Clinics Optimizing MEthadone Take-homes for opioid use disorder (COMET): Protocol for a stepped-wedge randomized trial to facilitate clinic level changes.
Regulatory changes made during the COVID-19 public health emergency (PHE) that relaxed criteria for take-home dosing (THD) of methadone offer an opportunity to improve quality of care with a lifesaving treatment. There is a pressing need for research to study the long-term effects of the new PHE THD rules and to test data-driven interventions to promote more effective adoption by opioid treatment programs (OTPs). We propose a two-phase project to develop and test a multidimensional intervention for OTPs that leverages information from large State administrative data. We propose a two-phased project to develop then test a multidimensional OTP intervention to address clinical decision making, regulatory confusion, legal liability concerns, capacity for clinical practice change, and financial barriers to THD. The intervention will include OTP THD specific dashboards drawn from multiple State databases. The approach will be informed by the Health Equity Implementation Framework (HEIF). In phase 1, we will employ an explanatory sequential mixed methods design to combine analysis of large state administrative databases-Medicaid, treatment registry, THD reporting-with qualitative interviews to develop and refine the intervention. In phase 2, we will conduct a stepped-wedge trial over three years with 36 OTPs randomized to 6 cohorts of a six-month clinic-level intervention. The trial will test intervention effects on OTP-level implementation outcomes and patient outcomes (1) THD use; 2) retention in care; and 3) adverse healthcare events). We will specifically examine intervention effects for Black and Latinx clients. A concurrent triangulation mixed methods design will be used: quantitative and qualitative data collection will occur concurrently and results will be integrated after analysis of each. We will employ generalized linear mixed models (GLMMs) in the analysis of stepped-wedge trials. The primary outcome will be weekly or greater THD. The semi-structured interviews will be transcribed and analyzed with Dedoose to identify key facilitators, barriers, and experiences according to HEIF constructs using directed content analysis. This multi-phase, embedded mixed methods project addresses a critical need to support long-term practice changes in methadone treatment for opioid use disorder following systemic changes emerging from the PHE-particularly for Black and Latinx individuals with opioid use disorder. By combining findings from analyses of large administrative data with lessons gleaned from qualitative interviews of OTPs that were flexible with THD and those that were not, we will build and test the intervention to coach clinics to increase flexibility with THD. The findings will inform policy at the local and national level.


Constitutive nuclear factor kappaB activity is required to elicit interferon-gamma-induced expression of chemokine CXC ligand 9 (CXCL9) and CXCL10 in human tumour cell lines.
CXC ligand 10 (CXCL10) and CXCL9 are chemoattractants for activated T cells and possess angiostatic activity. Both CXCL9 and CXCL10 have been considered as important components for the anti-tumour activities of interferon-gamma (IFNgamma) and interleukin-12 in animal models. In this article we show that the CXCL9 and CXCL10 genes in some types of human tumour cell lines are not inducible by IFNgamma and we describe experiments designed to explore the molecular mechanisms involved in this impaired induction. The human oral squamous carcinoma line Ca9-22 and the glioma line A172 failed to express CXCL9 and CXCL10 mRNAs in response to IFNgamma, whereas other carcinoma lines including HSC-2 did express these mRNAs. Production of these chemokine proteins was also impaired in Ca9-22 cells. The impaired expression was not due to any deficiency in the IFNgamma/signal transducer and activator of transcription 1 (STAT1)-dependent signalling pathway. Instead, analysis of nuclear factor kappaB (NF-kappaB) activity revealed that the constitutive low level of NF-kappaB activity, which is seen in cells that express these chemokines, was absent in Ca9-22 and A172 cells. Activation of NF-kappaB in Ca9-22 cells restored the expression of IFNgamma-stimulated CXCL9 and CXCL10 mRNAs. In contrast, inhibition of the constitutive NF-kappaB in HSC-2 cells by adenovirus-mediated gene transfer of a dominant-negative IkappaBalpha suppressed the IFNgamma-induced expression of the CXCL9 and CXCL10 mRNAs. These results indicate that constitutive NF-kappaB activity, which is often associated with tumour development, is required for the induced expression of CXCL9 and CXCL10 genes in human tumour cell lines in response to IFNgamma.


Prospective imaging study of asymptomatic patellar tendinopathy in elite junior basketball players.
To evaluate the ability of ultrasonography to predict eventual symptoms in an at-risk population, 52 elite junior basketball players' patellar tendons were studied at baseline and again 16 months later. The group consisted of 10 study tendons (ultrasonographically hypoechoic at baseline) and 42 control tendons (ultrasonographically normal at baseline). By design, all tendons were asymptomatic at baseline. No differences were noted between subjects and controls at baseline for age, height, weight, training hours, and vertical jump. Functional (P < 0.01) and symptomatic outcome (P < 0.05) were poorer for subjects' tendons than for controls. Relative risk for developing symptoms of jumper's knee was 4.2 times greater in case tendons than in control tendons. Men were more likely to develop ultrasonographic changes than women (P < 0.025), and they also had significantly increased training hours per week (P < 0.01) in the study period. Half (50%) of abnormal tendons in women became ultrasonographically normal in the study period. Our data suggest that presence of an ultrasonographic hypoechoic area is associated with a greater risk of developing jumper's knee symptoms. Ultrasonographic patellar tendon changes may resolve, but this is not necessary for an athlete to become asymptomatic. Qualitative or quantitative analysis of baseline ultrasonographic images revealed it was not possible to predict which tendons would develop symptoms or resolve ultrasonographically.


Diagnosing odontogenic sinusitis of endodontic origin: A multidisciplinary literature review.
Endodontic disease is one of the most common causes of bacterial odontogenic sinusitis (ODS). Diagnosing ODS of endodontic origin involves otolaryngologists confirming sinusitis, and dental specialists confirming endodontic sources. The purpose of this study was to conduct a multidisciplinary literature review to highlight clinical and microbiological features of ODS, and the most optimal diagnostic modalities to confirm endodontic disease. An extensive review of both medical and dental literature was performed by rhinologists, endodontists, and an infectious disease specialist. Frequencies of various clinical and microbiological features from ODS studies were collected, and averages were calculated. Different endodontic testing and imaging modalities were also evaluated on their abilities to confirm endodontic disease. ODS patients most often present with unilateral sinonasal symptoms for over 3 months, purulence on nasal endoscopy, and overt dental pathology on computed tomography (CT). Subjective foul smell, and maxillary sinus cultures demonstrating anaerobes and α-streptococci (viridans group) may be more specific to ODS. For endodontic evaluations, cold pulp testing and cone-beam CT imaging are most optimal for confirming pulpal and periapical disease. Diagnosing ODS requires collaboration between otolaryngologists and dental specialists. Clinicians should suspect ODS when patients present with unilateral sinonasal symptoms, especially foul smell. Patients will generally have purulent drainage on nasal endoscopy, and both sinus opacification and overt dental pathology on CT. However, some patients will have subtle or absent dental pathology on CT. For suspected endodontic disease, endodontists should be consulted for at least cold pulp testing, and ideally cone-beam CT.


Prognostic significance of previous tonsillectomy after radical cystectomy for bladder cancer.
Introduction: To examine whether previous tonsillectomy (TE) impacts on survival after radical cystectomy (RC) for bladder cancer (BC).Patients and Methods: A total of 320 patients were staged cM0 and underwent RC for BC between 2002 and 2013. We retrospectively investigated whether patients had undergone TE prior to RC. Chi-square/Fisher-Exact test was carried out to compare clinicopathological features between the TE- and non-TE-group. Kaplan-Meier analysis with log-rank test was used to estimate recurrence-free survival (RFS) and multivariable Cox-regression analysis of risk factors of recurrence. The median follow-up was 31 months (interquartile range: 9-54).Results: A history of TE was present in 18 of the 320 patients (5.6%). All TEs were performed for benign conditions. TE prior to RC was associated with a history of appendectomy (p = 0.045), lower age at RC (p = 0.029), tumor unifocality (p < 0.001), advanced histopathological tumor stage (p = 0.015), non-pure urothelial carcinoma (p = 0.025), lymphovascular invasion (p = 0.035) and receipt of palliative chemotherapy (p = 0.004). The 3-year RFS was 39.2% for patients with previous TE and 62.4% for those without (p = 0.008). In multivariable analysis, adjusted for all significant parameters of univariable analysis, lymph-node tumor involvement (p = 0.017), positive surgical margins (p = 0.047), tumor grade (p = 0.032), advanced tumor stage (≥pT3a; p = 0.049) and a history of TE (p = 0.021) remained independent prognosticators of recurrence.Conclusion: In this series, previous TE was an independent predictor of recurrence after RC for BC. Further studies are needed to assess whether TE induces immunological alterations that might exert adverse effects on cancer progression of patients with invasive BC.


Changes in size of the stretch reflex of cat and man attributed to aftereffects in muscle spindles.
1. This is a report of experiments carried out on the cat and on man, which demonstrate that conditioning of a muscle by contraction and movement can lead to changes in amplitude of stretch reflexes elicited in that muscle. 2. In triceps surae of the cat, the reflex response to a brief stretch was recorded after conditioning with a whole-muscle contraction followed by a pause at a length either 5 mm longer or shorter than the length at which the reflex was elicited. Following conditioning at the long length the reflex response was less than half as large as that following conditioning at the short length. 3. The changes in reflex amplitude could be correlated with an altered stretch responsiveness of muscle spindles in the soleus muscle. When the muscle had been held long during conditioning, a subsequent brief stretch applied at an intermediate length elicited fewer impulses in primary endings of spindles than after conditioning at a short length. 4. The same kind of experiment was then carried out on adult human subjects. When a tendon tap was applied to the Achilles tendon after a voluntary contraction and relaxation of triceps surae with the muscle at a long length, (foot dorsiflexed) the reflex was frequently less than half the size it had been after a contraction at a short length (foot plantarflexed). It was concluded that the same kind of spindle aftereffects as observed for cat soleus spindles were responsible for the changes in reflex amplitude. 5. It was found both in the cat and in human subjects that the changes in reflex amplitude after conditioning became progressively less as the test length was made longer. 6. The explanation put forward to account for these observations is that stable cross-bridges form between actin and myosin filaments of passive intrafusal (and extrafusal) fibers. When the muscle is shortened several seconds after a contraction at a long length, the intrafusal fibers, stiffened by the presence of cross-bridges, fall slack. Slack does not develop after a contraction at a short muscle length, as the fiber is stretched to the test length. Since any slack must first be taken up by the test stretch, there is a smaller afferent response and consequently a smaller reflex contraction in response to a tendon tap after conditioning at a long length.(ABSTRACT TRUNCATED AT 400 WORDS)


Clustering ionic flow blockade toggles with a mixture of HMMs.
Ionic current blockade signal processing, for use in nanopore detection, offers a promising new way to analyze single molecule properties with potential implications for DNA sequencing. The alpha-Hemolysin transmembrane channel interacts with a translocating molecule in a nontrivial way, frequently evidenced by a complex ionic flow blockade pattern with readily distinguishable modes of toggling. Effective processing of such signals requires developing machine learning methods capable of learning the various blockade modes for classification and knowledge discovery purposes. Here we propose a method aimed to improve our stochastic analysis capabilities to better understand the discriminatory capabilities of the observed the nanopore channel interactions with analyte. We tailored our memory-sparse distributed implementation of a Mixture of Hidden Markov Models (MHMMs) to the problem of channel current blockade clustering and associated analyte classification. By using probabilistic fully connected HMM profiles as mixture components we were able to cluster the various 9 base-pair hairpin channel blockades. We obtained very high Maximum a Posteriori (MAP) classification with a mixture of 12 different channel blockade profiles, each with 4 levels, a configuration that can be computed with sufficient speed for real-time experimental feedback. MAP classification performance depends on several factors such as the number of mixture components, the number of levels in each profile, and the duration of a channel blockade event. We distribute Baum-Welch Expectation Maximization (EM) algorithms running on our model in two ways. A distributed implementation of the MHMM data processing accelerates data clustering efforts. The second, simultanteous, strategy uses an EM checkpointing algorithm to lower the memory use and efficiently distribute the bulk of EM processing in processing large data sequences (such as for the progressive sums used in the HMM parameter estimates). The proposed distributed MHMM method has many appealing properties, such as precise classification of analyte in real-time scenarios, and the ability to incorporate new domain knowledge into a flexible, easily distributable, architecture. The distributed HMM provides a feature extraction that is equivalent to that of the sequential HMM with a speedup factor approximately equal to the number of independent CPUs operating on the data. The MHMM topology learns clusters existing within data samples via distributed HMM EM learning. A Java implementation of the MHMM algorithm is available at http://logos.cs.uno.edu/~achurban.


Evaluation of Micronutrients and Pro-Inflammatory Cytokines Levels in Nutritionally Deprived Children-A Tertiary Care Hospital-Based Study.
Severe acute malnutrition (SAM) is a significant public health problem in developing countries, including India, where a significant proportion of children suffer from malnutrition. This research aims to investigate the factors contributing to severe acute malnutrition (SAM). Additionally, the study seeks to explore the relationship between micronutrient levels and pro-inflammatory cytokines in SAM children with and without clinical complications. Furthermore, the effectiveness of antibiotic treatment in SAM children without complications is evaluated. The study involved three groups comprising 66 subjects each: a healthy control group, SAM children with complications, and SAM children without complications. Blood samples were collected, and various analyses were conducted, including biochemical, hematological, micronutrient, and pro-inflammatory marker quantification. The data were analyzed using SPSS version 22.0. The results indicate that the levels of IL-6, CRP, and TNF-α were significantly higher in the SAM group with complications compared to both the control group and the SAM group without complications. Zinc and copper levels were significantly lower in both SAM groups compared to the control group, and a negative correlation was observed between zinc levels and inflammatory markers. The study also assessed the efficacy of antibiotic treatment in SAM children without complications by comparing their weight, height, weight-for-height, and weight-for-age at baseline and after a 15-day follow-up period. Significant improvements in these parameters were observed in both the group receiving antibiotic treatment and the group not receiving antibiotic treatment. The findings suggest that a combination of antibiotic treatment and nutritional support can lead to significant clinical improvements in SAM children without complications. This study has important implications for the management and treatment of SAM in India and other developing countries.


Susceptibility of MED-Q1 and MED-Q3 Biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae) Populations to Essential and Seed Oils.
Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a major pest of many agricultural and ornamental crops in tropical and subtropical regions causing damages that result in important economic losses. Insecticides are commonly used in greenhouses or fields to control B. tabaci populations leading to rapid evolution of resistance that render treatments inefficient. Therefore, and for environmental and human health concerns, other approaches must be developed for this pest management. In the present study, we compare, using the leaf dip method, the toxicity of three essential oils (Cymbopogon citratus, Ocimum americanum, and Hyptis spicigera) and three seed oils (Lannea microcarpa, Lannea acida, and Carapa procera) with three chemical insecticides (acetamiprid, deltamethrin, and chlorpyrifos-ethyl) on adults. Two B. tabaci biotypes (MED-Q1 and MED-Q3) belonging to the Mediterranean species and collected in Burkina Faso were used. Essential oils were analyzed by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. We showed that these two biotypes have different levels of resistance to the three insecticides, MED-Q3 being more sensitive than MED-Q1. Moreover, they differ in the frequency of resistance alleles to insecticides, especially for organophosphates, as these alleles are almost fixed in MED-Q1. On the other hand, the two biotypes prove to be more susceptible to the plant extracts than to insecticides except for chlorpyrifos-ethyl, with essential oils that showed the highest insecticidal activities. Monoterpenes content were the most abundant and showed the highest insecticidal activities. Our results indicated that essential oils, but also seed oils, have the potential to constitute an alternative strategy of pest management.


A sustaining rod increases necrosis of loop ileostomies: a randomized controlled trial.
Defunctioning loop ileostomies (LI) are commonly used in colorectal surgery to reduce the potentially detrimental consequences of anastomotic leakages. However, stoma-related morbidity is high with up to 75% of patients having local complications. The aim of this study was to investigate the effect of a sustaining rod on the local complication rate. In this prospective, multi-center, randomized controlled trial, subjects were allocated to either a rod or a rod-less protocol (NCT00959738). The primary outcome was local morbidity as measured by a stoma specific morbidity score (SSMS) during the first 3 months postoperatively. Between August 2008 and July 2014, a total of 122 patients were enrolled in the study, of which 78 (63.8%) completed the study [44 (56.4%) rod, 34 (43.6%) rod-less]. There was no significant difference in the SSMS between the two groups. The incidence of necrosis or partial necrosis, however, was significantly increased in the rod group: 13 (29.5%) vs. 1 (2.9%) in the rod-less group (p < 0.01). The retraction rate did not differ significantly between the groups: two (4.5%) in the rod vs. five (14.7%) in the rod-less group (p = 0.13). High body mass index (BMI > 26) was associated with an odds ratio of 5 (p < 0.01) for severe stoma complications. A rod-less technique for loop ileostomies reduces the risk of stomal necrosis, with a high BMI being an independent risk factor for stomal complications.


An ex vivo assay to assess the potential of skin keratinocytes for wound epithelialization.
Wound closure following injury to the skin is a complex process involving both dermal contraction and keratinocyte migration. Murine models of wound healing are potentially useful because of the ability to determine protein function through gene manipulation. Owing to the dominant role of dermal contraction, the technical difficulties in preparing the wound site for morphologic studies, and the postnatal phenotypes altering the properties of transgenic skin, there are difficulties in assessing the epithelial contribution to wound closure in mouse skin. We describe a simple ex vivo assay utilizing explant culture that enables a quantitative assessment of the potential of mouse keratinocytes for wound epithelialization. In this assay, the behavior and properties of skin keratinocytes mimic well those that occur at the edge of skin wounds in situ, including a dependence upon connective tissue element(s), proliferation, and migration. The epithelial cell outgrowths emerging from skin explants can be studied in real-time or examined at specific time-points for markers of interest in the epithelialization process. The assay is quantitative and can successfully detect increases or decreases in epithelialization potential, and can be useful in the characterization of transgenic mouse models.


Comorbidity of Migraine.
Migraine is a common neurological disorder and can be severely disabling during attacks. The highest prevalence occurs between the ages of 25 and 55 years. Prior studies have found that migraine occurs together with other illnesses at a greater coincidental rate than is seen in the general population. These occurrences are called "comorbidities". To delineate the comorbidities of migraine is important, because it can help improve treatment strategies and the understanding of the possible pathophysiology of migraine.


Hybrid sponge comprised of galactosylated chitosan and hyaluronic acid mediates the co-culture of hepatocytes and endothelial cells.
When constructing an in vitro model of liver tissue to mimic the in vivo liver microenvironment, the major challenge is to preserve and maintain the hepatocyte phenotype. The aim of this study was to develop a novel intelligent hybrid sponge for use in a dense co-culture system designed to simulate the liver microenvironment. We prepared a galactosylated chitosan (GCs)/hyaluronic acid (HA) hybrid sponge using a freeze-drying technique for the co-culture of primary hepatocytes and endothelial cells. Subsequently, we investigated the biocompatibility of the GCs/HA scaffold with primary hepatocytes and endothelial cells in terms of cell attachment, morphology, bioactivity, and maintenance of specific liver functions. The GCs/HA-hybrid sponge demonstrated good biocompatibility not only with primary hepatocytes, but also with endothelial cells. In our model, primary hepatocytes exhibited superior bioactivity and higher levels of liver-specific functions in terms of hepatocyte-specific gene expression, urea production, and testosterone metabolism as compared to a monoculture system. We succeeded in constructing a liver tissue-like model using the GCs/HA-hybrid sponge. Therefore, we anticipate that GCs/HA-hybrid sponges may be a promising matrix for the co-culture of hepatocytes and endothelial cells in liver tissue engineering, and might be employed as a novel co-culture model for applications in toxicology and drug metabolism.


Developmental expression of cell recognition molecules in the mushroom body and antennal lobe of the locust Locusta migratoria.
We examined the development of olfactory neuropils in the hemimetabolous insect Locusta migratoria with an emphasis on the mushroom bodies, protocerebral integration centers implicated in memory formation. Using a marker of the cyclic adenosine monophosphate (cAMP) signaling cascade and lipophilic dye labeling, we obtained new insights into mushroom body organization by resolving previously unrecognized accessory lobelets arising from Class III Kenyon cells. We utilized antibodies against axonal guidance cues, such as the cell surface glycoproteins Semaphorin 1a (Sema 1a) and Fasciclin I (Fas I), as embryonic markers to compile a comprehensive atlas of mushroom body development. During embryogenesis, all neuropils of the olfactory pathway transiently expressed Sema 1a. The immunoreactivity was particularly strong in developing mushroom bodies. During late embryonic stages, Sema 1a expression in the mushroom bodies became restricted to a subset of Kenyon cells in the core region of the peduncle. Sema 1a was differentially sorted to the Kenyon cell axons and absent in the dendrites. In contrast to Drosophila, locust mushroom bodies and antennal lobes expressed Fas I, but not Fas II. While Fas I immunoreactivity was widely distributed in the midbrain during embryogenesis, labeling persisted into adulthood only in the mushroom bodies and antennal lobes. Kenyon cells proliferated throughout the larval stages. Their neurites retained the embryonic expression pattern of Sema 1a and Fas I, suggesting a role for these molecules in developmental mushroom body plasticity. Our study serves as an initial step toward functional analyses of Sema 1a and Fas I expression during locust mushroom body formation.


Soil radon measurements as a potential tracer of tectonic and volcanic activity.
In Earth Sciences there is a growing interest in studies concerning soil-radon activity, due to its potential as a tracer of numerous natural phenomena. Our work marks an advance in the comprehension of the interplay between tectonic activity, volcanic eruptions and gas release through faults. Soil-radon measurements, acquired on Mt. Etna volcano in 2009-2011, were analyzed. Our radon probe is sensitive to changes in both volcanic and seismic activity. Radon data were reviewed in light of the meteorological parameters. Soil samples were analyzed to characterize their uranium content. All data have been summarized in a physical model which identifies the radon sources, highlights the mechanism of radon transport and envisages how such a mechanism may change as a consequence of seismicity and volcanic events. In the NE of Etna, radon is released mainly from a depth of <1400 m, with an ascent speed of >50 m/day. Three periods of anomalous gas release were found (February 2010, January and February 2011). The trigger of the first anomaly was tectonic, while the second and third had a volcanic origin. These results mark a significant step towards a better understanding of the endogenous mechanisms that cause changes in soil-radon emission at active volcanoes.


Identification of Essential Genes for the Establishment of Spray-Induced Gene Silencing-Based Disease Control in Fusarium graminearum.
As resistance to chemical fungicides continues to increase inFusarium graminearum, there is a growing need to develop novel disease control strategies. To discover essential genes that could serve as new disease control targets, we selected essential gene candidates that had failed to be deleted in previous studies. Thirteen genes were confirmed to be essential, either by constructing conditional promoter replacement mutants or by employing a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-mediated editing strategy. We synthesized double-stranded RNAs (dsRNAs) targeting these essential genes and analyzed their protective effects in plants using a spray-induced gene silencing (SIGS) method. When dsRNAs targeting Fg10360, Fg13150, and Fg06123 were applied to detached barley leaves prior to fungal inoculation, disease lesions were greatly reduced. Our findings provide evidence of the potential of essential genes identified by a SIGS method to be effective targets for the control of fungal diseases.


Effects of concentrated arabinoxylan and β-glucan compared with refined wheat and whole grain rye on glucose and appetite in subjects with the metabolic syndrome: a randomized study.
Several studies emphasise that arabinoxylan and β-glucan have more beneficial effects on glucose metabolism than low-dietary fibre (DF) meals. Less attention has been paid to the effects of concentrated DF compared with whole grain. We compared the effects of DF and whole grain on glucose, hormone responses and appetite in subjects with the metabolic syndrome (MetS). Fifteen subjects with MetS participated in this acute, randomised, cross-over intervention study. The test breads provided 50 g of digestible carbohydrate: wheat bread with concentrated arabinoxylan (AX) or β-glucan (BG), rye bread with kernels (RK) and wheat bread (WB) as control. Blood samples were drawn for 270 min to determine glucose, insulin, glucagon-like peptide-1, glucose-dependent insulinotropic peptide (GIP) and ghrelin. Appetite score was addressed every 30 min. Ad libitum energy intake (EI) was measured 270 min after test meals. Compared with WB, BG and RK induced lower initial glycaemic responses (P<0.001), whereas AX only reduced the glucose peak value (P<0.001). RK reduced insulin (P<0.001) and GIP responses (P<0.001) compared with the other breads. BG lowered insulin responses more than AX (P<0.001). AX, BG and RK increased satiety feeling (P<0.001) more than WB, but did not differ significantly in terms of subsequent EI (P=0.089). BG and RK had beneficial impact on the glucose response, whereas AX had only effect on the postprandial glucose peak. The impact of the AX bread was influenced by higher protein content. Whether the metabolic effects of the breads are still present to mixed meals remains to be tested.


[Results of mandibular bone plastic repair with vascularized autotransplants].
A total of 96 vascularized flaps used in the reconstruction of the mandible in 91 patients are analyzed. In 82 cases the defects appeared after treatment of malignant and benign lesions of the mandible. In 18 patients the mandible was reconstructed using rib grafts. The lateral border of the scapula was used in 30 patients and fibular grafts in 12 patients. In 36 cases mandibular and soft tissue defects were corrected using vascularized latissmus dorsi flaps combined with the tenth or eleventh rib. The results were positive in 86-92% of the cases. The postoperative analysis shows excellent esthetic and functional results in all the patients who had primary plastic reconstruction of the mandible and the results were good in 36% of cases of secondary plastic reconstruction of the mandible. The donor sites in all cases showed no functional disturbances.


A human genome-wide library of local phylogeny predictions for whole-genome inference problems.
Many common inference problems in computational genetics depend on inferring aspects of the evolutionary history of a data set given a set of observed modern sequences. Detailed predictions of the full phylogenies are therefore of value in improving our ability to make further inferences about population history and sources of genetic variation. Making phylogenetic predictions on the scale needed for whole-genome analysis is, however, extremely computationally demanding. In order to facilitate phylogeny-based predictions on a genomic scale, we develop a library of maximum parsimony phylogenies within local regions spanning all autosomal human chromosomes based on Haplotype Map variation data. We demonstrate the utility of this library for population genetic inferences by examining a tree statistic we call 'imperfection,' which measures the reuse of variant sites within a phylogeny. This statistic is significantly predictive of recombination rate, shows additional regional and population-specific conservation, and allows us to identify outlier genes likely to have experienced unusual amounts of variation in recent human history. Recent theoretical advances in algorithms for phylogenetic tree reconstruction have made it possible to perform large-scale inferences of local maximum parsimony phylogenies from single nucleotide polymorphism (SNP) data. As results from the imperfection statistic demonstrate, phylogeny predictions encode substantial information useful for detecting genomic features and population history. This data set should serve as a platform for many kinds of inferences one may wish to make about human population history and genetic variation.


A Real-Time PCR Assay for Detection of Low Pneumocystis jirovecii Levels.
Here we report a new real-time PCR assay using SYBR Green which provides higher sensitivity for the specific detection of low levels of Pneumocystis jirovecii. To do so, two primer sets were designed, targeting the family of genes that code for the most abundant surface protein of Pneumocystis spp., namely the major surface glycoproteins (Msg), and the mitochondrial large subunit rRNA (mtLSUrRNA) multicopy gene, simultaneously detecting two regions. PCR methods are instrumental in detecting these low levels; however, current nested-PCR methods are time-consuming and complex. To validate our new real-time Msg-A/mtLSUrRNA PCR protocol, we compared it with nested-PCR based on the detection of Pneumocystis mitochondrial large subunit rRNA (mtLSUrRNA), one of the main targets used to detect this pathogen. All samples identified as positive by the nested-PCR method were found positive using our new real-time PCR protocol, which also detected P. jirovecii in three nasal aspirate samples that were negative for both rounds of nested-PCR. Furthermore, we read both rounds of the nested-PCR results for comparison and found that some samples with no PCR amplification, or with a feeble band in the first round, correlated with higher Ct values in our real-time Msg-A/mtLSUrRNA PCR. This finding demonstrates the ability of this new single-round protocol to detect low Pneumocystis levels. This new assay provides a valuable alternative for P. jirovecii detection, as it is both rapid and sensitive.


Radioautographic visualization of sulfur-35 disposition in the articular cartilage and bone of suckling rats following injection of labeled sodium sulfate.
The localization of sulfur-35 administered intraperitoneally as sodium sulfate to 7-day-old rats was determined by radioautography of sections of humeri and tibia-knee-femur combinations removed 24, 48, 96, 216, and 290 hours after injection of this isotope. Radioautography of sections of bone and cartilage that had been fixed in formalin indicated that the tracer isotope was present throughout the entire epiphysis. Its concentration therein was highest initially at the epiphyseal-diaphyseal junction where the more mature cartilage cells were present. By the 96th hour post injection the sulfur-35 had become more uniformly distributed in the epiphyses and an even distribution of it throughout the epiphyseal cartilage was almost attained by the 216th hour post injection. As centers of secondary ossification arose in the epiphyseal cartilage, the sulfur-35 appeared to diminish in concentration and disappear from these loci. However, radioautographs of cartilage fixed in formalin saturated with barium hydroxide, instead of in formalin only, disclosed the fact that the tracer isotope was still present in these loci. When bone and bone marrow were fixed in formalin the autographs indicated the presence of sulfur-35 primarily in the periosteum. Only a negligible amount appeared to be present in the bone shaft and marrow. However, when these tissues were fixed in formalin saturated with barium hydroxide it was possible to demonstrate the presence of the tracer isotope in both bone and bone marrow.


Effects of benzo-a-pyrene on oxytocin-induced Ca2+ oscillations in myometrial cells.
Benzo-a-pyrene (BaP) is a polycyclic aromatic hydrocarbon that exists as a major environmental pollutant. The effect of this carcinogen/mutagen upon myometrial Ca(2+) signaling in a human myometrial cell line (PHM1) was examined. Exposure of cells to BaP did not alter basal Ca(2+) levels or the inositol(1,4,5) trisphosphate-releasable Ca(2+) pool. However, BaP significantly decreased the initial oxytocin-induced Ca(2+) transient and the frequency of oxytocin-induced Ca(2+)oscillations as well as delayed their onset. To determine the specific effects of BaP, pharmacologic agents that target intracellular Ca(2+) homeostasis mechanisms were used. Genistein (a non-specific tyrosine kinase inhibitor) and AG1478 (an epidermal growth factor receptor blocker) markedly reduced the oxytocin-induced Ca(2+) oscillations in control, but had no effect in BaP treated cells. Addition of epidermal growth factor or serum before or after oxytocin restored the Ca(2+) oscillations in BaP treated cells to a level similar to control cells, while the K(+) channel blocker tetraethylammonium chloride, partially restored the Ca(2+) response. These data suggest that the tyrosine kinase pathway, which is part of the G-protein coupled receptor pathway response to oxytocin in PHM1 cells, is a target of BaP action and that EGF or serum can restore the oxytocin-induced Ca(2+) oscillations.


Photo-responsive liposomes composed of spiropyran-containing triazole-phosphatidylcholine: investigation of merocyanine-stacking effects on liposome-fiber assembly-transition.
A spiropyran-containing triazole-phosphatidylcholine (SPTPC) was synthesized through a copper-catalyzed azide alkyne cyclo-addition (CuAAC) reaction. In water, SPTPCs self-assembled and a spontaneous spiropyran-to-merocyanine (SP-to-MC) isomerization occurred, resulting in coexistence of liposomes and fibers, and switching from the spiropyran (SP) to the merocyanine (MC) isomeric structure induced a reversible transition between these molecular assemblies. Study of the self-assembly of SPTPCs and photo-induced liposome-fiber assembly-transition revealed that the presence of MC enabled additional inter-membrane interaction during self-assembly and that the MC-stacking effect was the driving force for the assembly-transition. Exposure to UV light induced switching from SP to MC, where the planar structure of MC and the confinement of MC led to enhanced MC-stacking. The effect of MC-stacking was both advantageous and disadvantageous: MC-stacking perturbed the hydrophobic phase in the bilayer membrane and facilitated the liposome-to-fiber transition, otherwise the MC-stacking retarded switching of MC to SP, and caused an incomplete recovery of MC to SP during fiber-to-liposome recovery, thus a fatigue of SP was induced by MC-stacking during the liposome-to-fiber transition cycle. To decrease the intermolecular interactions and suppress MC-stacking, photo-inert triazole-phosphatidylcholine (TPC) was incorporated to prepare two-component TPC/SPTPC-liposomes, which exhibited better recovery kinetics. The photo-adaptive behavior of TPC/SPTPC-liposomes confirmed the disturbance of bilayer membranes by inter-membrane MC-stacking and the formation of MCTPC-enriched phases in the bilayer membrane.


Aetiology of diarrhoea and virulence properties of diarrhoeagenic Escherichia coli among patients and healthy subjects in southeast Nigeria.
Diarrhoeal diseases are one of the most important causes of illness and death all over the world. In Nigeria, the aetiology of diarrhoeagenic bacteria and the virulence of various Escherichia coli pathotypes have not been well-studied because most currently-published data were from the southwestern axis of the country. In total, 520 stool samples were collected from infants, young children, and other age-groups with acute diarrhoeal diseases in Enugu and Onitsha, southeastern Nigeria. Stool samples were collected from 250 apparently-healthy individuals, with similar age distribution and locality, who were considered control subjects. The stool samples were screened for diarrhea-causing bacterial agents. E. coli strains were isolated from both the groups and were examined by polymerase chain reaction (PCR) for 16 virulence genes. Of the 520 stool samples in the diarrhoea group, 119 (44.74%) were E. coli. Fifty (49.02%) were enteropathogenic E. coli (EPEC), 22 (21.57%) were enterotoxigenic E. coli (ETEC) while 7.84% was enteroaggregative E. coli (EAEC). Sex had no effect on the distribution of diarrhoeagenic bacteria, except for EIEC. The E. coli strains isolated from the diarrhoea and healthy asymptomatic age-matched control groups examined by PCR for 16 virulence genes indicate that the detection of EAEC, ETEC, EPEC, and EIEC was significantly associated with diarrhoea (p=0.0002). The study confirmed that several bacterial pathogens, such as E. coli, play an important role in the aetiology of acute diarrhoea in southeastern Nigeria. A routine surveillance, especially for diarrhoeagenic E. coli, would be useful in identifying outbreaks and help identify the potential reservoirs and transmission routes.


Smoking as a confounder in ecologic correlations of cancer mortality rates with average county radon levels.
Cohen has reported a negative correlation between lung cancer mortality and average radon levels by county. In this paper, the correlation of U.S. county mortality rates for various types of cancers during the period 1970-1994 with Cohen's radon measurements is examined. In general, quantitatively similar, strongly negative correlations are found for cancers strongly linked to cigarette smoking, weaker negative correlations are found for cancers moderately increased by smoking, whereas no such correlation is found for cancers not linked to smoking. The results indicate that the negative trend previously reported for lung cancer can be largely accounted for by a negative correlation between smoking and radon levels across counties. Hence, the observed ecological correlation provides no substantial evidence for a protective effect of low level radon exposure.


Clinical and metabolic changes after conventional treatment of type 2 diabetic patients with chronic periodontitis.
The aim of this study was to compare the response to conventional periodontal treatment between patients with or without type 2 diabetes mellitus from a clinical and metabolic standpoint. A prospective, parallel, comparative longitudinal clinical study was performed between type 2 diabetics and non-diabetics with moderate generalized chronic periodontitis. The study period was 6 months. Conventional periodontal scaling and root planing were performed, and the response to this treatment was compared between the groups at 3 and 6 months, measuring the plaque index, bleeding on probing, probing depth, level of clinical attachment, and gingival recession. In the diabetic patients, the clinical response was related to measurements of HbA1c and glucose in blood at 3 and 6 months. An improvement in all clinical variables was observed, with no statistically significant differences between the groups, with the exception of probing depth (P <0.0207). The improvement observed in blood HbA1c levels confirmed a positive metabolic response to periodontal treatment, with a lower value for this variable at each measurement time. Both groups of patients showed a clinical improvement after basic non-surgical periodontal treatment. The diabetic patients showed improved metabolic control (lower HbA1c) at 3 and 6 months after periodontal treatment.


Biosynthesized silver nanorings as a highly efficient and selective electrocatalysts for CO2 reduction.
Inspiration from nature has driven the development and applications of greener inorganic nanomaterials prepared using biotemplates in the field of nanoscience. In this study, we report the superiority of using a biosynthesized silver nanoring material for CO formation in CO2 saturated KHCO3. Compared to bulk silver and free silver nanoparticles prepared by pure chemical reduction, this silver nanoring (assembled on tobacco mosaic virus coat protein) exhibits significantly enhanced activity and selectivity for the conversion of CO2 to CO. The highest CO faradaic efficiency reaches 95.0% at an overpotential of 910 mV. Additionally, the CO partial current density is 2.7-fold higher than that of the free silver nanoparticles. We believe that the improved catalytic performance is related to the structuring ligand effect of the protein. The numerous functional groups on the protein may tune the reaction activity by influencing the binding energies of the intermediate species from CO2 reduction or hydrogen evolution.


The Ca2+/CaMKK2 axis mediates the telbivudine induced upregulation of creatine kinase: Implications for mechanism of antiviral nucleoside analogs' side effect.
Telbivudine (LdT), a widely prescribed anti-hepatitis B virus (HBV) drug for the treatment of chronic Hepatitis B (CHB), causes adverse reactions ranging from creatine kinase (CK) elevation to myopathy. The purpose of this study was to explore the mechanism(s) of LdT induced CK elevation. The effects of LdT on mitochondrial morphology and proteins (TK2 and β-actin), oxidative stress, intracellular Ca2+ levels, Ca2+-related signaling pathway (CaMKK2/AMPK), and Ca2+-related biomarkers such as superoxide dismutase (SOD) and malondialdehyde (MDA) were assessed in human skeletal muscle cells (HSKMCs). The results showed that LdT induced a dose-dependent increase in CK activity in HSKMCs, without affecting mitochondrial morphology, and TK2 and β-actin protein levels, following 72 h of treatment. In addition, LdT increased Ca2+ production, ROS generation, MDA and lipid peroxide (LPO) levels, and activated the CaMKK2/AMPK signaling pathway. Moreover, these effects were attenuated by the BAPIA-AM (the calcium chelator). We also confirmed the presence of relevant markers (MDA, LPO, and SOD) in serum from CHB patients after LdT treatment, and found that CK was positively correlated with MDA and LPO, and negatively associated with SOD. These findings indicate that LdT induces CK elevation and oxidative stress associated with imbalance of intracellular Ca2+ in HSKMCs, suggesting that Ca2+/CaMKK2 axis imbalance may underlie human LdT-induced CK elevation. The present findings provide a solid basis for assessing the mechanism of drug-induced CK elevation, which can help develop new tools for the prevention and treatment of diseases associated with drug-induced CK elevation.


Rare variants and biological pathways identified in treatment-refractory depression.
Individuals diagnosed with major depressive disorder not responding to at least two adequate treatments are defined as treatment-refractory major depressive disorder (TR-MDD). Some TR-MDD patients have altered metabolic phenotypes that may be pharmacologically reversed. The characterization of these phenotypes and their underlying etiologies is paramount, particularly their genetic components. In this study, TR-MDD patients (n = 124) were recruited and metabolites were quantified in their cerebrospinal fluid (CSF) and peripheral blood. Three sub-categories of deficiencies were examined, namely 5-methyltetrahydrofolte (in CSF; n = 13), tetrahydrobiopterin (in CSF; n = 11), and abnormal acylcarnitine profiles (in peripheral blood; n = 8). Whole exome sequencing was performed on genomic DNA from the entire TR-MDD cohort and exonic variant allele frequencies for cases were compared to a control cohort (1:5 matching on ancestry). Low frequency, damaging alleles were identified and used for in silico pathway analyses. Three association signals for TR-MDD approached genome-wide significance on chromosomes 22, 7, and 3. Three risk-associated variants from a prior depression study were replicated. Relevant biological pathways were identified that contained an enrichment of rare, damaging variants in central nervous system (CNS)-specific pathways, including neurotransmitter receptors, potassium channels, and synapse transmission. Some TR-MDD patients had rare variants in genes that were previously associated with other psychiatric disorders, psychiatric endophenotypes, CNS structural defects, and CNS-related cellular and molecular functions. Exome analysis of metabolically phenotyped TR-MDD patients has identified potentially functional gene pathways and low frequency, deleterious gene variants for further investigation. Further studies in larger cohorts of biochemically phenotyped TR-MDD patients are desirable to extend and confirm these findings.


Robotic Intracorporeal Orthotopic Neobladder: Urodynamic Outcomes, Urinary Function, and Health-related Quality of Life.
Intracorporeal orthotopic neobladder (iONB) creation following robotic radical cystectomy is an emerging procedure and robust functional data are required. To evaluate urodynamic features of iONB and bladder cancer-specific and general health-related quality-of-life (HRQOL) outcomes. We retrospectively assessed 28 men who underwent iONB creation (January 2012 to October 2013) and compared results to a previously characterized cohort of 79 of open ONB procedures. iONB pressure-volume properties were characterized using multichannel urodynamics (UDS). The Bladder Cancer Index (BCI) questionnaire, modified with mucus- and pad-related questions, and the Short Form Health Survey (SF-36) were used to evaluate urinary function and HRQOL. ONB cohorts were compared for functional outcomes and BCI score. Multivariable linear regression was used to assess predictors of BCI score. The median follow-up was 9.4 mo for the iONB and 62.1 mo for the open ONB group (p<0.0001); ≥2-yr follow-up had been completed for one (4%) patient in the iONB group compared to 75 (95%) patients in the open ONB group (p<0.0001). In UDS tests, the iONB group had minimal postvoid residual volume, normal compliance, and a mean capacity of 514 cm(3) (range 339-1001). BCI mean scores for urinary function (p=0.58) and urinary bother (p=0.31) were comparable between the groups. The surgical approach was not associated with the BCI score on multivariable analysis. Rates of 24-h pad use were comparable between iONB and open ONB groups (pad-free 17% vs. 19%; ≤2 pads 84% vs. 79%), as reflected by total pad usage (p=0.1); pad size and daytime wetness were worse in the iONB group. The clean intermittent catheterization rate was 10.7% in the iONB and 6.3% in the open ONB group. Limitations include the retrospective comparison, small number of patients and short follow-up for the iONB group. iONB had adequate UDS characteristics and comparable bladder cancer-specific HRQOL scores to open ONB. However, pad size and daytime wetness were worse for iONB, albeit over significantly shorter follow-up. We demonstrate that the volumetric and pressure characteristics are acceptable for a neobladder created using an entirely robot-assisted laparoscopic technique after bladder removal for cancer. Urinary function and quality-of-life outcomes related to the robotic technique were compared to those for neobladders created via an open surgical technique. We found that urinary function and bother indices were comparable; however, the robotic group required larger incontinence pads that were wetter during the daytime. This may be explained by the significantly shorter duration of recovery after surgery in the robotic group.


CO2 angiography--a technique for vascular imaging in renal allograft dysfunction.
Use of iodinated contrast for vascular imaging can be associated with nephrotoxicity and hypersensitivity reactions. Renal injury following conventional angiography is more likely to manifest in the setting of preexisting renal dysfunction. In the setting of suboptimal renal allograft function, these considerations are particularly relevant. Recently, CO2 has received attention as a nontoxic, injectable, rapidly absorbed gas that is a cost-effective alternative to standard contrast agents in high-risk patients, such as renal transplant recipients. We report the clinical course of a patient with transplant renal artery stenosis and a serum creatinine of 2.8 mg/dl who has successfully undergone angiography and percutaneous transluminal angioplasty using CO2 as the sole contrast agent. This case illustrates the potential use for CO2 as a contrast agent for vascular imaging in patients with suboptimal renal function who require definitive vascular imaging or therapy.


Rapamycin inhibits telomerase activity by decreasing the hTERT mRNA level in endometrial cancer cells.
Rapamycin exerts its biological activity by inhibiting the kinase mammalian target of rapamycin (mTOR), which regulates important cellular processes such as control of cell cycle and cell size, translation initiation, and transcription. The ability of rapamycin to inhibit cancer cell proliferation has led to efforts to develop rapamycin and related mTOR inhibitors as anticancer agents. Some investigators have hypothesized that loss of the PTEN tumor suppressor may sensitize tumor cells to the antiproliferative activity of rapamycin because PTEN loss leads to activation of the mTOR pathway. Because PTEN loss is frequent in endometrial cancer, we have characterized the effect of rapamycin in endometrial cancer cells. We show that rapamycin in the nanomolar concentration range exerts a potent growth-inhibitory effect on endometrial cancer cells through induction of cell cycle arrest. This effect is independent of PTEN status because PTEN-positive ECC-1 cells are as sensitive to rapamycin as PTEN-null Ishikawa and Hec-1B cells, suggesting that rapamycin may be effective against a broad range of endometrial cancers. We also show that rapamycin rapidly inhibits telomerase activity by decreasing the mRNA level of hTERT, the catalytic subunit of telomerase. This implies that rapamycin leads to inhibition of hTERT gene transcription. We demonstrate that rapamycin inhibits phosphorylation of downstream targets of mTOR such as p70(S6K) kinase and 4E-BP1 translation repressor. This work suggests that rapamycin is a potentially useful targeted therapy for endometrial cancer and that loss of telomerase activity may be a good surrogate biomarker for assessing antitumor activity of rapamycin.


Feasibility of lower-limb muscle power training to enhance locomotor function poststroke.
Poststroke motor control is characterized by greatly reduced muscle power generation. To date, the extent to which muscle power limits walking performance or whether its remediation should be a primary component of locomotor rehabilitation has yet to be established. The purpose of this study was to examine the feasibility and the effects of Poststroke Optimization of Walking using Explosive Resistance training, an intervention aimed at improving poststroke muscular and locomotor function. Twelve subjects (6-60 mo poststroke) participated in 24 training sessions (3 sessions/wk for 8 wk). Exercises included leg press, calf raises, and jump training, all performed at high concentric velocity, as well as trials of fast walking. We measured self-selected and fastest comfortable walking speeds as well as knee extensor and plantar flexor strength and power at pretraining, posttraining, and 8 wk follow-up time points. In addition, we also performed a number of clinical assessments commonly used in poststroke rehabilitation trials. Following training, significant improvements in lower-limb muscle strength and power were realized and accompanied by improvements in self-selected as well as fastest comfortable walking speeds. No changes in clinical assessments resulted from training.


Distribution patterns of three long-horned beetles (Coleoptera: Cerambycidae) shortly after fire in boreal forest: adults colonizing stands versus progeny emerging from trees.
We identified the factors that affect the early colonization of burned stands by adults and the progeny surviving in fire-killed black spruce trees for three cerambycid beetles: Acmaeops proteus proteus (Kirby), Acmaeops pratensis (Laicharting), and Monochamus scutellatus scutellatus (Say) (Coleoptera: Cerambycidae) in the northern Canadian boreal forest. Furthermore, we measured if progeny emerging from burned trees was related to patterns of adults captured in traps the same year as the fire. Fire severity at the stand and landscape scales were the most important predictors for colonizing adults. Except for A. pratensis, thick-barked and lightly burned trees positively influenced the occurrence of surviving progeny at the tree level. Last-instar larvae of A. pratensis emerged from burned trees more often in severely burned landscapes. This may result from biotic interactions with intraguild species or predators. With the exception of A. pratensis, variables affecting the postfire abundance and occurrence pattern of adults were strikingly different from progeny emerging after fire. Progeny emerging from burned trees was almost exclusively related to tree- or stand level characteristics, whereas colonizing adults were correlated with variables measured at various spatial scales, and most often at the landscape scale. Moreover, A. proteus proteus and M. scutellatus scutellatus adults were more common in severely burned landscapes, although their progeny emerged more often in lightly or moderately burned trees. Host selection behavior within stands (e.g., host acceptance) by colonizing adults or host suitability for the larvae might have caused this discrepancy.


Chromium supplementation alters the performance and health of feedlot cattle during the receiving period and enhances their metabolic response to a lipopolysaccharide challenge.
Crossbred steers (n = 180; 230 ± 6 kg) were fed during a 56-d receiving period to determine if supplementing chromium (Cr; KemTRACE Chromium Propionate 0.04%, Kemin Industries) would improve feedlot performance and health of newly-received cattle. A completely randomized block design (36 pens; 9 pens/treatment; 5 steers/pen) was used. Chromium premixes were supplemented to add 0 (Con), 0.1, 0.2, or 0.3 mg/kg of Cr to the total diet on a DM basis. No differences were detected on d 0 or after the first 14 d on feed. From d 0 to d 28, DMI (P = 0.07) and ADG increased linearly (P = 0.04) as Cr concentrations increased. From d 0 to d 56, BW (P = 0.08) displayed a tendency to increase linearly, and consequently ADG and G:F increased linearly (P ≤ 0.05) as Cr concentrations increased. The number of steers treated at least once for respiratory symptoms tended (P = 0.07) to linearly decrease as Cr concentrations increased. Twenty additional steers (235 ± 4 kg) were fed 56 d to determine if supplementing Cr (Con or 0.2 mg/kg Cr) would alter the metabolic response of newly-received cattle to a lipopolysaccharide (LPS) challenge. Cattle were fitted with jugular catheters on d 52. On d 55, blood samples were collected at 0.5-h intervals from -2 to 8 h, and again at 24 h relative to a LPS challenge (0.5 μg/kg BW) at 0 h. Serum glucose, insulin, and NEFA concentrations were determined from blood samples. Steer BW was also measured at cannulation, and 24 h and 8 d post-LPS. Steer BW did not differ at cannulation (P = 0.37), but 24 h post-LPS, Cr-supplemented steers had lost less BW (P = 0.03). Pre-LPS glucose concentration did not differ (P = 0.97). Post-LPS, there was a time × treatment interaction (P < 0.01) such that glucose concentration peaked earlier (0.5 h) and at a greater concentrations in Cr-supplemented steers (P < 0.01). Insulin concentration did not differ between treatments pre- or post-LPS (P > 0.13). Concentration of NEFA did not differ pre-LPS (P = 0.54); but 0.5 h post-LPS Cr-treated steers produced a greater peak NEFA concentration (P < 0.04). Results of this study indicate that supplementation of Cr to the basal diet can have beneficial effects on the performance and health of newly-received steers. These data also suggest that supplementation of Cr attenuated BW loss and allowed for a quicker recovery after a LPS challenge.


Bacterial Methionine Metabolism Genes Influence Drosophila melanogaster Starvation Resistance.
Animal-associated microorganisms (microbiota) dramatically influence the nutritional and physiological traits of their hosts. To expand our understanding of such influences, we predicted bacterial genes that influence a quantitative animal trait by a comparative genomic approach, and we extended these predictions via mutant analysis. We focused on Drosophila melanogaster starvation resistance (SR). We first confirmed that D. melanogaster SR responds to the microbiota by demonstrating that bacterium-free flies have greater SR than flies bearing a standard 5-species microbial community, and we extended this analysis by revealing the species-specific influences of 38 genome-sequenced bacterial species on D. melanogaster SR. A subsequent metagenome-wide association analysis predicted bacterial genes with potential influence on D. melanogaster SR, among which were significant enrichments in bacterial genes for the metabolism of sulfur-containing amino acids and B vitamins. Dietary supplementation experiments established that the addition of methionine, but not B vitamins, to the diets significantly lowered D. melanogaster SR in a way that was additive, but not interactive, with the microbiota. A direct role for bacterial methionine metabolism genes in D. melanogaster SR was subsequently confirmed by analysis of flies that were reared individually with distinct methionine cycle Escherichia coli mutants. The correlated responses of D. melanogaster SR to bacterial methionine metabolism mutants and dietary modification are consistent with the established finding that bacteria can influence fly phenotypes through dietary modification, although we do not provide explicit evidence of this conclusion. Taken together, this work reveals that D. melanogaster SR is a microbiota-responsive trait, and specific bacterial genes underlie these influences.IMPORTANCE Extending descriptive studies of animal-associated microorganisms (microbiota) to define causal mechanistic bases for their influence on animal traits is an emerging imperative. In this study, we reveal that D. melanogaster starvation resistance (SR), a model quantitative trait in animal genetics, responds to the presence and identity of the microbiota. Using a predictive analysis, we reveal that the amino acid methionine has a key influence on D. melanogaster SR and show that bacterial methionine metabolism mutants alter normal patterns of SR in flies bearing the bacteria. Our data further suggest that these effects are additive, and we propose the untested hypothesis that, similar to bacterial effects on fruit fly triacylglyceride deposition, the bacterial influence may be through dietary modification. Together, these findings expand our understanding of the bacterial genetic basis for influence on a nutritionally relevant trait of a model animal host.


Estimating gene penetrance from family data.
Family data are useful for estimating disease risk in carriers of specific genotypes of a given gene (penetrance). Penetrance is frequently estimated assuming that relatives' phenotypes are independent, given their genotypes for the gene of interest. This assumption is unrealistic when multiple shared risk factors contribute to disease risk. In this setting, the phenotypes of relatives are correlated even after adjustment for the genotypes of any one gene (residual correlation). Many methods have been proposed to address this problem, but their performance has not been evaluated systematically. In simulations we generated genotypes for a rare (frequency 0.35%) allele of moderate penetrance, and a common (frequency 15%) allele of low penetrance, and then generated correlated disease survival times using the Clayton-Oakes copula model. We ascertained families using both population and clinic designs. We then compared the estimates of several methods to the optimal ones obtained from the model used to generate the data. We found that penetrance estimates for common low-risk genotypes were more robust to model misspecification than those for rare, moderate-risk genotypes. For the latter, penetrance estimates obtained ignoring residual disease correlation had large biases. Also biased were estimates based only on families that segregate the risk allele. In contrast, a method for accommodating phenotype correlation by assuming the presence of genetic heterogeneity performed nearly optimally, even when the survival data were coded as binary outcomes. We conclude that penetrance estimates that accommodate residual phenotype correlation (even only approximately) outperform those that ignore it, and that coding censored survival outcomes as binary does not substantially increase the mean-square error of the estimates, provided the censoring is not extensive.


The invariance of the total direct DNA strand break yield.
The invariance of the total direct strand break yield when DNA is irradiated by different types of particles and energies has been reported by previous works. This study is intended to explain the physical causes of this behavior. The GEANT4-DNA extension of the GEANT4 general purpose Monte Carlo simulation toolkit has been used to determine direct strand break yields induced by protons and alpha particles impacting on a B-DNA geometrical model, including five organization levels of the human genetic material. The linear energy transfer (LET) of such particles ranges from 4.8 keV/microm (10 MeV protons) to about 235 keV/microm (2 MeV alpha particles), at 5.225 pm depth (near the center of the region of interest). Direct total, single and double strand break probabilities have been determined in a liquid water homogeneous medium with a 1.06 g/cm3 density. The energetic spectra of single strand breaks (SSB), the number of energy deposition events, and the SSB/event ratio were determined. The target-hit probability was found to be independent of both the type and the energy of the incident particle, even if this latter is a secondary electron. This probability is determined by the geometrical properties of the system. The total strand break yield and the number of energy deposition events required to reach a certain absorbed dose were found nearly independent of the type and energy of the incident ion (proton or alpha). In contrast, the double strand break (DSB) yield was found strongly dependent on the LET of the incident radiation. The SSB generation process is homogeneous and independent of the LET of the particles involved, at least within the proton and alpha particle energy range here studied. The target-hit probability is only determined by the ratio between the total volume occupied by targets and that of the ROI where the radiation deposits its energy. The maximum separation distance between two adjacent SSBs to produce a DSB is the parameter that breaks the homogeneity of the target-hit process, making the DSB production process strongly heterogeneous.


Sensitivity of brachial versus femoral vein injection of agitated saline to detect right-to-left shunts with Transcranial Doppler.
Transcranial Doppler (TCD) can detect a right-to-left shunt (RLS) with high sensitivity but has a 5% chance of a false negative study. TCD is usually performed with injection of agitated saline into an arm vein. We compared the sensitivity of TCD performed from the brachial versus femoral veins. Patients presenting to the cardiac catheterization laboratory for percutaneous closure of a patent foramen ovale (PFO) were enrolled. Power M-mode Transcranial Doppler (Terumo 150 PMD) was conducted. After injection of a mixture of 8 cc of agitated saline, 0.5 cc of air, and 1 cc of blood into the brachial vein, embolic tracks were counted over the middle cerebral arteries. The degree of RLS was evaluated by TCD at rest, and with Valsalva at 40 mmHg aided by visual feedback with a manometer device. The test was repeated using femoral venous injections. Sixty five patients were enrolled, mean age 52, 43% male. TCD grades were significantly higher with femoral injections compared to brachial injections at rest (p<0.0001), and with the Valsalva maneuver (p<0.0001). The presence of a RLS was confirmed by intracardiac echocardiography (ICE) during cardiac catheterization in 62 (95.4%) patients. The sensitivity of TCD for detection of RLS is increased when agitated saline injections are performed through the femoral vein. In patients with a high clinical suspicion for RLS, low TCD grades obtained with traditional brachial venous access should be interpreted with caution. When possible, a repeat study using femoral venous access may be considered.


Biosynthesis of platelet-activating factor (PAF) in human polymorphonuclear leucocytes. The role of lyso-PAF disposal and free arachidonic acid.
Theophylline and 1-methyl-3-isobutylxanthine (MIX), compounds that block eicosanoid formation and modulate phospholipase A2 activity, inhibited in a dose-dependent manner the formation of both leukotriene B4 (LTB4) and platelet-activating factor (PAF) by human polymorphonuclear leucocytes (PMN) in response to ionophore A23187. Theophylline and MIX lacked any inhibitory effect on acetyl-CoA: lyso-PAF acetyltransferase activity, which is the rate-limiting step for PAF biosynthesis in PMN. The effect of theophylline and MIX on PAF formation could be reversed by incubating the cells in the presence of 1-10 microM exogenous lyso-PAF. Incubation of PMN homogenates in the presence of unsaturated non-esterified fatty acids resulted in dose-dependent inhibition of the acetyltransferase. This effect was linked to the presence of a free carboxyl group, since both arachidonic acid methyl ester and palmitoyl-arachidonoyl phosphatidylcholine lacked inhibitory activity. This inhibitory effect was also dependent on the number of double bonds, since arachidonic acid (C20:4) and eicosapentaenoic acid (C20:5) displayed maximal effect. Kinetic analysis showed that the effect of arachidonic acid was consistent with competitive inhibition, with a Ki value of about 19 microM. Oxidative metabolites of arachidonic acid showed a lesser inhibitory effect with the following order of potency: arachidonic acid greater than 15-HETE (15-hydroxy-6,8,11,14-eicosatetraenoic acid) greater than LTB4 greater than 5-HETE (5-hydroxy-6,8,11,14-eicosatetraenoic acid) greater than lipoxin A4. Examination of enzymes involved in CoA-dependent acylation revealed a low activity of both arachidonoyl-CoA synthetase and arachidonoyl-CoA: lyso-PAF arachidonoyltransferase. These data indicate a strong influence on PAF biosynthesis of the products of the phospholipase A2 reaction, with lyso-PAF disposal being a critical event for PAF formation, and unsaturated fatty acids acting as feed-back inhibitors. The conversion of arachidonic acid via oxidative metabolism into less active inhibitors of acetyl-CoA:lyso-PAF acetyltransferase seems to be an additional mechanism of modulation of this enzyme activity, linked to the function of lipoxygenases. Finally, the enzyme activities involved in arachidonoyl-CoA-dependent acylation of lyso-PAF show a low efficiency in capturing arachidonic acid.


Three Pathways of Seclusion Reduction Programs to Sustainability: Ten Years Follow Up in Psychiatry.
From 2004 onwards, above 50 seclusion reduction programs (SRP) were developed, implemented and evaluated in the Netherlands. However, little is known about their sustainability, as to which extent obtained reduction could be maintained. This study monitored three programs over ten years seeking to identify important factors contributing to this. We reviewed documents of three SRPs that received governmental funding to reduce seclusion. Next, we interviewed key figures from each institute, to investigate the SRP documents and their implementation in practice. We monitored the number of seclusion events and the number of seclusion days with the Argus rating scale over ten years in three separate phases: 2008-2010, 2011-2014 and 2015-2017. As we were interested in sustainability after the governmental funding ended in 2012, our focus was on the last phase. Although in different rate, all mental health institutes showed some decline in seclusion events during and immediately after the SRP. After end of funding one institute showed numbers going up and down. The second showed an increase in number of seclusion days. The third institute displayed a sustained and continuous reduction in use of seclusion, even several years after the received funding. This institute was the only one with an ongoing institutional SRP after the governmental funding. To sustain accomplished seclusion reduction, a continuous effort is needed for institutional awareness of the use of seclusion, even after successful implementation of SRPs. If not, successful SRPs implemented in psychiatry will easily relapse in traditional use of seclusion.


Quantification of hepatitis delta virus RNA in serum by consensus real-time PCR indicates different patterns of virological response to interferon therapy in chronically infected patients.
Hepatitis delta virus (HDV), in association with hepatitis B virus, is responsible for severe acute and chronic hepatitis. Treatment of the infection relies on the long-term administration of high doses of alpha interferon (IFN), and the treatment efficiency is monitored by the detection of anti-HDV immunoglobulin M and HDV genome in serum. Like the case for other chronic viral infections, HDV genome quantification in serum should be useful for the follow-up of infected patients. The aims of this study were to develop a quantitative assay for the detection of any type of HDV in serum and to evaluate the benefit of HDV RNA quantification for the follow-up of chronically infected patients receiving IFN. A real-time reverse transcription-PCR assay was developed to quantify the HDV RNA load in serum. Its efficacy was evaluated with 160 serum samples, 76 of which were collected from 11 chronically infected patients who were treated with pegylated IFN. The assay was sensitive (100 copies/ml of serum) and efficient for all HDV types, including type 3 and the recently described types 5, 6, and 7. The viral load determinations for treated patients allowed us to identify different profiles of virological responses to IFN therapy with more accuracy than that attainable with the qualitative approach. In conclusion, we have developed a quantitative HDV RNA assay for serum which is adapted to the follow-up of antiviral treatment for patients infected with any HDV type. The assay will help us to understand the natural history of HDV infection and to define guidelines for the management of chronic delta hepatitis.


Should endomyocardial biopsy be performed for detection of myocarditis? A decision analytic approach.
Performance of endomyocardial biopsy (EMB) to diagnose myocarditis in patients with dilated cardiomyopathy is controversial because of a lack of evidence favoring immunosuppressive therapy. In spite of advances in heart failure treatment, dilated cardiomyopathy carries a poor prognosis, and myocardial inflammation and viral infection are potential therapeutic targets. We used decision analysis to determine the efficacy (5-year risk reduction in mortality or transplantation) that a treatment for myocarditis would require to favor a biopsy-guided approach over conventional therapy. Literature-based estimates included prevalence of myocarditis among patients with dilated cardiomyopathy with or without borderline myocarditis (16% and 11%, respectively); probability of 5-year transplantation-free survival (55%); sensitivity (50% and 63%, respectively), specificity (95.4%), and mortality rate (0.4%) of EMB; side effects resulting in withdrawal of immunosuppressive treatment (4%); and a 6-month mortality rate for immunosuppressive treatment (0.1%). All estimates were varied to determine impact on model results (sensitivity analysis). A therapy that decreased the rate of death or transplantation by 12.7% and 7.1% for patients without or with borderline myocarditis, respectively, favored EMB. Sensitivity analysis indicated that therapeutic efficacy was influenced by myocarditis prevalence and biopsy-related death, but not by accuracy of biopsy or probability of immunosuppressive therapy side effects. Randomized trials powered to detect 7% and 25% reductions in death and transplantation would require 5790 and 380 end points, respectively. Decreasing the rate of death or transplantation by 7.1% offsets therapy side effects, EMB-related death, and inaccuracies in histologic diagnosis. Prospective randomized trials of treatments for myocarditis may be more feasible during periods of high prevalence or with more sensitive diagnostic techniques.


Combinatorial design of multimeric chelating peptoids for selective metal coordination.
Current methods for metal chelation are generally based on multidentate organic ligands, which are generated through cumbersome multistep synthetic processes that lack flexibility for systematically varying metal-binding motifs. Octadentate ligands incorporating hydroxypyridinone or catecholamide binding moieties onto a spermine scaffold are known to display some of the highest affinities towards f-elements. Enhancing binding affinity for specific lanthanide or actinide ions however, necessitates ligand architectures that allow for modular and high throughput synthesis. Here we introduce a high-throughput combinatorial library of 16 tetrameric N-substituted glycine oligomers (peptoids) containing hydroxypyridinone or catecholamide chelating units linked via an ethylenediamine bridge and, for comparison, we also synthesized the corresponding mixed ligands derived from the spermine scaffold: 3,4,3-LI(1,2-HOPO)2(CAM)2 and 3,4,3-LI(CAM)2(1,2-HOPO)2. Coordination-based luminescence studies were carried out with Eu3+ and Tb3+ to begin probing the properties of the new ligand architecture and revealed higher sensitization efficiency with the spermine scaffold as well as different spectroscopic features among the structural peptoid isomers. Solution thermodynamic properties of selected ligands revealed different coordination properties between the spermine and peptoid analogues with Eu3+ stability constants log β 110 ranging from 28.88 ± 3.45 to 43.97 ± 0.49. The general synthetic strategy presented here paves the way for precision design of new specific and versatile ligands, with a variety of applications tailored towards the use of f-elements, including separations, optical device optimization, and pharmaceutical development.


Impact of "+Contigo" training on the knowledge and attitudes of health care professionals about suicide.
to evaluate the results of "+Contigo" training, developed by nurses and directed at 66 health professionals of integrated school health teams in Primary Health Care. quantitative with data collection through the Suicide Behavior Attitude Questionnaire, administered before and after the training. significant increases were observed in suicide prevention knowledge and in changing attitudes of health professionals towards individuals with suicidal behavior. these results allow us to affirm that nurses hold scientific and pedagogical knowledge that grant them a privileged position in the health teams, to develop training aimed at health professionals involved in suicide prevention.


Mohs versus surgical excision in nonmelanoma skin cancers: does location matter?
Every year, nearly 1.2 million people are affected by nonmelanoma skin cancers (NMSCs) in the United States. Most published data focus on comparing the efficacy of Mohs micrographic surgery (MMS) versus traditional surgical excision (TSE) for NMSCs in H-zone lesions of the face. There is paucity of data regarding the 2 treatments in other areas such as the non-H-zone areas of the face, the trunk, and extremities. Our study focused on the efficacy of the 2 treatments in areas of the body where the skin was not of premium. A retrospective chart review was performed of patients with NMSCs treated with TSE at the West Los Angeles Veterans Affairs Hospital between 2000 and 2008. Patients with at least a 3-year follow-up were selected for the study. Institutional review board approval was obtained before commencement of the study. Age, sex, and race-matched patients were selected in the MMS group. Data collected included demographic data, tumor characteristics, surgical treatment, reconstructions, recurrence rates, complications, and follow-up course. Data were analyzed using SigmaStat 3.5. A total of 588 patients were treated for NMSCs at our institute between 2000 and 2008, of which 289 patients had non-H-zone, extremity, and trunk lesions. The follow-up period for these patients was at least 3 years. Average age of this group was 67.1 (11.4) with 89.9% being males. Age, sex, and race-matched group of 200 patients treated with MMS for NMSCs were randomly chosen from the same time range. Average size of lesions was 17.4 (16.9) mm in the TSE group and 1.1 (0.4) mm in the MMS group (P < 0.05). Primary reconstruction was performed in non-premium areas (ie, non-H-zone areas of the face, the trunk, and extremities) in 98.7% patients in the TSE group and 61.5% patients in the MMS group (P < 0.05). Secondary reconstructive rate was 1.3% in TSE compared to 37.5% in MMS. Overall recurrence rate was 4.8% (compared to 3% with MMS). Of the 29 patients who had recurrences within the TSE group, 27 were H-zone lesions and 2 were non-H-zone lesions. One of the primary goals of NMSC management is to treat the lesion with adequate oncologic margins, while preserving maximal function and cosmesis. Our data look at the non-premium areas to quantify the clinical efficacy of TSE versus MMS. The size of lesions treated by TSE was significantly larger than those treated by MMS in all areas of the body. The primary closure rates were significantly higher and secondary procedure rates significantly lower in the TSE group compared to the MMS group, in non-premium areas. Our data suggest that patients with NMSCs may be more effectively treated with TSE than MMS in non-premium areas of the body. Additional studies are ongoing, including economic modeling and cost analysis.


The evaluation of trustworthiness to identify health insurance fraud in dentistry.
According to the investigations of the U.S. Government Accountability Office (GAO), health insurance fraud has caused an enormous pecuniary loss in the U.S. In Taiwan, in dentistry the problem is getting worse if dentists (authorized entities) file fraudulent claims. Several methods have been developed to solve health insurance fraud; however, these methods are like a rule-based mechanism. Without exploring the behavior patterns, these methods are time-consuming and ineffective; in addition, they are inadequate for managing the fraudulent dentists. Based on social network theory, we develop an evaluation approach to solve the problem of cross-dentist fraud. The trustworthiness score of a dentist is calculated based upon the amount and type of dental operations performed on the same patient and the same tooth by that dentist and other dentists. The simulation provides the following evidence. (1) This specific type of fraud can be identified effectively using our evaluation approach. (2) A retrospective study for the claims is also performed. (3) The proposed method is effective in identifying the fraudulent dentists. We provide a new direction for investigating the genuineness of claims data. If the insurer can detect fraudulent dentists using the traditional method and the proposed method simultaneously, the detection will be more transparent and ultimately reduce the losses caused by fraudulent claims.


Automated detection of case clusters of waterborne acute gastroenteritis from health insurance data - pilot study in three French districts.
This pilot study was conducted to assess the utility of using a health insurance database for the automated detection of waterborne outbreaks of acute gastroenteritis (AGE). The weekly number of AGE cases for which the patient consulted a doctor (cAGE) was derived from this database for 1,543 towns in three French districts during the 2009-2012 period. The method we used is based on a spatial comparison of incidence rates and of their time trends between the target town and the district. Each municipality was tested, week by week, for the entire study period. Overall, 193 clusters were identified, 10% of the municipalities were involved in at least one cluster and less than 2% in several. We can infer that nationwide more than 1,000 clusters involving 30,000 cases of cAGE each year may be linked to tap water. The clusters discovered with this automated detection system will be reported to local operators for investigation of the situations at highest risk. This method will be compared with others before automated detection is implemented on a national level.


[Comparative analysis of existing criteria in the assessment of iodine-deficiency in dwellers of the Far North areas].
New standards of the thyroid volume proposed by F. Delange (1977) are now being introduced. In view of this, we compared incidence of goiter by F. Delange and by R. Gutekunst (1988) criteria. The examination covered 3920 schoolchildren in 12 settlements of the Tyumen region in 1996. Iodine deficiency was mild to moderate. The greatest deficiency was observed in the North. Ultrasonic investigation of the thyroid by standards of R. Gutekunst (1988) was also indicative of the greatest iodine deficiency in the North. In the Far North, Polar Urals there was severe goiter endemia, in the rest areas the endemy was moderate. By F. Delange (1977) criteria, mild and moderate endemy was in the Far North and Polar Urals, the endemy was absent in the rest areas. As shown by comparison of the palpation data to thyroid ultrasonography, iodinuria, R. Gutekunst criteria are more reliable whereas criteria of F. Delange provide overestimated data.


Management of neurotrauma by surgeons and orthopedists in a military operational setting.
There is a considerable discrepancy between the potential demand for neurosurgeons and the actual availability of such specialists not only in civilian settings but even more so in military operational environments. For this reason, the Department of Neurosurgery at the German Armed Forces Hospital in Ulm conducts courses for surgeons and orthopedists on the management of patients with neurotrauma. Twelve such courses have already taken place. Each course lasts 1 week. Participants can expect to gain the theoretical knowledge and practical skills they need to provide initial surgical care for patients with traumatic brain injuries and/or spinal trauma. Surgical techniques are practiced above all in pig and human cadavers. At the end of the course, participants with previous surgical knowledge should be able to independently perform a craniotomy, from the planning of the procedure to the closure of the wound. Former course participants have successfully used their neurosurgical knowledge in countries of deployment where they managed patients during teleconsultation sessions and helped repatriate, or even provided surgical treatment to, patients with traumatic brain injuries. In these situations, it was particularly helpful when the physician deployed abroad and the neurosurgeon in Germany knew each other personally. In the future, efforts will be made to combine telemedicine and neuronavigation in an attempt to further improve direct support for physicians under military deployment conditions.


Genotoxic damage in hospital workers exposed to ionizing radiation and metabolic gene polymorphisms.
Of all workers exposed globally to synthetic sources of radiation, medical personnel represent the largest group, but receive relatively low doses. Accidental or therapeutic acute radiation exposure of humans was observed to induce various forms of cytogenetic damage, including the possibility of increasing the incidence of micronuclei (MN) and chromosomal aberrations (CA). The aim of this study was to assess occupationally induced chromosomal damage in a large population of hospital workers exposed to low doses of ionizing radiation (IR). The cytokinesis-block MN and comet assays were used to examine peripheral blood lymphocytes (PBL) of 31 exposed workers to IR and 33 control subjects corresponding in gender, age, and smoking. Glutathione S-transferases (GSTM1, GSTT1, and GSTP1) are postulated to be involved in the detoxification of endogenous and exogenous genotoxicants. The association between these biomarkers and polymorphic genes of xenobiotic metabolizing enzymes was thus also assessed. MN frequency was significantly higher in the exposed subjects compared controls. Comet assay results showed a significant increase of tail length in workers exposed to IR. Data obtained suggest that GSTM1, GSTT1, and GSTP1 polymorphism do not modify significantly the genotoxic potential of IR. Therefore, the exposed medical personnel need to carefully apply radiation protection procedures and minimize, as low as possible, IR exposure to avoid possible genotoxic effects.


Effect of N3im-methyl-thyrotropin releasing hormone on the human pituitary-thyroid axis.
The N3im-methyl analogue of thyrotropin release hormone (methyl-TRH) was compared with TRH as a thyrotropin releaser in 30 euthyroid volunteers (ages 19-61 years). The mean TSH response to 100 mug of methyl-TRH was greater (P less than 0.005) than the TSH response to 500 mug of TRH from 10 min to 240 min after giving the releasing factors. The mean peak TSH (at 30 min), maximum deltaTSH, and integrated TSH response area were greater (P less than 0.005) after administration of methyl-TRH than after TRH. The TSH response to methyl-TRH was significantly greater (P less than 0.05) for the 11 females than for the 19 males in this study. The mean baseline TSH was correlated with the maximum deltaTSH (r = 0.72, P less than 0.01) after methyl-TRH stimulation. The mean serum T3 concentration after methyl-TRH was significantly elevated at 60 min, peaked at 210 min and remained significantly elevated at 240 min. The peak serum T3, maximum T3 and T3 response area were significantly greater (P less than 0.005) after giving methyl-TRH than after TRH. The methyl-TRH induced T3 response area was 1.4 times the TRH induced T3 response area. The serum T4 concentration after methyl-TRH was elevated at 90 min (P less than 0.005), reached a peak at 210 min, and at 240 min was still 1.25 times the mean baseline T4. The peak serum T4, maximum deltaT4 and T4 response area after methyl-TRH were significantly greater than after TRH. The methyl-TRH induced T4 response area was 1.4 times the TRH induced T4 response area. The data indicate that methyl-TRH is a more potent thyrotropin releaser than TRH. Since N3im-methyl-histidine has been found in the brain, the possibility that this methyl analogue of TRH is a physiologic thyrotropin releaser should be evaluated.


Paraneoplastic Neurologic Syndromes.
Progress is ongoing in understanding paraneoplastic neurologic disorders, with new syndromes and antibodies being described and more detailed evidence available to guide workup for diagnosis and treatment to improve outcomes. Many excellent reviews have summarized the molecular features of different antibodies, but this article emphasizes the clinical features of each syndrome that may help guide initial diagnosis and treatment, which often should occur before an antibody or cancer is found to confirm the diagnosis. Recent findings include updated diagnostic criteria with validated sensitivity and specificity, discovery of novel antibodies, and clinical findings that increase the likelihood of an underlying paraneoplastic disorder. Suggestive syndromes that have been recently identified include faciobrachial dystonic seizures and pilomotor auras in anti-leucine-rich glioma inactivated protein 1 encephalitis, extreme delta brush on EEG in N-methyl-d-aspartate (NMDA)-receptor encephalitis, déjà vu aura in anti-glutamic acid decarboxylase 65 (GAD65) encephalitis, and sleep disturbances in several disorders. In addition, there is confirmed utility of brain positron emission tomography (PET) and CSF markers, including carcinoembryonic antigen and oligoclonal bands, as well as improved tests for the presence of leptomeningeal cancer cells in CSF. Associations of cancer immunotherapies with paraneoplastic syndromes and herpes simplex virus encephalitis (and COVID-19) with NMDA-receptor encephalitis have been described. All neurologists should be aware of advances regarding paraneoplastic neurologic syndromes, as patients can present with a wide variety of neurologic symptoms and earlier diagnosis and treatment can improve outcomes.


A potential molecular pathogenesis of cardiac/laterality defects in Oculo-Facio-Cardio-Dental syndrome.
Pitx2 is the last effector of the left-right (LR) cascade known to date and plays a crucial role in the patterning of LR asymmetry. In Xenopus embryos, the expression of Pitx2 gene in the left lateral plate mesoderm (LPM) is directly regulated by Xnr1 signaling, which is mediated by Smads and FoxH1. Previous studies suggest that the suppression of Pitx2 gene in the left LPM is a potential cause of cardiac/laterality defects in Oculo-Facio-Cardio-Dental (OFCD) syndrome, which is known to be caused by mutations in BCL6 co-repressor (BCOR) gene. Recently, our work has revealed that the BCL6/BCOR complex blocks Notch-dependent transcriptional activity to protect the expression of Pitx2 in the left LPM from the inhibitory activity of Notch signaling. These studies indicated that uncontrolled Notch activity in the left LPM caused by dysfunction of BCOR may result in cardiac/laterality defects of OFCD syndrome. However, this Notch-dependent inhibitory mechanism of Pitx2 gene transcription still remains unknown. Here we report that transcriptional repressor ESR1, which acts downstream of Notch signaling, inhibits the expression of Pitx2 gene by binding to a left side-specific enhancer (ASE) region in Pitx2 gene and recruiting histone deacetylase 1 (HDAC1) to this region. Once HDAC1 is tethered, histone acetyltransferase p300 is no longer recruited to the Xnr1-dependent transcriptional complex on the ASE region, leading to the suppression of Pitx2 gene in the left LPM. The study presented here uncovers the regulatory mechanism of Pitx2 gene transcription which may contribute to an understanding of pathogenesis of OFCD syndrome.


Disseminated intravascular coagulation in hematologic malignancies.
Disseminated intravascular coagulation (DIC) significantly contributes to the bleeding and thrombotic complications in patients with hematologic malignancies. As shown in other cancer settings, an underlying condition of activation of the coagulation system leading to a prothrombotic state of chronic or subclinical DIC is detectable. A variety of disease- or treatment-related factors may affect this condition, enhancing the risk of thrombosis or of bleeding and further triggering mechanisms of DIC in this setting. An overt DIC is diagnosed in approximately 15% of patients with acute leukemia, and bleeding manifestations prevail over thrombosis, with the highest and most harmful clinical impact in acute promyelocytic leukemia (APL). Pathogenic mechanisms include a series of intrinsic properties of malignant cells, able to directly activate the coagulation system or to stimulate prothrombotic effects by the host cells. Moreover, chemotherapy or concomitant infections play an important concurrent role. In this review the coagulation abnormalities, clinical manifestations, and the presently known pathophysiologic mechanisms of DIC in patients with hematologic malignancies are discussed, focusing on the most extensively studied condition of APL. Current approaches and open issues for the management and treatment of these patients are also reviewed.


Comparison of blood-film microscopy, the OptiMAL dipstick, Rhodamine-123 fluorescence staining and PCR, for monitoring antimalarial treatment.
In an attempt to see if the OptiMAL dipstick (Flow Inc., Portland, OR) can be used to monitor antimalarial treatment, a pilot study of 17 patients with Plasmodium falciparum malaria, admitted to the Hospital for Tropical Diseases in London, U.K., was conducted. Sequential, follow-up, blood specimens were obtained from day 1 to day 3, 4 or 5 post-admission. Thin and thick films prepared from these samples were examined for the presence of malarial parasites, and the intensities of parasitaemia were estimated. In addition, each specimen was tested with the OptiMAL dipstick, Rhodamine-123 fluorescence staining and, on specimens collected on day 1 and the last follow-up before discharge, by a PCR-based test. The results showed that OptiMAL has good sensitivity for the initial diagnosis of P. falciparum malaria and also mirrors the decline in viability of the parasites on treatment, giving the potential to follow the efficacy of drug treatment. The results of the PCR-based tests were still positive when the blood film and OptiMAL result were negative. The OptiMAL dipstick compared well with blood-film microscopy for monitoring antimalarial treatment and could be a useful replacement for microscopy to monitor treatment in places where facilities for microscopy are either lacking or inadequate. In developed countries it could be a useful adjunct to blood-film microscopy, and it might permit a reduction in the duration of hospitalization and give an early warning of treatment failure.