Pulmonary functions in atopic and nonatopic asthmatic children.
we examined how lung function and certain clinical and laboratory characteristics in asthmatic children were changed according to skin test positivity to aeroallergens. a skin prick test was conducted using standardized extracts of 10 different allergens in 56 children with bronchial asthma, aged 5-15 years, in Dicle University Hospital. Lung function was measured by Microplus spirometer. among the 56 subjects, asthma was classified as mild in 16, moderate in 42 and severe in 3. At least one skin prick test was positive (monosensitized) in 35 subjects (62 %) and positive reactivity to two or more aeroallergens (polysensitized) was found in 17 subjects (30 %). Positive skin test reactions to aeroallergens were associated with a decrease (as percentage of the predicted decrease) in FEV1, FVC and PEF values. Significant differences were also found between prick test-positive and -negative asthmatics in duration of breastfeeding (8.5 5 months vs 15 7 months, respectively, p < 0.007), age at which cow's milk had been started (5.7 1.6 vs 10.5 5.4, p = 0.004); total serum IgE concentration (350 221 IU/ml vs 234 164 IU/ml, p = 0.02), age at onset of asthma symptoms (2.5 1.9 years vs 4.1 2.2 years) and number of asthma attacks per year (7.0 3.1 vs 5.2 3.5, p = 0.012). When one-way ANOVA and a post-Hoc test were used, asthma attacks were more frequent and severe and allergic conjunctivitis symptoms were more frequent in the polysensitized group than in the nonsensitized and monosensitized groups (p = 0.03). children with positive skin prick test results, especially those with combined sensitivity to dust mite, cat and dog, were at increase risk of more severe asthma.


Characterization of microglia/macrophages in gliomas developed in S-100β-v-erbB transgenic rats.
Glioma-infiltrating microglia/macrophages are referred to as tumor-associated macrophages (TAMs). Transgenic (TG) rats expressing v-erbB, which is a viral form of the epidermal growth factor receptor, under transcriptional regulation by the S100-β promoter, develop brain tumors. This study was designed to clarify the pathological characteristics of TAMs in these experimental tumors. We carried out immunohistochemical and morphometrical analyses of microglia/macrophages in brain tumors (5 malignant glioma, 4 anaplastic oligodendroglioma, 4 astrocytoma) that developed in TG rats. TAMs with ionized calcium-binding adaptor molecule 1 (Iba1) positivity and morphology of activated, non-phagocytic microglia increased within and around the tumors in malignant gliomas and anaplastic astrocytomas. The Iba1-positive TAMs of the tumor core were significantly more activated than Iba1-positive microglia of non-neoplastic brain tissue in intraparenchymal anaplastic oligodendrogliomas. Iba1 expression showed a significant positive correlation to Ki-67 expression in all the gliomas. Most TAMs showed no or little expression against CD68, CD163 or CD204, although CD204-positive TAMs were observed in necrosis as well as in the proliferating vascular wall. In conclusion, S-100β-v-erbB TG rats may serve as a useful animal model for further analysis of TAMs in terms of tumor cell proliferation, microvascular proliferation and phagocytosis, and as a tool for therapeutic use in malignant gliomas, although it should be noted that the polarization of TAMs toward the M2 phenotype remains unclear.


A new fluorimetric enzyme assay for the diagnosis of Niemann-Pick A/B, with specificity of natural sphingomyelinase substrate.
6-Hexadecanoylamino-4-methylumbelliferylphosphorylcholine (HMUPC) was shown to be a specific substrate for the determination of acid (lysosomal) sphingomyelinase (ASM; gene SMPD1). Fibroblasts (n = 27) and leukocytes (n = 8) from both the A and B types of Niemann-Pick disease showed < 6% and < 10% of mean normal ASM activity, respectively. Niemann-Pick A or B patients bearing the Q292K mutation had apparently normal ASM activity with our new artificial substrate. These patients with false-normal sphingomyelinase activity, however, could readily be detected by determining the extent of inhibition of enzymatic hydrolysis of the artificial substrate HMU-PC by an unlabelled natural substrate, in particular lysosphingomyelin. This approach is generally applicable. Our novel assay for ASM combines the ease of a rapid and robust enzyme assay using a fluorogenic substrate with the specificity of an ASM assay using a natural substrate. Such assays are obviously more convenient to the diagnostic laboratory, since radiolabelled substrates are not required.


A modified MethyLight assay predicts the clinical outcomes of anti-epidermal growth factor receptor treatment in metastatic colorectal cancer.
DNA methylation status correlates with clinical outcomes of anti-epidermal growth factor receptor (EGFR) treatment. There is a strong need to develop a simple assay for measuring DNA methylation status for the clinical application of drug selection based on it. In this study, we collected data from 186 patients with metastatic colorectal cancer (mCRC) who had previously received anti-EGFR treatment. We modified MethyLite to develop a novel assay to classify patients as having highly methylated colorectal cancer (HMCC) or low-methylated colorectal cancer (LMCC) based on the methylation status of 16 CpG sites of tumor-derived genomic DNA in the development cohort (n = 30). Clinical outcomes were then compared between the HMCC and LMCC groups in the validation cohort (n = 156). The results showed that HMCC had a significantly worse response rate (4.2% vs 33.3%; P = .004), progression-free survival (median: 2.5 vs 6.6 mo, P < .001, hazard ratio [HR] = 0.22), and overall survival (median: 5.6 vs 15.5 mo, P < .001, HR = 0.23) than did LMCC in patients with RAS wild-type mCRC who were refractory or intolerable to oxaliplatin- and irinotecan-based chemotherapy (n = 101). The DNA methylation status was an independent predictive factor and a more accurate biomarker than was the primary site of anti-EGFR treatment. In conclusion, our novel DNA methylation measurement assay based on MethyLight was simple and useful, suggesting its implementation as a complementary diagnostic tool in a clinical setting.


The binding of DNA to water soluble carbodiimide modified proteins.
Bovine serum albumin and human serum transferrin modified by the water soluble carbodiimides N-ethyl-N'-(3-dimethyl propylamino) carbodiimide (CDI) or N-ethyl-N'-(3-trimethyl propylammonium) carbodiimide (Me+CDI) to yield N-acylurea proteins bind pBR322 DNA reversibly showing electrostatic and non-electrostatic components in the binding energies (delta G overall). It is proposed that initially an electrostatic interaction arises from ion pair formation between the DNA phosphates and the N-acylurea entities. This is consolidated, in single stranded regions, by a second event in which it is suggested that the base guanine interacts with elements of the N-acylurea moieties through hydrogen bonding or a glyoxal-type addition.


[Genetics of ischemic stroke].
Stroke is one of the most widespread causes of mortality und disability worldwide. Around 80 % of strokes are ischemic and different forms of intracranial bleeding account for the remaining cases. Monogenic stroke disorders are rare but the diagnosis may lead to specific therapeutic consequences for the affected patients who are predominantly young. In common sporadic stroke, genetic factors play a role in the form of susceptibility genes. Their discovery may give rise to new therapeutic options in the future.


Aging of the circadian system in zebrafish and the effects of melatonin on sleep and cognitive performance.
Aging is a complex process involving intracellular changes and, notably, modifications in intercellular communications, required for coordinated responses to internal and external events. One of the potential reasons for such changes is an age-dependent failure of the integrating systems, including the circadian clock. Here we demonstrate that aging in a diurnal vertebrate, zebrafish (Danio rerio), is associated with major but selective circadian alterations. By 3-5 years of age, zebrafish have reduced amplitude and increased fragmentation of entrained circadian rhythms of activity, with fast desynchronization of the rhythms in the absence of environmental time cues. Aging in zebrafish is also associated with a reduction in the overall duration of nighttime sleep, followed by lower activity levels and a higher arousal threshold during the day. The production of the principal circadian hormone, melatonin, progressively declines during zebrafish aging. However, the ability of melatonin to acutely promote sleep and entrain circadian rhythms of activity remains robust until at least 4-5 years of age, consistent with the preserved levels of mRNA expression for melatonin receptors. Aged zebrafish have altered expression of the circadian genes zBmal1 and zPer1 but not zClock1. A lack of circadian time cues alters cognitive performance in aged more than in young zebrafish and this can be partially attenuated by daily melatonin administration. The advantages of zebrafish as a diurnal, small, prolific and genetically well-characterized vertebrate model provide new opportunities to clarify the intrinsic circadian factors involved in human aging and promote the search for prophylactic and treatment strategies.


A novel near-infrared fluorescent probe for H2O2 in alkaline environment and the application for H2O2 imaging in vitro and in vivo.
H2O2 as one of the most important ROS (Reactive Oxygen Species) has more attack activity to biomolecules such as DNA, RNA, protein and enzyme in alkaline environment and leads to a series of disease. However, no attention has been paid to the fluorescent detection of H2O2 in alkaline environment in the past. Herein, we reported the first ratiometric near-infrared fluorescent probe based on a boric acid derivative of Changsha near-infrared dye (CSBOH) for H2O2 detection in alkaline condition and the application for H2O2 imaging in vivo. ICT (intra-molecular charge transfer) mechanism was used in CSBOH to modulate the fluorescence change. The photophysical change of CSBOH was investigated by comparison with a phenol derivative of Changsha near-infrared dye (CSOH), a structural analogue bearing phenol group. In the presence of H2O2, CSBOH exhibited remarkably different fluorescence change at 650 nm and 720 nm when excited by 560 nm and 670 nm light respectively in alkaline buffer and showed high selectivity toward H2O2. Cellular experiments demonstrate that CSBOH can image endogenously generated H2O2 in macrophages and A431 cells. In vivo experiment demonstrates that both CSOH and CSBOH can be used for bio-imaging, and CSBOH can image H2O2 in living animal successfully.


Effect of aging and testing method on bond strength of CAD/CAM fiber-reinforced composite to dentin.
To evaluate and compare the outcomes of shear (S) and microtensile (μT) bond strength tests of CAD/CAM fiber-reinforced composite (FRC) to dentin. Aging with either fatigue or thermocycling were conducted for comparison with baseline microtensile group. CAD/CAM FRC (Trinia, Bicon LLC, Boston, USA) blocks were milled to 3-mm diameter cylinders for shear and to blocks (5×5×5mm) for μT. Sixty extracted human molars were flattened to obtain dentin surfaces and randomly divided in four groups (n=15): (1) SC: samples tested in shear 24h after bonding; (2) μTC: samples tested in μT 24h after bonding); (3) μTF: samples submitted to mechanical fatigue prior to μT test, and; (4) μTT: thermocycling prior to μT test. Bonding system was applied onto the FRC material (Cera-Resin Bond, CRB, Shofu Dental, Kyoto, Japan). A conventional three-step adhesive system (All-bond 3, Bisco, Schaumburg, USA) was use with a self-cure resin cement (C&B resin cement, Bisco, Schaumburg, USA). Bond strength tests were conducted at 0.75mm/min and data analyzed using Weibull distribution (p<0.05). Weibull contour plots showed a significantly lower characteristic strength (η) and Weibull modulus (m) for SC (η=6.9MPa and m=1.4) compared to μTC (η=20.9MPa and m=4.5). Fatigued and thermocycled μT groups presented significantly reduced characteristic strength (η=3.1MPa and η=4.1MPa, respectively) compared to μTC. Weibull modulus was significantly reduced only for SC and μTF groups compared μTC. Failure predominantly occurred at the cement/FRC interface. FRC bonded to dentin samples presented lower Weibull modulus and characteristic bond strength when immediately tested in shear compared to microtensile. Aging through thermocycling or mechanical fatigue significantly reduced the characteristic strength in microtensile testing, with the majority of failures emerging between restoration material and cement interface.


Flow-injection immuno-bioassay for interleukin-6 in humans based on gold nanoparticles modified screen-printed graphite electrodes.
A flow-injection electrochemical immunoassay system based on a disposable immunosensor for the determination of interleukin-6 (IL-6) was proposed. The immunosensor was prepared by entrapping horseradish peroxidase (HRP)-labeled IL-6 antibody into gold nanoparticles-modified composite membrane at a screen-printed graphite electrode. With a non-competitive immunoassay format, the immunosensor was inserted in the flow system with an injection of sample, and the injected sample containing IL-6 antigen was produced transparent immunoaffinity reaction with the immobilized HRP-labeled IL-6 antibody. The formed antigen-antibody complex inhibited partly the active center of HRP, and decreased the immobilized HRP to H2O2 reduction. The performance and factors influencing the performance of the immunosensor were investigated. Under optimal conditions, the current change obtained from the labeled HRP relative to thionine-H2O2 system was proportional to the IL-6 concentration in the range of 5-100 ng L(-1) with a detection limit of 1.0 ng L(-1) (at 3delta). The flow-injection immunoassay system could automatically control the incubation, washing and measurement steps with acceptable reproducibility and good stability. Moreover, the proposed immunosensors were used to analyze IL-6 in human serum specimens. Analytical results of clinical samples show the developed immunoassay has a promising alternative approach for detecting IL-6 in the clinical diagnosis.


Inorganic arsenic exposure affects pain behavior and inflammatory response in rat.
Inorganic arsenic (iAs) contamination of drinking water is a worldwide problem associated with an increased risk for the development of various types of cancer and noncancerous damage. In vitro studies have suggested that iAs can modulate the activity of macrophages producing an over-expression of cyclooxygenase-2 (COX-2) and resulting in an increase in prostaglandin E(2) (PGE(2)) concentrations in endothelial cells. These effects may lead to an in vivo enhancement of inflammatory and pain responses. Our aim was to determine the effect of a single dose of arsenic or subchronic exposure to arsenic on pain behavior and tissue inflammation in rats. Rats were given a single dose of sodium arsenite (0.1, 1 and 10 mg/kg i.p.) or submitted to subchronic exposure to arsenic added to the drinking water for 4 weeks (0.1, 1, 10 and 100 ppm). Inflammatory pain was assessed by using the formalin and tail-flick tests, while inflammation was evaluated with the carrageenan model. Arsenite did not induce pain or significant inflammation by itself. In contrast, arsenite in both single dose administration and subchronic exposure increased not only the inflammatory process and the underlying hyperalgesic pain, but also induced a decrease in the pain threshold. Alterations in pain processing were dependent on the arsenic dose and the length of exposure, and the underlying mechanism involved an increased release of local PGE(2). These results suggest that inorganic arsenic exposure enhances pain perception and exacerbates the pathological state of inflammatory diseases.


Medical Licensure Questions and Physician Reluctance to Seek Care for Mental Health Conditions.
To determine whether state medical licensure application questions (MLAQs) about mental health are related to physicians' reluctance to seek help for a mental health condition because of concerns about repercussions to their medical licensure. In 2016, we collected initial and renewal medical licensure application forms from 50 states and the District of Columbia. We coded MLAQs related to physicians' mental health as "consistent" if they inquired only about current impairment from a mental health condition or did not ask about mental health conditions. We obtained data on care-seeking attitudes for a mental health problem from a nationally representative convenience sample of 5829 physicians who completed a survey between August 28, 2014, and October 6, 2014. Analyses explored relationships between state of employment, MLAQs, and physicians' reluctance to seek formal medical care for treatment of a mental health condition because of concerns about repercussions to their medical licensure. We obtained initial licensure applications from 51 of 51 (100%) and renewal applications from 48 of 51 (94.1%) medical licensing boards. Only one-third of states currently have MLAQs about mental health on their initial and renewal application forms that are considered consistent. Nearly 40% of physicians (2325 of 5829) reported that they would be reluctant to seek formal medical care for treatment of a mental health condition because of concerns about repercussions to their medical licensure. Physicians working in a state in which neither the initial nor the renewal application was consistent were more likely to be reluctant to seek help (odds ratio, 1.21; 95% CI, 1.07-1.37; P=.002 vs both applications consistent). Our findings support that MLAQs regarding mental health conditions present a barrier to physicians seeking help.


Comparison of the genetic structures surrounding qnrA1 in Korean Enterobacter cloacae and Chinese Escherichia coli strains isolated in the early 2000s: evidence for qnrA mobilization via Inc HI2 type plasmid.
The flanking genetic structure of qnrA1 in Korean Enterobacter loacae was identical to that of the Chinese Escherichia coli strain, the first qnrA1-carrying strain reported in Asia. Analysis of restriction enzyme sites and Southern blot hybridization results showed that qnrA1 was transferred between E. cloacae and E. coli via Inc HI2 type plasmid.


Acellular Dermal Matrix Favorably Modulates the Healing Response after Surgery.
When first described for breast reconstruction, the presence of acellular dermal matrices was associated with increased seroma formation and infection. However, clinical safety data have gradually improved with surgeon experience to an acceptable outcomes profile of acellular dermal matrix-assisted reconstruction when compared to submuscular implant coverage. In fact, acellular dermal matrix use potentially decreases capsular contracture rates and facilitates expansion for staged prepectoral breast reconstruction. Because of new regulatory requirements, the collection of unbiased, well-powered premarket approval data summarizing long-term clinical outcomes will be essential over the coming years to understand the clinical performance of acellular dermal matrix use in breast reconstruction. Currently, the authors can highlight the physiologic benefits of acellular dermal matrix use in breast reconstruction by evaluating the components of surgical wound healing that are favorably augmented by the implanted collagen substrate. Acellular dermal matrix takes advantage of the wound healing cascade to incorporate into the patient's tissues-a process that requires a coordinated inflammatory infiltrate and angiogenesis. The presence of acellular dermal matrix augments and modulates the wound healing process to its advantage by simultaneously increasing the invasion of appropriate cellular constituents to facilitate expeditious healing and accelerate angiogenesis. In this article, the authors summarize the wound healing literature to demonstrate the mechanisms acellular dermal matrices use to biointegrate and the literature in which cellular constituents and soluble growth factors are up-regulated in the presence of acellular dermal matrix. Lastly, the authors use their experimental observations of acellular dermal matrix incorporation to corroborate the literature.


Total mesorectal excision with intraoperative assessment of internal anal sphincter innervation provides new insights into neurogenic incontinence.
The aim of this prospective study was to assess internal anal sphincter (IAS) innervation in patients undergoing total mesorectal excision (TME) by intraoperative neuromonitoring (IONM). Fourteen patients underwent TME. IONM was carried out through pelvic splanchnic nerve stimulation under continuous electromyography of the IAS. Anorectal function was assessed with the digital rectal examination scoring system and a standardized questionnaire. Nine of 11 patients who underwent low anterior resection had positive IONM results, with stimulation-induced increased IAS electromyographic amplitudes (median 0.23 μV (interquartile range [IQR] 0.05, 0.56) vs median 0.89 μV (IQR 0.64, 1.88), p < 0.001) after TME. The patients with the positive IONM results were continent after stoma closure. Of 2 patients with negative IONM results, 1 had fecal incontinence after closure of the defunctioning stoma and received a permanent sigmoidostomy. In the other patient the defunctioning stoma was deemed permanent due to decreased anal sphincter function. In 3 patients who underwent abdominoperineal excision, IONM assessed denervation of the IAS after performance of the abdominal part. This study demonstrated that IONM of IAS innervation in rectal cancer patients is feasible and may predict neurogenic fecal incontinence.


Changes in choroidal thickness after vitamin D supplementation in patients with vitamin D deficiency.
To determine the relationship between vitamin D (vit D) deficiency and choroidal thickness and to investigate changes in choroidal thickness after vit D treatment. Sixty-five patients diagnosed with vit D deficiency (group 1) and 60 healthy individuals with normal vit D levels (group 2) were included in the study. The choroidal thickness in the subfoveal, nasal, and temporal regions of the fovea were measured manually using enhanced depth imaging optical coherence tomography. The choroidal thickness in the patients in group 1 was re-evaluated after treatment with vit D. The groups were similar in terms of age and sex (p = 0.138 and p = 0.198, respectively). The subfoveal, temporal (500 and 1.500 µm), and nasal (500 and 1.500 µm) choroidal thickness values in group 1 were statistically significantly lower than those in group 2 (p < 0.001 for all). After vit D replacement therapy, there was a significant increase in the choroidal thicknesses compared with the pretreatment measurements (p < 0.001 for all). There was a positive correlation between vit D values and subfoveal, temporal (500 and 1.500 µm), and nasal (500 and 1.500 µm) choroidal thickness values (r = 0.558 and p < 0.001, r = 0.415 and p < 0.001, r = 0.352 and p < 0.001, r = 0.349 and p < 0.001, and r = 0.294 and p = 0.001, respectively). The choroidal thickness decreased in patients with vit D deficiency. The choroidal thickness values increased after vit D replacement therapy. Serum vit D levels should be assessed in studies evaluating choroidal thickness.


Proteomic identification of predictive tissue biomarkers of sensitive to neoadjuvant chemotherapy in squamous cervical cancer.
The regimens of neoadjuvant chemotherapy (NAC) in squamous cervical cancer (SCC) frequently use cisplatin combined with paclitaxel. Unfortunately, some cervical cancers show resistance to the principal chemotherapeutic agents in the treatment, decreasing the effectiveness of this therapy. The objective of this study was to search for predictive markers of response to NAC in patients with SCC. Two-dimensional gel electrophoresis (2-DE) accompanied by matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS) was used to analyze and identify differentially expressed proteins in ten cases of advanced cervical cancer patients receiving cisplatin-based NAC. Each of these patients received more than two cycles of NAC. Cell proliferation rate in cisplatin resistant human cervical cancer cell Hela/DDP and its parent cell Hela after treatment with Hsp70 inhibitor and/or cisplatin were tested by MTT assay. Twelve protein spots changed in abundance, quantitative comparison of spot volumes showed that seven protein spots were up-regulated and five spots were down-regulated in the NAC non-responders compared to the NAC responders. These proteins are involved in various cellular processes essential for cell metabolism, migration and apoptotic signal transduction. The high-fold changes proteins of stathmin1, Hsp70 and pyruvate kinase isoform M2 were validated by Western blot analysis. Over-expression of Hsp70 inhibits the efficacy of cisplatin. Hsp70 inhibitor enhanced the sensitivity to cisplatin in both Hela and Hela/DDP cells. The study found many candidate proteins involved in chemotherapy resistance/sensitivity, among them Hsp70 might be potential biomarkers to predict the efficacy of chemotherapy for SCC patients.


Evaluation of mebrofenin hepatoscintigraphy in neonatal-onset jaundice.
The prognosis of infants with prolonged neonatal jaundice is dependent on early diagnosis because of the need for prompt surgical management of biliary atresia. To evaluate the usefulness of 99 mTcm-trimethylbromo-iminodiacetic acid (TBIDA, mebrofenin) in the investigation of infantile jaundice. A retrospective study was undertaken of 58 patients with unexplained prolonged neonatal jaundice. Sixty-eight scans were reviewed. Mebrofenin scintigraphy confirmed the presence of a choledochal cyst in three of the four cases with that diagnosis. There were no false negative results in the nine patients with extrahepatic biliary atresia (EHBA). Three further infants had an incorrect histological diagnosis of EHBA. A gall bladder was identified by US in each case and in one of these, scintigraphy showed gut excretion. In the 16 patients with no gut excretion by 24 h, the final diagnoses were intrahepatic cholestasis (n = 7), Alagille's syndrome (n = 3), neonatal hepatitis (n = 3), alpha-1-antitrypsin deficiency (n = 2) and juvenile xanthogranuloma (n = 1). Seven infants had repeat scintigraphy after the administration of ursodeoxycholic acid (URSO). This changed five non-excretors with hepatitis into excretors. Two infants with hepatitis continued to show non-excretion after URSO, but a gallbladder was identified by US in both. Mebrofenin scintigraphy is accurate in confirming the presence of a choledochal cyst and in refuting the diagnosis of EHBA. While histology and scintigraphy are each 100 % sensitive for the diagnosis of EHBA, neither, individually, is accurate and the investigation of prolonged neonatal jaundice requires a multi-modality imaging strategy.


Synthesis of difuran derivatives and their biological activity.
Visnaginone and khellinone react with chloroacetamide and chloroacetonitrile in dimethyl formamide to give either the cyclised benzo[1,2-b : 5,4-b']difuran or the open form depending on the heating period. The antibacterial activity of the compounds was tested.


Unfinished agenda of the neonates in developing countries: magnitude of neonatal sepsis: systematic review and meta-analysis.
Neonatal sepsis is the major cause of mortality and morbidity globally, particularly in developing countries. Despite studies revealed the extent of neonatal sepsis in developing countries, the findings were inconclusive. Therefore, the main aim of this study was to determine the pooled prevalence of neonatal sepsis in developing countries. We used a systematic review and Meta-analysis study method. The reviewed studies were accessed through an electronic web-based search strategy from the electronic database (PUBMED), advanced google scholar, different journal sites. The data extraction was done by two researchers using a data extraction table and the disparity between data extractors was resolved by the third researcher. The analysis was done using STATA version 11. The I2 test was used to assess heterogeneity across studies. The Funnel plot, Begg's test, and Egger's test were used to check for publication bias. The random-effect model was used to determine the pooled effect size. All studies related to neonatal sepsis which fulfill the inclusion criteria were considered into this study. The quality of each study was checked using the Newcastle-Ottawa Scale and studies graded low score were excluded from the study. At the end, 36 articles fit with our study objectives. Studies conducted in Ethiopia were significant the source of heterogeneity of the study with a coefficient = 90, P-value = 0.025. The overall pooled prevalence of the study was 29.92%. The limitations of this study would be the authors were only used articles reported in the English language, and publication bias. The pooled prevalence of neonatal sepsis was found to be high which accounted for a third of the neonates. Despite countries have established possible prevention and treatment mechanisms, neonatal sepsis is the major public health problem in lower and middle-income countries till now.


Structure based design, synthesis and biological evaluation of amino phosphonate derivatives as human glucokinase activators.
Glucokinase (GK) is a potential therapeutic target of type 2 diabetes and GK activators (GKAs) represent a promising class of small organic molecules which enhance GK activity. Based on the configuration and conformation of the allosteric site of GK, we have designed a novel class of amino phosphonate derivatives in order to develop potent GKAs. The QSAR model developed using numerous descriptors revealed its potential with the best effective statistical values of RMSE=1.52 and r2=0.30. Moreover, application of this model on the present test set GKAs proved to be worthy to predict their activities as a better linear relationship was observed with RMSE=0.14 and r2=0.88. ADME studies and Lipinski filters encouraged them as safer therapeutics. The molecular dynamics and docking studies against the GK allosteric site revealed that all GKAs bind with best affinities and the complexes are strengthened by H-bonding, phosphonate salt bridges, hydrophobic and arene cat ionic interactions. Finally, in vitro evaluation with human liver GK revealed their potential to increase the GK activity by different folds. The results from QSAR, ADME, molecular docking and in vitro assays strongly suggested that the present molecules could be used as effective and safer therapeutics to control and manage type 2 diabetes.


In the ovariectomized rat, tamoxifen conserves bone similarly in parathyroid-intact and parathyroidectomized animals.
To examine the ability of tamoxifen (TAM) to conserve bone in the estrogen-deficient ovariectomized (OVX) rat in the presence and absence of parathyroid hormone (PTH) six groups of rats with 45Ca-labeled bones were studied for 12 weeks. Rats were OVX, parathyroidectomized (PTX), or given sham operations and treated with TAM (10 mg/kg body wt./wk subcutaneously) or TAM-vehicle. Treatments were: group 1 = Sham-OVX; group 2 = Sham-OVX + TAM; group 3 = OVX; group 4 = OVX + TAM; group 5 = OVX + PTX; and group 6 = OVX + PTX + TAM. To monitor bone resorption serial measurements of urinary hydroxyproline and 45Ca excretion were made during the study. Ovariectomy raised these markers of bone breakdown and caused significant osteopenia, whereas TAM prevented ovariectomy increasing urinary hydroxyproline or 45Ca and conserved bone. Final total body calcium values (TBCa) in groups 1-6, respectively, were (mg +/- SD): 3240 +/- 300; 3260 +/- 289; 2750 +/- 231; 3212 +/- 312; 2742 +/- 199; and 3387 +/- 252. Thus ovariectomy reduced TBCa similarly in the presence and absence of the parathyroids (p < 0.001). In contrast TAM fully protected both PT-intact and PTX rats from the osteopenic effect of ovariectomy, despite the fact that PTX rats had a lower rate of bone turnover than PT-intact rats. However, TAM-treated OVX rats had shorter femora than OVX rats given TAM-vehicle, suggesting that TAM suppresses growth of the long bones to some degree in estrogen-deficient animals.(ABSTRACT TRUNCATED AT 250 WORDS)


Esophageal hematoma after atrial fibrillation ablation: incidence, clinical features, and sequelae of esophageal injury of a different sort.
Esophageal hematoma recently has been reported as a form of esophageal injury after atrial fibrillation (AF) ablation, attributed to the use of transesophageal echocardiography (TEE). We sought to determine the incidence, clinical features, and sequelae of this form of esophageal injury. This was a prospective series of 1110 AF ablation procedures performed under general anesthesia (GA) over 9 years. TEE was inserted after induction of GA to exclude left atrial appendage thrombus, define cardiac function, and guide transseptal puncture. The procedural incidence of esophageal hematoma was 0.27% (3/1110 procedures, mortality 0%). Odonyphagia, regurgitation, and hoarseness were the predominant symptoms, with an onset within 12 hours. There was absence of fever and neurological symptoms. Chest computed tomography excluded atrio-esophageal fistula and was diagnostic of esophageal hematoma localized to either the upper esophagus or extending the length of the mid and lower esophagus; endoscopy confirmed the diagnosis. Management was conservative in all cases comprising of ceasing oral intake and anticoagulation. Long term sequelae included esophageal stricture formation requiring dilatation, persistent esophageal dysmotility (mid esophageal hematoma), and vocal cord paralysis, resulting in hoarse voice (upper esophageal hematoma). Esophageal hematoma is a rare but important differential diagnosis for esophageal injury after TEE-guided AF ablation under GA, and can result in significant patient morbidity. Key clinical features differentiate presentation of esophageal hematoma from that of an atrio-esophageal fistula.


MR perfusion imaging by alternate slab width inversion recovery arterial spin labeling (AIRASL): a technique with higher signal-to-noise ratio at 3.0 T.
To propose a new arterial spin labeling (ASL) perfusion-imaging method (alternate slab width inversion recovery ASL: AIRASL) that takes advantage of the qualities of 3.0 T. AIRASL utilizes alternate slab width IR pulses for labeling blood to obtain a higher signal-to-noise ratio (SNR). Numerical simulations were used to evaluate perfusion signals. In vivo studies were performed to show the feasibility of AIRASL on five healthy subjects. We performed a statistical analysis of the differences in perfusion SNR measurements between flow-sensitive alternating inversion recovery (FAIR) and AIRASL. In signal simulation, the signal obtained by AIRASL at 3.0 and 1.5 T was 1.14 and 0.85%, respectively, whereas the signal obtained by FAIR at 3.0 and 1.5 T was 0.57 and 0.47%, respectively. In an in vivo study, the SNR of FAIR (3.0 T) and FAIR (1.5 T) were 1.73 ± 0.49 and 1.02 ± 0.20, respectively, whereas the SNRs of AIRASL (3.0 T) and AIRASL (1.5 T) were 3.93 ± 1.65 and 1.34 ± 0.31, respectively. SNR in AIRASL at 3.0 T was significantly greater than that in FAIR at 3.0 T. The most significant potential advantage of AIRASL is its high SNR, which takes advantage of the qualities of 3.0 T. This sequence can be easily applied in the clinical setting and will enable ASL to become more relevant for clinical application.


Powering up microbiome-microglia interactions.
Despite extensive evidence implicating the microbiota in regulating the immune system, the precise mechanisms underlying microbial control of microglial maturation remain unclear. In a methodological tour de force, Erny et al. (2021) identify acetate as an essential microbiota-derived molecule driving microglial metabolic pathways and functions during healthy and diseased states.


Clinical implications of cardiovascular outcome trials in type 2 diabetes.
Cardiovascular disease (CVD) is the main reason for premature death in patients with type 2 diabetes. Hyperglycemia, the hallmark of diabetes, has long been considered the link between diabetes and CVD, and many trials focused on preventing CVD manifestations by means of tight glucose control. However, diabetes is a multifactorial disease in which, e. g., insulin resistance, endothelial dysfunction, and factors such as hypertension and dyslipidemia contribute. Thus, treatment needs to be multifactorial and take cardiovascular aspects into account. Newer classes of drugs, originally launched for glucose lowering, among them dipeptidyl-peptidase (DPP)-4 inhibitors, sodium-glucose cotransporter (SGLT)-2 inhibitors, and glucagon-like peptide (GLP)-1 receptor agonists, have been studied in large cardiovascular outcome trials (CVOT). Several SGLT-2 inhibitors and GLP-1 receptor agonists are associated with a reduction of cardiovascular events (cardiovascular death, nonfatal myocardial infarction, and nonfatal stroke). Although the mechanisms behind the effects are not fully understood, an important reason for the benefits of SGLT-2 inhibitors seems be a reduction in heart failure, while GLP-1 receptor agonists may retard the development of the atherosclerotic vascular disease or may be effective by stabilizing plaques. The outcomes of these studies have been taken into account in recently issued guidelines and an important task for diabetologists, cardiologists, and general practitioners is to incorporate the findings of these trials into clinical practice.


Risk factors for repeat abortion and implications for addressing unintended pregnancy in Vietnam.
To determine predictors of repeat abortion in 3 provinces in Vietnam. In a cross-sectional study between August and December 2011, women who underwent abortion were interviewed after the procedure in 62 public health facilities in Hanoi, Khanh Hoa, and Ho Chi Minh City (HCMC). Information on sociodemographic factors, contraceptive and reproductive history and intentions, and opinions and experience of abortion services was collected. The primary outcome was repeat (≥2) abortions. Overall, 1224 women were interviewed: 534 from Hanoi, 163 from Khanh Hoa, and 527 from HCMC. The mean age and parity of the respondents were 29 years and 1.8, respectively, and 79.6% were married. Approximately half of the respondents were not using contraception before pregnancy. The prevalence of repeat abortion was 31.7%. In multivariate models, significant predictors of repeat abortion included living in Hanoi, higher parity, age 35 years or older, and having 2 or more daughters (versus 1) or no sons (versus 1) after controlling for parity (all P < 0.05). Repeat abortion remains high in Vietnam, fueled partly by inadequate contraceptive use. Son preference seems to be an important predictor of repeat abortion. Strengthening post-abortion contraceptive counseling and promoting long-acting contraceptive methods are essential to reduce repeat abortion.


Disrupted in schizophrenia 1 (DISC1): subcellular targeting and induction of ring mitochondria.
Several independent studies have identified Disrupted In Schizophrenia 1 (DISC1) as a potential susceptibility factor in the pathogenesis of schizophrenia and severe recurrent major depression. To identify potential mechanisms by which DISC1 may influence development of psychiatric illness, we investigated the cellular consequences of recombinant DISC1 expression in COS-7 cells. We show that the N-terminal head domain is sufficient for DISC1 mitochondrial and nuclear targeting, while sequence from the C-terminus facilitates centrosomal association. Loss of C-terminal sequence alters DISC1 subcellular distribution, significantly increasing nuclear localization. DISC1 over-expression produces striking mitochondrial reorganization in some cells, with formation of mitochondrial ring-like structures, indicating a potential involvement of DISC1 in mitochondrial fusion and/or fission.


Radiometric analysis of oxidative reactions in aromatization by placental microsomes. Presence of differential isotope effects.
In order to study the initial as well as the final steps in the aromatization of androgens to estrogens, high-specific activity [19-C3H3]androstenedione and testosterone were synthesized. Incubations of [19-C3H3]androstenedione with human placental microsomes resulted in the generation of [3H]water, as a result of the dual hydroxylation at C-19, and [3H]formic acid reflecting final aromatization. After an initial lag in the production of [3H]formic acid, the two radiolabeled products were formed linearly with time at a ratio of 2 to 1 under subsaturating conditions and 2.2 to 1 when saturating levels of substrate were present. Incubation of a mixture of [19-C3H3]- and [4-14C]androstenedione with human placental microsomes yielded 19-hydroxy- and 19-oxoandrostenedione, respectively, products of one and two hydroxylations at C-19. The isotope ratios of these derivatives revealed the presence of a tritium isotope effect in the first but not in the second hydroxylation at that site. When [19-C3H2]- and [4-14C]19-hydroxyandrostenedione were used as the substrate, the isotope ratio of the isolated 19-oxoandrostenedione showed no evidence of any isotope effect in its formation. Thus, the second hydroxylation at C-19 exhibits no isotope effect irrespective of whether androstenedione or 19-hydroxyandrostenedione are the substrates, and therefore, a concerted process and catalytic commitment are not responsible for the difference in isotope effects between the first and second C-19 hydroxylation by the placental aromatase complex. Radiometric kinetic analysis employing [19-C3H3]- and [1 beta,2 beta-3H]androstenedione as the comparative substrates provided evidence that the isotope effect is exerted solely through the Vmax component of the reaction. The distinction between the successive hydroxylations at C-19 in the aromatization sequence suggests, but does not prove, that different mechanisms, and hence different catalytic sites, may be involved in these steps.


Accumulation of guanosine tetraphosphate induced by polymixin and gramicidin in Escherichia coli.
The effects of two polypeptide antibiotics, polymixin B and gramicidin S, on the intracellular pool size and turnover of guanosine tetraphosphate (ppGpp) were analyzed in stringent (relA+) and relaxed (relA) strains of Escherichia coli. When either one of these two drugs was added to stringent bacteria cultures at a final concentration that blocked protein and RNA synthesis, ppGpp was found to accumulate. Under similar conditions of inhibition of macromolecular synthesis, ppGpp also appeared to accumulate in relaxed bacteria. Moreover, in either type of strain, no significant accumulation of guanosine pentaphosphate (pppGpp) could be detected upon drug treatment. It was, therefore, concluded that polymixin and gramicidin elicit ppGpp accumulation through a mechanism independent of the relA gene product and, consequently, quite distinct from the stringent control system triggered by amino acid starvation. Further experiments performed by using tetracycline as an inhibitor of ppGpp synthesis, showed that the increase in the level of this nucleotide induced by drug action was due, in fact, to a strong restriction of its degradation rate.


Serial interferon-γ release assay in children with latent tuberculosis infection and children with tuberculosis.
Interferon-γ (IFN-γ) release assay (IGRA) is used for diagnosis of latent tuberculosis infection (LTBI), and for serial testing of active tuberculosis (TB). The aim of this study was to evaluate the results of IGRA for diagnosis and treatment monitoring of children with LTBI and children with TB. IGRA was performed in BCG vaccinated children before and six months after the beginning of treatment. A total of 59 BCG vaccinated children aged 4-18 years were investigated due to exposure to active TB. The participants were divided into two groups: Group 1, children with LTBI (N = 41), and Group 2, children with TB (N = 18). IGRA (QuantiFERON-TB Gold In-Tube) was performed twice, i.e., before treatment and at the end of prophylaxis and therapy. There was no significant difference in IFN-γ concentrations between Group 1 and Group 2 subjects either before or after the treatment. Difference between pre-treatment and post-treatment IFN-γ concentrations compared in either Group 1 or Group 2 was not statistically significant. During follow-up, children with LTBI did not develop active TB. In addition, in children with TB, signs and symptoms of TB improved with anti-TB therapy. This study showed that the concentrations of IFN-γ did not differ in children with LTBI and TB either before or at the end of treatment. IGRA may remain positive over a long period of time. It seems that IGRA is not useful for monitoring treatment of children with LTBI and children with TB.


Quality of Life in Mucormycosis Patients Post Discharge: A Pilot Study Using a New MQOL-36 Questionairre.
To evaluate the Quality of Life in Mucormycosis patients in our hospital using a new MQOL36 questionnaire post discharge. 37 cases between Apr 2021 to July 2021 were included in the study with a minimum follow up period of 4 weeks. They were administered Mucormycosis quality-of-life questionnaire (MQOL-36) either in person or via telephonic interview and answers were recorded into digital questionnaire modulated by us using a digital data recording app-KoBoCollect. Their demographic, clinical, imaging, histopathological and treatment data was retrieved and analysed. Most of the patients reported their health to be good with only 1 patient who had extensive cranial involvement reporting as poor and still undergoing repeated surgeries. 46.67% had no nasal complaints, 20% had nasal obstruction, 13.33% had nasal discharge with 2 patients complaining of crusting and 1 of whistling sound at quiet respiration. Most worrying factor was financial condition with 33.33% patients financially distraught and 43.33% being affected but barely managing at present. Only 2 patients reported no economic impact. Mucormycosis in COVID-19 has changed the face of otorhinolaryngology as we know it-while treating the disease is important, treating the post operative aftermath also becomes equally important. The MQOL-36 produces a quality-of-life profile scoring an individual's perception of quality of life in the following domains: Physical, Psychological, Level of Independence, Social Relationships and Environment. Monitoring the quality of life in patients post discharge could help us manage the enormous morbidity associated with the disease.


The relatedness of phonological processes of a hearing-impaired child.
This is a study, in the generative framework, of the phonology of a 6-year-old, hard-of-hearing child. The study has two main results: (1) the phonological substitutions and delections of this hearing imparied child are basically the same in kind as those found in the speech of younger normals and (2) the phonological processes of the child's system fit into groups of processes, each group operationalizing some phonetic preference of the child. Implications for a general theory of phonologica acquistion are discussed.


Elements in normal and cirrhotic human liver tissue. II. Potassium, sulphur, chlorine and bromine measured by X-ray fluorescence spectrometry.
Elements (K, S, Cl, Br) were measured by X-ray fluorescence spectrometry in normal liver tissue obtained at autopsy from 74 subjects (44 male, 30 female), median age 62 years (range 20-87), and in cirrhotic liver tissue from 27 subjects (14 alcoholic, 13 non-alcoholic cirrhosis). The element content (median and 5-95 percentile interval) in normal livers in mmol/kg dry tissue was: K, 198.75 (150.95-256.42); S, 147.60 (73.86-233.97); Cl, 98.73 (69.51-163.55); Br, 0.1101 (0.0701-0.4205). None of the elements displayed any sex difference. There were correlations between the content of Br and Cl (r = 0.26, P less than 0.05) and Br and K (r = 0.24, p less than 0.05). In both groups of cirrhotic livers, K, S and Cl content was within the normal range. Median Br content was above normal both in alcoholic (0.2265 mmol/kg, P less than 0.01) and non-alcoholic cirrhotic livers (0.2228 mmol/kg, P less than 0.01).


Direct mapping of local redox current density on a monolith electrode by laser scanning.
An optical method of mapping local redox reaction over a monolith electrode using simple laser scanning is described. As the optical signal is linearly proportional to the maximum redox current that is measured concomitantly by voltammetry, the optical signal quantitatively maps the local redox current density distribution. The method is demonstrated on two types of reactions: (1) a reversible reaction where the redox moieties are ionic, and (2) an irreversible reaction on two different types of enzymes immobilized on the electrode where the reaction moieties are nonionic. To demonstrate the scanning capability, the local redox behavior on a "V-shaped" electrode is studied where the local length scale and, hence, the local current density, is nonuniform. The ability to measure the current density distribution by this method will pave the way for multianalyte analysis on a monolith electrode using a standard three-electrode configuration. The method is called Scanning Electrometer for Electrical Double-layer (SEED).


Molecular microbial and chemical investigation of the bioremediation of two-phase olive mill waste using laboratory-scale bioreactors.
Two-phase olive mill waste (TPOMW) is a semisolid effluent that is rich in contaminating polyphenols and is produced in large amounts by the industry of olive oil production. Laboratory-scale bioreactors were used to investigate the biodegradation of TPOMW by its indigenous microbiota. The effect of nutrient addition (inorganic N and P) and aeration of the bioreactors was studied. Microbial changes were investigated by PCR-temperature time gradient electrophoresis (TTGE) and following the dynamics of polar lipid fatty acids (PLFA). The greatest decrease in the polyphenolic and organic matter contents of bioreactors was concomitant with an increase in the PLFA fungal/bacterial ratio. Amplicon sequences of nuclear ribosomal internal transcribed spacer region (ITS) and 16S rDNA allowed identification of fungal and bacterial types, respectively, by comparative DNA sequence analyses. Predominant fungi identified included members of the genera Penicillium, Candida, Geotrichum, Pichia, Cladosporium, and Aschochyta. A total of 14 bacterial genera were detected, with a dominance of organisms that have previously been associated with plant material. Overall, this work highlights that indigenous microbiota within the bioreactors through stimulation of the fungal fraction, is able to degrade the polyphenolic content without the inoculation of specific microorganisms.


Radioactive Iodine-Refractory Differentiated Thyroid Cancer in the Elderly.
Most common thyroid cancers are differentiated thyroid cancers (DTCs) and have papillary, follicular, or Hürthle cell morphology. Papillary thyroid carcinoma (PTC) is the most common malignant tumor of the thyroid gland. The incidence of DTC increases with age. While most of the patients with DTC have an excellent prognosis, the outcome can be poor when diagnosed in elderly patients. Current treatment approach for DTC includes surgery, thyroid-stimulating hormone (TSH) suppression, radioactive iodine, external beam radiotherapy, or systemic treatments such as kinase inhibitors. Radioactive iodine therapy (RAI) is the primary first-line systemic treatment for advanced DTC. However, during the course of treatment, the tumor may become refractory to RAI. Elderly patients are more likely to be diagnosed with advanced disease that can be refractory to RAI. The advent of TKIs (tyrosine kinase inhibitors) and their usage in RAI refractory disease has shown improved progression-free survival. These agents are, however, associated with increased toxicity. The variable nature of disease and toxicity associated with the systemic therapy makes it important to have an individualized approach to management, especially in the elderly population who can be more susceptible to toxicities.


MRFy: Remote Homology Detection for Beta-Structural Proteins Using Markov Random Fields and Stochastic Search.
We introduce MRFy, a tool for protein remote homology detection that captures beta-strand dependencies in the Markov random field. Over a set of 11 SCOP beta-structural superfamilies, MRFy shows a 14 percent improvement in mean Area Under the Curve for the motif recognition problem as compared to HMMER, 25 percent improvement as compared to RAPTOR, 14 percent improvement as compared to HHPred, and a 18 percent improvement as compared to CNFPred and RaptorX. MRFy was implemented in the Haskell functional programming language, and parallelizes well on multi-core systems. MRFy is available, as source code as well as an executable, from http://mrfy.cs.tufts.edu/.


The impact of height and body mass index on the risk of testicular cancer in 600,000 Norwegian men.
The present study aimed at exploring the relations between body mass index (BMI) and stature and testicular cancer in a huge Norwegian cohort with measured height and weight. Height and weight were measured in 600,000 Norwegian men aged 14-44 years during 1963-2001. Results from parts of the study cohort have been reported previously. During follow-up, 1,357 testicular cancers were registered. Relative risks (RRs) of testicular cancer were estimated using Cox proportional hazards regression. The risk of testicular cancer decreased with adult BMI. Compared with men with normal BMI, overweight and obese men had a relative risk of cancer of 0.89 (95% CI: 0.77-1.03) and 0.83 (95% CI: 0.58-1.17). The relative risk of testicular cancer per unit increase in BMI was 0.97 (95% CI: 0.95-1.00). The risk of testicular cancer was not associated with adolescent BMI. A moderate increase in risk of seminomas was seen with increasing adult height. Compared with men with height 170-79 cm, men with height 180 cm and above had a relative risk of 1.17 (95% CI: 1.00-1.37).


Effects of naled and dichlorvos on growth and production of Luteoskyrin byPenicillium islandicum.
Two organophosphorus insecticides, 1, 2-dibromo -2, 2-dichloroethyl dimethyl phosphate (naled) and dimethyl 2, 2 -dichlorovinyl phosphate (dichlorvos) were used for investigation of their effects on growth and production of luteoskyrin mycotoxin byPenicillium islandicum in various cultural media at 25°C for 30 days or 60 days. When the concentration of naled and dichlorvos in Czapek solution broth reached 5mg/50mL, growth and production of luteoskyrin by the fungus was completely inhibited. In unpolished rice medium, 15mg/50g of naled was required to retard fungal growth, while the concentration to inhibit the biosynthesis of luteoskyrin was 10mg/50g. On the other hand, if the medium contained dichlorvos at the level of 30mg/50g, the ability to produce luteoskyrin byP islandicum was significantly reduced. In the unhulled rice case, both naled and dichlorvos at the concentration of 15mg/50g were necessary to retard the fungal growth, and 1 mg/50g of each compound exhibits its ability to inhibit the toxin production. Furthermore, it was also found when the cultural medium contained only small amounts of naled and dichlorvos [0.5mg/50g (mL)] the capability to synthesize luteoskyrin by the fungus was drastically reduced. These data strongly suggest that both naled and dichlorvos have similar ability to inhibit luteoskyrin biosynthesis byP islandicum and are also able to retard the fungal growth.


Molecular characterization of Cry1F resistance in fall armyworm, Spodoptera frugiperda from Brazil.
Fall armyworm, Spodoptera frugiperda (J.E. Smith) is a major lepidopteran pest of maize in Brazil and its control particularly relies on the use of genetically engineered crops expressing Bacillus thuringiensis (Bt) toxins such as Cry1F. However, control failures compromising the efficacy of this technology have been reported in many regions in Brazil, but the mechanism of Cry1F resistance in Brazilian fall armyworm populations remained elusive. Here we investigated the molecular mechanism of Cry1F resistance in two field-collected strains of S. frugiperda from Brazil exhibiting high levels of Cry1F resistance. We first rigorously evaluated several candidate reference genes for normalization of gene expression data across strains, larval instars and gut tissues, and identified ribosomal proteins L10, L17 and RPS3A to be most suitable. We then investigated the expression pattern of ten potential Bt toxin receptors/enzymes in both neonates and 2nd instar gut tissue of Cry1F resistant fall armyworm strains compared to a susceptible strain. Next we sequenced the ATP-dependent Binding Cassette subfamily C2 gene (ABCC2) and identified three mutated sites present in ABCC2 of both Cry1F resistant strains: two of them, a GY deletion (positions 788-789) and a P799 K/R amino acid substitution, located in a conserved region of ABCC2 extracellular loop 4 (EC4) and another amino acid substitution, G1088D, but in a less conserved region. We further characterized the role of the novel mutations present in EC4 by functionally expressing both wild type and mutated ABCC2 transporters in insect cell lines, and confirmed a critical role of both sites for Cry1F binding by cell viability assays. Finally, we assessed the frequency of the mutant alleles by pooled population sequencing and pyrosequencing in 40 fall armyworm populations collected from maize fields in different regions in Brazil. We found that the GY deletion being present at high frequency. However we also observed many rare alleles which disrupt residues between sites 783-799, and their diversity and abundance in field collected populations lends further support to the importance of the EC4 domain for Cry1F toxicity.


Biological properties and clinical application of propolis. XI. Histophathological analysis after intravenous application of ethanol extract of poprolis (EEP).
Mice were given i.v. injections of water solutions of ethanol extract of propolis (EEP) over a varying period of time. Some of the animals were sacrificed immediately following the last injection and some within 2--4 weeks after the last injections. Internal organ samples were taken and underwent a microscopic examination. Insignificant pathological changes in the liver were observed. These changes were transient and regressed within 2--4 weeks after the application of EEP.


Reduced cytokine production by glycogen-elicited peritoneal cells from diabetic rats.
IL-1beta, TNF-alpha, cytokine-induced neutrophil chemoattractant-2alpha/beta, and IL-10 measurements were performed in elicited peritoneal cells from control, diabetic, and insulin-treated diabetic rats. Production/liberation of these cytokines was decreased in elicited peritoneal cells from diabetic rats. These changes were abolished by insulin treatment of diabetic rats. The alterations observed might be involved in the impaired inflammatory response and high occurrence of apoptosis observed in neutrophils under diabetic states.


The relationship between the density of Fasciola hepatica miracidia and the net rate of miracidial infections in Lymnaea truncatula.
Experimental studies on the dynamics of infection of Lymnaea truncatula by the miracidia of Fasciola hepatica show that there is a linear relationship between the net rate of attachment of miracidia to snail epithelium and the density of infective stages. The probability of a snail remaining uninfected after exposure to a single miracidium varies with the size class of the snail. Under the conditions of the experiment (temperature, 15 degrees C; exposure period, 30 min; water volume, 8 ml), the probability was 0.46-0.48, 0.27-0.26 and 0.13-0.19 for snail size classes, 0.5-2.9, 3.0-5.9 and 6.0-8.9 mm respectively.


Recovery Curves for Pediatric Burn Survivors: Advances in Patient-Oriented Outcomes.
Patient-reported outcomes serving as benchmarks for recovery of pediatric burn survivors are lacking, and new approaches using longitudinal cohorts for monitoring their expected recovery based on statistical models are needed for patient management during the early years following the burn. To describe multidimensional patient-reported outcomes among pediatric burn survivors younger than 5 years to establish benchmarks using recovery curve methods. Prospective cohort study of pediatric burn survivors younger than 5 years at 12 burn centers. Age-matched nonburned reference groups were studied to define expected results in normal growth and development. The Burn Outcomes Questionnaire for children aged 0 to 5 years (BOQ0-5) was administered to parents of children who had burns and were younger than 5 years. Mixed models were used to generate 48-month recovery curves for each of the 10 BOQ0-5 domains. The study was conducted between January 1999 and December 2008. The 10 BOQ0-5 domains including play, language, fine motor skills, gross motor skills, emotional behavior, family functioning, pain/itching, appearance, satisfaction with care, and worry/concern up to 48 months after burn injury. A total of 336 pediatric burn survivors younger than 5 years (mean [SD] age, 2.0 [1.2] years; 58.4% male; 60.2% white, 18.6% black, and 12.0% Hispanic) and 285 age-matched nonburned controls (mean [SD] age, 2.4 [1.3] years; 51.1% male; 67.1% white, 8.9% black, and 15.0% Hispanic) completed the study. Predicted scores improved exponentially over time for 5 of the BOQ0-5 domains (predicted scores at 1 month vs 24 months: play, 48.6 vs 52.1 [P = .03]; language, 49.2 vs 54.4 [P < .001]; gross motor skills, 48.7 vs 53.0 [P = .002]; pain/itching, 15.8 vs 33.5 [P < .001]; and worry/concern, 31.6 vs 44.9 [P < .001]). Pediatric burn survivors had higher scores in language, emotional behavior, and family functioning domains compared with healthy children in later months. This study demonstrates significant deficits in multiple functional domains across pediatric burn survivors compared with controls. Recovery curves can be used to recognize deviation from the expected course and tailor care to patient needs.


How do you measure workforce integrity?
Behavioral hallmarks such as honesty, high ethical standards, and maintained moral principles define workforce integrity. Here, learn methods for integrating them into management practices.


Grade inflation in the internal medicine clerkship: a national survey.
Grade inflation is a growing concern, but the degree to which it continues to exist in 3rd-year internal medicine (IM) clerkships is unknown. The authors sought to determine the degree to which grade inflation is perceived to exist in IM clerkships in North American medical schools. A national survey of all Clerkship Directors in Internal Medicine members was administered in 2009. The authors assessed key aspects of grading. Response rate was 64%. Fifty-five percent of respondents agreed that grade inflation exists in the Internal Medicine clerkship at their school. Seventy-eight percent reported it as a serious/somewhat serious problem, and 38% noted students have passed the IM clerkship at their school who should have failed. A majority of clerkship directors report that grade inflation still exists. In addition, many note students who passed despite the clerkship director believing they should have failed. Interventions should be developed to address both of these problems.


On-farm evaluation of models to predict herbage intake of dairy cows grazing temperate semi-natural grasslands.
The objective of the present on-farm study was to evaluate the adequacy of existing models in predicting the pasture herbage DM intake (PDMI) of lactating dairy cows grazing semi-natural grasslands. The prediction adequacy of 13 empirical and semi-mechanistic models, which were predominantly developed to represent stall-fed cows or cows grazing high-quality pastures, were evaluated using the mean bias, relative prediction error (RPE), and partitioning of mean square error of prediction, where models with an RPE ≤ 20% were considered adequate. The reference dataset comprised n = 233 individual animal observations from nine commercial farms in South Germany with a mean milk production, DM intake, and PDMI (arithmetic means ± one SD) of 24 kg/d, (±5.6), 21 kg/d (±3.2), and 12 kg/d (±5.1), respectively. Despite their adaptation to grazing conditions, the behaviour-based and semi-mechanistic grazing-based models had the lowest prediction adequacy among the evaluated models. Their underlying empirical equations likely did not fit the grazing and production conditions of low-input farms using semi-natural grasslands for grazing. The semi-mechanistic stall-based model Mertens II with slight modifications achieved the highest and a satisfactory modelling performance (RPE = 13.4%) when evaluated based on the mean observed PDMI, i.e., averaged across animals per farm and period (n = 28). It also allowed for the adequate prediction of PDMI on individual cows (RPE = 18.5%) that were fed < 4.8 kg DM of supplement feed per day. Nevertheless, when used to predict PDMI of individual animals receiving a high supplementation level, the model Mertens II also did not meet the threshold for an acceptable adequacy (RPE = 24.7%). It was concluded that this lack of prediction adequacy for animals receiving greater levels of supplementation was due to a lack of modelling precision, which mainly could be related to inter-animal and methodological limitations such as the lack of individually measured supplement feed intake for some cows. The latter limitation is a trade-off of the on-farm research approach of the present study, which was chosen to represent the range in feed intake of dairy cows across the diverse low-input farming systems using semi-natural grasslands for grazing.


Effectiveness of different accelerated partial breast irradiation techniques for the treatment of breast cancer patients: Systematic review using indirect comparisons of randomized clinical trials.
This systematic review was conducted to compare the effectiveness of different accelerated partial breast irradiation (APBI) techniques for the treatment of breast cancer patients. Numerous (APBI) techniques are available for clinical practice. Systematic review of randomized controlled trials of APBI versus whole breast irradiation (WBI). The data from APBI studies were extracted for the analyses. Indirect comparisons were used to compare different APBI techniques. Ten studies fulfilled the inclusion criteria. A total of 4343 patients were included, most of them with tumor stage T1-T2 and N0. Regarding APBI techniques, six trials used external beam radiation therapy; one intraoperative electrons; one intraoperative low-energy photons; one brachytherapy; and one external beam radiation therapy or brachytherapy. The indirect comparisons related to 5-years local control and 5-years overall survival were not significantly different between APBI techniques. Based on indirect comparisons, no differences in clinical outcomes were observed among diverse APBI techniques in published clinical trials that formally compared WBI to APBI. However wide confidence intervals and high risk of inconsistency precluded a sound conclusion. Further head-to-head clinical trials comparing different APBI techniques are required to confirm our findings. Studies comparing different techniques using individual participant data and/or real-life data from population-based studies/registries could also provide more robust results.


Retreatment of Recurrent or Second Primary Head and Neck Cancer After Prior Radiation: Executive Summary of the American Radium Society Appropriate Use Criteria.
Retreatment of recurrent or second primary head and neck cancers occurring in a previously irradiated field is complex. Few guidelines exist to support practice. We performed an updated literature search of peer-reviewed journals in a systematic fashion. Search terms, key questions, and associated clinical case variants were formed by panel consensus. The literature search informed the committee during a blinded vote on the appropriateness of treatment options via the modified Delphi method. The final number of citations retained for review was 274. These informed 5 key questions, which focused on patient selection, adjuvant reirradiation, definitive reirradiation, stereotactic body radiation, and reirradiation to treat nonsquamous cancer. Results of the consensus voting are presented along with discussion of the most current evidence. This provides updated evidence-based recommendations and guidelines for the retreatment of recurrent or second primary cancer of the head and neck.


Lumbar and femoral bone density in perimenopausal women with irregular cycles.
The aim of the study was to evaluate the impact of irregular cycles on bone mass in perimenopausal women. University clinic. 118 healthy women aged between 45 and 55 years were divided into three groups according to the previous year's menstrual history (premenopause: regular cycles; perimenopause: irregular cycles; early postmenopause: 1-3 years since the last menstrual cycle). Among the perimenopausal group, only women presenting intervals without menstruation longer than 3 months during the year of the study were selected. Bone mineral density of the lumbar spine and of the proximal femur was measured by dual energy X-ray absorptiometry (DEXA) at the start of the study and after 12 months. The serum estradiol level was significantly lower in postmenopausal women and the FSH serum level showed a significant increase in perimenopausal and postmenopausal women in comparison with premenopausal controls. Bone density at L2-L4, femoral neck, greater trochanter, and total femur did not show significant differences at the beginning of the between-group study. Only the density in Ward's triangle was significantly lower in perimenopausal and postmenopausal women. After twelve months the densities of L2-L4, greater trochanter, Ward's triangle, and total femur showed a significant decrease in postmenopausal women in comparison. The perimenopausal group showed a significant decrease in total femur and Ward's triangle density in comparison with premenopausal. The occurrence of marked irregularity of menstrual cycles preceding menopause can influence bone mass, in particular at the femoral site.


Intrathecal carbachol and clonidine produce a synergistic antiallodynic effect in rats with a nerve ligation injury.
Antiallodynic effects have been demonstrated after intrathecal administration of alpha-2 adrenoceptor agonists and cholinesterase inhibitors in rats. Intrathecal carbachol also increases the activity of cholinergic receptor system at the spinal level. However, there is no study regarding the antagonism of carbachol on touch-evoked allodynia and the interaction with clonidine. This study examines the intrathecal interaction between two drugs in a rat model of nerve ligation injury. Rats were prepared with tight ligation of the left L5-6 spinal nerves and chronic intrathecal catheter implantation. Tactile allodynia was measured by using application of von Frey filaments to the lesioned hindpaw. Carbachol (0.3-10 microg) and clonidine (1-30 microg) were administered to obtain the dose-response curves and the 50% effective dose (ED(50)) for each drug. Fractions of ED(50) values (1/2, 1/4, 1/8, and 1/16) were administered intrathecally to establish the ED(50) of the carbachol-clonidine combination. Isobolographic and fractional analyses of drug interaction were performed. Intrathecal carbachol and clonidine alone produced a dose-dependent reduction of tactile allodynia without severe motor weakness or sedation. A carbachol-clonidine combination produced a dose-dependent increase in withdrawal threshold of the lesioned hindpaw with a reduced incidence and magnitude of side effects. Both analyses revealed a synergistic interaction after the coadministration of carbachol and clonidine. This study indicates that carbachol, like clonidine, provides a moderate antagonism on touch-evoked allodynia at the spinal level. The results suggest that intrathecally administered carbachol is synergistic when combined with clonidine.


A green chemical oligomerization of phloroglucinol induced by plasma as novel α-glucosidase inhibitors.
A new facile method was developed for simple green synthesis of methylene-bridged phloroglucinol oligomers using nonthermal dielectric barrier discharge (DBD) plasma in methanolic solution. The chemical structures of these newly generated oligomers 2-5 were determined by interpretation of the spectroscopic data, and the inhibitory activity toward α-glucosidase of all isolates was evaluated. The unusual phloroglcuinol pentamer 5 connected by four methylene linkages showed a much higher potential inhibitory effect against α-glucosidase than the other generated oligomers 2-4 and appeared to be a promising lead for development as a potential antidiabetic agent. Abbreviations: T2DM, type2 diabetes mellitus; DBD, dielectric barrier discharge; HPLC, high-performance liquid chromatography; IC50, 50% inhibition concentration; NMR, nuclear magnetic resonance; FABMS, fastatom bombardment mass spectrometry.


First clinical experience with Excimer assisted high flow bypass surgery of the brain.
A new type of revascularization of the brain is used in patients with brain ischaemia, caused by an occlusion of one or both internal carotid arteries, and in patients in whom the internal carotid artery has been deliberately occluded for the treatment of a giant aneurysm of the internal carotid artery. A so-called high flow extra-intracranial bypass operation is performed. An arterial or venous transplant is interposed between a branch of the external carotid artery or the external carotid artery itself and the intracranial portion of the internal carotid artery. The anastomosis with the intracranial portion of the internal carotid artery is made without temporary occlusion of the recipient artery and with the aid of the newly developed Excimer laser assisted anastomosis technique. The results of animal experiments and of a clinical series of 9 patients are reported.


[Prevalence of diabetes mellitus and cardiovascular disease among immigrants from Turkey and Morocco and the indigenous Dutch population].
To gain insight into the prevalence of diabetes mellitus and cardiovascular disease among Turkish and Moroccan immigrants in the Netherlands. Secondary analysis of data from a cross-sectional study in 1999/'00. The prevalence of self-reported diabetes and cardiovascular disease was studied in a sample of 743 Turkish, 641 Moroccan and 537 ethnic Dutch persons aged 35 to 74 years from the population of Amsterdam, the Netherlands, using an oral questionnaire. Ethnic differences in the prevalence of diabetes and cardiovascular disease were studied by logistic regression analysis controlling for age, sex and educational level. Further, in the case of diabetes, we examined whether or not the differences could be explained by overweight and physical inactivity, and in the case of cardiovascular disease we investigated the relationship with smoking and diabetes. In addition, the association between diabetes and cardiovascular disease was investigated. The Turkish and Moroccan population reported diabetes more often than did the ethnic Dutch (12.3, 12.4 and 3.0%, respectively). The differences were still present after controlling for sex, age, educational level, overweight and physical inactivity. Cardiovascular disease was more prevalent among the Turks (10.6%) than among the indigenous Dutch population (5.0%), but this was not true for the Moroccans (5.4%). The difference between Turkish and ethnic Dutch people still persisted after controlling for educational level, overweight, physical inactivity, smoking and diabetes. The association between diabetes and cardiovascular disease was consistent for all groups studied. The prevalence of diabetes was higher among Turkish and Moroccan immigrants than among the indigenous Dutch population. Cardiovascular disease was more prevalent among the Turks, but not among the Moroccans. The association between diabetes and cardiovascular disease was consistent for all three ethnic groups.


A shell-less hen's egg test as infection model to determine the biocompatibility and antimicrobial efficacy of drugs and drug formulations against Pseudomonas aeruginosa.
A shell-less hen's egg based infection test with Pseudomonas aeruginosa was established to investigate the antimicrobial efficacy of drugs and drug formulations close to the in vivo situation. The test system using preincubated fertilized chicken eggs transferred in petri dishes was optimized with respect to the controlled local application of liquid materials and bacteria as well as the bacterial cultivation conditions. The applicability of the ex ovo infection model was confirmed with antimicrobial susceptibility tests using tobramycin, ciprofloxacin and meropenem. The validity of the ex ovo data was demonstrated by correlation with in vitro data of the CellTiter®-Blue and the microplate laser nephelometry assay. Real-time imaging of the progress of infection and the efficacy of the treatment could be realized by the MolecuLight i:X™ technique. Furthermore, in a proof-of-concept efficacy, biocompatibility and even the presence of irritants were determined side-by-side using commercial ophthalmics. In conclusion, this egg based infection model could bridge the gap between in vitro and in vivo models for the evaluation of antimicrobial susceptibility to reduce animal tests according to the 3R concept.


[Psychiatric and psychosomatic consulation-liaison. An overview].
A high rate of psychiatric comorbidity is found in patients with somatic diseases in general hospitals. Reasons for psychiatric disorders in somatically ill patients are variable and can lead to or be a result of the physical illness or just occur coincidentally. Consultation-liaison services (C/L) assist the physician and the treatment team on internal surgical wards in general hospitals in paying attention to and caring for somatic psychiatric comorbidities with diagnostic, therapeutic and, if appropriate, secondary preventative methods. In this article an overview of the development of C/L psychiatry is given. Furthermore, the importance of particular psychiatric disorders seen by psychiatric C/L services and their treatment is described. Finally, specific aspects of service delivery with regard to collaborative care between general hospital physicians and private practitioners are discussed.


A genetic and biochemical analysis of a polymorphism of bovine alpha S2-casein.
Using gel electrophoresis a genetic polymorphism of alpha S2-casein (Cn) was discovered in individual milk samples from 2 bovine breeds of the eastern part of France (Vosgienne and Montbéliarde). The 3 observed phenotypes (Plate 1) are determined by 2 co-dominant alleles at an autosomal locus. The alpha S2-Cn A variant was the only one known up to now in European breeds (reference variant) and alpha S2-Cn D is a new variant, whose bands overlap the beta-casein A band at pH 8.6, and migrate faster than alpha S2-Cn A at pH 3.0. The sequence of the polypeptide chain alpha S2-Cn D differs from that of alpha S2-Cn A by the deletion of a very acidic nonapeptide, which includes a cluster of 3 phosphoseryl residues. Due to the characteristics of the reference sequence, this deletion cannot be exactly located but it involves residues 50-58, or 51-59, or 52-60. A genetic analysis shows that locus alpha S2-Cn is closely linked to the cluster alpha S1-Cn--beta-Cn--kappa-Cn. The 4 casein species are thus synthesized by 4 closely linked loci.


Glucagon-Like Peptide-1 Receptor Agonists in Type 2 Diabetes Mellitus and Cardiovascular Disease: The Past, Present, and Future.
Type 2 diabetes mellitus (T2DM) is associated with high cardiovascular morbidity and mortality, and cardiovascular diseases are the leading causes of death and disability in people with T2DM. Unfortunately, therapies strictly aimed at glycemic control have poorly contributed to a significant reduction in the risk of cardiovascular events. On the other hand, randomized controlled trials have shown that five glucagon-like peptide-1 receptor agonists (GLP-1 RAs) and one exendin-based GLP-1 RA reduced atherosclerotic cardiovascular events in patients with diabetes at high cardiovascular risk. Furthermore, a meta-analysis including these six agents showed a reduction in major adverse cardiovascular events as well as all-cause mortality compared with placebo, regardless of structural homology. Evidence has also shown that some drugs in this class have beneficial effects on renal outcomes, such as preventing the onset of macroalbuminuria. In addition to lowering blood pressure, these drugs also favorably impacted on body weight in large randomized controlled trials as in real-world studies, a result considered a priority in T2DM management; these and other factors may justify the benefits of GLP-1 RAs upon the cardiovascular system, regardless of glycemic control. Finally, studies showed safety with a low risk of hypoglycemia and no increase in pancreatitis events. Given these benefits, GLP-1 RAs were preferentially endorsed in the guidelines of the European and American societies for patients with these conditions. This narrative review provides a current and comprehensive overview of GLP-1 RAs as cardiovascular and renal protective agents, far beyond their use as glucose-lowering drugs, supporting their effectiveness in treating patients with T2DM at high cardiovascular risk.


Osmo-ionic regulation and carbonic anhydrase, Na+/K+-ATPase and V-H+-ATPase activities in gills of the ancient freshwater crustacean Aegla schmitti (Anomura) exposed to high salinities.
Aeglidae anomuran crabs originated in the sea, but invaded and diversified in southern South American freshwater (FW) streams. We here aimed at examining their tolerance of increased salinity, after a long time of evolution in FW (~33 million years). Aegla schmitti were exposed to FW and dilute seawater of salinities 15, 20, and 25‰ for 1, 5 and 10 days. Mortality in 35‰ was also assessed. Hemolymph osmolality, Na+, K+, Cl-, and Mg2+ ions, and hydration levels of the abdominal muscle were assayed. The activities of the Carbonic Anhydrase (CA), Na+/K+-ATPase (NKA) and V-H+-ATPase (VHA) were also assayed in the gills. A. schmitti preserves osmoregulatory mechanisms of its marine ancestors. It is able to survive in high salinities (25‰) for at least 10 days. Mortality in 35‰ was of 56% after 1 day, and of 100% after 7 days. In 25‰, NaCl is apparently hyporegulated at all times, while hemolymph osmolality rises after 5 days. CA and NKA activities remained unchanged in all experimental conditions, while VHA activity decreased after 10 days in 25‰. Hemolymph NaCl data was compatible with either hyporegulation and/or putative influx of NaCl into cells for regulatory volume increase (RVI). Further studies should deepen the understanding of the roles of low permeabilities and saturation of high affinity uptake systems in truly FW decapods, in their responses to high salinities. Moreover, the fate of extracellular NaCl as secretion in true hypo-regulation and/or influx into cells for RVI should also be investigated.


Development of progesterone-facilitated lordosis in female guinea pigs: relationship to neural estrogen and progestin receptors.
Ovariectomized (OVX), 20-day-old female guinea pigs did not exhibit lordosis following treatment with estradiol benzoate (EB) and progesterone. Behavioral responsiveness to EB and progesterone developed abruptly; by 30 days of age, responses typical of adult females (greater than 9 weeks of age) were observed. In vitro assays of neural steroid receptors were performed to test the hypothesis that a deficiency in the concentration of hypothalamic and/or preoptic area estrogen and/or progestin receptors contributes to the lack of progesterone-facilitated lordosis in juvenile (20-day-old) females. Assay of cytosol progestin receptors in the mediobasal hypothalamus (MBH) and preoptic area (POA) of immature and adult females 40 h after injection of EB confirmed a previous report that the concentration of estradiol-induced cytosol progestin receptors in the MBH of juvenile females was slightly lower than that observed in adults. Furthermore, MBH cell nuclear accumulation of progestin receptors after progesterone injection was markedly lower (i.e. 42%) in immature, as compared to adult females. The concentrations of cytosol estrogen receptors in the MBH and POA did not differ between immature and adult OVX guinea pigs. Also, cell nuclear estrogen receptor accumulation in the MBH after estradiol injection was comparable in immature and adult females. These data suggest that a deficiency in cell nuclear progestin receptor accumulation in the MBH may contribute to the absence of progesterone-facilitated lordosis in estradiol-primed, immature female guinea pigs. This age-related difference in cell nuclear progestin receptor accumulation may be due, in part, to reduced concentrations of estradiol-induced cytosol progestin receptors.(ABSTRACT TRUNCATED AT 250 WORDS)


Antimicrobial central venous catheters do not reduce infections in pre-term babies.
The studyGilbert R, Brown M, Rainford N et al. Antimicrobial-impregnated central venous catheters for prevention of neonatal bloodstream infection (PREVAIL): an open-label, parallel-group, pragmatic, randomised controlled trial. Lancet Child Adolesc Health 2019;3:381-90.The study was funded by the NIHR Health Technology Assessment programme (project number 12/167/02).To read the full NIHR Signal, go to https://discover.dc.nihr.ac.uk/content/signal-000782/antimicrobial-central-venous-catheters-for-pre-term-babies-do-not-reduce-infections.


Allergies in Germany -- prevalence and perception by the public.
During the recent decades allergies have become more frequent all over the world. However, it is unclear how important the topic of allergies is for the general German population and how appropriately patients with allergies are treated. A telephone survey was performed on a representative random sample of n = 1,004 adults in Germany. The survey was performed by the Forsa Institute for Social Research and Statistical Analysis, Berlin, Germany, in the period from 31 January to 2 February 2012. Of the interviewees 52% responded that the topic of allergies concerned them; in 33% actually an allergy had been diagnosed by a physician. The proportion of allergies in the population correlated with the level of school education and was higher among people with a higher educational status. No differences in allergy rates were found between Eastern and Western Germany. Among allergic persons, 53% reported to be burdened by their allergy, 48% suffered from impaired performance because of their allergic symptoms. Among people suffering from pollen allergy, only 28% received sublingual immune therapy, with which 70% were satisfied. While 58% practiced self-medication, only 21% of the allergic persons were treated with anti-allergic drugs during their allergy flares. Allergic diseases are a common, often burdensome problem in the German population, but nevertheless the medical treatment of people affected is still insufficient. The proportion of patients receiving sublingual immune therapy as causal treatment is comparatively low. Active steps are needed to improve the utilization behavior of patients, e. g. to take advice of an allergy specialist.


Pharmaceutical development of a parenteral lyophilized formulation of the investigational antitumor neuropeptide antagonist [Arg6, D-Trp7,9, MePhe8]-Substance P [6-11].
The aim of this study was to develop a stable parenteral dosage form for the investigational cytotoxic drug [Arg6, D-Trp79,MePhe8]-Substance P [6-11] (Substance P Antagonist G; Antagonist G). Antagonist G bulk drug was structurally and analytically characterized. The drug exhibits excellent aqueous solubility, although relatively poor aqueous stability characteristics. Lyophilization was, therefore, selected as the manufacturing process. Differential scanning calorimetry studies were conducted to determine the freeze-drying cycle parameters which resulted in a stable, lyophilized formulation of Antagonist G. The prototype, containing 50 mg Antagonist G per vial, was found to be the optimal formulation in terms of solubility, length of the freeze-drying cycle, stability, and dosage requirements in the planned phase I clinical trials. Quality control of the freeze-dried formulation showed that the manufacturing process does not change the integrity of Antagonist G. Shelf life studies demonstrated that the formulation is stable for at least 3 years, when stored at 2-8 degrees C in a dark environment. Oxidative degradation products of Antagonist G were isolated and structurally characterized by mass spectrometry, nuclear magnetic resonance spectroscopy, and infrared spectroscopy.


MK-801 induced amnesia for the elevated plus-maze in mice.
MK-801, a non-competitive NMDA receptor antagonist, has been shown to have amnesic properties in animal models. The purpose of the present study was to examine potential amnesic effects of MK-801 in mice using the modified elevated plus-maze paradigm. An animal was placed on the distal end of an open arm, and the transfer latency, i.e. the time in which it moves to the enclosed arm, was measured. Four different experimental schedules (i.e. the combination of the treatment and the testing) were used: MK-801 (0.075, 0.15, 0.25 and 0.4 mg/kg or saline) were given (a) 30 min before the acquisition session, (b) immediately after the acquisition session, (c) 60 min after the acquisition session, and (d) 30 min before the retention session. The retention session always followed 24 h after the acquisition session. Analysis of data showed a significant shortening of the transfer latency in saline-treated animals during the retention session. Further, MK-801 (at the dose range of 0.15--0.4 mg/kg) administered before and immediately after the acquisition session as well as before the retention session prolonged the transfer latency during the retention session. In fact, transfer latencies in MK-801 treated mice did not differ from those measured during the acquisition session. Thus, prolongation of the transfer latency in MK-801-treated mice indicates deficits in 'memorization' processes. On the contrary, MK-801 given 60 min after the acquisition session failed to increase the transfer latency, which suggests that the memory trace was sufficiently consolidated at this time. Based on the present results, the glutamatergic NMDA receptor mechanisms play an important role in a spatial orientation of mice placed on the elevated plus-maze.


Distribution of endocytosed molecules to intracellular acidic environments correlates with immunotoxin activity.
We have investigated the internalization to low pH intracellular compartments of transferrin (Tfn), diphtheria toxin (DT) and of anti-cell surface antibodies (MAb) by a cytofluorometric assay based on low pH quenching of fluorescein (FITC) emission. FITC-labelled Tfn, anti-CD3, anti-CD5 and anti-Thy 1.2 MAb internalization resulted in a progressively lower FITC quenching effect. Following internalization, a distinction could be made between molecules that enter low pH compartments without undergoing intracellular degradation (e.g., Tfn, anti-CD3 MAb) and molecules that are internalized through low pH organelles and are then degraded within the cell (e.g., DT). A strict correlation was observed between quenching of internalized FITC-protein fluorescent emission and the cytotoxic activity of DT-based immunotoxins (IT).


Diet therapy in gastrointestinal disease: a commentary.
Dietary regimens in gastrointestinal disease can be divided into two categories: First, those of proven value include: (a) Disaccharide elimination for disaccharidase deficiency and exclusion of monosaccharides for sugar malabsorption; (b) gluten-free diet for celiac/sprue; (c) elimination of certain allergens because of food allergies; (d) protein restriction for portal systemic encephalopathy; (e) low-carbohydrate diet for dumping syndrome; (f) low-fiber diet for diarrheal syndromes; and (g) low-fat diet for steatorrhea. Second, controversial diets include a bland diet for acid-peptic disorders, a high-fiber diet for colonic disorders, and a low-fat diet for gallbladder disease. It is important to separate facts from fancy in the dietary management of patients with gastrointestinal disease and base the recommendations for a particular diet on available objective evidence, not on traditional or fashionable trends.


Fueling ab initio folding with marine metagenomics enables structure and function predictions of new protein families.
The ocean microbiome represents one of the largest microbiomes and produces nearly half of the primary energy on the planet through photosynthesis or chemosynthesis. Using recent advances in marine genomics, we explore new applications of oceanic metagenomes for protein structure and function prediction. By processing 1.3 TB of high-quality reads from the Tara Oceans data, we obtain 97 million non-redundant genes. Of the 5721 Pfam families that lack experimental structures, 2801 have at least one member associated with the oceanic metagenomics dataset. We apply C-QUARK, a deep-learning contact-guided ab initio structure prediction pipeline, to model 27 families, where 20 are predicted to have a reliable fold with estimated template modeling score (TM-score) at least 0.5. Detailed analyses reveal that the abundance of microbial genera in the ocean is highly correlated to the frequency of occurrence in the modeled Pfam families, suggesting the significant role of the Tara Oceans genomes in the contact-map prediction and subsequent ab initio folding simulations. Of interesting note, PF15461, which has a majority of members coming from ocean-related bacteria, is identified as an important photosynthetic protein by structure-based function annotations. The pipeline is extended to a set of 417 Pfam families, built on the combination of Tara with other metagenomics datasets, which results in 235 families with an estimated TM-score over 0.5. These results demonstrate a new avenue to improve the capacity of protein structure and function modeling through marine metagenomics, especially for difficult proteins with few homologous sequences.


An evaluation of the durability of apex locator insulated probes after autoclaving.
The purpose of this study was to evaluate the durability of insulated probes designed to be used with the Endocater Apex Locator when put through multiple cleaning and autoclaving cycles. The manufacturer has reported that the plastic coating on the probes is reliable with normal use and sterilizable up to a maximum of three cycles. The quality of the seal provided against electrical conductivity by the coating was evaluated using 60 probes of various sizes (10, 15, 20, 25, 40) before and after autoclaving. Visual and electrical field methods were used to detect any leakage in the insulated coating. Defects were detected electronically before they could be detected visually. Statistically significant differences in millimeters of coating lost were found between the original evaluation and after each autoclaving cycle. The size 40 probes lost statistically significantly more coating than did any of the other sizes. The loss of coating seemed to be related to the millimeter markings. The quality of the probes as they came from the manufacturer was excellent; no defects were found in the coating prior to autoclaving.


Single nucleotide polymorphisms of IGFBP-3 gene and lung cancer risk in a Korean population.
Insulin-like growth factor binding protein-3 (IGFBP-3) is a putative tumor suppressor and inhibits the mitogenic and antiapoptotic activity of insulin-like growth factor (IGF) by blocking the binding to its receptors or by IGF-independent manner. Epidemiological studies have suggested that serum level of IGFBP-3 is associated with lung cancer risk. The present study was conducted to evaluate the association between single nucleotide polymorphisms (SNPs) of the IGFBP-3 gene and susceptibility to lung cancer and the effect of the SNPs on the serum levels of IGFBP-3, total IGF-I and free IGF-I in a Korean population. After a preliminary study for polymorphism screening of the promoter region of the IGFBP-3 gene in randomly selected 41 lung cancer patients, two polymorphisms (-1590 C>A; -202 A>C) were identified with PCR-based restriction fragment length polymorphism (PCR-RFLP) and SNaPshot primer extension assay in 415 Korean lung cancer patients and 415 matched normal controls. Although the genotype and allele frequency distribution of each SNP did not differ significantly between cases and controls in the single-locus analysis, we found that the polymorphisms of -1590A and -202C were associated with increased risk for lung cancer in females (-1590 C>A; adjusted OR=2.11, 95% CI=1.08-4.12, -202 A>C; adjusted OR=1.96, 95% CI=1.02-3.75, respectively) and never-smokers (-1590 C>A; adjusted OR=2.06, 95% CI=1.12-3.78, -202 A>C; adjusted OR=1.99, 95% CI=1.10-3.62, respectively) in a dominant model after stratification for gender and smoking history. Haplotype analysis showed that carriers of A-C had significantly higher risk for lung cancer in females (dominant aOR=2.05, 95% CI=1.14-3.68) and in never-smokers (codominant aOR=1.71, 95% CI=1.11-2.64 and dominant aOR=1.99, 95% CI=1.17-3.40). Functional analysis in H1703 cell line using DNA fragments containing A-C haplotype of the promoter region showed significantly decreased transcriptional activity. Furthermore, there was a negative correlation between the number of polymorphic alleles in -1590/-202 loci and serum levels of IGFBP-3 in lung cancer patients, but it was not statistically significant in the test for linear trend. These results suggest that IGFBP-3 promoter polymorphisms of -1590 C>A and -202 A>C might be a genetic risk factor for lung cancer by means of decreased IGFBP-3 expression among females or never-smoking Koreans.