Variations in Healthcare Transition Preparation Among Youth With Chronic Conditions.
Youth with special healthcare needs have low rates of healthcare transition services, which can affect lifelong functioning and quality of life. This study examines the variations in receipt of healthcare transition services among youth with special healthcare needs. Data from the 2016-2018 National Survey of Children's Health (N=102,341) were analyzed in 2021. Receipt of healthcare transition services by youth with select health conditions was compared with youth with other special healthcare needs. Bivariate and multivariable analyses assessed the associations between the receipt of healthcare transition services, sociodemographic characteristics, and health conditions. Among youth with special healthcare needs, the prevalence of receiving healthcare transition services was lowest among youth with speech or other language disorders (8.5%), intellectual disabilities (9.4%), and autism spectrum disorder (11.1%). Low prevalence of receiving healthcare transition services was also observed for youth with developmental delays (12.6%), learning disabilities (14.2%), and behavior or conduct problems (15.5%). Youth with developmental delays (AOR=0.70, 95% CI=0.52, 0.95), intellectual disabilities (AOR=0.45, 95% CI=0.26, 0.78), learning disabilities (AOR=0.77, 95% CI=0.60, 0.99), autism spectrum disorder (AOR=0.60, 95% CI=0.41, 0.86), and speech or other language disorders (AOR=0.48, 95% CI=0.32, 0.72) had lower odds of receiving healthcare transition services than youth with other special healthcare needs. Findings suggest that the receipt of healthcare transition services varies substantially by the type of chronic health condition and highlight the need for increased healthcare transition services for youth with special healthcare needs, especially for youth with neurodevelopmental disabilities and speech or other language disorders.


Fat Mass is Associated with Subclinical Left Ventricular Systolic Dysfunction in Patients with Type 2 Diabetes Mellitus Without Established Cardiovascular Diseases.
Left ventricular global longitudinal strain (GLS) is considered to be the first marker of diabetes mellitus-related subclinical cardiac dysfunction, but whether it is attributable to fat mass and distribution remains uncertain. In this study, we explored whether fat mass, especially fat mass in the android area, is associated with subclinical systolic dysfunction before the onset of cardiac disease. We conducted a single-center prospective cross-sectional study between November 2021 and August 2022 on inpatients of the Department of Endocrinology, Nanjing Drum Tower Hospital. We included 150 patients aged 18-70 years with no signs, symptoms, or history of clinical cardiac disease. Patients were evaluated with speckle tracking echocardiography and dual energy X-ray absorptiometry. The cutoff values for subclinical systolic dysfunction were set at a global longitudinal strain (GLS) < 18%. After adjusting for sex and age, patients with GLS < 18% had a higher mean (± standard deviation) fat mass index (8.06 ± 2.39 vs. 7.10 ± 2.09 kg/m2, p = 0.02), higher mean trunk fat mass (14.9 ± 4.9 vs. 12.8 ± 4.3 kg, p = 0.01), and higher android fat mass (2.57 ± 1.02 vs. 2.18 ± 0.86 kg, p = 0.02) than those in the GLS ≥ 18%. Partial correlation analysis showed that the fat mass index, truck fat mass, and android fat mass were negatively correlated with GLS after adjusting for sex and age (all p < 0.05). Adjusted for traditional cardiovascular metabolic factors, fat mass index (odds ratio [OR] 1.27, 95% confidence interval [CI] 1.05-1.55, p = 0.02), trunk fat mass (OR 1.13, 95% CI 1.03-1.24, p = 0.01), and android fat mass (OR 1.77, 95% CI 1.16-2.82, p = 0.01) were independent risk factors for GLS < 18%. Among patients with type 2 diabetes mellitus without established clinical cardiac disease, fat mass, especially android fat mass, was associated with subclinical systolic dysfunction independently of age and sex.


Effect of MX-68 on airway inflammation and hyperresponsiveness in mice and guinea-pigs.
MX-68 is a newly synthesized antifolate, which is a derivative of methotrexate (MTX). In this paper, the effect of MX-68 on allergic airway responses in mice and guinea-pigs was studied. In the first experiment, antigen-induced airway inflammation and airway hyperresponsiveness (AHR) to acetylcholine in mice were examined and compared with the effects of classical antifolate methotrexate and prednisolone. Mice were sensitized with ovalbumin as an antigen and challenged with ovalbumin inhalation three times. After the last inhalation, AHR and airway inflammation were observed. An increase in Th2 cytokines (IL-4 and IL-5) and a decrease in a Th1 cytokine (IFN-gamma) in the bronchoalveolar lavage fluid (BALF), as well as an elevation of the immunoglobulin level in serum, were observed in sensitized mice. Oral administration of MX-68 had no effect on changes of body weight, but prednisolone reduced body weight during the experiment. The antigen-induced AHR and increases in the number of eosinophils and lymphocytes in BALF were significantly inhibited by MX-68. MX-68 interfered with the elevation of IL-4 and IL-5 levels in BALF, but had no effect on the decrease in IFN-gamma. Moreover, MX-68 significantly inhibited the elevation of serum IgE and IgG levels. In the guinea-pig model for bronchial asthma, biphasic increases in airway resistance (immediate asthmatic response, IAR, and late asthmatic response, LAR), as well as accumulated inflammatory cells in BALF, were observed after repeated antigen challenge. These asthmatic responses and inflammatory signs were significantly decreased by administration of MX-68. These results suggest that MX-68 obviously has an anti-inflammatory effect in an animal model of asthma and would be useful clinically for the treatment of bronchial asthma.


A hazard associated with improper connection of the Bain breathing circuit.
A case of cardiac arrest is presented which was caused by improper connection of a modified Mapleson D circuit (Bain breathing circuit). Excessive enternal deadspace was created by interchanging the gas inflow line and the attachment for an airway pressure manometer. This resulted in a marked respiratory acidosis, clinically undetected until the concomitant hypoxia produced a severe cardiac depression and arrest. The sequence of events was reproduced in a dog under comparable anaesthetic conditions. In order to avoid this error it is recommended to permanently fuse the connecting piece placed in the circuit with the tubing leading to the airway pressure manometer of the respirator.


Prognostic index in prediction of mortality from infective endocarditis.
Out of 176 patients with infective endocarditis complicating rheumatic (120) and congenital heart disease (38), mortality occurred in 35 patients (19.9%). Presence of leukocytosis, heart failure, major embolisation and isolation of certain organisms including Staphylococci, beta-haemolytic Streptococci, Pseudomonas and Klebsiella were ominous with higher mortality rates compared with those when they were absent (P less than 0.02). Stepwise logistic multiple regression was then applied and the four most important independent variables were identified. A prognostic index for the prediction of mortality for infective endocarditis was then constructed by the summation of the regression coefficients. By applying this index, patients with infective endocarditis could be divided into subgroups with increasing proportional mortality from 5.8 to 83.3%. It provides an objective assessment of the risk patients with infective endocarditis, and a more reliable evaluation of benefit of any new treatment regimen, including cardiac surgery, during the acute stage.


The clinical, cardiac, renal, arterial and neurohormonal effects of omapatrilat, a vasopeptidase inhibitor, in patients with chronic heart failure.
We sought to examine the effects of long-term vasopeptidase inhibition in patients with heart failure. The long-term effects of omapatrilat, an agent that inhibits both neutral endopeptidase and angiotensin-converting enzyme, on clinical status, neurohormonal indexes and left ventricular function in patients with chronic heart failure (CHF) have not been previously documented. Forty-eight patients in New York Heart Association functional class II or III, with left ventricular ejection fraction (LVEF)< or =40% and in sinus rhythm were randomized to a dose-ranging pilot study of omapatrilat for 12 weeks. Measurements were performed at baseline and 12 weeks. There was an improvement in functional status, as reported by the patient (p<0.001) and physician (p<0.001) at 12 weeks. Dose-dependent improvements in LVEF (p<0.001) and LV end-systolic wall stress (sigma) (p<0.05) were seen, together with a reduction in systolic blood pressure (p<0.05). There was evidence of a natriuretic effect (p<0.001), and total blood volume decreased (p<0.05). Omapatrilat induced an increase in postdose plasma atrial natriuretic peptide levels (p<0.01) in the high dose groups, with a reduction in predose plasma brain natriuretic peptide (p<0.001) and epinephrine (p<0.01) levels after 12 weeks of therapy. Omapatrilat was well tolerated. The sustained hemodynamic, neurohumoral and renal effects of omapatrilat, together with improved functional status, suggest that vasopeptidase inhibition has potential as a new therapeutic modality for the treatment of CHF.


Endothelial response to cardiopulmonary bypass surgery.
The vascular endothelium has been shown to actively participate in maintaining normal cardiovascular homeostasis by influencing the regulation of membrane permeability, lipid transport, vasomotor tone, coagulation, fibrinolysis, and inflammation. Endothelial cells are very responsive to a wide range of local and systemic stimuli that occur during cardiopulmonary bypass (CPB) operation. Major pathologic conditions result from impaired vascular function secondary to CPB, including vasospasm, coagulopathy, and widespread neutrophil adhesion secondary to a systemic inflammatory response. Additionally, more chronic responses to endothelial cell injury include the development of intimal hyperplasia and arteriosclerosis, both of which limit the long-term success of coronary artery bypass grafting. Because of the increasingly recognized role of the endothelium in the maintenance of normal cardiovascular function, this article will review the normal structure and function of the endothelium, as well as the major pathologic conditions that result in response to CPB. Potential treatments to counteract endothelial cell dysfunction secondary to CPB are under active investigation. Strategies may be directed toward blocking single cytokines, integrins, or adhesion molecules involved in endothelial dysfunction or, alternatively, toward targeting a molecular event that governs the expression of these proinflammatory, procoagulant, and vasoactive genes. In our laboratory, we have used both strategies to study the pathologic response to CPB. We blocked neutrophil adhesion in subhuman primates with a monoclonal antibody. Alternatively, we targeted the transcriptional activation of multiple genes involved in the endothelial cell's response to CPB. Although both therapies help elucidate the multiple, redundant pathways involved in the pathologic response to CPB, it is through molecular biology that we are beginning to understand the mechanics of transcriptional control and translational expression that occurs in the endothelial cell in response to CPB. This knowledge will allow the development of therapies that inhibit not a single cytokine or adhesion molecule, but rather an array of substances that result in the endothelial cell's pathologic response to CPB.


The effect of feature image on sensitivity of the statistical analysis in the pipeline of a tractography atlas-based analysis.
Tractography atlas-based analysis (TABS) is a new diffusion tensor image (DTI) statistical analysis method for detecting and understanding voxel-wise white matter properties along a fiber tract. An important requisite for accurate and sensitive TABS is the availability of a deformation field that is able to register DTI in native space to standard space. Here, three different feature images including the fractional anisotropy (FA) image, T1 weighted image, and the maximum eigenvalue of the Hessian of the FA (hFA) image were used to calculate the deformation fields between individual space and population space. Our results showed that when the FA image was a feature image, the tensor template had the highest consistency with each subject for scalar and vector information. Additionally, to demonstrate the sensitivity and specificity of the TABS method with different feature images, we detected a gender difference along the corpus callosum. A significant difference between the male and female group in diffusion measurement appeared predominantly in the right corpus callosum only when FA was the feature image. Our results demonstrated that the FA image as a feature image was more accurate with respect to the underlying tensor information and had more accurate analysis results with the TABS method.


New Genera and Species Records of Nicaraguan Eumolpinae (Coleoptera: Chrysomelidae) Including a New Species in a New Generic Record for Central America.
In this work, we have examined a large sample of Eumolpinae leaf beetles from Nicaragua and found 18 species reported for the first time in this country, including the new species Caryonoda funebris n. sp., which also represents a new genus record for Central America, and two genera of Typophorini not reported from Nicaragua so far: Paria LeConte, 1858 and Tijucana Bechyné, 1957. Apart from the description of the new species and taxonomic commentaries on each of the new country records, we also illustrate these species along with drawings of male genitalia and spermathecae when available to assist the interpretation of our taxonomic decisions in the future. We take the opportunity in this work to formalize the combination of Chrysodina cupriceps Lefèvre, 1877 as Chrysodinopsis cupriceps (Lefèvre) n. comb.


Genetic differences in hypothalamic-pituitary-adrenal axis responsiveness to acute ethanol and acute ethanol withdrawal.
There is a growing body of evidence suggesting that corticosteroids contribute to the increased neural excitability observed during ethanol withdrawal. In the present study, this was further investigated using mouse strains which differ in ethanol withdrawal severity. DBA/2 (DBA) mice were found to display more severe acute ethanol withdrawal seizures than C57BL/6 (C57) mice. Additionally, DBA mice showed a greater stress response than C57 mice, as measured by higher plasma concentrations of adrenocorticotropic hormone (ACTH) and corticosterone, to an acute dose of ethanol. Mimicking withdrawal plasma corticosterone levels by administering corticosterone to ethanol-naive mice resulted in increases in handling-induced convulsions in the range observed during withdrawal. There did not appear to be a strain difference in sensitivity to the excitatory effects of corticosterone. In summary, the greater stress response to ethanol by DBA mice may account, in part, for the more severe ethanol withdrawal syndrome of this strain.


Contrasting mechanisms of hypercalcemia in patients with early and advanced humoral hypercalcemia of malignancy.
The mechanisms of hypercalcemia were assessed in 15 patients with humoral hypercalcemia of malignancy (HHM) who had tumors at various stages of progression. In patients with early tumors, bone biopsies were generally normal and the hypercalcemia was due to an elevation in renal tubular resorption of calcium. Conversely, osteoclastic resorption was markedly increased in patients with advanced tumors, particularly those in whom the biopsies were obtained postmortem. Osteoclast surface (Oc.S) correlated positively with the stage of tumor progression (r = 0.80, p less than 0.002), degree of immobility (r = 0.87, p less than 0.002), and level of urinary cyclic AMP excretion (r = 0.60, p less than 0.02). When compared with a group of ambulant patients with primary hyperparathyroidism (HPT), osteoblast surface (Ob.S%) in HHM was depressed (median and range): 1.2% (0-11.6%) versus 5.3% (1.1-32.0%) (p less than 0.001). However, a relatively low Ob.S (4%) and raised Oc.S (43.5%) were also seen in an immobilized patient with severe HPT. These data suggest that the PTH-related peptides currently invoked in the pathogenesis of HHM may initially cause hypercalcemia by enhancing renal tubular calcium resorption. The increase in osteoclastic activity and depression of osteoblastic activity that subsequently occurs is probably due to the combined effects of immobilization and higher circulating levels of PTHrP on the skeleton. However, the release of other bone-resorbing factors by the tumor, which have a depressant effect on osteoblastic activity, remains possible.


Mice cloned from olfactory sensory neurons.
Cloning by nuclear transplantation has been successfully carried out in various mammals, including mice. Until now mice have not been cloned from post-mitotic cells such as neurons. Here, we have generated fertile mouse clones derived by transferring the nuclei of post-mitotic, olfactory sensory neurons into oocytes. These results indicate that the genome of a post-mitotic, terminally differentiated neuron can re-enter the cell cycle and be reprogrammed to a state of totipotency after nuclear transfer. Moreover, the pattern of odorant receptor gene expression and the organization of odorant receptor genes in cloned mice was indistinguishable from wild-type animals, indicating that irreversible changes to the DNA of olfactory neurons do not accompany receptor gene choice.


Microdose intravaginal levonorgestrel contraception: a multicentre clinical trial. IV. Bleeding patterns. World Health Organization. Task Force on Long-Acting Systemic Agents for Fertility Regulation.
A multicentred clinical trial was conducted in 12 countries to assess the contraceptive efficacy and side effects of a low-dose levonorgestrel-releasing vaginal ring. Contraceptive efficacy and side effects, expulsions and removals, and the relation between pregnancy rate and admission body weight of the women have been reported previously. The effect of ring use on vaginal bleeding patterns as observed from daily menstrual diaries kept by the women is the object of this paper. A total of 1005 women were fitted with the ring and 702 of them provided a menstrual diary suitable for analysis. The description of the bleeding patterns is made using a 90-day reference period approach and following the guidelines published by WHO. In addition the longest bleeding/spotting episode and longest bleeding/spotting-free interval are calculated over the entire diary length. Half of the ring users have vaginal bleeding patterns similar to normal untreated patterns. A quarter experience irregular bleeding, 10% have prolonged cycles and 10% have shortened cycles. Over one year of continuous use, there appears to be no significant trend in these patterns. Users with the worst bleeding patterns tend to discontinue first during the clinical trial and are influenced by their more recent experience of vaginal bleeding irregularities. Life-table analysis of discontinuation rates gives an underestimation of the true incidence of bleeding irregularities. Some comments are made on the limitations of the methods of analysis of menstrual diaries.


The prevalence of brain lesions after in utero surgery for twin-to-twin transfusion syndrome on third-trimester MRI: a retrospective cohort study.
Due to the increased risk of antenatal brain lesions, we offer a third-trimester magnetic resonance imaging (MRI) scan to all patients who underwent an in utero intervention for twin-twin transfusion syndrome (TTTS). However, the usefulness of such a policy has not been demonstrated yet. Therefore, we determined the prevalence of antenatal brain lesions detected on third-trimester MRI and the proportion of lesions detected exclusively on MRI. We conducted a retrospective cohort study of monochorionic diamniotic twin pregnancies complicated by TTTS that underwent laser coagulation of the vascular anastomoses or fetal reduction by umbilical cord occlusion between 2010 and 2017. We reviewed the third-trimester MRI findings and compared those with the prenatal ultrasonography. Of the 141 patients treated with laser coagulation and 17 managed by cord occlusion, 112/141 (79%) and 15/17 (88%) patients reached 28 weeks. Of those, 69/112 (62%) and 11/15 (73%) underwent an MRI between 28 and 32 weeks. After laser coagulation, MRI detected an antenatal brain lesion in 6 of 69 pregnancies (9%) or in 6 of 125 fetuses (5%). In 4 cases (67%), the lesion was detected only on MRI. In the 11 patients treated with cord occlusion, no brain lesions were diagnosed. The prevalence of brain lesions detected by third-trimester MRI is higher compared to prenatal ultrasonography alone, making MRI a useful adjunct to detect antenatal brain lesions in twin pregnancies after in utero treatment for TTTS. • In utero interventions for twin-to-twin transfusion syndrome (TTTS) do not prevent the occurrence of antenatal brain lesions. • Fetal magnetic resonance imaging (MRI) has high accuracy in detecting anomalies of cortical development and can be a useful adjunct to ultrasonography in diagnosing certain brain abnormalities. • After laser coagulation of the anastomoses for TTTS, third-trimester MRI diagnosed a brain lesion that was not detected earlier on ultrasound scan in 6% of pregnancies.


Modified Girdlestone arthroplasty and hip arthrodesis using the Ilizarov external fixator as a salvage method in the management of severely infected total hip replacement.
Resection arthroplasty or hip arthrodesis after total hip replacement (THR) can be used to salvage the limb in case with deep infection and severe bone loss. The Ilizarov fixator provides stability, axial correction, weight-bearing and good fusion rates. We retrospectively assessed the outcomes of 37 patients with severe periprosthetic infection after THR treated between 1999 and 2011. The treatment included implant removal, debridement and a modified Girdestone arthroplasty (29 cases) or hip arthrodesis (seven cases) using the Ilizarov fixator. The Ilizarov fixation continued from 45 to 50 days in the modified arthroplasty group and 90 days in the arthrodesis group. One case was treated using the conventional resection arthroplasty bilaterally. Eighteen months after treatment, infection control was seen in 97.3% cases. Six hips were fused as one patient died in this group. Limb length discrepancy (LLD) averaged 5.5 cm. The Harris hip score ranged from 35 to 92 points. Hip joint motion ranged from 10° to 30° in the modified arthroplasty group. All subjects could walk independently or using support aids. No subluxation or LLD progression was observed. The modified Girdlestone arthroplasty and hip arthrodesis using the Ilizarov apparatus results in sufficient ability for ambulation and good infection control in cases of failed THR associated with severe infection.


Association between diabetes mellitus and chronic rhinosinusitis with nasal polyps: A population-based cross-sectional study.
To investigate the association between physician-diagnosed diabetes mellitus (DM) and chronic rhinosinusitis (CRS) phenotypes in a national population-based study. Retrospective cross-sectional study. Population-based survey data were collected by the Korean National Health and Nutrition Survey between January 2008 and December 2012. A total of 34 670 participants aged over 19 years were enrolled in the Korea National Health and Nutrition Examination Surveys from 2008 to 2012. The relationship of CRS prevalence, with and without nasal polyps, with physician-diagnosed DM and non-DM were assessed. Differences in sinonasal symptoms between patients with and without DM were analysed in this cross-sectional study. A significant association was observed between DM and CRS with nasal polyps after adjustment for multiple variables. No substantial association was observed between DM and CRS without nasal polyps. Among patients with CRS, olfactory dysfunction for >3 months was significantly more frequent in the DM group than in the non-DM group. We demonstrated significant associations between DM and CRS with nasal polyps and olfactory dysfunction among patients with CRS in a large national clinical cohort study. The direct mechanism of the association between DM and CRS with nasal polyps should be further investigated to clarify the pathogenesis of CRS with nasal polyps.


Prospective Multicenter Study Validate a Prediction Model for Surgery Uptake Among Women with Atypical Breast Lesions.
Diagnosis of atypical breast lesions (ABLs) leads to unnecessary surgery in 75-90% of women. We have previously developed a model including age, complete radiological target excision after biopsy, and focus size that predicts the probability of cancer at surgery. The present study aimed to validate this model in a prospective multicenter setting. Women with a recently diagnosed ABL on image-guided biopsy were recruited in 18 centers, before wire-guided localized excisional lumpectomy. Primary outcome was the negative predictive value (NPV) of the model. The NOMAT model could be used in 287 of the 300 patients included (195 with ADH). At surgery, 12 invasive (all grade 1), and 43 in situ carcinomas were identified (all ABL: 55/287, 19%; ADH only: 49/195, 25%). The area under the receiving operating characteristics curve of the model was 0.64 (95% CI 0.58-0.69) for all ABL, and 0.63 for ADH only (95% CI 0.56-0.70). For the pre-specified threshold of 20% predicted probability of cancer, NPV was 82% (77-87%) for all ABL, and 77% (95% CI 71-83%) for patients with ADH. At a 10% threshold, NPV was 89% (84-94%) for all ABL, and 85% (95% CI 78--92%) for the ADH. At this threshold, 58% of the whole ABL population (and 54% of ADH patients) could have avoided surgery with only 2 missed invasive cancers. The NOMAT model could be useful to avoid unnecessary surgery among women with ABL, including for patients with ADH. NCT02523612.


Quantification of Chemotaxis or Respiratory Burst Using Ex Vivo Culture-Derived Murine Neutrophils.
Two critical functional responses of neutrophils are chemotaxis, a response driven by concentration gradients of chemokines released by infected or inflamed tissues, and production of reactive oxygen species (ROS), molecules essential to their capacity to kill pathogens. Assays to accurately test each response have been important to assess efficacies of pharmaceuticals predicted to block recruitment of neutrophils or attenuate their ROS production. Identified antagonists to neutrophil functions may help to reduce tissue damage following inflammation. Described are detailed assays to test these functions, along with steps to generate neutrophils from ex vivo-cultured murine bone marrow that produce robust responses in either assay. The first function protocol details a quantitative assay for chemotaxis that involves culture plates with dual chamber wells that separate cells from a chemokine with small pore-sized membranes. Quantitative measurements of cell numbers in the chemokine-containing chamber are performed with either fluorescence or luminescence detection reagents, which provide signals directly proportional to the numbers of migrated cells. Multiwell plates are used for rapidly testing a variety of conditions and/or chemoattractants. Described in the second function protocol is an assay to measure ROS produced by stimulated neutrophils, again using a multiwell platform for rapid, quantitative measurements of several conditions simultaneously.


Prediction of Individual Analgesic Response to Intravenous Lidocaine in Painful Diabetic Peripheral Neuropathy: A Randomized, Placebo-controlled, Crossover Trial.
Intravenous lidocaine can alleviate painful diabetic peripheral neuropathy (DPN) in some patients. Whether quantitative sensory testing (QST) can identify treatment responders has not been prospectively tested. This was a prospective, randomized, double-blind, crossover, placebo-controlled trial comparing intravenous lidocaine to normal saline (placebo) for painful DPN. Thirty-four participants with painful DPN were enrolled and administered intravenous lidocaine (5 mg/kg ideal body weight) or placebo as a 40-minute infusion, after a battery of QST parameters were tested on the dorsal foot, with a 3-week washout period between infusions. Thirty-one participants completed both study sessions and were included in the final analysis. Lidocaine resulted in a 51% pain reduction 60 to 120 minutes after infusion initiation, as assessed on a 0 to 10 numerical rating scale, while placebo resulted in a 33.5% pain reduction (difference=17.6%, 95% confidence interval [CI], 1.9%-33.3%, P=0.03). Neither mechanical pain threshold, heat pain threshold, or any of the other measured QST parameters predicted the response to treatment. Lidocaine administration reduced mean Neuropathic Pain Symptom Inventory paresthesia/dysesthesia scores when compared with placebo by 1.29 points (95% CI, -2.03 to -0.55, P=0.001), and paroxysmal pain scores by 0.84 points (95% CI, -1.62 to -0.56, P=0.04), without significant changes in burning, pressing or evoked pain subscores. While some participants reported therapeutic benefit from lidocaine administration, QST measures alone were not predictive of response to treatment. Further studies, powered to test more complex phenotypic interactions, are required to identify reliable predictors of response to pharmacotherapy in patients with DPN.


Identification and tissue distribution of odorant binding protein genes in the beet armyworm, Spodoptera exigua.
Odorant binding proteins (OBPs) contribute to the remarkable sensitivity of the insect's olfactory system and play an important role in insect chemical communication. In this study, we identified 11 putative cDNAs encoding OBPs (namely SexiOBP1-11) from the antennal full length cDNA library of the beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) and examined their expression profiles in different adult body tissues (antennae, heads, thoraxes, abdomens, legs and wings) by real-time quantitative PCR (qPCR). All SexiOBPs had the characteristic typical features of the OBP family, with the exception of SexiOBP11, which lacked the predicted signal peptide sequence at the N-terminus. qPCR revealed that all of these genes were highly transcribed in the antennae. SexiOBP1-4 and SexiOBP10 were dominantly restricted to antennae. Within antennae, SexiOBP2-4 and SexiOBP10 exhibited female-biased expression patterns, while the expression of SexiOBP7 was male-biased, indicating that they might be involved in interacting with sex pheromones. In general, these OBPs were mainly expressed in chemosensory-specific tissues, although some displayed non-chemosensory or ubiquitous tissue expression. The data is helpful for further determining the potential physiological functions of S. exigua OBPs, and paves the way towards a better understanding of the chemosensory perception of this pest, which may help to uncover new targets for behavioral interference used as a control strategy.


Prosthetic metals impair murine immune response and cytokine release in vivo and in vitro.
This study was designed to investigate whether prosthetic metals adversely affect immune responses and the release of immunoregulatory cytokines in vivo and in vitro. Titanium and cobalt-chromium alloy were injected into the peritoneal cavity of female mice. At 5, 8, and 12 weeks after the injection, the levels of cobalt and chromium in the blood were significantly increased compared with the levels in control mice; the level of titanium was not significantly changed until 12 weeks. The release of interleukin-2 was significantly inhibited by cobalt-chromium particles after 3 weeks; titanium particles did not have the same effect until 8 and 12 weeks. The release of interleukin-4 was significantly inhibited by cobalt-chromium particles after 3 weeks but was not significantly inhibited by titanium particles until 12 weeks. The release of interferon-gamma was significantly inhibited by cobalt-chromium particles only at 12 weeks and was not inhibited by titanium particles. The proliferation of T cells was significantly inhibited by cobalt-chromium particles at 3 weeks and by titanium particles at 8 and 12 weeks, and the proliferation of B cells was significantly inhibited by cobalt-chromium particles after 3 weeks but was not inhibited by titanium particles. The production of immunoglobulin by lipopolysaccharide-stimulated B cells was also significantly reduced by cobalt-chromium particles after 3 weeks and by titanium particles at 8 and 12 weeks. The cytokine release by lymphocytes, proliferation of T and B cells, and immunoglobulin production by B cells were also significantly inhibited by titanium and cobalt-chromium particles, as well as by titanium, cobalt, and chromium ions in vitro, whereas these metals are not cytotoxic to murine lymphocytes in vitro. The data indicate that the metal-induced immunosuppression may be another important factor in the development of implant-associated infection in patients with a prosthesis.


Use of topically applied silver sulphadiazine plus cerium nitrate in major burns.
All patients hospitalized between May 1987 and June 1988 suffering from burns covering over 50 per cent of the body surface area were treated by topical application of a cream containing cerium nitrate (0.05 M) and silver sulphadiazine (0.03 M) (CN + SSD). Eleven patients were included in this series, with a mean age of 35 years (range 22-65), a mean total burn size of 78 per cent (range 50-96 per cent) and full skin thickness covering a mean of 48 per cent (range 10-91 per cent). Eight patients survived (73 per cent) (mean age 36 years; mean total burn surface 73 per cent; mean full skin thickness burn surface, 38 per cent). These results are far better than those obtained in our Unit where a survival rate of 34 per cent was obtained in a comparable series of patients treated before 1987. Sixty positive blood cultures were obtained, which included a large variety of organisms with a slight predominance of Staph. aureus, Candida albicans and Ps. aeruginosa. Wound cultures were positive in 72 per cent of swabs and showed a predominance of Ps. aeruginosa (59 per cent of all the strains isolated). Even if CN + SSD appears in this series not to be very efficient in preventing wound colonization and septic complications, it permitted a very high survival rate in the treated patients, taking into account the extreme severity of the injuries. This beneficial effect is probably the consequence of the protective action of the yellow-green eschar formed by CN + SSD.(ABSTRACT TRUNCATED AT 250 WORDS)


Fabrication of chitosan nanoparticles with phosphatidylcholine for improved sustain release, basolateral secretion, and transport of lutein in Caco-2 cells.
Biopolymers-based nanoparticles delivery emerged alternatively to improve nutraceuticals and drug bioavailability. The intestinal physiology suggested a prerequisite of lipid moiety for carotenoid absorption. This study aimed to fabricate chitosan-based nanoparticles with phosphatidylcholine (PC) to enhance lutein bioavailability. Lutein encapsulated chitosan nanoparticles with PC (LCNPC) or without PC (LCN) were assessed for bioaccessibility, sustain release, cellular uptake/internalization, and basolateral secretion of lutein in Caco-2 cells. Standard lutein mixed micelles (LMM), and micelles derived through in vitro digestion of green leafy vegetables (GMM) treated as controls. The LCNPC showed reduced particle size, higher colloidal stability, homogeneous dispersion, and suitable for oral administration compared to LCN. The cellular uptake of lutein (20 h) in LCNPC was higher than LCN, LMM, and GMM, respectively. Interestingly, lutein uptake was maximum at 8 h in LMM and gradually decreased against sustain-release response in LCNPC and LCN, whereas considerably low lutein uptake from GMM at all time points. Further, LCNPC significantly increased basolateral secretion of triglyceride (TG) and positively correlated enhanced lutein uptake/internalization process than LCN and micelles. Also, LCNPC demonstrated the upregulation of endocytosis, paracellular, scavenger receptor class B type 1 (SRB-1), and peroxisome proliferator-activated receptor gamma (PPARγ) mediated lutein transport mechanism. These results suggested that fabrication of biopolymer-based nanoparticles with PC could provide greater insight to improve lutein bioavailability at enterocyte levels, to avoid age-related macular degeneration and other chronic diseases.


Psychosocial issues for children and adolescents with chronic illness: self-esteem, school functioning and sports participation.
Self-esteem, school functioning, and sports participation are among the most significant psychosocial issues that affect children and adolescents with chronic illness. Although these capacities are essential components of development for all children, they present special concerns for children with limitations of health. Parents, teachers, and coaches play important roles in providing normalizing and gratifying opportunities for children who struggle to be competent and accepted by their peers. Much can be done to provide chronically ill children with experiences and support that will allow them to grow up happier, feel better about themselves, and enjoy more success.


Antioxidant Activity of Ruthenium Cyclopentadienyl Complexes Bearing Succinimidato and Phthalimidato Ligands.
In these studies, we investigated the antioxidant activity of three ruthenium cyclopentadienyl complexes bearing different imidato ligands: (η5-cyclopentadienyl)Ru(CO)2-N-methoxysuccinimidato (1), (η5-cyclopentadienyl)Ru(CO)2-N-ethoxysuccinimidato (2), and (η5-cyclopentadienyl)Ru(CO)2-N-phthalimidato (3). We studied the effects of ruthenium complexes 1-3 at a low concentration of 50 µM on the viability and the cell cycle of peripheral blood mononuclear cells (PBMCs) and HL-60 leukemic cells exposed to oxidative stress induced by hydrogen peroxide (H2O2). Moreover, we examined the influence of these complexes on DNA oxidative damage, the level of reactive oxygen species (ROS), and superoxide dismutase (SOD) activity. We have observed that ruthenium complexes 1-3 increase the viability of both normal and cancer cells decreased by H2O2 and also alter the HL-60 cell cycle arrested by H2O2 in the sub-G1 phase. In addition, we have shown that ruthenium complexes reduce the levels of ROS and oxidative DNA damage in both cell types. They also restore SOD activity reduced by H2O2. Our results indicate that ruthenium complexes 1-3 bearing succinimidato and phthalimidato ligands have antioxidant activity without cytotoxic effect at low concentrations. For this reason, the ruthenium complexes studied by us should be considered interesting molecules with clinical potential that require further detailed research.


Acute subarachnoid haemorrhage: 3D time-of-flight MR angiography versus intra-arterial digital angiography.
To evaluate the efficacy and reliability of 3D time-of-flight MR angiography (TOF MRA) as a noninvasive procedure, 27 patients with acute subarachnoid haemorrhage (SAH) were studied with MRA immediately before or after intra-arterial digital subtraction angiography (DSA).3DTOF MRA was performed with an axial slab of 60 mm centered on the circle of Willis and isotropic voxels. DSA showed 22 aneurysms and 1 dural arteriovenous fistula in 21 patients; the aneurysms ranged in size from 2 to 8mm. MRA failed to show 2 small aneurysms, at the origin of the posterior and anterior communicating arteries. The 3D display of the intracranial vessels obtained with maximum intensity projection (MIP) or targetted MIP sometimes rendered the aneurysms better than DSA. However, due to its high spatial resolution, DSA more clearly defined the overall anatomy of the walls of the normal and abnormal vessels.


Antiproliferative, antiangiogenic and apoptotic effect of new hybrids of quinazoline-4(3H)-ones and sulfachloropyridazine.
Three new sets of quinazolinones bearing sulfachloropyridazine 4a-f, 6a-i and 8a-i were designed and synthesized. All the synthesized compounds were screened for their in vitro cytotoxicity against a panel of 60 cancer cell lines. The most potent compounds 4b, 4d, 6f, 6g, 8c, 8f and 8g were evaluated as VEGFR-2 inhibitors. Compounds 8f, 8c and 6f were the most active with IC50 = 66 ± 0.002, 108 ± 0.004 and 146 ± 0.006 nM, respectively. Compound 8f showed also moderate inhibition against PDGFR (IC50 = 180 ± 0.009 nM), EGFR (IC50 = 98 ± 0.004 nM), FGFR-1 (IC50 = 82 ± 0.004 nM) and ability to reduce migration of cells in wound healing assay. Compound 8f showed cell cycle arrest at S-phase and induced early and late apoptosis in Annexien V-FITC assay. In addition, compound 8f increased the level of caspase-3 and up regulate Bax expression and down regulate Bcl-2 in UO-31 cells. The cytotoxicity of compounds 6f, 6g and 8f against UO-31 and melanoma cells was slightly affected by combination with γ-radiation. Also, compound 8f showed low toxicity against human normal renal (RPTEC) cell line. Docking studies of the most potent compounds 4b, 4d, 6f, 6g, 8c, 8f and 8g were performed to have more insights on their binging mode within VEGFR-2 active site.


Pediatric vasculitis.
The primary systemic vasculitides in childhood are quite rare except for Henoch-Schönlein purpura and Kawasaki disease. These are usually self-limited diseases. The etiopathogenesis for most of them is not clearly understood. However, it is usually thought that environmental triggers (mostly infectious) evoke an aggravated inflammatory response in susceptible individuals. The classification criteria for most of them were recently validated for children. Clinical manifestations are determined by the organs affected, reflecting the involved vessels on a background of constitutional symptoms. For treatment, the main data are adapted from adult studies, especially for rarer vasculitides in childhood. In this review, the recent classification criteria, current pathophysiologic pathways, and treatment protocols are summarized.


[Analysis of the quality of surgical treatment of colorectal cancer, in 2008. A national study].
A national study conducted for the Spanish Association of Surgeons with the aim of analysing the surgical treatment of colorectal cancer (CRC) in Spain and to compare it with scientific literature. A multicentre, descriptive, prospective and longitudinal study of patients with CRC who were treated by elective surgery. A total of 50 hospitals in 15 Autonomous Regions took part, with 496 treated cases in 2008. A total of 88 variables were collected. The median age was 72 years, increase in ASA III patients; correct preoperative studies, 4% with no staging in the rectum. There was a tendency not to use the colon cleansing or to do it only one day. The percentage of complications is within the ranges in the literature, with the exception of surgical wound infections (19%). Mean of resected lymph nodes: 13.2; 4.3% no mesorectal resection. Mechanical anastomosis: 80.8%, 65.9% of the operations performed by a colorectal surgeon. Preoperative radiotherapy in 43.5% of rectal cancers. Chemotherapy: 32.9%. Laparoscopy: 35.1% of cases, conversion rate 13.8%. Use of antibiotics: 37.1%, blood transfusion: 20.6% and parenteral nutrition: 26.5%. Surgical treatment of CRC in Spain has a level of quality and peri-operative results similar to the rest of Europe. Compared to previous studies, it was observed that there were advances in preparation of the patient, preoperative studies, imaging techniques, and improvements in surgical techniques with adoption of mesorectal excision, appropriate lymphadenectomies and preservation of sphincters. There are areas for improvement, such as a reduction in surgical wound infections, increase use of protective stoma, appropriate use of antibiotics, parenteral nutrition or neoadjuvants and complete colonoscopies.


Conserved epitopes with high HLA-I population coverage are targets of CD8+ T cells associated with high IFN-γ responses against all dengue virus serotypes.
Cytotoxic CD8+ T cells are key for immune protection against viral infections. The breadth and cross-reactivity of these responses are important against rapidly mutating RNA viruses, such as dengue (DENV), yet how viral diversity affect T cell responses and their cross-reactivity against multiple variants of the virus remains poorly defined. In this study, an integrated analysis was performed to map experimentally validated CD8+ T cell epitopes onto the distribution of DENV genome sequences across the 4 serotypes worldwide. Despite the higher viral diversity observed within HLA-I restricted epitopes, mapping of 609 experimentally validated epitopes sequences on 3985 full-length viral genomes revealed 19 highly conserved epitopes across the four serotypes within the immunogenic regions of NS3, NS4B and NS5. These conserved epitopes were associated with a higher magnitude of IFN-γ response when compared to non-conserved epitopes and were restricted to 13 HLA class I genotypes, hence providing high coverage among human populations. Phylogeographic analyses showed that these epitopes are largely conserved in most of the endemic regions of the world, and with only some of these epitopes presenting distinct mutated variants circulating in South America and Asia.This study provides evidence for the existence of highly immunogenic and conserved epitopes across serotypes, which may impact design of new universal T-cell-inducing vaccine candidates that minimise detrimental effects of viral diversification and at the same time induce responses to a broad human population.


Low quality diet and challenging temperatures affect vital rates, but not thermal tolerance in a tropical insect expanding its diet to an exotic plant.
Determining responses of organisms to changing temperatures is a research priority, as global warming threatens populations and ecosystems worldwide. Upper thermal limits are frequently measured as the critical thermal maximum (CTmax), a quick bioassay where organisms are exposed to increasing temperatures until individuals are not able to perform basic motor activities such as walking or flying. A more informative approach to understand organism responses to global warming is to evaluate how vital rates, such as growth or survival, change with temperatures. The main objectives of this study are: (1) to determine if factors affecting insect vital rates such as diet quality, developmental temperatures or acclimation also affect CTmax and (2) to determine if vital rates of different life stages (i.e., insect larvae or adults) display different responses to temperature changes. If different life stages have particular thermal requirements, this may indicate different susceptibility to global warming. This study focuses on Cephaloleia belti (Coleoptera: Chrysomelidae), a tropical insect currently expanding its diet to an exotic host plant. We determined how high and low-quality diets (i.e., native vs novel host), as well as exposure temperatures affect CTmax of adult beetles. We also estimated larval and adult survival when feeding on high and low quality host plants, when exposed to temperatures typical of the elevational distribution of this species, or when exposed to projected temperatures in 100 years. We did not detect an effect of diet quality or acclimation on CTmax. However, larvae and adults had different thermal requirements. CTmax is not affected by previous diet or acclimation as an adult. We propose that to understand processes involved in the adaptation and persistence of ectotherm populations in a warming world, studies must explore responses beyond CTmax, and focus on the response of vital rates to changing temperatures.


Gestational trophoblastic disease and associated factors among women experiencing first trimester pregnancy loss at a regional referral hospital in central Tanzania: a cross-sectional study.
Gestational trophoblastic diseases (GTDs) may follow any form of pregnancy or a pregnancy loss. Early detection of GTDs is important, as some benign forms of the disease may progress into a chemoresistant and metastatic disease. This study aimed at determining the frequency of GTDs among women experiencing first trimester pregnancy loss and the associated patients' characteristics. This was a cross-sectional study that included 200 conveniently sampled women who experienced first trimester pregnancy loss from January to December 2019 at a Regional Referral Hospital in central Tanzania. The specimen obtained from products of conception were collected, formalin-fixed and paraffin-embedded and submitted for histopathological evaluation, for which haematoxylin and eosin stain was used. Data were analysed using SPSS version 23.0. The χ2 test was used to determine the association between categorical variables. p-Values ˂0.05 were considered statistically significant. Among 200 study participants, the overall frequency of GTDs was 42 (21%). Among those with GTDs, the most common histopathological diagnosis was partial hydatidiform mole (18 [42.9%]), followed by complete hydatidiform mole (17 [40.5%]) and choriocarcinoma (7 [16.5%]). In the studied participants, only increased human chorionic gonadotropin hormone levels were found to be statistically significantly associated with GTDs (p=0.000). Results from this study suggest that routine histopathological evaluation of the products of conception is recommended in order to allow early detection of GTDs, including choriocarcinoma, which usually carries a poor prognosis. The histopathological reporting of choriocarcinoma among first trimester products of conception from Tanzania is novel.


Comparing efficacy of three methods of tranexamic acid administration in reducing hemoglobin drop following total knee arthroplasty.
The ideal method of providing tranexamic acid (TXA) for decreasing hemoglobin drop after TKA is still controversial. In this clinical trial, 200 patients were randomly allocated to four groups. In group 1,500 mg TXA was administered intravenously. In group 2, the joint irrigated with 3 g of TXA in 100 cc of saline. In group 3, 1.5 g of TXA was injected through the drain. Group 4 did not take TXA. Albeit all methods had a statistical effect on hemoglobin drop, drainage and number of transfused units when compared to controls, but intravenous injection of TXA seems to be much more effective in terms of reducing hemoglobin drop and transfused units; and what's more TXA injection by drain is more effective regarding to reducing postoperative drainage.


T2 signal intensity and volume abnormalities of hippocampal subregions in patients with amnestic mild cognitive impairment by magnetic resonance imaging.
The volumetry of the hippocampal subregion may provide additional information in the early investigation of amnestic mild cognitive impairment (aMCI) and the T2 signal intensity (T2-SI) of the hippocampal subregion has not been well studied quantitatively by magnetic resonance imaging (MRI) in aMCI. Using combined MRI-based hippocampal volumetry and T2-SI at the levels of the whole hippocampus and hippocampal subregion, 18 patients with aMCI and 18 age-matched controls were investigated. Significantly lower left whole hippocampal and hippocampal head volumes and higher T2-SI in the bilateral whole hippocampus and hippocampal head were shown, whereas atrophy of the right whole hippocampus and hippocampal subregion was not significant in aMCI. Additionally, correlations were found among the hippocampal volume, T2-SI and Mini-Mental State Examination (MMSE) scores for aMCI in the whole hippocampus and some hippocampal subregions and an almost perfect correlation was found between T2-SI of the left hippocampal head and MMSE scores regarding aMCI (r = -0.831, P = 0.000). Abnormalities of the hippocampal volume and T2-SI were documented in aMCI, whereas T2-SI was implied to be more susceptible than the volume in the pathohistological progression in aMCI. Additionally, T2-SI in the left hippocampal head may be a potential biomarker to facilitate the early diagnosis of aMCI.


Induced Hepatic Glutathione and Metabolomic Alterations Following Mixed Pesticide and Fertilizer Exposures in Juvenile Leopard Frogs (Lithobates sphenocephala).
The increasing use of agrochemicals, alone and in combination, has been implicated as a potential causative factor in the decline of amphibians worldwide. Fertilizers and pesticides are frequently combined into single-use tank mixtures for agricultural applications to decrease costs while meeting the food demands of a growing human population. Limited data are available on the effects of increased nitrogen levels in nontarget species, such as amphibians, and therefore investigating alterations in the nitrogen cycle and its impacts on amphibians needs to be considered in best management practices going forward. The objective of the present study was to elucidate the impact of fertilizer (urea) and herbicide (atrazine and/or alachlor) tank mixtures on the hepatic metabolome of juvenile leopard frogs as well as to investigate alterations in oxidative stress by relating these changes to glutathione (GSH) levels. Herbicide exposure only moderately increased this parameter in amphibians, however, urea alone and in combination with either atrazine or alachlor statistically elevated GSH levels. Interestingly, urea also inhibited pesticide uptake: calculated bioconcentration factors were greatly decreased for atrazine and alachlor when urea was present in the exposure mixture. Metabolomic profiling identified fluxes in hepatic metabolites that are involved in GSH and carbohydrate metabolic processes as well as altered intermediates in the urea cycle. Ultimately, understanding the biological impacts of nitrogenous fertilizers alone and in combination with pesticide exposure will inform best management practices to conserve declining amphibian populations worldwide. Environ Toxicol Chem 2022;41:122-133. © 2021 SETAC.


Functional characterization of the human atrial essential myosin light chain (hALC-1) in a transgenic rat model.
Most patients with hypertrophic cardiomyopathy and congenital heart diseases express the atrial essential myosin light chains (ALC-1) in their ventricles, partially replacing the ventricular essential light chains (VLC-1). This VLC-1/ALC-1 isoform shift is correlated with an increase in cross-bridge cycling kinetics as measured using skinned fibers from the hypertrophied ventricles of human hearts. To study the functional importance of hALC-1 in the intact perfused heart, we generated a transgenic rat model (TGR) overexpressing hALC-1 in the heart. Twelve-week-old TGR rats expressed 17 +/- 4 microg hALC-1 per mg of whole SDS-soluble protein. Their perfused heart contractility parameters were evaluated using the Langendorff preparation. Expression of hALC-1 was accompanied by statistically significant improvements (P<0.001) in the contractile parameters of the hearts of the TGR compared to the age matched control (WKY) animals, represented by increases from 20.8 +/- 2.3 to 45.1 +/- 3.6 mmHg/g heart weight in the developed left ventricular pressure, 1,035.7 +/- 89.8 to 2,181 +/- 135.4 mmHg/s in the contraction rate, and 713 +/- 60.2 to 1,364 +/- 137.4 mmHg/s in the relaxation rate in the WKY and the TGR groups respectively. Characterizing the functional effects of hALC-1 at the whole organ level represents a step towards gene therapy of heart failure.


Multi-step neoplastic transformation of normal human fibroblasts by Co-60 gamma rays and Ha-ras oncogenes.
As reported previously (Namba et al., 1985; Namba, 1985), normal human fibroblasts were transformed into immortal cells with abnormal karyotypes by Co-60 gamma-ray irradiation. These immortally transformed cells (KMST-6) showed no clonability in soft agar and were not tumorigenic. However, by treatment with Ha-ras oncogenes derived from a human lung carcinoma or Harvey murine sarcoma virus, the KMST-6 cells acquired elevated clonability in soft agar and transplantability in nude mice. All the tumors produced grew progressively without showing regression and killed the mice. The tumors were also serially transplantable into other mice. The Ha-ras oncogene alone did not convert normal human fibroblasts into either immortal or tumorigenic cells. Our current data suggest that gamma rays worked as an initiator of carcinogenesis in normal human cells, giving rise to chromosome aberrations and immortality, and the Ha-ras oncogene played a role in the progression of the immortally transformed cell population to a neoplastic one showing enhanced colony formation in soft agar and tumorigenicity in nude mice.


Screening of interleukin 17F gene polymorphisms and eight subgingival pathogens in chronic periodontitis in Libyan patients.
Background: Chronic periodontitis (CP) is triggered by periodontal pathogens influenced by genetic and environmental factors. Recent studies have suggested that anti-inflammatory cytokines such as interleukin 17 (IL-17) play a prominent role in the pathogenesis of CP.Aim: This study aimed to investigate the association between eight sub-gingival pathogens and interleukin 17F (IL-17F) gene single nucleotide polymorphisms with CP among Libyans.Materials and Methods: A case-control study was conducted on 100 individuals between the ages of 25-65 years. Species-specific 16S rRNA primers for each pathogen were used in a multiplex PCR reaction to detect sub-gingival pathogens from a paper point sample. DNA was also extracted from buccal swab samples and IL-17F polymorphisms were detected by Sanger sequencing.Results: A highly significant association between the seven sub-gingival pathogens and CP, (p-value 0.0001) and a high prevalence of P. intermedia (100%), T. forsythia (96%), T. denticola and E. corrodens (92%), P. gingivalis (82%), C. rectus (74%), P. nigrescens (72%), A. actinomvcetcmcomitans (40%) were found in the case group compared with control group. A novel variant in the c. *34 G>A in IL-17F gene caused a change in glutamic amino acid to lysine amino acid, position on chromosome number (6) in the third exon, mRNA/genomic position 597, found in 14.6% of CP patients (p-value = 0.010) while the IL-17F (rs763780) SNP showed no association with CP (p-value = 0.334).Conclusion: P. intermedia appear as keystone pathogen for CP in the Libyan population. A novel variant in the IL-7F gene may be related to the severity of CP.


Attitudes towards patient gender among psychiatric hospital staff: results of a case study with focus groups.
There is an increasing awareness of gender-related issues in psychiatry. However, empirical findings on attitudes of psychiatric staff towards patient gender are limited. Gender-related issues are particularly relevant in the debate about mixed versus segregated sex wards, yet while the appropriateness of mixed-sex wards is questioned in Great Britain this is not the case in Germany. To investigate attitudes of psychiatric staff towards both patient gender and mixed versus segregated sex wards, we conducted a case study using focus groups with members of professional teams. We evaluated the transition process from two single-sex wards to two mixed-sex wards in a 330-bed psychiatric hospital in a rural area in south Germany. Staff described female patients as more externally oriented, motivating of others, demanding, and even sexually aggressive. Male patients, on the other hand, were described as more quiet, modest, or lazy. Furthermore, participants described the mixing process as a positive development whereas they did not see a need for gender-separated wards in order to protect vulnerable female patients. Some gender descriptions by professionals are "reversed" in comparison with gender stereotypes supposed to be present in wider society. The perception of crossed gender norms may affect staff attitudes towards the vulnerability of female patients in psychiatric settings and the provision of single-sex wards in in-patient psychiatric care. Practical implications are discussed against the background of a high rate of female patients with sexual abuse histories.


Use of the microalgae Thalassiosira weissflogii to assess water toxicity in the Suape industrial-port complex of Pernambuco, Brazil.
Microalgae are among the organisms that are most frequently recommended to assess water toxicity levels. They provide qualitative and quantitative population changes and have become important in environmental monitoring. The aim of the present study was to assess the sensitivity of the microalgae Thalassiosira weissflogii to water toxicity in the Suape industrial-port complex, and to potassium dichromate in the months of July, September and November of 2010 and January and September of 2011, as well as to compare the microalgae results to those obtained in tests with the sea urchin Lytechinus variegatus. The chronic toxicity tests with both species followed standardized methods. Once the exposure period was completed, the population increase of the algae was assessed using cell density and growth rate data and the percentage of L. variegatus pluteus larvae revealing normal and aberrant development was determined. The algal growth and percentage of pluteus results demonstrated that toxicity in the studied area varied according to the month. The present study confirmed that the best timeframe to conclude tests with the microalgae T. weissflogii was 72 h. The microalgae cell density results after 72 h were sufficient to assess the toxicity of surface water samples and were more sensible than the growth rates. The microalgae T. weissflogii was a sensitive test-organism for seawater toxicity.


Wrist fusion versus limited carpal fusion in advanced Kienbock's disease.
We treated 18 patients with advanced Kienbock's disease surgically. Six had total wrist fusions and 12 had limited carpal fusions. The average age was 39.6 yrs and the average follow up was 61.8 months and 66.8 months respectively. The visual analogue pain scores, the patient satisfaction scores and the SF 12 were better in the total wrist fusion group. The DASH (Disabilities of the Arm, Shoulder and Hand) scores, the range of movement and the grip strengths were better in the limited carpal fusions group but this was not statistically significant. Four patients with limited carpal fusions had a non-union that required revising. We believe that total wrist fusion should be offered earlier to patients with advanced stages of the disease, as there are less surgical failures, more satisfied patients, better post operative pain scores and consistent long-term results with less potential for further deterioration with time as compared to other treatment methods.


Thyroid adenylate cyclase stimulating immunoglobulins in thyroid diseases.
The occurrence of serum immunoglobulins with capacity to stimulate thyroid adenylate cyclase (TSAb) was studied in seventy-two healthy volunteers and 120 unselected patients with various thyroid diseases. A high frequency of TSAb (82.5%, P less than 0.00006) was found in Graves' disease, while TSAb was present only in 13--20% of serum from patients with nontoxic nodular goitre, nontoxic diffuse goitre, toxic adenoma, toxic nodular goitre and myxoedema. These patients had low level of TSAb compared to patients with Graves' disease. In patients with Graves' disease there was no correlation between the level of TSAb and hormonal status except serum triiodothyronine (rs = 0.29, P less than 0.05), and no relation with eye involvement or presence of microsomal thyroid antibodies was found. The results indicate that the human thyroid adenylate cyclase assay system with 1 hour incubation periods is a sensitive method for detection of immunoglobulins with TSH-like capacity to stimulate the thyroid gland.


Intracavernous Alprostadil Alfadex--an effective and well tolerated treatment for erectile dysfunction. Results of a long-term European study.
Long-term efficacy and safety of Alprostadil-Alfadex (EDEX/VIRIDAL) in intracavernous self-injection therapy for chronical erectile failure was investigated in a four year running multicenter European trial. Of the 16,886 protocolled injections 93% (15,713) resulted in rigid erections followed by successful sexual intercourse. Reported side effects by patients were prolonged erections > 6 h only occurring during the first year in 1.2% (2 out of 162), painful erections in 29% (47 out of 162) during the first year and decreasing to 12.1% by year 4, hematomas, neither requiring therapeutic measures nor impeding sexual performance in 33.3% (54 out of 162) in year 1 with a decrease to 12.1% by year 4, fibrotic penile alterations such as nodules, plaques or deviations in 11.7% (19 out of 162) with spontaneous healing in 48% (9 out of 19). Of the 162 patients involved in this trial 54 completed the 4 y. Of the 54 completers 91.4% considered the tolerability good or very good and were satisfied or very satisfied with self-injection therapy with Alprostadil-Alfadex. The respective rates of the female partners were 51.7% very satisfied and 39.7% satisfied. These data of the world-wide longest running prospective trial with a vasoactive drug in self-injection therapy provided impressive proof that Alprostadil-Alfadex represents a very effective and safe treatment for erectile dysfunction of both psychogenic and organogenic origin.


Histochemical and ultrastructural modifications of mice endometrium, vagina and pituitary following zeranol treatment.
The histochemical and ultrastructural changes produced by zeranol treatment on the endometrium and the gonadotropic FSH cells of the adenohypophysis have been investigated in mice at the prepuberal and virginal stages. Modifications similar to those induced by estrogen treatment were observed. It is concluded that both estrogens and zeranol share the same activity on the tissues examined.


Safety and immunogenicity of tyrosinase DNA vaccines in patients with melanoma.
Immunity to self antigens on cancer is constrained by tolerance/ignorance. DNA vaccines encoding xenogeneic differentiation antigens, such as tyrosinase (TYR), mediate tumor protection and regression in implantable mouse models, and dogs with spontaneous melanoma. We conducted a trial of mouse and human TYR DNA vaccines in stage III/IV melanoma patients. Eighteen human leukocyte antigen (HLA)-A*0201(+) melanoma patients were randomized as follows: one group received three mouse TYR DNA injections followed by three human TYR DNA injections; the other group received the same vaccines in opposite sequence. The study was conducted at three dose levels: 100, 500, and 1,500 microg DNA/injection, administered intramuscularly (IM) every 3 weeks. Most toxicities were grade 1 injection site reactions. Seven patients developed CD8(+) T-cell responses, defined by a >3 SD increase in baseline reactivity to TYR peptide in tetramer or intracellular cytokine staining (ICS) assays. There was found to be no relationship between dose, assigned schedule, and T-cell response. At a median of 42 months follow-up, median survival has not been reached. Mouse and human TYR DNA vaccines were found safe and induced CD8(+) T-cell responses in 7 of 18 patients. T cells recognizing a native TYR peptide had a phenotype consistent with that of effector memory cells.


Two possible orientations of the HI-6 molecule in the reactivation of organophosphate-inhibited acetylcholinesterase.
The inhibition of acetylcholinesterase (AChE) by organophosphorus compounds (OPs) causes acute toxicity or death of the intoxicated individual. One group of these compounds, the OP nerve agents, pose an increasing threat in the world due to their possible use in the battlefield or terrorist acts. Antidotes containing oxime compounds to reactivate the inhibited enzyme are highly valued for treatment against OP poisoning. One of these reactivators, HI-6, was shown to be significantly more effective in treating soman toxicity than other oximes, such as 2-PAM, TMB4, and obidoxime. However, HI-6 was less effective in reactivating AChE inhibited by the OP pesticide, paraoxon. In this study, the mechanism for HI-6-induced reactivation of OP-AChE conjugates was investigated using mouse mutant AChEs inhibited with different OPs including organophosphate paraoxon, and several methylphosphonates. Results indicate that the HI-6 molecule may assume two different orientations in the reactivation of AChE inhibited by organophosphate and Sp methylphosphonates. These conclusions were further corroborated by reactivation studies using an analog of HI-6 in which the bispyridinium moieties are linked by a methylene bridge rather than an ether oxygen.


A bibliometric analysis of the application of solar energy to the organic Rankine cycle.
Solar Thermal is one of many heat sources that support the organic Rankine cycle (ORC) system. The advantage of using solar thermal as a heat source when combined with the ORC system is that it can provide affordable energy supplies in remote areas and it is suitable in disaster territories. Growth in solar-ORC research is not only on a lab/workshop scale but also on an industrial scale. Most articles refer to methods and tools in the design process, combining heat and improvement of the cycle, working fluids selection, and case studies based on primary or secondary data. The paper reports the newest trends on the research of solar application on ORC using bibliometric analysis. The study of bibliometric using VOSviewer was recognized as a captivating method in the literature that allows examining the scientific progress of a particular topic. Moreover, a qualitative approach can be performed by analyzing the keywords contained in different documents to identify a circumstance that features the research trend and to understand future viewpoints.


Not all that glistens is gold: civilian white phosphorus burn injuries.
White phosphorus is an incendiary agent used in particular types of ammunition. Exposure to phosphorus-containing compounds may cause severe burn injuries. Systemic effects may be fatal for the casualty even though only a small surface of the body is burned. We report 4 cases of white phosphorus burn injuries due to an exceptional accident mechanism. All casualties were holidaymakers in a coastal area at the Baltic Sea and confused white phosphorus ammunition residues with amber on a beachwalk. The supposed treasure was taken into the pants pocket. The phosphorus substances ignited spontaneously when they dried because of the body heat. Injury patterns combined simultaneous full thickness burn injuries of both hands with a burn injury of one thigh or the abdominal region. The acute treatment and the controversy surrounding decontamination of such patients are discussed. In addition, pathology of tissue damage and systemic injury are elucidated.


Colloidal silver ingestion with copper and caeruloplasmin deficiency.
The copper concentration in serum can be affected by the presence of other trace elements such as silver. Low serum copper may result in decreased caeruloplasmin synthesis. We report the case of a 59-year-old woman, who was admitted to hospital with acute psychosis and who had been ingesting chronically, colloidal silver.


Quenching of chlorophyll fluorescence induced by silver nanoparticles.
The interaction between chlorophyll (Chl) and silver nanoparticles (AgNPs) was evaluated by analyzing the optical behavior of Chl molecules surrounded by different concentrations of AgNPs (10, 60, and 100nm of diameter). UV-Vis absorption, steady state and time-resolved fluorescence measurements were performed for Chl in the presence and absence of these nanoparticles. AgNPs strongly suppressed the Chl fluorescence intensity at 678nm. The Stern-Volmer constant (KSV) showed that fluorescence suppression is driven by the dynamic quenching process. In particular, KSV was nanoparticle size-dependent with an exponential decrease as a function of the nanoparticle diameter. Finally, changes in the Chl fluorescence lifetime in the presence of nanoparticles demonstrated that the fluorescence quenching may be induced by the excited electron transfer from the Chl molecules to the metal nanoparticles.


Platelet life span in pregnancies resulting in small-for-gestational age infants.
Platelet number and life span were determined in the last trimester of pregnancy in 22 women who were delivered of small-for-gestational age (SGA) infants and in 21 women with infants having normal birth weights. Platelet life span was determined by means of a nonradioisotopic method involving the use of acetylsalicylic acid. Mean platelet life span was 7.2 days in women with SGA infants, which is significantly shorter than the 9.2 days found in the control group. The mean platelet count in the SGA group was 235 x 10(9)/L, whereas in the control group it was 208 x 10(9)/L. These data indicate the existence of a compensated thrombocytolytic state in pregnancies with insufficient fetal growth. This could be explained by an increased platelet consumption in the uteroplacental arterial bed.


Implementing Geospatial Science and Technology to Get to Zero New HIV Infections.
Tremendous advancements have been made in HIV treatment and prevention during the last 40 years that zero new HIV cases has become an attainable goal declared by international agencies. However, new cases of HIV infection persist. The emerging field of geospatial science is positioned to play key role in the reduction of continued HIV incidence through technology-driven interventions and innovative research that gives insights into at-risk populations. As these methods become more utilized, findings consistently show the important role of location and environment plays in HIV incidence and treatment adherence. This includes distance to HIV provider, locations of where HIV transmissions occurs compared to where people with HIV reside, and how geospatial technology has been leveraged to identify unique insights among varying groups of those at increased risk for HIV, among others. Given these insights, leveraging geospatial technology would play a prominent role in achieving zero new cases of HIV infections.


Orexin actions in hypothalamic paraventricular nucleus: physiological consequences and cellular correlates.
Orexinergic neurons originating in the perifornical, lateral hypothalamus project to numerous brain sites including neuroendocrine centers known to be important in the physiologic response to stress. Those projections suggest an action of endogenous orexin on adrenocorticotropin (ACTH) release, either by neuromodulatory effects in the paraventricular nucleus (PVN), or by neuroendocrine actions in the pituitary gland following release into the median eminence. We sought to determine if exogenously applied orexin A might act in the brain to alter ACTH release and to determine if a site of action in the hypothalamic paraventricular nucleus could be identified. Cerebroventricular administration of orexin A in conscious male rats resulted in a dose-related elevation in circulating ACTH levels. At 30 min post-infusion, ACTH levels were elevated 2.5-fold by the low dose of orexin A (0.3 nmol), 5.7-fold by the middle dose tested (1.0 nmol), and 7.5-fold by the highest dose tested (3.0 nmol). Pretreatment with a CRH-antagonist (i.v.) blocked the ability of i.c.v. administered orexin A to activate the hypothalamo-pituitary-adrenal (HPA) axis. Bath application of orexin A in hypothalamic slice preparations resulted in depolarizations (8.0+/-0.6 mV), accompanied by increases in spike frequency in identified magno- and parvocellular neurons in the PVN. Our data suggest a potential role for endogenous orexin in the hypothalamic regulation of stress hormone secretion.


Quality of perinatal death registration. A study in Hainaut, Belgium.
The quality of national perinatal mortality statistics was evaluated from a survey in nine maternity hospitals in Hainaut, Belgium (total births: 7862). The overall completeness of perinatal death registration was 86%. Under-registration was especially frequent in low birth weight babies. In 69% of cases, the birth weight value reported on death certificates was in exact agreement with the value in hospital records. Using detailed categories of causes, there was, in 37% of cases, agreement between the underlying cause on death certificates and the main cause identified in hospital records. Using gross categories of causes, the level of agreement was 56%. Disagreement was mostly due to the lack of specificity of the underlying cause on death certificates. The authors suggest ways to improve the quality of registration.


Purification and characterization of an anti-sticking factor from goat epididymal plasma that inhibits sperm--glass and sperm--sperm adhesions.
An anti-sticking factor (ASF-I) that showed high affinity for inhibiting adhesion of spermatozoa to glass was isolated from goat epididymal plasma and characterized. The factor was purified approx. 5600-fold and showed a single protein band when examined by non-denaturation and SDS-polyacrylamide gel electrophoresis. The molecular mass and S20w value of ASF-I were approx. 47 kDa and 4.25 S. ASF-I at a concentration of 1 nM showed nearly maximal anti-sticking activity when approx. 60% of the intact spermatozoa were prevented from adhesion to glass and it showed a high degree of protein specificity. Studies with trypsin and glycosidases demonstrated that both the sugar and protein parts of the molecule are essential for its anti-sticking activity. Evidence has been presented to support the view that the outer surface of sperm possesses specific ASF-I receptors that bind to 125I-labelled ASF and mediate cell adhesion to glass. ASF-I also showed high affinity for inhibiting agglutination of corpus-epididymal spermatozoa. The ASF activity was found to be distributed in all the tissues tested and its specific activity was markedly higher in blood plasma than in the tissues. The results suggest that ASF may play an important biological role by serving as a specific inhibitor of cell-substratum and cell-cell adhesions.


Encephalitis with coinfection by Jamestown canyon virus (JCV) and varicella zoster virus (VZV).
We present the case of a 59-year-old Midwestern farmer who presented with altered mental status, dysarthria, urinary incontinence, and a right-sided L5 dermatomal rash; he had recently received a course of oral corticosteroids for treatment of radicular low back pain. Lumbar puncture revealed the presence of varicella zoster virus (VZV) and IgM antibodies against a California-group encephalitis virus, later confirmed as Jamestown Canyon virus (JCV). Unfortunately, the patient's health declined despite aggressive treatment, developing progressive subarachnoid hemorrhage. He died after withdrawal of supportive care following 3 weeks in the intensive care unit. To our knowledge, this is the first documented case of encephalitis associated with coinfection by VZV and JCV. While the relative contributions of these viral pathogens to the patient's illness are difficult to ascertain, the clinical features of this case are consistent with co-pathogenesis, possibly driven by antecedent corticosteroid use. This case highlights the emerging role of viral coinfections in the etiology of viral illnesses.


Gender and stress: differential psychophysiological reactivity to stress reexposure in the laboratory.
Research has shown that women are more prone to the development of depression and anxiety disorders throughout their lifetimes. Stress reactivity and adaptation to repeated stressors have been linked to depression and anxiety, but studies examining gender differences in psychophysiological responses to repeated stressors are very limited. This study examined gender differences in response to initial and repeated exposure to a laboratory stressor as well as potential mechanisms for these differences. Participants viewed a Holocaust video on two occasions with a 2-day interval between sessions. Self reported negative affect and cardiovascular reactivity were recorded at both sessions. Although gender differences were not found following initial exposure, women exhibited significantly greater heart rate (HR) and negative affect (NA) reactivity to the second exposure as compared to men. Women also reported significantly greater intrusive thoughts and avoidance after the first exposure than men, but these were not found to be significant mediators. The findings indicate that women may be more vulnerable to repeated stress exposures compared to men suggesting sensitization. The implications of our findings and suggestions for future research are discussed.


Inflammatory panel cytokines are elevated in the neocortex of late-stage Alzheimer's disease but not Lewy body dementias.
Chronically dysregulated neuroinflammation has been implicated in neurodegenerative dementias, with separate studies reporting increased brain levels of inflammatory mediators and gliosis in Alzheimer's disease (AD) as well as in Lewy body dementias (LBD). However, it is unclear whether the nature and extent of neuroinflammatory responses in LBD are comparable to those in AD. In this study, we performed head-to-head measurements of a panel of cytokines in the post-mortem neocortex of AD versus the two major clinical subtypes of LBD, namely, dementia with Lewy bodies (DLB) and Parkinson's disease dementia (PDD). Post-mortem tissues from the mid-temporal cortex (Brodmann area 21) of a cohort of neuropathologically well-defined AD, PDD and DLB patients were processed and measured for a comprehensive range of cytokines (IL-1α, IL-1Ra, IL-8, IL-10, IL-12p70, IL-13, IFN-γ, GM-CSF and FGF-2) using a multiplex immunoassay platform. Associations between inflammation markers and neuropathological measures of neuritic plaques, neurofibrillary tangles as well as Lewy bodies were also performed. We found IL-1α, IFN-γ, GM-CSF and IL-13 to be elevated in the mid-temporal cortex of AD patients. In contrast, none of the measured cytokines were significantly altered in either DLB or PDD. Similar cytokine changes were observed in two other neocortical areas of AD patients. Furthermore, increases of IL-1α, IFN-γ, GM-CSF, IL-10 and IL-13 associated with moderate-to-severe neurofibrillary tangle burden, but not with neuritic plaques or Lewy bodies. Our findings of elevated neocortical pro- and anti-inflammatory cytokines in AD, but not in DLB or PDD, suggest that neuroinflammatory responses are strongly linked to neurofibrillary tangle burden, which is higher in AD compared to LBD. In conclusion, neuroinflammation may not play a prominent role in the pathophysiology of late-stage LBD.


Açai (Euterpe oleracea Mart.) feeding attenuates dimethylhydrazine-induced rat colon carcinogenesis.
This study investigated the protective effect of spray-dried açaí powder (AP) intake on colon carcinogenesis induced by 1,2-dimethylhydrazine (DMH) in male Wistar rats. After 4 weeks of DMH administrations, the groups were fed with standard diet, a diet containing 2.5% or 5.0% AP or a diet containing 0.2% N-acetylcysteine (NAC) for 10 weeks, using aberrant crypt foci (ACF) as the endpoint. Additionally, two groups were fed with standard diet or a diet containing 5.0% AP for 20 weeks, using colon tumors as the endpoint. In ACF assay, a reduction in the number of aberrant crypts (ACs) and ACF (1-3 AC) were observed in the groups fed with 5.0% AP (37% AC and 47% ACF inhibition, p=0.036) and 0.2% NAC (39% AC and 41% ACF inhibition, p=0.042). In tumor assay, a reduction in the number of invasive tumors (p<0.005) and tumor multiplicity (p=0.001) was observed in the group fed with 5.0% AP. Also, a reduction in tumor Ki-67 cell proliferation (p=0.003) and net growth index (p=0.001) was observed in the group fed with 5.0% AP. Therefore the findings of this study indicate that AP feeding may reduce the development of chemically-induced rat colon carcinogenesis.


Analysis of antigenic diversity among human cytomegaloviruses by kinetic neutralization tests with high-titered rabbit antisera.
Neutralizing antisera to human cytomegalovirus were produced in rabbits with alkaline-buffered extracts of infected cell cultures. The antibody activity was complement dependent and associated primarily with the 7S immunoglobulin fraction. Antisera with homologous K values greater than 10.00 were shown to be suitable for neutralization kinetic studies and were so used to examine the antigenic relatedness of strains AD169, Davis, Esp, C-87, Kerr, and Towne. Based upon the degree of relationship as determined by normalized K values, antisera to the Davis or AD169 strains discriminated three antigenic groups, and an antiserum to strain Esp discriminated two groups.


Nuclear hormone and peptide hormone therapeutics for NAFLD and NASH.
Non-alcoholic steatohepatitis (NASH) is a spectrum of histological liver pathologies ranging from hepatocyte fat accumulation, hepatocellular ballooning, lobular inflammation, and pericellular fibrosis. Based on early investigations, it was discovered that visceral fat accumulation, hepatic insulin resistance, and atherogenic dyslipidemia are pathological triggers for NASH progression. As these pathogenic features are common with obesity, type 2 diabetes (T2D), and atherosclerosis, therapies that target dysregulated core metabolic pathways may hold promise for treating NASH, particularly as first-line treatments. In this review, the latest clinical data on nuclear hormone- and peptide hormone-based drug candidates for NASH are reviewed and contextualized, culminating with a discovery research perspective on emerging combinatorial therapeutic approaches that merge nuclear and peptide strategies. Several drug candidates targeting the metabolic complications of NASH have shown promise in early clinical trials, albeit with unique benefits and challenges, but questions remain regarding their translation to larger and longer clinical trials, as well as their utility in a more diseased patient population. Promising polypharmacological approaches can potentially overcome some of these perceived challenges, as has been suggested in preclinical models, but deeper characterizations are required to fully evaluate these opportunities.


Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 1: Alternative mobile phases and fine tuning of the separation.
Cation exchange chromatography (CEX) of therapeutic monoclonal antibodies is generally performed with either salt gradient (MES buffer + NaCl) or using commercial pH gradient buffer. The goal of this study was to find out some alternative buffer systems for CEX separation of mAbs, which may offer alternative selectivity, while maintaining similar peak shapes. Among the new buffers that were tested, (N-morpholino)ethanesulfonic acid (MES) / 1,3-diamino-2-propanol (DAP), and citric acid / 2-(cyclohexylamino)ethanesulfonic acid (CHES) systems were particularly promising, especially when combining them with a moderate salt gradient of NaCl. This two buffer system provides an equivalent or slightly better separation than the standard, mobile phases for therapeutic mAbs. It was also demonstrated that working with salt-mediated pH gradients, allows to extend the possibilities in method development, since the concentration of salt in the mobile phase has a significant impact on selectivity. Using HPLC modeling software (Drylab), it was possible to successfully develop CEX methods for authentic mAb samples within only 6 h, by optimizing the gradient steepness and salt concentration in the B eluent.


Femtosecond Spectroscopy and Nonlinear Optical Properties of aza-BODIPY Derivatives in Solution.
The fast relaxation processes in the excited electronic states of functionalized aza-boron-dipyrromethene (aza-BODIPY) derivatives (1-4) were investigated in liquid media at room temperature, including the linear photophysical, photochemical, and nonlinear optical (NLO) properties. Optical gain was revealed for nonfluorescent derivatives 3 and 4 in the near infrared (NIR) spectral range under femtosecond excitation. The values of two-photon absorption (2PA) and excited-state absorption (ESA) cross-sections were obtained for 1-4 in dichloromethane using femtosecond Z-scans, and the role of bromine substituents in the molecular structures of 2 and 4 is discussed. The nature of the excited states involved in electronic transitions of these dyes was investigated using quantum-chemical TD-DFT calculations, and the obtained spectral parameters are in reasonable agreement with the experimental data. Significant 2PA (maxima cross-sections ∼2000 GM), and large ESA cross-sections ∼10-20 m2 of these new aza-BODIPY derivatives 1-4 along with their measured high photostability reveal their potential for photonic applications in general and optical limiting in particular.


MicroRNA-224-5p nanoparticles balance homeostasis via inhibiting cartilage degeneration and synovial inflammation for synergistic alleviation of osteoarthritis.
MicroRNAs play a crucial role in regulating cartilage extracellular matrix (ECM) metabolism and are being explored as potential therapeutic targets for osteoarthritis (OA). The present study indicated that microRNA-224-5p (miR-224-5p) could balance the homeostasis of OA via regulating cartilage degradation and synovium inflammatory simultaneously. Multifunctional polyamidoamine dendrimer with amino acids used as efficient vector to deliver miR-224-5p. The vector could condense miR-224-5p into transfected nanoparticles, which showed higher cellular uptake and transfection efficiency compared to lipofectamine 3000, and also protected miR-224-5p from RNase degradation. After treatment with the nanoparticles, the chondrocytes showed an increase in autophagy rate and ECM anabolic components, as evidenced by the upregulation of autophagy-related proteins and OA-related anabolic mediators. This led to a corresponding inhibition of cell apoptosis and ECM catabolic proteases, ultimately resulting in the alleviation of ECM degradation. In addition, miR-224-5p also inhibited human umbilical vein endothelial cells angiogenesis and fibroblast-like synoviocytes inflammatory hyperplasia. Integrating the above synergistic effects of miR-224-5p in regulating homeostasis, intra-articular injection of nanoparticles performed outstanding therapeutic effect by reducing articular space width narrowing, osteophyte formation, subchondral bone sclerosis and inhibiting synovial hypertrophy and proliferation in the established mouse OA model. The present study provides a new therapy target and an efficient intra-articular delivery method for improving OA therapy. STATEMENT OF SIGNIFICANCE: Osteoarthritis (OA) is the most prevalent joint disease worldwide. Gene therapy, which involves delivering microRNAs, has the potential to treat OA. In this study, we demonstrated that miR-224-5p can simultaneously regulate cartilage degradation and synovium inflammation, thereby restoring homeostasis in OA gene therapy. Moreover, compared to traditional transfection reagents such as lipofectamine 3000, G5-AHP showed better efficacy in both microRNA transfection and protection against degradation due to its specific surface structure. In summary, G5-AHP/miR-224-5p was developed to meet the clinical needs of OA patients and the high requirement of gene transfection efficiency, providing a promising paradigm for the future application and development of gene therapy.


Prenylated phenylpropanoid compounds from the stem bark of Illicium burmanicum.
Ten new prenylated phenylpropanoid compounds, burmanicumols A-G (1-6) and illifrognones D-G (7-10), and seven known prenylated phenylpropanoid compounds, were isolated from the stem bark of Illicium burmanicum. The structures of the new compounds were determined on the basis of 1D and 2D NMR, HRESIMS, and CD experiments. Compounds 3, 8, 10, and 14 were moderately potent inhibitors of the NF-κB pathway, with IC50 values ranging from 10.31 to 35.16μg/mL tripterygium glycosides and pavlin were used as the positive controls.


Estrogen withdrawal-induced human breast cancer tumour regression in nude mice is prevented by Bcl-2.
We recently showed that estrogen induces expression of the anti-apoptotic protein, Bcl-2 in MCF-7 human breast cancer cells. Since estrogen-dependent breast tumours can regress following estrogen withdrawal, we hypothesized that stable Bcl-2 expression would prevent estrogen-withdrawal induced regression of MCF-7 tumours. We therefore established tumours in ovariectomized female nude mice implanted with an estrogen-release pellet using untransfected MCF-7 cells or MCF-7 cells stably transfected with a Bcl-2 cDNA sense or antisense expression vector. All tumours grew at similar rates indicating that Bcl-2 levels have no effect on tumour formation. After removal of the estrogen pellet, Bcl-2 antisense tumours and untransfected MCF-7 tumours regressed means of 49% and 52%, respectively, after estrogen pellet removal whereas Bcl-2 sense tumours were significantly stabilized. Regressing tumours displayed characteristics of apoptotic cells. These results show that Bcl-2 can prevent hormone-dependent breast tumour regression and are consistent with the notion that decreased Bcl-2 levels following estrogen withdrawal renders hormone-dependent breast tumour cells sensitive to apoptotic regression.


Reactive architecture profiling with a methyl acyl phosphate electrophile.
Activity-based protein profiling has facilitated the study of the activity of enzymes in proteomes, inhibitor development, and identification of enzymes that share mechanistic and active-site architectural features. Since methyl acyl phosphate monoesters act as electrostatically selective anionic electrophiles for the covalent modification of nucleophiles that reside adjacent to cationic sites in proteins, we synthesized methyl hex-5-ynoyl phosphate (MHP) to broadly target such protein architectures. After treating the soluble proteome of Paucimonas lemoignei with MHP, biotinylating the resulting acylated proteins using click chemistry, enriching the protein adducts using streptavidin, and analyzing the proteins by LC-MS/MS, a set of 240 enzymes and 132 non-enzyme proteins were identified for a wide spectrum of biological processes and from all 7 enzyme classes. Among those enzymes identified, β-hydroxybutyrate dehydrogenase (PlHBDH) and CTP synthase (E. coli orthologue, EcCTPS) were purified as recombinant enzymes and their rates of inactivation and sites of modification by MHP and methyl acetyl phosphate (MAP) were characterized. MHP reacted more slowly with these proteins than MAP but exhibited greater specificity, despite its lack of multiple binding determinants. Generally, MAP modified more surface residues than MHP. MHP specifically modified Ser 146, Lys 156, and Lys 163 at the active site of PlHBDH. MHP and MAP modified numerous residues of EcCTPS with CTP furnishing the greatest level of protection against MHP- and MAP-dependent modification and inactivation, respectively, followed by ATP and glutamine. Overall, MHP served as an effective probe to identify proteins that are potentially amenable to inhibition by methyl acyl phosphates.


Segmentation of brain MRI in young children.
This article deals with an automatic tissue segmentation of brain magnetic resonance imaging (MRI) in young children. We examine the suitability of state-of-the-art methods developed for the adult brain when applied to the segmentation of the brain MRI in young children. We develop a method of creation of a population-specific atlas in young children using a single manual segmentation. The method is based on nonlinear propagation of the segmentation into population and subsequent affine alignment into a reference space and averaging. Using this approach, we significantly improve the performance of the popular expectation-maximization algorithm on brain MRI in young children. The method can be used for building probabilistic atlases with any number of structures. We compare resulting algorithm with nonrigid registration-based label propagation. Finally, both methods are used to measure the volume of seven brain structures and measure the growth between 1 and 2 years of age.