Impact of South American heroin on the US heroin market 1993-2004.
The past two decades have seen an increase in heroin-related morbidity and mortality in the United States. We report on trends in US heroin retail price and purity, including the effect of entry of Colombian-sourced heroin on the US heroin market. The average standardized price ($/mg-pure) and purity (% by weight) of heroin from 1993 to 2004 was from obtained from US Drug Enforcement Agency retail purchase data for 20 metropolitan statistical areas. Univariate statistics, robust Ordinary Least Squares regression and mixed fixed and random effect growth curve models were used to predict the price and purity data in each metropolitan statistical area over time. Over the 12 study years, heroin price decreased 62%. The median percentage of all heroin samples that are of South American origin increased an absolute 7% per year. Multivariate models suggest percent South American heroin is a significant predictor of lower heroin price and higher purity adjusting for time and demographics. These analyses reveal trends to historically low-cost heroin in many US cities. These changes correspond to the entrance into and rapid domination of the US heroin market by Colombian-sourced heroin. The implications of these changes are discussed.


COVID-induced sovereign risk in the euro area: When did the ECB stop the spread?
This paper studies how the announcement of the ECB's monetary policies stopped the spread of the COVID-19 pandemic to the European sovereign debt market. We show that up to March 9, the occurrence of new cases in euro area countries had a sizeable and persistent effect on 10-year sovereign bond spreads relative to Germany: 10 new confirmed cases per million people were accompanied by an immediate spread increase of 0.03 percentage points (ppt) that lasted 5 days, for a total increase of 0.35 ppt. For periods afterwards, the effect falls to near zero and is not significant. We interpret this change as an indicator of the success of the ECB's March 12 press conference, despite the "we are not here to close spreads" controversy. Our results hold for the stock market, providing further evidence of the effectiveness of the ECB's March 12 announcements in stopping the financial turmoil. A counterfactual analysis shows that without the shift in the sensitivity of sovereign bond markets to COVID-19, spreads would have surged to 4.2% in France, 12.5% in Spain, and 19.5% in Italy by March 18, when the ECB's Pandemic Emergency Purchase Programme was finally announced.


Symptomatic arachnoid cyst of the left frontal convexity presenting with memory disturbance--case report.
A 48-year-old female presented with vertiginous feeling and behavior disturbance. Computed tomography showed an arachnoid cyst on the left cerebral convexity. Single photon emission computed tomography revealed decreased cerebral blood flow (CBF) in the left frontal lobe. The Wechsler Memory Scale-Revised test demonstrated memory dysfunction. The arachnoid cyst was partially removed. Disturbances in CBF and behavior disappeared postoperatively. Local ischemia induced by compression due to arachnoid cyst may cause memory dysfunction and behavior disturbance. Neuropsychometric examination is useful for the evaluation of such symptoms.


Temporal characteristics of velopharyngeal function in children.
This investigation was designed to examine the performance of children with normal speech on temporal aspects of aerodynamic tasks related to velopharyngeal closure. The investigation was a descriptive evaluation of variability in aerodynamic features related to velopharyngeal function during multiple repetitions of the word "hamper." Children without speech or velopharyngeal difficulties were seen in an experimental laboratory setting for the evaluation procedures. Twenty-seven subjects were recruited for the experiment. Three subjects were rejected because of behavioral difficulties, and the remaining 24 subjects were subdivided into 4 groups of 6 children (3 males and 3 females) aged 3, 6, 9, and 12 years. The children, who were from local schools and day care centers, volunteered to participate in the experiment. All of the children had age-appropriate speech, language, and hearing abilities, as determined by screening tests administered by one of the examiners (L.T.). Mean and variability of pressure-flow measures of peak intraoral air pressure and peak nasal airflow and the temporal measures accompanying each air pressure or airflow pulse were evaluated for the age groups of children examined in the experiment. The aerodynamic procedures employed to evaluate velopharyngeal closure during speech were reliable for use with young children. There was a numerical trend toward decreased duration of the temporal parameters with increasing age. Thus, children demonstrated durational values similar to those previously reported for normal-speaking adults. In general, peak oral air pressure and nasal airflow values were like those of previous investigations and demonstrated low variability across all age groups of children tested. The data from the present investigation provide a preliminary base for comparison of temporal features of velopharyngeal closure for the aerodynamic evaluation of children with impaired velopharyngeal function.


Long-term results of gamma knife radiosurgery for foramen magnum meningiomas.
Outcomes of 37 patients of foramen magnum meningioma (FMM) were evaluated, and the related literature was reviewed to determine the efficacy of Gamma Knife radiosurgery (GKRS) for treating patients with FMM. We present the largest series reported from a single institution with the longest follow-up to date. The database of patients who underwent GKRS for FMM between 2007 and 2019 was evaluated retrospectively. A total of 37 patients with radiological and pathological features consistent with FMM were included in this series. Thirty-three patients were female, and 4 were male. The median age was 58 years (range, 23-74 years). The most common symptom at diagnosis was headache (64.9%). Twelve patients had a history of microsurgical resection. The median duration from the initial onset of symptoms to GKRS was 12 months (range 1-140 months). Among the 37 tumors, eight (21.6%) were located ventrally, 24 (64.9%) laterally, and five (13.5%) dorsally. The median target volume was 3.30 cm3 (range, 0.6-17.6 cm3). Thirty-five patients (95%) were treated with single fraction GKRS, and two patients (5%) were treated with hypofractionated GKRS. The median clinical follow-up was 80 months (range, 18-151 months), while the median radiological follow-up was 84 months (range, 18-144 months). At the last clinical follow-up after GKRS, 27 patients (73%) had improved symptoms, and none had worsened pre-GKRS symptoms. At the last radiological follow-up after GKRS, 23 tumors (62.2%) remained stable, 13 (35.1%) decreased in size, and 1 (2.7%) increased in size. Tumor control, including stable and regressed tumors, was achieved in 97.3% of patients. Our cohort demonstrates that GKRS is an effective and safe treatment for patients with either primary or recurrent/residual FMM.


Regulation of microRNA expression and abundance during lymphopoiesis.
Although the cellular concentration of miRNAs is critical to their function, how miRNA expression and abundance are regulated during ontogeny is unclear. We applied miRNA-, mRNA-, and ChIP-Seq to characterize the microRNome during lymphopoiesis within the context of the transcriptome and epigenome. We show that lymphocyte-specific miRNAs are either tightly controlled by polycomb group-mediated H3K27me3 or maintained in a semi-activated epigenetic state prior to full expression. Because of miRNA biogenesis, the cellular concentration of mature miRNAs does not typically reflect transcriptional changes. However, we uncover a subset of miRNAs for which abundance is dictated by miRNA gene expression. We confirm that concentration of 5p and 3p miRNA strands depends largely on free energy properties of miRNA duplexes. Unexpectedly, we also find that miRNA strand accumulation can be developmentally regulated. Our data provide a comprehensive map of immunity's microRNome and reveal the underlying epigenetic and transcriptional forces that shape miRNA homeostasis.


Cluster headache: conventional pharmacological management.
Cluster headache pain is very intense, usually increases in intensity very rapidly from onset, and attacks are often frequent. These clinical features result in significant therapeutic challenges. The most effective pharmacological treatment options for acute cluster attack include subcutaneous sumatriptan, 100% oxygen, and intranasal zolmitriptan. Subcutaneous or intramuscular dihydroergotamine and intranasal sumatriptan are additional options. Transitional therapy is applicable mainly for patients with high-frequency (>2 attacks per day) episodic cluster headache, and options include short courses of high-dose oral corticosteroids, dihydroergotamine, and occipital nerve blocks with local anesthetic and steroids. Prophylactic therapy is important both for episodic and chronic cluster headache, and the main options are verapamil and lithium. Verapamil is drug of first choice but may cause cardiac arrhythmias, and periodic electrocardiograms (EKGs) during dose escalation are important. Many other drugs are also in current use, but there is an insufficient evidence base to recommend them.


Childhood cancer data initiative: Status report.
In March 2023, over 800 researchers, clinicians, patients, survivors, and advocates from the pediatric oncology community met to discuss the progress of the National Cancer Institute's Childhood Cancer Data Initiative. We present here the status of the initiative's efforts in building its data ecosystem and updates on key programs, especially the Molecular Characterization Initiative and the planned Coordinated National Initiative for Rare Cancers in Children and Young Adults. These activities aim to improve access to childhood cancer data, foster collaborations, facilitate integrative data analysis, and expand access to molecular characterization, ultimately leading to the development of innovative therapeutic approaches.


Expression of metallothionein type 2 and 3 genes in Prosopis glandulosa leaves treated with copper.
For a better understanding of the strategies that are used by Prosopis glandulosa in heavy metal tolerance, the present study evaluated the gene expression of three metallothioneins (MTs; PgMt2-1, PgMt2, and PgMt3) in plants exposed to sub-lethal concentrations of copper. The PgMt2-1, PgMt2, and PgMt3 sequences were homologous to the MT type 2 (isoform 1), Mt2, and Mt3 sequences of other plant species found in GenBank. A reverse transcriptase-polymerase chain reaction showed that treatment with 100 mM Cu2+ induced a significant increase in PgMt2 and PgMt3 expression during the first 4 h of exposure compared to that of PgMt2-1. However, after 8 h of exposure, the expression levels of PgMt2 and PgMt3 were significantly lower than those of PgMt2-1. PgMt transcript levels only increased significantly during the first hour after exposure to copper, suggesting that PgMts could play a key role in the plant's detoxification mechanism. However, additional studies are required to confirm MTs as a mechanism of heavy metal tolerance and accumulation in this species.


Use of tumor necrosis factor alpha inhibitors in hepatitis B surface antigen-positive patients: a literature review and potential mechanisms of action.
As a class, tumor necrosis factor (TNF)-alpha inhibitors have provided clinicians significant control over chronic inflammatory diseases. With their widespread use has come the emergence of new side effects such as the reactivation of latent infections. One such infection that may reactivate is the hepatitis B virus (HBV). It is currently unknown if HBV reactivation is a class effect or attributable to a particular TNF-alpha inhibitor. To answer this question, a comprehensive literature review to identify trends in related cases was performed. A systemic literature review was performed using the PubMed and Medline databases (1996 to January 2010) searching for the index term "Hepatitis B" combined with the terms "tumor necrosis factor," "TNF-alpha inhibitors," "etanercept," "adalimumab," "certolizumab," and "golimumab." All relevant articles in English were reviewed, and secondary references of interest were also retrieved. Thirty-five cases with hepatitis B surface antigen (HBsAg) positivity known prior to initiation of TNF-alpha inhibitors were identified. Infliximab was used in 17 cases, etanercept in 12 cases, and adalimumab in 6 cases. All six cases of clinically symptomatic hepatitis were associated with infliximab therapy. Infliximab was associated with the most cases of greater than 2-fold increase in alanine aminotransferase (six of nine cases) and greater than 1,000-fold increase in HBV DNA load (three of four). The two deaths reported occurred with infliximab therapy. Potential mechanisms of action for the reported observations include differences in molecular design, route of administration, and potency in clearing TNF-alpha. In patients with a positive HBsAg prior to starting a TNF-alpha inhibitor, infliximab has the most reported cases associated with HBV reactivation. While such reactivation may be due to a variety of reasons, clinicians prescribing TNF-alpha inhibitors to HBsAg-positive patients should consider prophylactic antiviral therapy and close monitoring for any clinical or serological evidence of hepatitis.


How do novel molecular genetic markers influence treatment decisions in acute myeloid leukemia?
Acute myeloid leukemia (AML) is the most common acute leukemia diagnosed in adults, and the majority of patients with AML die from relapsed disease. Although many studies over the past 4 decades have identified disease alleles in AML, recent genome-wide and candidate gene studies have identified additional recurrent somatic mutations in AML patients with biologic, clinical, and therapeutic importance. Herein we review our current understanding of the molecular pathogenesis of AML and discuss how mutational profiling can be used to refine prognostication in AML and to inform therapeutic approaches. We also review the current challenges in translating genomic studies to the clinical setting, which remains a significant challenge and an urgent priority.


Self-assembly of mesoporous CuO nanosheets-CNT 3D-network composites for lithium-ion batteries.
A facile, flexible and large-scale technique was proposed to prepare a CuO-CNT 3D-network composite with the aid of electrostatic interactions in aqueous solution. The composite greatly improves the electrochemical performance. At a rate of 0.1 C, the cycling discharge capacity of the optimal composite is more than 2.3 times of that of unmodified mesoporous CuO nanosheets as the active material in an anode after 40 cycles.


Participatory identification of learning objectives in eight medical schools in Vietnam.
The advantages of outcome-based education for medical students have been recognised for several years and in several countries. Until recently in Vietnam, as in many countries, the curriculum for medical doctors was a broad framework that individual teachers filled in according to their own ideas and experience. During the past few years, the main eight medical schools in Vietnam have worked together to develop detailed learning objectives within the framework from the Ministries of Health and of Education and Training. The process was planned in an innovative and participatory way that involved more than one thousand teachers and other experts and resulted in a book listing the expected outcome in the form of the knowledge, attitudes and skills expected of a medical doctor graduating from any medical school in Vietnam. The process of identifying the objectives was followed by revision of the curriculum to be more community-oriented and to include more focus on teaching of skills, and by production of teaching, learning and assessment materials to fit the new curriculum. The process is described as an example of a comprehensive and bottom-up approach to curriculum development that could provide an example for other disciplines and schools in Vietnam or elsewhere.


Novel accelerometer realized by a polarization-maintaining photonic crystal fiber for railway monitoring applications.
In this paper, we present a novel accelerometer based on the Sagnac interferometer configuration using a polarization-maintaining photonic crystal fiber (PM-PCF), which has a sensitivity of ~8 pm/G, and a resonant frequency exceeding 2.5 kHz. The proposed accelerometer is capable of functioning with a constant sensitivity in a large frequency range from 0 to 1 kHz which is much wider than many FBG-based accelerometers. Experimental results obtained from a field test in railway monitoring, demonstrate a broader frequency range for the proposed accelerometer compared to that of the FBG based accelerometer and is comparable to the conventional piezoelectric sensor. The abrupt change in the acceleration measured by the sensor aids in locating any defect or crack present on the railway track. To the best of our knowledge, this is the first demonstration of an accelerometer based on a fiber interferometer aimed for the railway industry. The proposed accelerometer operating at high accelerations (>40 G) and capable of functioning at a broad frequency range, shows significant potential in being used in applications which require detection of strong and fast vibrations, especially in structural health monitoring of trains and railway tracks in real time.


[Leptospirosis. Serological study of an epidemic outbrake in the province of Camagüey].
A brief historical review of human leptospirosis up to 1972 in Cuba is made. Serological studies were made among sugar cane workers from the Army of Working Youth (EJT) during an epidemic leptospirosis outbreak at the Camgüey province in 1973. Results of those studies are presented. The outbreak was due to several serotypes of Leptospira. Other works of the author as well as several reports obtained from specialized medical literature are compared.


The relative age effect in selection to international team matches in Norwegian handball.
Many elite sport organisations have introduced structured talent identification and development (TID) initiatives in youth sports to better facilitate elite sport performance. However, selection mechanisms for TID programmes (e.g., junior international team) are biased towards relatively older athletes and limited studies exist with Scandinavian contexts. Therefore, the aim of this study was to explore the relative age effect (RAE) in youth, junior and senior male and female international team selections among Norwegian handball players (n = 657). A Chi-square goodness-of-fit test assessed whether a skewed birthdate distribution occurred at the youth, junior and senior international team levels and odds-ratios were calculated for RAE distribution. Moreover, a Kruskal-Wallis test was used to assess differences between the number of international youth, junior and senior level appearances by birth quartiles. Significant uneven birth date distributions were shown for youth (χ2(7) = female 40.383 and male 105.716, p <0.001) and junior (χ2(7) = female 27.427 and male 30.948, p <0.001) international players, favouring the relatively older player (odds-ratio of 1.9-8.3). At the senior level, no uneven distribution was identified. The comparison of the number of matches in each age category and the quartile of birth showed a difference in the women's youth category, where players in quartile 8 had a significantly lower number of appearances compared to quartile 1. The results form part of a growing body of knowledge about selection mechanisms in sport, which favours relatively older athletes within Norwegian Handball. Such findings are important for policy and practice for informing TID programmes for inclusive selection opportunities for all players.


Using EEG spatial correlation, cross frequency energy, and wavelet coefficients for the prediction of Freezing of Gait in Parkinson's Disease patients.
Parkinson's Disease (PD) patients with Freezing of Gait (FOG) often experience sudden and unpredictable failure in their ability to start or continue walking, making it potentially a dangerous symptom. Emerging knowledge about brain connectivity is leading to new insights into the pathophysiology of FOG and has suggested that electroencephalogram (EEG) may offer a novel technique for understanding and predicting FOG. In this study we have integrated spatial, spectral, and temporal features of the EEG signals utilizing wavelet coefficients as our input for the Multilayer Perceptron Neural Network and k-Nearest Neighbor classifier. This approach allowed us to predict transition from walking to freezing with 87 % sensitivity and 73 % accuracy. This preliminary data affirms the functional breakdown between areas in the brain during FOG and suggests that EEG offers potential as a therapeutic strategy in advanced PD.


Stability and change of attachment at 14, 24, and 58 months of age: behavior, representation, and life events.
Stability and change of attachment was examined longitudinally in a group of 48 children at 14, 24, and 58 months of age. Whereas attachment classifications showed stability between 14 and 24 months, lack of stability was the case between either 14 or 24 months and 58 months. Mothers of children who did not exhibit stability of attachment reported more negative and less positive life events. No correspondence was found between attachment classification at 14 or 24 months and mental representations at 58 months. However there was agreement between representational and behavioral measures of attachment at 58 months, where mental representations of attachment appeared to mirror and complement behavioral classification of the attachment relationship with the mother. Evidence for continuity of attachment from infancy to childhood internal representations was found only in a subgroup of children who showed stability of secure attachment between infancy and 58 months of age. These children exhibited greater Emotional Openness in the SAT at 4.5 years. Findings are discussed in terms of different positions in attachment theory and research regarding the stability and change of attachment relationships.


[Ulcerous disease of endocrinal origin caused by a non-gastric substance].
The purpose of this paper is to report on a form of duodenal ulceration associated with a major hypersecretion that we described in 1984. This entity belongs to the group of endocrine duodenal ulcer diatheses of which the Zollinger-Ellison Syndrome (ZES) is the prototype. We have recently in 1989 reported a series of patients with clinical and pathologic features indistinguishable from those of the Zollinger-Ellison syndrome but without hypergastrinemia. We have also found that the plasma and the tumor tissues of these patients contains a potent non-gastrin secretagogue.


Cytogenetic evidences on the evolutionary relationships between the tetraploids of the section Rhizomatosae and related diploid species (Arachis, Leguminosae).
Rhizomatosae is a taxonomic section of the South American genus Arachis, whose diagnostic character is the presence of rhizomes in all its species. This section is of particular evolutionary interest because it has three polyploid (A. pseudovillosa, A. nitida and A. glabrata, 2n = 4x = 40) and only one diploid (A. burkartii, 2n = 2x = 20) species. The phylogenetic relationships of these species as well as the polyploidy nature and the origin of the tetraploids are still controversial. The present study provides an exhaustive analysis of the karyotypes of all rhizomatous species and six closely related diploid species of the sections Erectoides and Procumbentes by cytogenetic mapping of DAPI/CMA heterochromatin bands and 5S and 18-26S rDNA loci. Chromosome banding showed variation in the DAPI heterochromatin distribution pattern, which, together with the number and distribution of rDNA loci, allowed the characterization of all species studied here. The bulk of chromosomal markers suggest that the three rhizomatous tetraploid species constitute a natural group and may have at least one common diploid ancestor. The cytogenetic data of the diploid species analyzed evidenced that the only rhizomatous diploid species-A. burkartii-has a karyotype pattern different from those of the rhizomatous tetraploids, showing that it is not likely the genome donor of the tetraploids and the non-monophyletic nature of the section Rhizomatosae. Thus, the tetraploid species should be excluded from the R genome, which should remain exclusively for A. burkartii. Instead, the karyotype features of these tetraploids are compatible with those of different species of the sections Erectoides and Procumbentes (E genome species), suggesting the hypothesis of multiple origins of these tetraploids. In addition, the polyploid nature and the group of diploid species closer to the tetraploids are discussed.


[Rate of dental treatment needs: the case of the Xakriaba Indians].
The present study was developed among Xakriabas Indians in Minas Gerais and aimed at identifying their needs for dental treatment so that patients could be scheduled for treatment. Native health agents were trained by dental-surgeons for conducting the survey in household visits. The total Indian population is of 6.498 inhabitants, 3.686 of these living within the territory of the "Base Mata Fome", where 61% of the population was examined. Results showed that the population is mainly young, a great number of individuals presented no need for treatment and dental caries was the most frequent problem. The survey is expected to help in the organization of a schedule that allows giving priority to the more serious cases and promoting equity.


Acquired Cystic Disease-associated Renal Cell Carcinoma (ACD-RCC): A Multiinstitutional Study of 40 Cases With Clinical Follow-up.
The incidence of renal cell carcinoma (RCC) is known to be higher in patients with end-stage renal disease, including those with acquired cystic kidney disease due to dialysis. Acquired cystic disease (ACD)-associated RCC was recently incorporated into the 2016 WHO Classification of Tumors of the Urinary System and Male Genital Tract as a distinct entity and is reportedly the most common RCC arising in end-stage renal disease. In this study, we sought to further describe clinicopathologic findings in a large series of ACD-RCC, emphasizing histologic features, immunophenotype, clinical outcome, and patterns of disease spread. We collected 40 previously unpublished cases of ACD-RCC with mean clinical follow-up of 27 months (median, 19 mo; range, 1 to 126 mo). Mean tumor size was 2.7 cm (median, 2.4 cm), and 32 tumors (80%) were confined to the kidney (pT stage less than pT3a). International Society of Urological Pathology grade was 3 in 37 cases (92.5%), grade 2 in 1 case (2.5%), and grade 4 in 2 cases (5%). Architectural variability among ACD-RCC was common, as 39 cases (98%) showed varying combinations of tubular, cystic, solid, and/or papillary growth. ACD-RCC frequently occurred in association with other renal tumor subtypes within the same kidney, including papillary RCC (14 patients), papillary adenomas (7 cases), clear cell papillary RCC (5 cases), clear cell RCC (1 case), and RCC, unclassified type (1 case). A previously undescribed pattern of perinephric and renal sinus adipose tissue involvement by dilated epithelial cysts with minimal or absent intervening capsule or renal parenchyma was identified in 20 cases (50%); these cysts were part of the tumor itself in 5 cases (25%) and were part of the non-neoplastic acquired cystic change in the background kidney in the remaining 15 cases (75%). Of the 24 cases (60%) with tissue available for immunohistochemical stains, 19 (79%) were positive for PAX8, 20 (83%) showed negative to patchy expression of cytokeratin 7, and 24 (100%) were both positive for AMACR and negative for CD117. Fumarate hydratase expression was retained in all tumors, including those with nuclear features resembling fumarate hydratase-deficient RCCs. Of the 36 patients (90%) with available follow-up information, 4 (11%) experienced adverse events: 2 patients developed a local recurrence, 1 patient experienced multiple visceral metastases and subsequently died of disease, and 1 patient developed metastases to regional lymph nodes only. One local recurrence and the lymph node only metastasis both had an unusual, exclusively cystic pattern of growth. In summary, we present the largest clinicopathologic series of ACD-RCC to date and describe previously unreported cystic patterns of local soft tissue involvement and recurrence/metastases.


Quebec employs user-pay philosophy in launching drug-insurance plan.
The Quebec government has introduced a new provincial drug-insurance scheme that by January 1997 will provide coverage to all Quebecers. The first phase of the measure, which was instituted for reasons that were partly financial and partly social, came into effect in August, to the chagrin of the province's welfare recipients and elderly. They are the first to pay 25% of the cost of prescriptions under the new copayment plan.


Fast spin of the young extrasolar planet β Pictoris b.
The spin of a planet arises from the accretion of angular momentum during its formation, but the details of this process are still unclear. In the Solar System, the equatorial rotation velocities and, consequently, spin angular momenta of most of the planets increase with planetary mass; the exceptions to this trend are Mercury and Venus, which, since formation, have significantly spun down because of tidal interactions. Here we report near-infrared spectroscopic observations, at a resolving power of 100,000, of the young extrasolar gas giant planet β Pictoris b (refs 7, 8). The absorption signal from carbon monoxide in the planet's thermal spectrum is found to be blueshifted with respect to that from the parent star by approximately 15 kilometres per second, consistent with a circular orbit. The combined line profile exhibits a rotational broadening of about 25 kilometres per second, meaning that β Pictoris b spins significantly faster than any planet in the Solar System, in line with the extrapolation of the known trend in spin velocity with planet mass.


Alveolar cleft closure by osseodistraction: pitfalls and troubleshooting.
Segmental maxillary osseodistraction is a proper method to close alveolar gaps in patients with clefts of lip, palate, and alveolus. The technique is mainly used in very wide clefts after failure of conventional bone grafting procedures. The aim of the study was to analyze problems that may occur during the distraction process. Patients with uncommon wide alveolar clefts or recurrent oronasal fistulas and patients with bilateral clefts of lip, plate, and alveolus and an additional vertical deficit of the posterior cleft segment underwent distraction procedures. The patients were subdivided according to the vector of distraction: linear (following the dental arch) and vertical. The devices for horizontal distraction were tooth-borne and manufactured to the specific clinical situation. For vertical distraction, standard devices were used. Of this sample, patients with complications occurring during or after surgery and during the distraction period were selected as the study participants. On the basis of the data collected on these complications, the need for additional bone grafting procedures was evaluated. Results suggest that modifications in preoperative planning and the construction of the distraction devices may be necessary to prevent unexpected problems in segmental maxillary distraction procedures. Considerations concerning the placement of distraction devices are discussed.


The Hard-To-Close Window of T-Type Calcium Channels.
T-Type calcium channels (TTCCs) are key regulators of membrane excitability, which is the reason why TTCC pharmacology is subject to intensive research in the neurological and cardiovascular fields. TTCCs also play a role in cancer physiology, and pharmacological blockers such as tetralols and dihydroquinazolines (DHQs) reduce the viability of cancer cells in vitro and slow tumor growth in murine xenografts. However, the available compounds are better suited to blocking TTCCs in excitable membranes rather than TTCCs contributing window currents at steady potentials. Consistently, tetralols and dihydroquinazolines exhibit cytostatic/cytotoxic activities at higher concentrations than those required for TTCC blockade, which may involve off-target effects. Gene silencing experiments highlight the targetability of TTCCs, but further pharmacological research is required for TTCC blockade to become a chemotherapeutic option.


The European Thoracic Surgery Database project: modelling the risk of in-hospital death following lung resection.
To identify pre-operative factors associated with in-hospital mortality following lung resection and to construct a risk model that could be used prospectively to inform decisions and retrospectively to enable fair comparisons of outcomes. Data were submitted to the European Thoracic Surgery Database from 27 units in 14 countries. We analysed data concerning all patients that had a lung resection. Logistic regression was used with a random sample of 60% of cases to identify pre-operative factors associated with in-hospital mortality and to build a model of risk. The resulting model was tested on the remaining 40% of patients. A second model based on age and ppoFEV1% was developed for risk of in-hospital death amongst tumour resection patients. Of the 3426 adult patients that had a first lung resection for whom mortality data were available, 66 died within the same hospital admission. Within the data used for model development, dyspnoea (according to the Medical Research Council classification), ASA (American Society of Anaesthesiologists) score, class of procedure and age were found to be significantly associated with in-hospital death in a multivariate analysis. The logistic model developed on these data displayed predictive value when tested on the remaining data. Two models of the risk of in-hospital death amongst adult patients undergoing lung resection have been developed. The models show predictive value and can be used to discern between high-risk and low-risk patients. Amongst the test data, the model developed for all diagnoses performed well at low risk, underestimated mortality at medium risk and overestimated mortality at high risk. The second model for resection of lung neoplasms was developed after establishing the performance of the first model and so could not be tested robustly. That said, we were encouraged by its performance over the entire range of estimated risk. The first of these two models could be regarded as an evaluation based on clinically available criteria while the second uses data obtained from objective measurement. We are optimistic that further model development and testing will provide a tool suitable for case mix adjustment.


Quantum mechanical force field for hydrogen fluoride with explicit electronic polarization.
The explicit polarization (X-Pol) theory is a fragment-based quantum chemical method that explicitly models the internal electronic polarization and intermolecular interactions of a chemical system. X-Pol theory provides a framework to construct a quantum mechanical force field, which we have extended to liquid hydrogen fluoride (HF) in this work. The parameterization, called XPHF, is built upon the same formalism introduced for the XP3P model of liquid water, which is based on the polarized molecular orbital (PMO) semiempirical quantum chemistry method and the dipole-preserving polarization consistent point charge model. We introduce a fluorine parameter set for PMO, and find good agreement for various gas-phase results of small HF clusters compared to experiments and ab initio calculations at the M06-2X/MG3S level of theory. In addition, the XPHF model shows reasonable agreement with experiments for a variety of structural and thermodynamic properties in the liquid state, including radial distribution functions, interaction energies, diffusion coefficients, and densities at various state points.


Histone methylation makes its mark on longevity.
How long organisms live is not entirely written in their genes. Recent findings reveal that epigenetic factors that regulate histone methylation, a type of chromatin modification, can affect lifespan. The reversible nature of chromatin modifications suggests that therapeutic targeting of chromatin regulators could be used to extend lifespan and healthspan. This review describes the epigenetic regulation of lifespan in diverse model organisms, focusing on the role and mode of action of chromatin regulators that affect two epigenetic marks, trimethylated lysine 4 of histone H3 (H3K4me3) and trimethylated lysine 27 of histone H3 (H3K27me3), in longevity.


[Enzyme histochemical characteristics of prostaglandin synthetase, adenylate cyclase and cAMP in a focus of inflammation in chronic tonsillitis].
Upon enzyme histochemical evaluation of palatine tonsils removed from 80 patients with chronic tonsillitis it was established that there is a close correlation between activity of prostaglandin synthetase (PG-synthetase), adenylate cyclase (AC), cellular cAMP, pattern of inflammation and the size of streptococcal foci. Enhanced enzymatic activity, elevated content of lymphoid and connective tissue cAMP are associated with minor interstitial streptococcal foci, being a frequent finding in mild inflammation. Low cellular cAMP in high PG-synthetase and AC activity occurs in mild inflammation and large-sized interstitial streptococcal foci. When both enzymatic activity and cellular cAMP are low, torpid inflammation indicates lack of tissue response to microbial invasion resulting in emergence of interstitial streptococcal colonies. In view of the essential role of mediator effects in the inflammation focus, high activity of tonsil PG-synthetase should be considered a major indicator of activated chronic tonsillitis.


Growth factor-like action of lysophosphatidic acid on human B lymphoblasts.
Proliferation and immunoglobulin secretion of B lymphocytes are regulated by specific antigens and numerous accessory immunomodulatory factors. Lysophosphatidic acid (LPA) is a glycerophospholipid mediator that is released from activated blood platelets, attains high levels in serum, and exerts potent stimulatory effects on, e.g., neutrophils, monocytes, and T lymphocytes. LPA is also generated by a secretory, cytokine-inducible phospholipase A2 present in high concentrations in inflammatory exudates and septic states. We investigated effects of LPA on human Epstein-Barr virus-immortalized B lymphoblasts, a model for immunoglobulin-secreting B cells. Intracellular Ca2+ was determined with fura 2 and the formation of inositol 1,4, 5-trisphosphate by anion-exchange chromatography. LPA stimulated an increase in inositol 1,4,5-trisphosphate levels and induced a transient rise in intracellular free Ca2+ concentration from 105 +/- 17 to 226 +/- 21 nM. This Ca2+ signal resulted from Ca2+ mobilization and Ca2+ influx and was subject to homologous desensitization. Pertussis toxin inhibited these responses by approximately 70%. Furthermore, LPA stimulated a 27.5% increase in guanosine 5'-O-(3-thiotriphosphate) binding to permeabilized B lymphoblasts, which suggests the direct activation of pertussis toxin-sensitive G proteins by LPA. LPA stimulated a strong increase in the specific phosphorylation of the mitogen-activated protein kinase (immunoblot analysis) that was prevented by the MEK inhibitor PD-98059. Finally, LPA triggered a 2-fold increase in DNA synthesis ([3H]thymidine incorporation) and a 2-fold increase in B lymphoblast number and evoked a 20- to 50-fold increase in immunoglobulin formation. By RT-PCR we detected specific mRNA transcripts for the recently cloned human LPA receptor. Thus our data suggest that LPA behaves as a B cell growth factor.


Multi-omics analysis of ten carbon nanomaterials effects highlights cell type specific patterns of molecular regulation and adaptation.
New strategies to characterize the effects of engineered nanomaterials (ENMs) based on omics technologies are emerging. However, given the intricate interplay of multiple regulatory layers, the study of a single molecular species in exposed biological systems might not allow the needed granularity to successfully identify the pathways of toxicity (PoT) and, hence, portraying adverse outcome pathways (AOPs). Moreover, the intrinsic diversity of different cell types composing the exposed organs and tissues in living organisms poses a problem when transferring in vivo experimentation into cell-based in vitro systems. To overcome these limitations, we have profiled genome-wide DNA methylation, mRNA and microRNA expression in three human cell lines representative of relevant cell types of the respiratory system, A549, BEAS-2B and THP-1, exposed to a low dose of ten carbon nanomaterials (CNMs) for 48 h. We applied advanced data integration and modelling techniques in order to build comprehensive regulatory and functional maps of the CNM effects in each cell type. We observed that different cell types respond differently to the same CNM exposure even at concentrations exerting similar phenotypic effects. Furthermore, we linked patterns of genomic and epigenomic regulation to intrinsic properties of CNM. Interestingly, DNA methylation and microRNA expression only partially explain the mechanism of action (MOA) of CNMs. Taken together, our results strongly support the implementation of approaches based on multi-omics screenings on multiple tissues/cell types, along with systems biology-based multi-variate data modelling, in order to build more accurate AOPs.


Clarifying the role of psychological pain in the risks of suicidal ideation and suicidal acts among patients with major depressive episodes.
The role of psychological pain in the risk of suicide was explored using a three-dimensional psychological pain model (pain arousal, painful feelings, pain avoidance). The sample consisted of 111 outpatients with major depressive episodes, including 28 individuals with suicidal histories. They completed the Chinese version of the Beck Scale for Suicide Ideation (BSI), the Beck Depression Inventory (BDI), the Psychache Scale, and the three-dimensional Psychological Pain Scale (TDPPS). A structured clinical interview was conducted to assess the history of suicidal acts. Significant correlations were found among BDI, BSI, and TDPPS scores (p < .01). Stepwise regression analyses showed that only pain avoidance scores significantly predicted suicide ideation at one's worst point (β = .79, p < .001) and suicidal acts (β = .46, p < .001). Pain avoidance was also a better predictor of current suicidal ideation (β = .37, p = .001) than were BDI scores (β = .31, p < .01). Increased levels of pain avoidance during a major depressive episode may be a dominant component of the motivation for suicide. Future clinical assessments for populations at high risk of suicide should include measures of psychological pain to reduce the incidence of suicide.


Fastigial nucleus cardiovascular response and brain stem lesions in the beagle.
Changes in the excitatory cardiovascular response (heart rate, arterial blood pressure, left ventricular pressure, and LV dP/dt as an index of myocardial contractility) resulting from electrical stimulation of the cerebellar fastigial nucleus (FN) were recorded after placement of DC or radio-frequency lesions or after microinjections of kainic acid into brain stem areas that receive FN projections and have been shown to be involved in central cardiovascular control. FN-induced increases in heart rate, blood pressure, and contractility were reduced or abolished by lesions made in the restiform body or the A5 area, which is homologous to the catecholamine-containing region in cats and rats. Lesions in the paramedian reticular nucleus, rostral and caudal to obex, failed to reduce the FN cardiovascular response. Nucleus of the solitary tract lesions augmented the FN pressor response and tachycardia. Kainic acid (1 microliter of 100 mM solution) caused profound depression of heart rate, blood pressure, and contractility and reduced or eliminated the FN-induced cardiovascular response when injected into the A5 area, previously identified by the pressor response following electrical stimulation. We concluded from these observations that a descending fastigiobulbar sympathoexcitatory pathway courses through a previously identified A5 pressor area that is also capable of a depressor response when the cell bodies alone are activated.


The impact of acute PAH exposure on the toadfish glucocorticoid stress response.
The objective of the present study was to determine whether the polycyclic aromatic hydrocarbons (PAHs) associated with the Deepwater Horizon (DWH) oil spill impacted the stress response of teleost fish. The hypothesis was that intraperitoneal (IP) treatment with PAHs associated with the DWH oil spill or waterborne exposure to DWH oil high energy water-accommodated fraction (HEWAF) would result in the downregulation of the stress response of Gulf toadfish, Opsanus beta, a benthic marine teleost fish that resides in the Gulf of Mexico. In vivo plasma cortisol levels and adrenocorticotropic hormone (ACTH)-mediated cortisol secretion by in vitro isolated kidney tissue were measured. Toadfish at rest IP-treated with naphthalene had higher plasma cortisol compared to fluorene-treated and control fish; phenanthrene-treated fish tended to have higher plasma cortisol levels that fluorene-treated and controls. When subjected to an additional crowding stress, naphthalene and phenanthrene-treated fish were no longer able to mount a stress response compared to fluorene-treated and control fish, suggesting exhaustion of the stress response. Supporting this in vivo data, there tended to be less cortisol released by the kidney in vitro from naphthalene and phenanthrene-treated fish in response to ACTH compared to controls. In contrast, toadfish at rest exposed to 3% Slick A HEWAF did not have significantly different plasma cortisol levels compared to controls. But, exposed fish did have significantly less cortisol released by the kidney in vitro in response to ACTH. When toadfish were subjected to an additional stress, there were no significant differences in plasma cortisol or ACTH, suggesting the action of a secondary secretagogue to maintain plasma cortisol in vivo. Combined, these data suggest that in response to acute PAH exposure, there may be internalization or downregulation of the melanocortin 2 receptor (MC2R) that mediates the action of ACTH.


"Pseudotumor" of the stomach. Endoscopical and operative findings.
Persistent pathological changes at endoscopy are reported in three patients. At explorative laparotomy the changes were seen to be caused by neighbouring organs (splenic artery, pancreas and gallbladder). A correct diagnosis of "pseudotumor" of the stomach is necessary to avoid unwarranted surgery or unnecessary biopsies.


Determination of serologic and colostral response in late-gestation cows vaccinated with a Mycoplasma bovis bacterin.
To determine whether vaccinating cows during late gestation against Mycoplasma bovis will result in adequate concentrations of M bovis-specific IgG(1) in serum, colostrum, and milk. 78 dairy cows. Serum samples were obtained 60 and 39 days prior to expected parturition in vaccinated and control cows from a single herd. Serum and colostrum samples were also obtained at parturition. Milk samples were obtained 7 to 14 days after parturition. Samples were analyzed for anti-M bovis IgG(1) concentrations. Prior to vaccination, control and vaccinated cows had similar anti-M bovis IgG(1) concentrations. After initial vaccination and subsequent booster and at parturition, there was a significant difference between the 2 groups, with vaccinated cows having higher IgG concentrations. Colostrum from vaccinated cows had higher anti-M bovis IgG(1) concentrations, compared with control cows; however, IgG(1) concentrations in milk did not differ between the 2 groups. Vaccination of late-gestation cows resulted in increased concentrations of anti-M bovis IgG(1) in colostrum. However, ingestion of colostrum by calves may not guarantee protection against M bovis infection.


[Susceptibility of the parasitoid Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) to Beauveria bassiana under laboratory conditions].
The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is the most important coffee pest worldwide. Beauveria bassiana is a generalist entomopathogenic fungus widely used by coffee farmers to control this pest and Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) is an African endoparasitoid of H. hampei adults, recently imported to several Latin American and Caribbean countries to aid in the coffee berry borer control. The objective of this study was to determine if B. bassiana is detrimental to P. coffea. The susceptibility of the parasitoid was evaluated in terms of adult survivorship, mean lethal concentration (LC50), mean lethal time (LT50), reproduction and immature mortality. The main effect of the fungus resulted in reduction of adult longevity and mortality of 100% for immature stages of this parasitoid. The LC50 for adults was 0.11% equivalent to 9.53 x 10(7) conidia/ml of B. bassiana and a LT50 of 29.4h, equivalent to reduction of 22% of its normal longevity as an adult. P. coffea was capable of disseminating spores of B. bassiana to non-infected H. hampei adults, which could indirectly cause the death of its own progeny. These results could be valuable when considering the use of both organisms in the field, especially in an integrated pest management program.


Cultural selection and biased transformation: two dynamics of cultural evolution.
Here, I discuss two broad versions of human cultural evolution which currently exist in the literature and which emphasize different underlying dynamics. One, which originates in population-genetic-style modelling, emphasizes how cultural selection causes some cultural variants to be favoured and gradually increase in frequency over others. The other, which draws more from cognitive science, holds that cultural change is driven by the biased transformation of cultural variants by individuals in non-random and consistent directions. Despite claims that cultural evolution is characterized by one or the other of these dynamics, these are neither mutually exclusive nor a dichotomy. Different domains of human culture are likely to be more or less strongly weighted towards cultural selection or biased transformation. Identifying cultural dynamics in real-world cultural data is challenging given that they can generate the same population-level patterns, such as directional change or cross-cultural stability, and the same cognitive and emotional mechanisms may underlie both cultural selection and biased transformation. Nevertheless, fine-grained historical analysis and laboratory experiments, combined with formal models to generate quantitative predictions, offer the best way of distinguishing them. This article is part of the theme issue 'Foundations of cultural evolution'.


Exploring the therapeutic mechanisms of Cassia glauca in diabetes mellitus through network pharmacology, molecular docking and molecular dynamics.
Cassia glauca is reported as anti-diabetic medicinal plant and is also used as an ethnomedicine. However, its mode of action as an anti-diabetic agent has not been clearly elucidated. Hence, the present study investigated the probable mechanism of action of C. glauca to manage diabetes mellitus via network pharmacology and molecular docking and simulations studies. The reported bioactives from C. glauca were retrieved from an open-source database, i.e. ChEBI, and their targets were predicted using SwissTargetPrediction. The proteins involved in the pathogenesis of diabetes were identified from the therapeutic target database. The targets involved in diabetes were enriched in STRING, and the pathways involved in diabetes were identified concerning the KEGG. Cytoscape was used to construct the network among bioactives, proteins, and probably regulated pathways, which were analyzed based on edge count. Similarly, molecular docking was performed using the Glide module of the Schrodinger suite against majorly targeted proteins with their respective ligands. Additionally, the drug-likeness score and ADMET profile of the individual bioactives were predicted using MolSoft and admetSAR2.0 respectively. The stability of these complexes were further studied via molecular dynamics simulations and binding energy calculations. Twenty-three bio-actives were retrieved from the ChEBI database in which cassiarin B was predicted to modulate the highest number of proteins involved in diabetes mellitus. Similarly, GO analysis identified the PI3K-Akt signaling pathway to be primarily regulated by modulating the highest number of gene. Likewise, aldose reductase (AKR1B1) was majorly targeted via the bioactives of C. glauca. Similarly, docking study revealed methyl-3,5-di-O-caffeoylquinate (docking score -9.209) to possess the highest binding affinity with AKR1B1. Additionally, drug-likeness prediction identified cassiaoccidentalin B to possess the highest drug-likeness score, i.e. 0.84. The molecular dynamics simulations and the MMGBSA indicate high stability and greater binding energy for the methyl-3,5-di-O-caffeoylquinate (ΔG bind = -40.33 ± 6.69 kcal mol-1) with AKR1B1, thus complementing results from other experiments. The study identified cassiarin B, cassiaoccidentalin B, and cinnamtannin A2 as lead hits for the anti-diabetic activity of C. glauca. Further, the PI3K-Akt and AKR1B1 were traced as majorly modulated pathway and target, respectively.


Machine Learning Analysis of Longevity-Associated Gene Expression Landscapes in Mammals.
One of the important questions in aging research is how differences in transcriptomics are associated with the longevity of various species. Unfortunately, at the level of individual genes, the links between expression in different organs and maximum lifespan (MLS) are yet to be fully understood. Analyses are complicated further by the fact that MLS is highly associated with other confounding factors (metabolic rate, gestation period, body mass, etc.) and that linear models may be limiting. Using gene expression from 41 mammalian species, across five organs, we constructed gene-centric regression models associating gene expression with MLS and other species traits. Additionally, we used SHapley Additive exPlanations and Bayesian networks to investigate the non-linear nature of the interrelations between the genes predicted to be determinants of species MLS. Our results revealed that expression patterns correlate with MLS, some across organs, and others in an organ-specific manner. The combination of methods employed revealed gene signatures formed by only a few genes that are highly predictive towards MLS, which could be used to identify novel longevity regulator candidates in mammals.


Joint-venture, capitation model can strengthen market share.
As health maintenance organizations (HMOs) and preferred provider organizations (PPOs) control an increasingly larger percentage of the healthcare market, many hospitals are entering capitation contracts to capture and preserve market share. A joint-venture and capitation model may provide an additional mechanism for large referral centers and community hospitals to jointly maintain or increase market share through improved service delivery. While the model includes many benefits, hospitals should thoroughly consider ramifications. Only through substantial institutional commitment, careful planning, and ongoing management will the model prove successful.


Optimal resistance exercise training parameters for stroke recovery: A protocol for a systematic review.
Stroke impacts nearly 14 million people annually. Muscle strength and physical function are often affected by stroke and important determinants of stroke recovery. Resistance exercise training (RT) has been shown to improve muscle strength, but RT prescriptions may be suboptimal for other aspects of stroke recovery. Parameters such as frequency, intensity, type, and duration may influence the effectiveness of RT interventions but have not been systematically evaluated. 1) To determine the effects of RT on stroke recovery, and 2) to examine the influence of RT parameters on intervention effects. Randomized controlled trials examining the effects of RT will be eligible for this systematic review if they: 1) included only adults with stroke or transient ischemic attack, 2) compared RT to no exercise or usual care, and 3) did not apply a co-intervention. Eight databases (MEDLINE, EMBASE, EMCARE, AMED, PsychINFO, CINAHL, SPORTDiscus, and Web of Science) and 2 clinical trials registries (ClinicalTrials.gov and the WHO International Clinical Trials Registry Platform) will be searched from inception. Two independent pairs of authors will compare titles, abstracts, and full-text reports against the eligibility criteria. Conflicts will be resolved by consensus or third author. The construct of interest is stroke recovery. An advisory group of clinicians, researchers, and partners with lived experience of stroke will be consulted to determine specific outcome measures of interest, and to rank their relative importance. We expect to include measures of physical function, strength, cognition, and quality of life. Random-effects meta-analyses will be used to pool results for each outcome across studies, and RT parameters (frequency, intensity, type, and duration) will be used as covariates in meta-regression analyses. The results of this review will inform the optimal RT prescription parameters for promoting stroke recovery.


The effects of fipronil on emotional and cognitive behaviors in mammals.
Fipronil is a phenylpyrazole insecticide that is widely used as a pesticide and a veterinary drug, although studies suggest that it could be toxic to mammals. The objectives of this study were to examine the pharmacokinetic profile of fipronil in mice, dogs, and cats, and to evaluate its effects on emotional and cognitive behaviors of dogs and cats using the data obtained from mice. The assessment of in vivo kinetics of fipronil was conducted in mice and dogs. We also performed behavioral tests (elevated plus-maze and Y-maze) and measured the levels of neurotransmitters in mice exposed to fipronil. In addition, the in vitro metabolism of fipronil were evaluated using liver microsomes of rats, mice, dogs, and cats. The results revealed that fipronil is distributed throughout the body (blood, brain, adipose tissue, and liver) of mice after dermal application. It was metabolized to fipronil sulfone primarily in the liver. The data on kinetics show that both fipronil and fipronil sulfone have a longer half-life in dogs and cats than in mice. The behavioral tests indicated that fipronil and fipronil sulfone could affect emotional and cognitive behaviors and alter the levels of neurotransmitters (dopamine in the striatum and serotonin in the hippocampus) in mice. Furthermore, we found that dogs and cats have a low ability to metabolize fipronil than mice and rats. However, further comprehensive studies are needed to determine whether fipronil affects the emotional and cognitive behaviors when administered to dogs and cats. To the best of our knowledge, this is the first study to examine the pharmacokinetic data and verify the effects of fipronil on emotional and cognitive behaviors of dogs and cats using the data obtained from mice.


Phospholipid binding and the activation of group IA secreted phospholipase A2.
Equilibrium dialysis was used to study the binding of two nonhydrolyzable, short chain phospholipid analogues to the secreted group IA phospholipase A(2) (PLA(2)), which has been shown to contain several phospholipid binding sites that dramatically affect activity. This study provides new insight into how these activations occur. One analogue contained a phosphorylethanolamine (DiC(6)SNPE) headgroup, while the other contained a phosphorylcholine (DiC(6)SNPC) headgroup. Using phospholipase D, we incorporated tritium into each analogue. No binding of DiC(6)SNPE to PLA(2) was observed under submicellar conditions. Addition of submicellar amounts of Triton X-100 resulted in a linear nonsaturating response to lipid concentration, suggestive of premicellar aggregation of the DiC(6)SNPE with Triton X-100 and PLA(2). Binding of DiC(6)SNPE when presented as Triton X-100 mixed micelles saturated at 0.93 binding sites per PLA(2) with a K(D) of 38 microM. Addition of sphingomyelin, a potent activator of PLA(2) hydrolysis of phosphorylethanolamine containing compounds, resulted in a 13-fold decrease in the K(D), to 2.8 microM. This suggests that changes in the catalytic site binding affinity contribute to "phosphatidylcholine activation". Binding of DiC(6)SNPC with 2.0 mM Triton X-100 showed positive cooperativity (Hill coefficient of 1.7), which saturated at 2.0 binding sites per PLA(2). No binding of either analogue was observed when the catalytic site was alkylated with p-bromophenacyl bromide. Since p-bromophenacyl bromide does not physically block the phosphatidylcholine activator site, this indicates that the two phosphatidylcholine binding sites interact. The binding studies show that DiC(6)SNPC binds cooperatively to two sites on group IA PLA(2), while DiC(6)SNPE binds to only one site.


Combined effects of varicocele and cell phones on semen and hormonal parameters.
The objective of this study was to evaluate if there is a combined effect of varicocele and cell phone storage in trousers pockets on semen and hormonal parameters. A retrospective analysis of 468 men attending an infertility clinic from 1993-2007 was performed. Varicoceles were determined by clinical examination and patients were questioned on cell phone usage and storage fashion. Semen samples were analyzed according to the World Health Organization (WHO) guidelines of 1999. Serum testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were assessed. There was a significant effect of cell phone storage in trousers pockets and varicocele in multivariate analysis (both p < 0.001). Varicocele showed an effect on sperm concentration (p = 0.003), LH (p = 0.014) and testosterone (p = 0.003). Compared to grade 1, grade 2 varicoceles showed a difference in sperm concentration (p = 0.004). Regarding testosterone differences were shown for grade 3 versus grade 1 (p = 0.002) and grade 3 compared to grade 2 (p = 0.003). Cell phone storage in trousers pockets showed an influence on the percentage of normal sperm morphology and LH (both p < 0.001). Varicocele and cell phone storage in trousers pockets did not show a combined effect (p = 0.76). This analysis showed an inverse relation between sperm concentration and degree of varicocele, with lower concentrations in higher grade varicoceles. Testosterone was significantly higher in higher grade varicoceles, which could reflect a compensatory mechanism to the impaired testicular function. Cell phone storage in trousers pockets showed an effect on LH and sperm morphology. A combined effect of varicocele and cell phone storage in trousers pockets was not detected.


Controlling pH-Sensitive Chemical Reactions Pathways with Light - a Tale of Two Photobases: an Arrhenius and a Brønsted.
Light-gated chemical reactions allow spatial and temporal control of chemical processes. Here, we suggest a new system for controlling pH-sensitive processes with light using two photobases of Arrhenius and Brønsted types. Only after light excitation do Arrhenius photobases undergo hydroxide ion dissociation, while Brønsted photobases capture a proton. However, none can be used alone to reversibly control pH due to the limitations arising from excessively fast or overly slow photoreaction timescales. We show here that combining the two types of photobases allows light-triggered and reversible pH control. We show an application of this method in directing the pH-dependent reaction pathways of the organic dye Alizarin Red S simply by switching between different wavelengths of light, i.e., irradiating each photobase separately. The concept of a light-controlled system shown here of a sophisticated interplay between two photobases can be integrated into various smart functional and dynamic systems.


Culture and phenotypic characterization of a Wolbachia pipientis isolate.
The recent isolation of Wolbachia pipientis in the continuous cell line Aa23, established from eggs of a strain of the Asian tiger mosquito Aedes albopictus, allowed us to perform extensive characterization of the isolate. Bacterial growth could be obtained in C6/36, another A. albopictus cell line, at 28 degrees C and in a human embryonic lung fibroblast monolayer at 28 and 37 degrees C, confirming that its host cell range is broader than was initially thought. The bacteria were best visualized by Diff-Quik and May-Grünwald-Giemsa staining. Proteins from 213 to 18 kDa with two major protein bands of 65 and 25 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By Western blotting with specific polyclonal mouse and rabbit antisera, dominant immunoreactive antigens were found at approximately 100, 80, and 30 kDa. The genome size was calculated to be 1,790 +/- 17 kb by pulsed-field gel electrophoresis. The sequence of the citrate synthase gene (gltA) of W. pipientis was determined by gene walking. Its position in the phylogenetic tree constructed with gltA confirmed that found in a phylogenetic tree constructed with 16S rRNA genes and that it belongs in the alpha subgroup of the class Proteobacteria and that it is closely related to but independent from the genera Ehrlichia, Anaplasma, and Neorickettsia.


Autoimmunity: Rationalizing possible pathways from initiation to disease.
Any physiological system that has as its output an activity that is biodestructive and ridding must have a way of distinguishing the host (self) from that which is other (nonself). The setting in which autoimmunity can be analyzed depends, in part and unavoidably, on the way in which the normal self (S)-nonself (NS) discrimination is accomplished. Any discussion of autoimmunity should include one's view of this latter. To this end, a pathway for the normal S-NS discrimination will be proposed. Then, a mechanism for the determination of effector class will be considered as autoimmune disease is consequent to it. Experiments challenging both the proposed model of normal behavior, as well as that of the extrapolation to autoimmunity, will be cited along with a discussion of some of the elements which, if rendered defective, would result in autoimmunity. The goal is to see how far this particular abstraction based largely on the logic of evolutionary biology can meaningfully guide understanding of the disease.


Investigation of the staggered herringbone mixer with a simple analytical model.
An analytical model is presented for the three-dimensional flow in the recently introduced staggered herringbone mixer for microchannels. In this model, the flow in the cross-section of the channel is treated as a lid-driven cavity flow. The model is shown to reproduce the advection patterns that were observed experimentally in the staggered herringbone mixer. The model is then used to study the quality of mixing in this flow as a function of geometry. Analysis is performed with Poincaré maps, mixing simulations, and residence time distributions. A range of optimal geometries is identified.


Toward next generation plasma profiling via heat-induced epitope retrieval and array-based assays.
There is a need for high throughput methods for screening patient samples in the quest for potential biomarkers for diagnostics and patient care. Here, we used a combination of undirected target selection, antibody suspension bead arrays, and heat-induced epitope retrieval to allow for protein profiling of human plasma in a novel and systematic manner. Several antibodies were found to reveal altered protein profiles upon epitope retrieval at elevated temperatures with limits of detection improving into lower ng/ml ranges. In a study based on prostate cancer patients, several proteins with differential profiles were discovered and subsequently validated in an independent cohort. For one of the potential biomarkers, the human carnosine dipeptidase 1 protein (CNDP1), the differences were determined to be related to the glycosylation status of the targeted protein. The study shows a path of pursuit for large scale screening of biobank repositories in a flexible and proteome-wide fashion by utilizing heat-induced epitope retrieval and using an antibody suspension bead array format.


Prostate cancer: prediction of biochemical failure after external-beam radiation therapy--Kattan nomogram and endorectal MR imaging estimation of tumor volume.
To determine whether magnetic resonance (MR) imaging and MR spectroscopic imaging findings can improve predictions made with the Kattan nomogram for radiation therapy. The institutional review board approved this retrospective HIPAA-compliant study. Ninety-nine men who underwent endorectal MR and MR spectroscopy before external-beam radiation therapy for prostate cancer (January 1998 to June 2007) were included. Linear predictors were calculated with input variables from the study sample and the Kattan original coefficients. The linear predictor is a single weighted value that combines information of all predictor variables in a model, where the weight of each value is its association with the outcome. Two radiologists independently reviewed all MR images to determine extent of disease; a third independent reader resolved discrepancies. Biochemical failure was defined as a serum prostate-specific antigen level of 2 ng/mL (2 μg/L) or more above nadir. Cox proportional hazard models were used to determine the probabilities of treatment failure (biochemical failure) in 5 years. One model included only the Kattan nomogram data; the other also incorporated imaging findings. The discrimination performance of all models was determined with receiver operating characteristics (ROC) curve analyses. These analyses were followed by an assessment of net risk reclassification. The areas under the ROC curve for the Kattan nomogram and the model incorporating MR imaging findings were 61.1% (95% confidence interval: 58.1%, 64.0%) and 78.0% (95% confidence interval: 75.7%, 80.4%), respectively. Comparison of performance showed that the model with imaging findings performed significantly better than did the model with clinical variables alone (P < .001). Overall, the addition of imaging findings led to an improvement in risk classification of about 28%, ranging from approximately a minimum of 16% to a maximum of 39%, depending on the risk change considered important. MR imaging data improve the prediction of biochemical failure with the Kattan nomogram after external-beam radiation therapy for prostate cancer. The number needed to image to improve the prediction of biochemical failure in one patient ranged from three to six.


Control of cell division in Streptococcus pneumoniae by the conserved Ser/Thr protein kinase StkP.
How the human pathogen Streptococcus pneumoniae coordinates cell-wall synthesis during growth and division to achieve its characteristic oval shape is poorly understood. The conserved eukaryotic-type Ser/Thr kinase of S. pneumoniae, StkP, previously was reported to phosphorylate the cell-division protein DivIVA. Consistent with a role in cell division, GFP-StkP and its cognate phosphatase, GFP-PhpP, both localize to the division site. StkP localization depends on its penicillin-binding protein and Ser/Thr-associated domains that likely sense uncross-linked peptidoglycan, because StkP and PhpP delocalize in the presence of antibiotics that target the latest stages of cell-wall biosynthesis and in cells that have stopped dividing. Time-lapse microscopy shows that StkP displays an intermediate timing of recruitment to midcell: StkP arrives shortly after FtsA but before DivIVA. Furthermore, StkP remains at midcell longer than FtsA, until division is complete. Cells mutated for stkP are perturbed in cell-wall synthesis and display elongated morphologies with multiple, often unconstricted, FtsA and DivIVA rings. The data show that StkP plays an important role in regulating cell-wall synthesis and controls correct septum progression and closure. Overall, our results indicate that StkP signals information about the cell-wall status to key cell-division proteins and in this way acts as a regulator of cell division.


An improved clinically relevant sepsis model in the conscious rat.
To develop an improved small animal experimental paradigm that more closely mimics human sepsis. Prospective, randomized, controlled animal study. Medical school research laboratory. Male Sprague-Dawley rats (280-320 g). We monitored the hemodynamic, hematologic, and biochemical consequences of abdominal sepsis produced by intraperitoneal implantation of a fibrin clot containing Escherichia coli in conscious, antibiotic-treated, rats. Similar to human sepsis, the implanted, infected clot (LD50 = 5-7 x 10(8) colony forming units/mL, n = 6) elevated cardiac index (>7% vs. sterile clot, p < .05, at 4 hrs), whereas mean arterial pressure and heart rate remained unaffected. The total peripheral resistance index and stroke volume index tended to decrease and increase, respectively. In contrast, an intravenous bolus injection of endotoxin (LD50 of E. coli lipopolysaccharide = 5.6 mg/kg, n = 7), the most commonly used sepsis model, induced profound hypodynamic responses manifested by a 27% decrease (vs. endotoxin vehicle, p < .01) in cardiac index, a 28% increase in the total peripheral resistance index (p < .01), and a 33% decrease in the stroke volume index (P < .01). The infectious peritonitis model also displayed dose-dependent thrombocytopenia (<61%, p < .05), leukopenia (<60%, p < .05), and mortality rate (50% at 5-7 x 10(8) colony forming units/mL, p < .05) with a minimally elevated serum tumor necrosis factor-alpha level (145 vs. 12 +/- 6 pg/mL in controls, p < .05). This rodent model of antibiotic-treated, intra-abdominal infection features key characteristics of clinical sepsis. Although the hyperdynamic response observed in septic patients undergoing resuscitation was not clearly elicited, this paradigm better mimics clinical sepsis compared with the commonly used endotoxin model. Thus, utilization of this paradigm may provide additional opportunities to explore mechanisms of sepsis and to examine novel therapeutics.


Spring phenology drives range shifts in a migratory Arctic ungulate with key implications for the future.
Annual variation in phenology can have profound effects on the behavior of animals. As climate change advances spring phenology in ecosystems around the globe, it is becoming increasingly important to understand how animals respond to variation in the timing of seasonal events and how their responses may shift in the future. We investigated the influence of spring phenology on the behavior of migratory, barren-ground caribou (Rangifer tarandus), a species that has evolved to cope with short Arctic summers. Specifically, we examined the effect of spring snow melt and vegetation growth on the current and potential future space-use patterns of the Porcupine Caribou Herd (PCH), which exhibits large, inter-annual shifts in their calving and post-calving distributions across the U.S.-Canadian border. We quantified PCH selection for snow melt and vegetation phenology using machine learning models, determined how selection resulted in annual shifts in space-use, and then projected future distributions based on climate-driven phenology models. Caribou exhibited strong, scale-dependent selection for both snow melt and vegetation growth. During the calving season, caribou selected areas at finer scales where the snow had melted and vegetation was greening, but within broader landscapes that were still brown or snow covered. During the post-calving season, they selected vegetation with intermediate biomass expected to have high forage quality. Annual variation in spring phenology predicted major shifts in PCH space-use. In years with early spring phenology, PCH predominately used habitat in Alaska, while in years with late phenology, they spent more time in Yukon. Future climate conditions were projected to advance spring phenology, shifting PCH calving and post-calving distributions further west into Alaska. Our results demonstrate that caribou selection for habitat in specific phenological stages drive dramatic shifts in annual space-use patterns, and will likely affect future distributions, underscoring the importance of maintaining sufficient suitable habitat to allow for behavioral plasticity.


Three-dimensional imaging of trabecular bone using the computer numerically controlled milling technique.
Although various techniques exist for high-resolution, three-dimensional imaging of trabecular bone, a common limitation is that resolution depends on specimen size. Most techniques also have limited availability due to their expense and complexity. We therefore developed a simple, accurate technique that has a resolution that is independent of specimen size. Thin layers are serially removed from an embedded bone specimen using a computer numerically controlled (CNC) milling machine, and each exposed cross section is imaged using a low-magnification digital camera. Precise positioning of the specimen under the camera is achieved using the programmable feature of the CNC milling machine. Large specimens are imaged without loss of resolution by moving the specimen under the camera such that an array of field-of-views spans the full cross section. The images from each field-of-view are easily assembled and registered in the postprocessing. High-contrast sections are achieved by staining the bone black with silver nitrate and embedding it in whitened methylmethacrylate. Due to the high contrast nature and high resolution of the images, thresholding at a single value yielded excellent predictions of morphological parameters such as bone volume fraction (mean +/- SD percent error = 0.70 +/- 4.28%). The main limitations of this fully automated "CNC milling technique" are that the specimen is destroyed and the process is relatively slow. However, because of its accuracy, independence of image resolution from specimen size, and ease of implementation, this new technique is an excellent method for ex situ imaging of trabecular architecture, particularly when high resolution is required.


Prevention of antibiotic resistance - an epidemiological scoping review to identify research categories and knowledge gaps.
Antibiotics have become the cornerstone for the treatment of infectious diseases and contributed significantly to the dramatic global health development during the last 70 years. Millions of people now survive what were previously life-threatening infections. But antibiotics are finite resources and misuse has led to antibiotic resistance and reduced efficacy within just a few years of introduction of each new antibiotic. The World Health Organization rates antibiotic resistance as a 'global security threat' impacting on global health, food security and development and as important as terrorism and climate change. This paper explores, through a scoping review of the literature published during the past 20 years, the magnitude of peer-reviewed and grey literature that addresses antibiotic resistance and specifically the extent to which "prevention" has been at the core. The ultimate aim is to identify know-do gaps and strategies to prevent ABR. The review covers four main data bases, Web of Science, Medline, Scopus and Ebsco searched for 2000-17. The broader research field "antibiotic OR antimicrobial resistance" gave 431,335 hits. Narrowing the search criteria to "Prevention of antibiotic OR antimicrobial resistance" resulted in 1062 remaining titles. Of these, 622 were unique titles. After screening of the 622 titles for relevance, 420 abstracts were read, and of these 282 papers were read in full. An additional 53 references were identified from these papers, and 64 published during 2018 and 2019 were also included. The final scoping review database thus consisted of 399 papers. A thematic structure emerged when categorizing articles in different subject areas, serving as a proxy for interest expressed from the research community. The research area has been an evolving one with about half of the 399 papers published during the past four years of the study period. Epidemiological modelling needs strengthening and there is a need for more and better surveillance systems, especially in lower- and middle-income countries. There is a wealth of information on the local and national uses and misuses of antibiotics. Educational and stewardship programmes basically lack evidence. Several studies address knowledge of the public and prescribers. The lessons for policy are conveyed in many alarming reports from national and international organizations. Descriptive rather than theoretical ambitions have characterized the literature. If we want to better understand and explain the antibiotic situation from a behavioural perspective, the required approaches are lacking. A framework for an epidemiological causal web behind ABR is suggested and may serve to identify entry points for potential interventions.


A new technic for segmental spinal osteosynthesis using the posterior approach.
The authors have designed a new method of instrumentation aimed at obtaining surgical fixation of the scoliotic curve without any postoperative external immobilisation. It is particularly strong and rigid and allows adequate reduction of the curve. This technique avoids the sublaminar space and thus prevents excessive blood loss and diminishes the danger of cord damage. The instrumentation is made of two parallel rough cylindrical rods inserted independently in the convexity and concavity of the curve. If necessary, they can be bent pre-operatively. They are attached to hooks placed on the laminae or pedicles, which are locked by bolts, thus allowing progressive straightening of the curve. They are joined by two transverse bars, one above and one below, to provide better rigidity to the device and to allow correction of rotation. The parts of the vertebrae left free by the device are denuded to allow the addition of grafts. Laboratory tests have demonstrated that this type of fixator is more rigid than the Harrington or Luque rods. Fifteen patients, either idiopathic or paralytic cases, were operated on without any neurological impairment. No loss of correction was observed since the hooks have been locked.


Single-particle tracking reveals that free ribosomal subunits are not excluded from the Escherichia coli nucleoid.
Biochemical and genetic data show that ribosomes closely follow RNA polymerases that are transcribing protein-coding genes in bacteria. At the same time, electron and fluorescence microscopy have revealed that ribosomes are excluded from the Escherichia coli nucleoid, which seems to be inconsistent with fast translation initiation on nascent mRNA transcripts. The apparent paradox can be reconciled if translation of nascent mRNAs can start throughout the nucleoid before they relocate to the periphery. However, this mechanism requires that free ribosomal subunits are not excluded from the nucleoid. Here, we use single-particle tracking in living E. coli cells to determine the fractions of free ribosomal subunits, classify individual subunits as free or mRNA-bound, and quantify the degree of exclusion of bound and free subunits separately. We show that free subunits are not excluded from the nucleoid. This finding strongly suggests that translation of nascent mRNAs can start throughout the nucleoid, which reconciles the spatial separation of DNA and ribosomes with cotranscriptional translation. We also show that, after translation inhibition, free subunit precursors are partially excluded from the compacted nucleoid. This finding indicates that it is active translation that normally allows ribosomal subunits to assemble on nascent mRNAs throughout the nucleoid and that the effects of translation inhibitors are enhanced by the limited access of ribosomal subunits to nascent mRNAs in the compacted nucleoid.


Observed parental responsiveness/warmth and children's coping: cross-sectional and prospective relations in a family depression preventive intervention.
The current study examined concurrent and prospective relations between observed parenting behaviors and children's coping strategies in the context of a preventive intervention designed to change both parenting and children's use of secondary control coping. Questionnaires and direct observations were obtained from parents with a history of depression (N = 180) and their children (ages 9-15 years) at baseline, 6-month (after completion of the intervention), and 18-month follow-up assessments. Cross-sectional analyses indicate that baseline observed parental responsiveness/warmth was significantly associated with composite parent/child reports of children's baseline primary control, secondary control, and disengagement coping. Using a mixed effects model, prospective mediational analyses indicate that intervention-driven improvements in observed parental responsiveness/warmth from baseline to 6 months significantly accounted for increases in children's use of secondary control coping strategies from baseline to the 18-month follow-up assessment. No significant mediation effects emerged for primary control coping or disengagement coping. The present findings suggest that it may be possible to improve children's coping strategies not only through targeted interventions, but also indirectly by improving responsive and warm parenting behaviors. Limitations and strengths are noted and implications for future research are outlined.


Glutamate immunoreactivity in the tiger salamander retina differentiates between GABA-immunoreactive and glycine-immunoreactive amacrine cells.
A higher proportion of retinal amacrine cells, largely thought to be inhibitory, in tiger salamander retina are immunoreactive (+) for the excitatory amino acid glutamate than for either of the inhibitory amino acids, GABA and glycine. The colocalizations of glutamate immunoreactivity, GABA immunoreactivity and glycine immunoreactivity were studied in the proximal retina of tiger salamander using postembedding immunocytochemical methods in consecutive semithin sections. A considerable degree of colocalization was observed between glutamate immunoreactivity and GABA immunoreactivity and to a lesser extent with glycine immunoreactivity. No glutamate-immunoreactive cells were single labelled in INL3 (proximal inner nuclear), 12% were single labelled in INL2 (mid INL; these are bipolar cells), 62% were singled labelled in the ganglion cell layer (GCL) and likely to be ganglion cells. Single-labelled glycine-immunoreactive cells comprised 16% in INL3, 2%; INL2 and GCL, 0%). Glutamate immunoreactivity co-localized four times as often with GABA immunoreactivity than glycine immunoreactivity in INL3 (77% versus 19%), comparably in INL2 (37% and 32%) and six times as often in the GCL (29% versus 5%). A computerized image analysis of the relative intensities of glutamate immunoreactivity in GABA-immunoreactive/glutamate-immunoreactive and glycine-immunoreactive/glutamate-immunoreactive double labelled amacrine cells in INL3 revealed that all glycine-immunoreactive cells contained low intensities of glutamate immunoreactivity; whereas 35% of GABA-positive cells had medium to high intensities of glutamate immunoreactivity. There was a significant correlation (r = 0.44; p < 0.01) between the intensities of glutamate immunoreactivity and GABA immunoreactivity in GABA-immunoreactive/glutamate-immunoreactive amacrine cells. The common colocalization of glutamate immunoreactivity with GABA immunoreactivity and the positive correlation of their relative intensities suggests that glutamate is likely to serve as a precursor for GABA synthesis. However, intense glutamate immunoreactivity in 5% of the amacrine cells suggests a transmitter function. Glycine-immunoreactive amacrine cells contained little or no glutamate immunoreactivity, suggesting a minimal role for glutamate as a neurotransmitter in these cells. The major distribution of intense glutamate immunoreactivity is in bipolar cells and ganglion cells, consistent with data suggesting that glutamate is the transmitter in the retinal through pathway.


Solvent effects in polyelectrolyte adsorption: computer simulations with explicit and implicit solvent.
The adsorption of strongly charged polyelectrolyte chains to an oppositely charged planar surface is studied using computer simulation. In addition to an explicit solvent model, two implicit solvent models are considered: one where the solvent induces an implicit Lennard-Jones (ILJ) interaction between polymer sites and one where the solvent induces a many body interaction that depends on the solvent accessible surface area (SASA) of the monomers. Molecular and Brownian dynamics simulations are reported for the explicit and implicit solvent models, respectively. All three models give similar results for the adsorption of the chains in good solvent. The electrostatic attraction between the surface and the polymers is not sufficient to drive the strong adsorption that is seen in experiments. In poor solvents, the models give different results for the adsorption excess and the mechanism for polyelectrolyte adsorption. With explicit solvent, thick adsorbed layers are formed at both charged and neutral surfaces. With the SASA model, adsorbed layers are formed on the charged but not on the neutral surface. With the ILJ model, adsorbed layers are not formed on any surfaces. The results show that the solvent plays a dominant role in the adsorption of polyelectrolytes under poor solvent conditions and that many-body solvent effects have a qualitative effect on the adsorption characteristics and mechanism. In particular, SASA and depletion effects could possibly play an important role; the former can be incorporated in the SASA model, but the latter cannot. The results suggest that accurate computational models for polymer adsorption under poor solvent conditions must incorporate the solvent explicitly.


PSI-BLAST-ISS: an intermediate sequence search tool for estimation of the position-specific alignment reliability.
Protein sequence alignments have become indispensable for virtually any evolutionary, structural or functional study involving proteins. Modern sequence search and comparison methods combined with rapidly increasing sequence data often can reliably match even distantly related proteins that share little sequence similarity. However, even highly significant matches generally may have incorrectly aligned regions. Therefore when exact residue correspondence is used to transfer biological information from one aligned sequence to another, it is critical to know which alignment regions are reliable and which may contain alignment errors. PSI-BLAST-ISS is a standalone Unix-based tool designed to delineate reliable regions of sequence alignments as well as to suggest potential variants in unreliable regions. The region-specific reliability is assessed by producing multiple sequence alignments in different sequence contexts followed by the analysis of the consistency of alignment variants. The PSI-BLAST-ISS output enables the user to simultaneously analyze alignment reliability between query and multiple homologous sequences. In addition, PSI-BLAST-ISS can be used to detect distantly related homologous proteins. The software is freely available at: http://www.ibt.lt/bioinformatics/iss. PSI-BLAST-ISS is an effective reliability assessment tool that can be useful in applications such as comparative modelling or analysis of individual sequence regions. It favorably compares with the existing similar software both in the performance and functional features.


Early first-line treatment response and subsequent disability worsening in relapsing-remitting multiple sclerosis.
Treatment success in relapsing-remitting multiple sclerosis (RRMS) is generally determined using relapse frequency and magnetic resonance imaging (MRI) activity in the first 6 or 12 months on treatment. The association of these definitions of short-term treatment success with disability worsening and disease activity in the longer term is unclear. In this study, we investigated risk factors associated with early first-line treatment failure in RRMS, and the association of early treatment failure with subsequent disability worsening or "no evidence of disease activity" (NEDA-3) status. We used data from CombiRx (clinicaltrials.gov identifier NCT00211887) to investigate risk factors associated with early treatment failure, and the association of early treatment failure at 6 and 12 months with subsequent disability worsening or NEDA-3 at 36 months. CombiRx included 1008 treatment-naïve participants with RRMS, who were randomly assigned to treatment with glatiramer acetate, interferon beta, or the combination of both. Early treatment failure at 6 or 12 months by several definitions was associated with NEDA-3 failure at 36 months, but not with subsequent disability worsening at 36 months. Expanded Disability Status Scale (EDSS) was the only baseline characteristic associated with the risk of disability worsening at 36 months. Approximately 70% of NEDA-3 failures occurred due to MRI activity, and <10% occurred due to EDSS worsening. Our investigation shows that current definitions of early treatment failure in RRMS are unrelated to patient-relevant disability worsening at 36 months of follow-up. Further research into useful definitions of treatment success and failure in RRMS is needed.


American Association of Clinical Endocrinology Clinical Practice Guideline: Developing a Diabetes Mellitus Comprehensive Care Plan-2022 Update.
The objective of this clinical practice guideline is to provide updated and new evidence-based recommendations for the comprehensive care of persons with diabetes mellitus to clinicians, diabetes-care teams, other health care professionals and stakeholders, and individuals with diabetes and their caregivers. The American Association of Clinical Endocrinology selected a task force of medical experts and staff who updated and assessed clinical questions and recommendations from the prior 2015 version of this guideline and conducted literature searches for relevant scientific papers published from January 1, 2015, through May 15, 2022. Selected studies from results of literature searches composed the evidence base to update 2015 recommendations as well as to develop new recommendations based on review of clinical evidence, current practice, expertise, and consensus, according to established American Association of Clinical Endocrinology protocol for guideline development. This guideline includes 170 updated and new evidence-based clinical practice recommendations for the comprehensive care of persons with diabetes. Recommendations are divided into four sections: (1) screening, diagnosis, glycemic targets, and glycemic monitoring; (2) comorbidities and complications, including obesity and management with lifestyle, nutrition, and bariatric surgery, hypertension, dyslipidemia, retinopathy, neuropathy, diabetic kidney disease, and cardiovascular disease; (3) management of prediabetes, type 2 diabetes with antihyperglycemic pharmacotherapy and glycemic targets, type 1 diabetes with insulin therapy, hypoglycemia, hospitalized persons, and women with diabetes in pregnancy; (4) education and new topics regarding diabetes and infertility, nutritional supplements, secondary diabetes, social determinants of health, and virtual care, as well as updated recommendations on cancer risk, nonpharmacologic components of pediatric care plans, depression, education and team approach, occupational risk, role of sleep medicine, and vaccinations in persons with diabetes. This updated clinical practice guideline provides evidence-based recommendations to assist with person-centered, team-based clinical decision-making to improve the care of persons with diabetes mellitus.


Therapeutic transdifferentiation of human fibroblasts into endothelial cells using forced expression of lineage-specific transcription factors.
Transdifferentiation is the direct conversion from one somatic cell type into another desired somatic cell type. This reprogramming method offers an attractive approach for regenerative medicine. Here, we demonstrate that neonatal fibroblasts can be transdifferentiated into endothelial cells using only four endothelial transcription factors, namely, ETV2, FLI1, GATA2, and KLF4. We observed a significant up-regulation of endothelial genes including KDR, CD31, CD144, and vWF in human neonatal foreskin (BJ) fibroblasts infected with the lentiviral construct encoding the open reading frame of the four transcription factors. We observed morphological changes in BJ fibroblasts from the fibroblastic spindle shape into a more endothelial-like cobblestone structures. Fluorescence-activated cell sorting analysis revealed that ~16% of the infected cells with the lentiviral constructs encoding 4F expressed CD31. The sorted cells were allowed to expand for 2 weeks and these cells were immunostained and found to express endothelial markers CD31. The induced endothelial cells also incorporated fluorescence-labeled acetylated low-density lipoprotein and efficiently formed capillary-like networks when seeded on Matrigel. These results suggested that the induced endothelial cells were functional in vitro. Taken together, we successfully demonstrated the direct conversion of human neonatal fibroblasts into endothelial cells by transduction of lentiviral constructs encoding endothelial lineage-specific transcription factors ETV2, FLI1, GATA2, and KLF4. The directed differentiation of fibroblasts into endothelial cells may have significant utility in diseases characterized by fibrosis and loss of microvasculature.