us with a set of physically identical choices that maximized reward and information, but could not be explained by simple stimulus-response rules. Contrasting rule-based choices with these residual choices revealed that following rules (1) decreased the energetic cost of decision-making; and (2) expanded rule-relevant coding dimensions and compressed rule-irrelevant ones. Together, these results suggest that we use rules, in part, because they reduce the costs of decision-making through a distributed representational warping in decision-making circuits.


Trafficking and signaling of G protein-coupled receptors in the nervous system: implications for disease and therapy.
G protein-coupled receptors (GPCRs) are expressed throughout the nervous system where they regulate multiple physiological processes, participate in neurological diseases, and are major targets for therapy. Given that many GPCRs respond to neurotransmitters and hormones that are present in the extracellular fluid and which do not readily cross the plasma membrane, receptor trafficking to and from the plasma membrane is a critically important determinant of cellular responsiveness. Moreover, trafficking of GPCRs throughout the endosomal system can initiate signaling events that are mechanistically and functionally distinct from those operating at the plasma membrane. This review discusses recent advances in the relationship between signaling and trafficking of GPCRs in the nervous system. It summarizes how receptor modifications influence trafficking, discusses mechanisms that regulate GPCR trafficking to and from the plasma membrane, reviews the relationship between trafficking and signaling, and considers the implications of GPCR trafficking to drug development.


A higher-level classification of the Pannonian and western Pontic steppe grasslands (Central and Eastern Europe).
What are the main floristic patterns in the Pannonian and western Pontic steppe grasslands? What are the diagnostic species of the major subdivisions of the class Festuco-Brometea (temperate Euro-Siberian dry and semi-dry grasslands)? Carpathian Basin (E Austria, SE Czech Republic, Slovakia, Hungary, Romania, Slovenia, N Croatia and N Serbia), Ukraine, S Poland and the Bryansk region of W Russia. We applied a geographically stratified resampling to a large set of relevés containing at least one indicator species of steppe grasslands. The resulting data set of 17 993 relevés was classified using the TWINSPAN algorithm. We identified groups of clusters that corresponded to the class Festuco-Brometea. After excluding relevés not belonging to our target class, we applied a consensus of three fidelity measures, also taking into account external knowledge, to establish the diagnostic species of the orders of the class. The original TWINSPAN divisions were revised on the basis of these diagnostic species. The TWINSPAN classification revealed soil moisture as the most important environmental factor. Eight out of 16 TWINSPAN groups corresponded to Festuco-Brometea. A total of 80, 32 and 58 species were accepted as diagnostic for the orders Brometalia erecti, Festucetalia valesiacae and Stipo-Festucetalia pallentis, respectively. In the further subdivision of the orders, soil conditions, geographic distribution and altitude could be identified as factors driving the major floristic patterns. We propose the following classification of the Festuco-Brometea in our study area: (1) Brometalia erecti (semi-dry grasslands) with Scabioso ochroleucae-Poion angustifoliae (steppe meadows of the forest zone of E Europe) and Cirsio-Brachypodion pinnati (meadow steppes on deep soils in the forest-steppe zone of E Central and E Europe); (2) Festucetalia valesiacae (grass steppes) with Festucion valesiacae (grass steppes on less developed soils in the forest-steppe zone of E Central and E Europe) and Stipion lessingianae (grass steppes in the steppe zone); (3) Stipo-Festucetalia pallentis (rocky steppes) with Asplenio septentrionalis-Festucion pallentis (rocky steppes on siliceous and intermediate soils), Bromo-Festucion pallentis (thermophilous rocky steppes on calcareous soils), Diantho-Seslerion (dealpine Sesleria caerulea grasslands of the Western Carpathians) and Seslerion rigidae (dealpine Sesleria rigida grasslands of the Romanian Carpathians).


Resistant Starch Regulates Gut Microbiota: Structure, Biochemistry and Cell Signalling.
Starch is one of the most popular nutritional sources for both human and animals. Due to the variation of its nutritional traits and biochemical specificities, starch has been classified into rapidly digestible, slowly digestible and resistant starch. Resistant starch has its own unique chemical structure, and various forms of resistant starch are commercially available. It has been found being a multiple-functional regulator for treating metabolic dysfunction. Different functions of resistant starch such as modulation of the gut microbiota, gut peptides, circulating growth factors, circulating inflammatory mediators have been characterized by animal studies and clinical trials. In this mini-review, recent remarkable progress in resistant starch on gut microbiota, particularly the effect of structure, biochemistry and cell signaling on nutrition has been summarized, with highlights on its regulatory effect on gut microbiota.


Preparation, immunogenicity, and protective efficacy, in a murine model, of a conjugate vaccine composed of the polysaccharide moiety of the lipopolysaccharide of Vibrio cholerae O139 bound to tetanus toxoid.
The epidemic and pandemic potential of Vibrio cholerae O139 is such that a vaccine against this newly emerged serogroup of V. cholerae is required. A conjugate made of the polysaccharide moiety (O-specific polysaccharide plus core) of the lipopolysaccharide (LPS) of V. cholerae O139 (pmLPS) was prepared by derivatization of the pmLPS with adipic acid dihydrazide and coupling to tetanus toxoid (TT) by carbodiimide-mediated condensation. The immunologic properties of the conjugate were tested using BALB/c mice injected subcutaneously three times at 2 weeks interval and then a fourth time 4 weeks later. Mice were bled 7 days after each injection and then once each month for the following 6 months. LPS and TT antibody levels were determined by enzyme-linked immunosorbent assay using immunoplates coated with either O139 LPS or TT. Both pmLPS and pmLPS-TT conjugate elicited low levels of immunoglobulin M (IgM), peaking 5 weeks after the first immunization. The conjugate elicited high levels of IgG antibodies, peaking 3 months after the first immunization and declining slowly during the following 5 months. TT alone, or as a component of conjugate, induced mostly IgG antibodies. Antibodies elicited by the conjugate recognized both capsular polysaccharide and LPS from V. cholerae O139 and were vibriocidal. They were also protective in the neonatal mouse model of cholera infection. The conjugation of the O139 pmLPS, therefore, enhanced its immunogenicity and conferred T-dependent properties to this polysaccharide.


Factor analyses differentiate clinical phenotypes of idiopathic and interferon-alpha-induced depression.
The discovery that prolonged administration of interferon-alpha (a pro-inflammatory cytokine) readily precipitates depressive symptoms has played a key role in development of the inflammation theory of major depressive disorder (MDD). However, it remains unclear whether the clinical phenotype of patients with inflammation-associated depression significantly overlaps with, or can be distinguished from that of patients with 'idiopathic' depression. Here we explored the Hamilton depression scale factor structure of 172 patients undergoing interferon-alpha treatment for hepatitis-C at the point of transition to a depressive episode of DSM IV defined major depression severity. The resulting factor structure was first compared with a model derived from 6 previous studies of 'idiopathic' MDD (Cole et al., 2004). This confirmatory factor analysis revealed that the factor structure of HAMD scores in our interferon-alpha treated cohort did not plausibly fit that previously described for 'idiopathic' MDD. Instead, subsequent exploratory factor analysis revealed a distinct four factor model with a novel primary factor grouping cognitive symptoms of depression and anxiety (HAMD items 1, 2, 9, 10, 11, 15). The second sleep disorder factor (items 4, 5, 6) replicated previous findings in 'idiopathic' depression. A third and unique factor grouped somatic symptoms and function (items 7, 12, 13, 14 and item 1). The final factor (also common in idiopathic depression studies), grouped gastrointestinal symptoms and weight loss (items 12 and 16). Severe depression items (3, 8, and 17) were excluded from analysis due to very low variance. At transition, interferon-alpha induced major depressive episodes therefore appears to have more associated anxiety features that covary with depressed mood than classical or 'idiopathic' MDD and a low likelihood of severe features such as suicidal ideation. Identification of this clinical phenotype may help identify patients with an inflammatory depression etiology and support the development of more effective and personalized therapies.


Constraint-induced movement therapy for motor recovery after stroke.
A new therapeutic approach to rehabilitation of movement after stroke, termed Constraint-Induced (CI) Movement Therapy, has been derived from basic research with monkeys given somatosensory deafferentation. CI consists of a family of therapies; their common element is that they induce stroke patients to greatly increase the use of a more affected upper extremity for many hours a day over a 10-14 consecutive-day period. These therapies have significantly improved quality of movement and substantially increased amount of use of a more affected extremity in the activities of daily living in the life situation. The purpose of this paper is to describe the protocol used by the investigative team that developed the family of CI therapies and examined them as an effective rehabilitation approach.


Insights into the light-driven hydrogen evolution reaction of mesoporous graphitic carbon nitride decorated with Pt or Ru nanoparticles.
Ru or Pt nanoparticles have been prepared following the organometallic approach and deposited onto the surface of mesoporous graphitic carbon nitride (mpg-CN). Three different Ru-based samples have also been compared to investigate the effect of 4-phenylpyridine as a stabilizing agent. The photocatalytic performance towards the hydrogen evolution reaction (HER) has been tested showing that all hybrid systems clearly outperform the photocatalytic activity of bare mpg-CN. In particular, Pt-decorated mpg-CN yields the largest H2 production upon visible-light irradiation (870 μmol h-1 g-1, TOF = 14.1 h-1, TON = 339 after 24 h) when compared with the Ru-based samples (137-155 μmol h-1 g-1, TOFs between 2.3-2.7 h-1, TONs between 54-57 after 24 h). Long-term photochemical tests (up to 65 h irradiation) show also an improved stability of the Pt-based samples over the Ru counterpart. Photophysical experiments aimed at rationalizing the photocatalytic performance of the different hybrid systems elucidate that the enhanced activity of the Pt-decorated mpg-CN over the Ru-based analogues arises from improved electron transfer kinetics from mpg-CN to the metal nanoparticles.


Insulin increases the cell number of the inner cell mass and stimulates morphological development of mouse blastocysts in vitro.
Previous studies showed that insulin promotes cell proliferation and morphological development of preimplantation mouse embryos. In this report, the receptor responsible for these actions and the cell populations that are affected were investigated. Insulin's 9% stimulation of blastocyst cell number was entirely due to a 23% increase in ICM cell number with an EC50 of 0.54 pM. This and the similar degrees of stimulation of immunosurgically isolated ICMs by both physiological and supraphysiological insulin concentrations suggest that insulin receptors are present on the ICM and respond to exogenous insulin transcytosed through the TE to promote expansion of the ICM cell numbers. In morphological studies, insulin increased the number of blastocysts and decreased the number of morulae by 10% after 54 h culture from 2-cell embryos with EC50s of about 0.95 pM. The equivalence of these EC50s suggests mediation of insulin's stimulation of blastocyst formation via insulin receptors which are functionally expressed around the time of compaction at the 8-cell stage. These results support our hypothesis that insulin has an important role in the regulation of growth during preimplantation development.


Thyroid gland ectopias.
Failures of descent of the medial anlage of the thyroid and incomplete obliteration of its vertical tract lead to midline or near-midline ectopias such as lingual thyroid and thyroglossal cysts. Each poses special diagnostic and therapeutic considerations. "Ectopias" of the thyroid gland lateral to the carotid artery and jugular vein, however, cannot be readily explained by current embryological information. In these instances, and especially for intranodal thyroid tissue, a metastasis from an occult thyroid primary is the mandatory first consideration.


Ethanol may suppress Wnt/beta-catenin signaling on human bone marrow stroma cells: a preliminary study.
Ethanol and glucocorticoids are risk factors associated with osteonecrosis. Previous reports suggest ethanol and glucocorticoids induce adipogenesis, decrease osteogenesis in bone marrow stroma cells, and produce intracellular lipid deposits resulting in death of osteocytes. The Wnt/beta-catenin signal pathway is involved in the regulation of homeostasis of bone and we presume glucocorticoids and ethanol may induce osteonecrosis in humans through a similar mechanism as in rodents. We hypothesized (1) ethanol, like glucocorticoids, decreases osteogenesis and increases adipogenesis through the Wnt/beta-catenin signaling pathway in human bone marrow stromal cells; and (2) ethanol decreases intranuclear translocation of beta-catenin. We found both dexamethasone and ethanol decrease the gene and protein expression of osteogenesis and increase that of adipogenesis through Wnt signaling-related genes by semiquantitative and quantitative polymerase chain reaction and Western blot. Ethanol hampered intranuclear translocation of beta-catenin by immunofluorescence analysis. The data suggest the Wnt/beta-catenin signaling pathway may be associated with ethanol-induced osteonecrosis.


Increased Platelet-Monocyte Interaction in Stable COPD in the Absence of Platelet Hyper-Reactivity.
Chronic obstructive pulmonary disease (COPD) is well known for its cardiovascular co-morbidities. Increased platelet-monocyte interaction is found in COPD and may reflect altered platelet function and a potential role for anti-platelet therapy. The objectives were to investigate platelet-monocyte interaction, platelet activation and reactivity and plasmatic coagulation in stable COPD. Platelet-monocyte interaction and platelet activation were determined by flow cytometry in 30 stable COPD patients and 25 controls. Platelet activation was measured by binding of fibrinogen to the activated fibrinogen receptor and platelet P-selectin expression at baseline and after platelet stimulation with platelet agonists. Plasmatic coagulation was measured by D-dimer and thrombin generation. Platelet-monocyte interaction was increased in stable COPD (median fluorescence intensity of platelet CD61 was 19.8 [IQR 14.0-33.2] vs. 10.0 [IQR 8.7-16.7], p = 0.002). In contrast, platelet activation and reactivity, reflected by fibrinogen binding and P-selectin expression, were the same in both groups. Plasma P-selectin and interleukin-6 were increased in COPD (p = 0.01 and p = 0.02, respectively), whereas soluble fibrinogen, D-dimer and thrombin generation were similar. Increased platelet-monocyte interaction was found in the absence of platelet hyper-reactivity and activation of plasmatic coagulation in stable COPD. Future clinical evaluation of the effects of different anti-platelet drugs in COPD is warranted, as anti-platelet therapy may interfere with platelet-monocyte interaction.


The therapeutic potential of erythropoietin receptor transgenes.
Allogeneic bone marrow transplantation is an effective curative therapy for both malignant and heritable diseases. The use of genetically altered autologous hematopoietic stem cells (HSC) is being increasingly investigated as a treatment for a variety of non-malignant but significantly morbid diseases, including hemoglobinopathies, immunodeficiencies and autoimmune diseases. Other hematopoietic cells capable of proliferation, such as antigen-specific T cells and dendritic cells, have also been used for adoptive immunotherapy. Genetic procedures to modify these various therapeutic cells so that they can be selectively amplified either in vitro or in vivo could enhance their efficacy. For example, HSC that contain a gene that confers a survival, selection or growth advantage may enhance their engraftment. Such enhancement could be expected to reduce graft failures and the intensity of the required conditioning regimen, thereby decreasing the toxicities of transplantation. In this review, the functions of cytokine receptor transgenes coding for erythropoietin receptors (EpoR) are analyzed. The characteristics of these transgenic cells and animals are discussed with regard to the possible therapeutic use of EpoR transgenes in the transplantation of hematopoietic cells.


Outside the Box: Working With Wildlife in Biocontainment.
Research with captive wildlife in Animal Biosafety Level 2 (ABSL2) and 3 (ABSL3) facilities is becoming increasingly necessary as emerging and re-emerging diseases involving wildlife have increasing impacts on human, animal, and environmental health. Utilizing wildlife species in a research facility often requires outside the box thinking with specialized knowledge, practices, facilities, and equipment. The USGS National Wildlife Health Center (NWHC) houses an ABSL3 facility dedicated to understanding wildlife diseases and developing tools to mitigate their impacts on animal and human health. This review presents considerations for utilizing captive wildlife for infectious disease studies, including, husbandry, animal welfare, veterinary care, and biosafety. Examples are drawn from primary literature review and collective 40-year experience of the NWHC. Working with wildlife in ABSL2 and ABSL3 facilities differs from laboratory animals in that typical laboratory housing systems, husbandry practices, and biosafety practices are not designed for work with wildlife. This requires thoughtful adaptation of standard equipment and practices, invention of customized solutions and development of appropriate enrichment plans using the natural history of the species and the microbiological characteristics of introduced and native pathogens. Ultimately, this task requires critical risk assessment, understanding of the physical and psychological needs of diverse species, creativity, innovation, and flexibility. Finally, continual reassessment and improvement are imperative in this constantly changing specialty area of infectious disease and environmental hazard research.


The role of perceived barriers and objectively measured physical activity in adults aged 65-100.
to test the predictive utility of perceived barriers to objectively measured physical activity levels in a stratified sample of older adults when accounting for social-cognitive determinants proposed by the Theory of Planned Behaviour (TPB), and economic and demographic factors. data were analysed from the Physical Activity Cohort Scotland survey, a representative and stratified (65-80 and 80+ years; deprived and affluent) sample of 584 community-dwelling older people, resident in Tayside, Scotland. Physical activity was measured objectively by accelerometry. perceived barriers clustered around the areas of poor health, lack of interest, lack of safety and lack of access. Perceived poor health and lack of interest, but not lack of access or concerns about personal safety, predicted physical activity after controlling for demographic, economic and TPB variables. perceived person-related barriers (poor health and lack of interest) seem to be more strongly associated with physical activity levels than perceived environmental barriers (safety and access) in a large sample of older adults. Perceived barriers are modifiable and may be a target for future interventions.


Fcγ receptors and immunomodulatory antibodies in cancer.
The discovery of both cytotoxic T lymphocyte-associated antigen 4 (CTLA4) and programmed cell death protein 1 (PD1) as negative regulators of antitumour immunity led to the development of numerous immunomodulatory antibodies as cancer treatments. Preclinical studies have demonstrated that the efficacy of immunoglobulin G (IgG)-based therapies depends not only on their ability to block or engage their targets but also on the antibody's constant region (Fc) and its interactions with Fcγ receptors (FcγRs). Fc-FcγR interactions are essential for the activity of tumour-targeting antibodies, such as rituximab, trastuzumab and cetuximab, where the killing of tumour cells occurs at least in part due to these mechanisms. However, our understanding of these interactions in the context of immunomodulatory antibodies designed to boost antitumour immunity remains less explored. In this Review, we discuss our current understanding of the contribution of FcγRs to the in vivo activity of immunomodulatory antibodies and the challenges of translating results from preclinical models into the clinic. In addition, we review the impact of genetic variability of human FcγRs on the activity of therapeutic antibodies and how antibody engineering is being utilized to develop the next generation of cancer immunotherapies.


Microencapsulation of Enteric Bacteriophages in a pH-Responsive Solid Oral Dosage Formulation Using a Scalable Membrane Emulsification Process.
A scalable low-shear membrane emulsification process was used to produce microencapsulated Escherichia coli-phages in a solid oral dosage form. Uniform pH-responsive composite microparticles (mean size ~100 µm) composed of Eudragit® S100 and alginate were produced. The internal microstructure of the gelled microcapsules was studied using ion-milling and imaging, which showed that the microparticles had a solid internal core. The microencapsulation process significantly protected phages upon prolonged exposure to a simulated gastric acidic environment. Encapsulated phages that had been pre-exposed to simulated gastric acid were added to actively growing bacterial cells using in vitro cell cultures and were found to be effective in killing E. coli. Encapsulated phages were also shown to be effective in killing actively growing E. coli in the presence of human epithelial cells. Confocal microscopy images showed that the morphology of encapsulated phage-treated epithelial cells was considerably better than controls without phage treatment. The encapsulated phages were stable during refrigerated storage over a four-week period. The process of membrane emulsification is highly scalable and is a promising route to produce industrial quantities of pH-responsive oral solid dosage forms suitable for delivering high titres of viable phages to the gastrointestinal tract.


Present and New Treatment Strategies in the Management of Glaucoma.
Glaucoma is a neurodegenerative disease characterized by retinal ganglion cell (RGC) death and axonal loss. It remains a major cause of blindness worldwide. All current modalities of treatment are focused on lowering intraocular pressure (IOP), and it is evident that increased IOP is an important risk factor for progression of the disease. However, it is clear that a significant number of glaucoma patients show disease progression despite of pressure lowering treatments. Much attention has been given to the development of neuroprotective treatment strategies, but the identification of such has been hampered by lack of understanding of the etiology of glaucoma. Hence, in spite of many attempts no neuroprotective drug has yet been clinically approved. Even though neuroprotection is without doubt an important treatment strategy, many glaucoma subjects are diagnosed after substantial loss of RGCs. In this matter, recent approaches aim to rescue RGCs and regenerate axons in order to restore visual function in glaucoma. The present review seeks to provide an overview of the present and new treatment strategies in the management of glaucoma. The treatment strategies are divided into current available glaucoma medications, new pressure lowering targets, prospective neuroprotective interventions, and finally possible neuroregenrative strategies.


Siberian larch (Larix sibirica Ledeb.) mitochondrial genome assembled using both short and long nucleotide sequence reads is currently the largest known mitogenome.
Plant mitochondrial genomes (mitogenomes) can be structurally complex while their size can vary from ~ 222 Kbp in Brassica napus to 11.3 Mbp in Silene conica. To date, in comparison with the number of plant species, only a few plant mitogenomes have been sequenced and released, particularly for conifers (the Pinaceae family). Conifers cover an ancient group of land plants that includes about 600 species, and which are of great ecological and economical value. Among them, Siberian larch (Larix sibirica Ledeb.) represents one of the keystone species in Siberian boreal forests. Yet, despite its importance for evolutionary and population studies, the mitogenome of Siberian larch has not yet been assembled and studied. Two sources of DNA sequences were used to search for mitochondrial DNA (mtDNA) sequences: mtDNA enriched samples and nucleotide reads generated in the de novo whole genome sequencing project, respectively. The assembly of the Siberian larch mitogenome contained nine contigs, with the shortest and the largest contigs being 24,767 bp and 4,008,762 bp, respectively. The total size of the genome was estimated at 11.7 Mbp. In total, 40 protein-coding, 34 tRNA, and 3 rRNA genes and numerous repetitive elements (REs) were annotated in this mitogenome. In total, 864 C-to-U RNA editing sites were found for 38 out of 40 protein-coding genes. The immense size of this genome, currently the largest reported, can be partly explained by variable numbers of mobile genetic elements, and introns, but unlikely by plasmid-related sequences. We found few plasmid-like insertions representing only 0.11% of the entire Siberian larch mitogenome. Our study showed that the size of the Siberian larch mitogenome is much larger than in other so far studied Gymnosperms, and in the same range as for the annual flowering plant Silene conica (11.3 Mbp). Similar to other species, the Siberian larch mitogenome contains relatively few genes, and despite its huge size, the repeated and low complexity regions cover only 14.46% of the mitogenome sequence.


Biophysical characterization of the calmodulin-like domain of Plasmodium falciparum calcium dependent protein kinase 3.
Calcium dependent protein kinases are unique to plants and certain parasites and comprise an N-terminal segment and a kinase domain that is regulated by a C-terminal calcium binding domain. Since the proteins are not found in man they are potential drug targets. We have characterized the calcium binding lobes of the regulatory domain of calcium dependent protein kinase 3 from the malaria parasite Plasmodium falciparum. Despite being structurally similar, the two lobes differ in several other regards. While the monomeric N-terminal lobe changes its structure in response to calcium binding and shows global dynamics on the sub-millisecond time-scale both in its apo and calcium bound states, the C-terminal lobe could not be prepared calcium-free and forms dimers in solution. If our results can be generalized to the full-length protein, they suggest that the C-terminal lobe is calcium bound even at basal levels and that activation is caused by the structural reorganization associated with binding of a single calcium ion to the N-terminal lobe.


Multitype Bellman-Harris branching model provides biological predictors of early stages of adult hippocampal neurogenesis.
Adult hippocampal neurogenesis, the process of formation of new neurons, occurs throughout life in the hippocampus. New neurons have been associated with learning and memory as well as mood control, and impaired neurogenesis has been linked to depression, schizophrenia, autism and cognitive decline during aging. Thus, understanding the biological properties of adult neurogenesis has important implications for human health. Computational models of neurogenesis have attempted to derive biologically relevant knowledge, hard to achieve using experimentation. However, the majority of the computational studies have predominantly focused on the late stages of neurogenesis, when newborn neurons integrate into hippocampal circuitry. Little is known about the early stages that regulate proliferation, differentiation, and survival of neural stem cells and their immediate progeny. Here, based on the branching process theory and biological evidence, we developed a computational model that represents the early stage hippocampal neurogenic cascade and allows prediction of the overall efficiency of neurogenesis in both normal and diseased conditions. Using this stochastic model with a simulation program, we derived the equilibrium distribution of cell population and simulated the progression of the neurogenic cascade. Using BrdU pulse-and-chase experiment to label proliferating cells and their progeny in vivo, we quantified labeled newborn cells and fit the model on the experimental data. Our simulation results reveal unknown but meaningful biological parameters, among which the most critical ones are apoptotic rates at different stages of the neurogenic cascade: apoptotic rates reach maximum at the stage of neuroblasts; the probability of neuroprogenitor cell renewal is low; the neuroblast stage has the highest temporal variance within the cell types of the neurogenic cascade, while the apoptotic stage is short. At a practical level, the stochastic model and simulation framework we developed will enable us to predict overall efficiency of hippocampal neurogenesis in both normal and diseased conditions. It can also generate predictions of the behavior of the neurogenic system under perturbations such as increase or decrease of apoptosis due to disease or treatment.


ccm1 cell autonomously regulates endothelial cellular morphogenesis and vascular tubulogenesis in zebrafish.
Cerebral cavernous malformations (CCMs) are a prevalent class of vascular anomalies characterized by thin-walled clusters of malformed blood vessels in the brain. Heritable forms are caused by mutations in CCM1, CCM2 and CCM3, but despite the importance of these factors in vascular biology, an understanding of their molecular and cellular functions remains elusive. Here we describe the characterization of a zebrafish embryonic model of CCM. Loss of ccm1 in zebrafish embryos leads to severe and progressive dilation of major vessels, despite normal endothelial cell fate and number. Vascular dilation in ccm1 mutants is accompanied by progressive spreading of endothelial cells and thinning of vessel walls despite ultrastructurally normal cell-cell contacts. Zebrafish ccm2 mutants display comparable vascular defects. Finally, we show that ccm1 function is cell autonomous, suggesting that it is endothelial cellular morphogenesis that is regulated by CCM proteins during development and pathogenesis.


Effect of low-dose doxycycline on serum oxidative status, gingival antioxidant levels, and alveolar bone loss in experimental periodontitis in rats.
Subantibiotic doses of doxycycline (low-dose doxycycline [LDD]) have been widely used in periodontal treatment for enzymatic inhibition and related anti-inflammatory properties. The aim of the present study is to verify the possible effects of LDD on oxidative stress in relation to periodontal attachment loss associated with ligature-induced experimental periodontal disease in rats. Thirty female Wistar albino rats were divided into three study groups as follows: 1) control (C) rats; 2) rats with experimental periodontitis (PED); and 3) rats with PED that were treated with doxycycline (PED + LDD). PED was induced by placing ligatures around the cervix of the maxillary second molars for 21 days. The PED + LDD group was treated orally with doxycycline (6 mg/kg) for 21 days after the ligature was placed. After 21 days, the rats were euthanized, and samples of the right maxilla were defleshed and used for histologic and morphometric analyses. The gingival tissue of the left maxilla was used for the analysis of lipid peroxidation (malondialdehyde [MDA]) and antioxidant enzymes (catalase, glutathione peroxidase, and superoxide dismutase). Levels of serum total antioxidant status (TAS)/total oxidant status (TOS) and oxidative stress index (OSI) were also analyzed. Alveolar bone loss was significantly higher in the PED group compared with the PED + LDD and C groups (P <0.05). Doxycycline exhibited the most prominent inhibition on gingival tissue levels of MDA and antioxidant enzymes (P <0.05). Doxycycline also significantly reduced TOS and OSI levels (P <0.05) but increased the TAS level. Doxycycline helps to prevent periodontal tissue breakdown by inhibiting local and systemic oxidative stress.


Vitamin C supplementation of the maternal diet reduces the rate of malformation in the offspring of diabetic rats.
An excess of reactive oxygen species (ROS) has been associated with the increased rate of congenital malformations in experimental diabetic pregnancy. Previous in vitro and in vivo studies show that antioxidants can protect the embryonic development in a diabetic environment. In the present investigation we examined the antiteratogenic capacity of vitamin C, an antioxidative agent not previously evaluated as a dietary supplement in diabetic pregnancy. Normal and streptozotocin diabetic rats were either fed a standard diet or a diet enriched with 0.9, 1.8 or 4% sodium ascorbate throughout pregnancy. On gestational day 20, the litters of normal and diabetic rats without vitamin C supplement contained 9 and 12% early resorptions, 2 and 17% late resorptions and 1 and 27% malformations, respectively. Vitamin C treatment reduced the rates of late resorptions and malformations in the diabetic groups in proportion to the dose administered. Thus, in the diabetic group with 4% ascorbate treatment we found unchanged numbers of early resorptions, but only 7% late resorptions (p < 0.05 vs untreated diabetic pregnancy) and 8% malformations (p < 0.05 vs untreated diabetic pregnancy). Maternal diabetes did not alter tissue levels of ascorbic acid in the fetuses at term, whereas vitamin C treatment caused accumulation of ascorbic acid in the placenta, maternal and fetal liver. Vitamin C supplementation yielded increased alpha-tocopherol concentration in the placenta and caused a reduction of the high concentrations of thiobarbituric acid reactive substances (TBARS) in serum of pregnant diabetic rats. Vitamin C treatment reduces the rates of congenital malformations and late resorptions, thereby supporting that ROS are involved in the embryonic dysmorphogenesis of diabetic pregnancy.


First report on mould and mycotoxin contamination of pistachios sampled in Algeria.
Mycotoxin contamination of pistachios represents a serious food safety hazard. The aim of this study was to evaluate fungal contamination and aflatoxin (AF) and ochratoxin A (OTA) occurrence in pistachio sampled in Algeria and to study the mycotoxigenic capacities of the isolates. A total of 31 pistachio samples were collected from retail outlets from different regions of Algeria. The most frequently found fungi were Penicillium spp. (38%), Aspergillus section Nigri (30%) and A. flavus (22%). A total of 56.5% of A. flavus isolates were able to produce AFB(1) and AFB(2). No A. section Nigri uniseriate isolate was OTA producer, whereas OTA production capacity was detected in 33.3% of the A. section Nigri biseriate. At least one of the potentially ochratoxigenic species was found in 64.5% of samples. Despite the high number of pistachio samples containing AFs and OTA-producing isolates, only two samples contained AFs (always below the EU maximum tolerable level) and only one sample showed OTA contamination. This is the first report on the occurrence of toxigenic moulds and mycotoxins in pistachios from Algerian market.