Unilateral acute retinal necrosis occurring 2 years after herpes simplex type 1 encephalitis.
Acute retinal necrosis (ARN) syndrome is known to occur occasionally in association with or shortly after herpetic encephalitis. We describe a patient with unilateral ARN occurring 2 years after herpes simplex virus (HSV) type 1 encephalitis. A 49-year-old man presented with unilateral visual loss. He had a history of HSV-1 encephalitis 2 years previously, and had been successfully treated without a subsequent recurrence. The ophthalmologic findings were all consistent with the ARN syndrome. The same type 1 HSV was identified from ocular fluid as was identified from the patient's cerebrospinal fluid 2 years ago. There was no evidence of a recurrence of encephalitis. This case suggests that following encephalitis retinal neurons may function as a reservoir for latent HSV-1, that can be reactivated to cause ARN in situ several years later.


The Cultivation of Pure Altruism via Gratitude: A Functional MRI Study of Change with Gratitude Practice.
Gratitude is an emotion and a trait linked to well-being and better health, and welcoming benefits to oneself is instrumentally valuable. However, theoretical and empirical work highlights that gratitude is more fully understood as an intrinsically valuable moral emotion. To understand the role of neural reward systems in the association between gratitude and altruistic motivations we tested two hypotheses: First, whether self-reported propensity toward gratitude relates to fMRI-derived indicators of "pure altruism," operationalized as the neural valuation of passive, private transfers to a charity versus to oneself. In young adult female participants, self-reported gratitude and altruism were associated with "neural pure altruism" in ventromedial prefrontal cortex (VMPFC) and nucleus accumbens. Second, whether neural pure altruism can be increased through practicing gratitude. In a double-blind study, we randomly assigned participants to either a gratitude-journal or active-neutral control journal group for 3 weeks. Relative to pre-test levels, gratitude journaling increased the neural pure altruism response in the VMPFC. We posit that as a context-dependent value-sensitive cortical region, the VMPFC supports change with gratitude practice, a change that is larger for benefits to others versus oneself.


Comparing two instructional methods of role playing and lecture on primigravida females, decision about type of delivery.
Each mother has the legal right to decide about her delivery, but this decision should be made based on scientific knowledge. Instructions during pregnancy help to choose the proper type of delivery. This study conducted aimed to compare two instructional methods of role playing and lecture on primigravida decision about type of delivery. In this single-blind clinical trial 67 primigravida, 34-36 week were selected using multi-stage sampling and assigned into two groups randomly. Decision-making (before, 2-week after, and at admission in maternity department) was tested by a questionnaire. In role-playing group, advantages and disadvantages of two type delivery were presented by role-playing in 90-min by three scenarios. In lecture group, it was also presented in a 90-min lecture. Data were analyzed by mean difference test, Fisher test, independent and paired t-test. Two groups showed a significant difference in terms of decision at admission to maternity department (P = 0.000). 75% of lecture group and 100% of role-playing group selected normal delivery. Postintervention knowledge score in lecture group was 18 ± 5.3 and in role-playing group 17.1 ± 4.0. Percent of change in knowledge scores in two groups was significant (P = 0.001). Participants' attitude, before and after the intervention, in both groups was significant (P < 0.05). Mean difference of pre- and post-test in relation to two groups' knowledge and attitude scores was not significant (P > 0.05). In this research, lecture was more effective in raising knowledge level, and role playing was more effective in raising decision to vaginal delivery and reducing elective caesarean section. It is therefore suggested to use both teaching methods altogether for pregnant women to decrease the rate of unnecessary cesarean.


Nutrient intake and dental health in school children.
Diet refers to the customary allowance of food and drink taken by a person from day to day. The diet may affect dental caries by reacting with the enamel surface and by serving as a substrate for cariogenic microorganisms. This study examined the role of specific nutrients in the caries experience of elementary school children. 120 children aged between 6-11, who attended the Pedodontics Department of Marmara University, Faculty of Dentistry for routine dental treatment received a dental examination, parent interview, and a 3-day diet diary. The diary was completed adequately for 100 children (83%). The average daily intake of nutrients were computed and total sugar consumption and Ca/P ratio were calculated. Each child's gender, age, fluoride history, socio-economic state of the family, previous dental experience, brushing habit, and nutrient intake were examined as independent variables. No association was apparent between dental caries and the intake of specific nutrients or Ca/P ratio, except with total sugar consumption.


Rosiglitazone affects lumen formation in MDCKII cell through regulating apico-basal polarity.
This study aimed to investigate the potential mechanisms underlying the effects of Rosiglitazone on the apico-basal polarity in renal epithelial cells. 3D-MDCK model was used to study the lumen formation and localization of polarity proteins at the early stage of the establishment of the apico-basal polarity. The calcium switch model, immunofluorescence staining and measurement of transmembrane electrical impedance are employed to investigate the epithelial apico-basal polarity including the development and maintenance of apical domains and the formation of tight junction. MDCKII cells were cultured with 20 uM rosiglitazone or DMSO. Results showed Rosiglitazone reduced the percentage of single central lumen cysts, but the percentage of multiple lumen cysts increased. At the early stage of MDCKII cysts (2-5 cells), Rosiglitazone induced mislocalization of apical and basolateral membrane proteins. In the repolarization process of MDCKII cell induced by a calcium switch (CS), Rosiglitazone delayed the apical membrane domain development in the early phase of cell polarization; while during the maintenance phase of cell polarity, the apical domain retention was significantly affected by Rosiglitazone. Rosiglitazone significantly delayed the formation of tight junctions (TJs); 24 h after CS, however, there were no apparent differences between control group and Rosiglitazone group; the development of transepithelial electrical resistance (TER) was significantly disturbed in Rosiglitazone group. This study shows Rosiglitazone may affect the development and maintenance of apical domains and the formation of TJs disturbs apical protein delivery to the plasma membrane, eventually leading to the abnormal apico-basal polarity, which affects lumen formation in MDCKII cells.


Modeling Optimal Cutoffs for the Brazilian Household Food Insecurity Measurement Scale in a Nationwide Representative Sample.
Background: This is the second part of a model-based approach to examine the suitability of the current cutoffs applied to the raw score of the Brazilian Household Food Insecurity Measurement Scale [Escala Brasileira de Insegurança Alimentar (EBIA)]. The approach allows identification of homogeneous groups who correspond to severity levels of food insecurity (FI) and, by extension, discriminant cutoffs able to accurately distinguish these groups.Objective: This study aims to examine whether the model-based approach for identifying optimal cutoffs first implemented in a local sample is replicated in a countrywide representative sample.Methods: Data were derived from the Brazilian National Household Sample Survey of 2013 (n = 116,543 households). Latent class factor analysis (LCFA) models from 2 to 5 classes were applied to the scale's items to identify the number of underlying FI latent classes. Next, identification of optimal cutoffs on the overall raw score was ascertained from these identified classes. Analyses were conducted in the aggregate data and by macroregions. Finally, model-based classifications (latent classes and groupings identified thereafter) were contrasted to the traditionally used classification.Results: LCFA identified 4 homogeneous groups with a very high degree of class separation (entropy = 0.934-0.975). The following cutoffs were identified in the aggregate data: between 1 and 2 (1/2), 5 and 6 (5/6), and 10 and 11 (10/11) in households with children and/or adolescents <18 y of age (score range: 0-14), and 1/2, between 4 and 5 (4/5), and between 6 and 7 (6/7) in adult-only households (range: 0-8). With minor variations, the same cutoffs were also identified in the macroregions. Although our findings confirm, in general, the classification currently used, the limit of 1/2 (compared with 0/1) for separating the milder from the baseline category emerged consistently in all analyses.Conclusions: Nationwide findings corroborate previous local evidence that households with an overall score of 1 are more akin to those scoring negative on all items. These results may contribute to guide experts' and policymakers' decisions on the most appropriate EBIA cutoffs.


The use of honey in button battery ingestions: a systematic review.
Button battery (BB) ingestions may cause severe and possibly fatal complications, especially if the battery is located in the esophagus. The application of oral honey has recently been proposed by the National Capital Poison Center in the USA and in an ESPGHAN position paper in Europe, but clinical trials and experimental studies are limited. The goal of this systematic review was to analyze the evidence for this approach. A systematic review of clinical trials and experimental studies on the oral application of honey after BB ingestion in children was performed. Inclusion criteria according to the PICO format were patient age 0-18 years, ingestion of BB, oral administration of honey or other substances, all in vivo and in vitro studies, as well as reported complication rate, esophageal injury, and mortality. A manual search in the databases MEDLINE, Web of Science and Cochrane was performed to identify relevant search terms to form the following queries and to construct the extensive search. Furthermore, the search was extended by using snowballing on the reports reference lists. The review is registered at Research Registry. The identifying number is reviewregistry1581. We found four publications that investigated the effects of honey after button battery ingestion. Three of these presented experimental in vitro and in vivo results and one reported a clinical retrospective study of 8 patients. Follow up studies are required to further elucidate the effectiveness of the treatment with honey. The time intervals in which the use of honey is effective is not clear. Furthermore, a physiological model is needed for in vitro testing, preferably mimicking peristalsis and dynamic flow of the applied substances. However, since it is easy to apply and of minimal risk in patients over one year of age, honey should be considered a possible treatment option during the interval between presentation and endoscopic removal of the retained BB. https://www.researchregistry.com/browse-the-registry#registryofsystematicreviewsmeta-analyses/registryofsystematicreviewsmeta-analysesdetails/643e9df96750410027ee11b0/, identifier: reviewregistry1581.


Multicentricity in Different Molecular Subtypes of Breast Cancer: A Cross-sectional Study in Isfahan.
Breast cancer is the most common cancer leading to death in women. Women with multicentric breast cancer were reported more likely to have poor prognosis. Here, we decided to study and compare the frequency distribution of multicentricity in different subtypes of breast cancer. This is a cross-sectional study that was performed in 2019-20 on medical records and breast pathology reports of 250 patients who undergone mastectomy due to breast cancer. Demographic data of all patients including age, along with other medical data such as menstruation condition, breast cancer grade, multicentricity status, stage, and expression of estrogen receptor (ER), progesterone (PR), and human epidermal growth factor receptor 2 (HER2) receptors were collected from medical records. Samples were divided into four subtypes of Luminal B, Luminal A, HER2 expressing, and basal-like. The mean age of patients was 50.21 ± 11.15 years. Ninety-five patients (38%) had multicentricity and HER2 expressing (48.5%) and Luminal A (41.4%) were most common in patients with multicentricity. In addition, basal-like group presented with least multicentricity (13.5%) among the subtypes (P = 0.008). We also showed significant increased chances of multicentricity in Luminal B (odds ratio [OR] = 3.782) (P = 0.033), Luminal A (OR = 5.164) (P = 0.002), and HER2-expressing group (OR = 5.393) (P = 0.011). Taken together, we showed significantly increased chances of multicentricity in patients with HER2-expression, Luminal A, and Luminal B groups compared to basal-like group or triple negative. These results were in line with most previous studies; however, we showed higher rates of multicentricity among our population compared to some previous reports.


Evaluation of the anticoagulants EDTA and citrate, theophylline, adenosine, and dipyridamole (CTAD) for assessing platelet activation on the ADVIA 120 hematology system.
Monitoring of platelet activation by the ADVIA 120 Hematology System requires an anticoagulant and protocol that ensures that platelets are sphered and their activation status is not altered artifactually in vitro. Blood from healthy controls was collected into tripotassium EDTA; citrate, theophylline, adenosine, and dipyridamole (CTAD); or a combination of both (E/C) and stored at ambient temperature or at 4 degrees C (E/C only) and then analyzed between 0 and 180 min later on the ADVIA 120. In addition, immunofluorescent flow cytometry was used to identify activated platelets and platelet-leukocyte aggregates. In blood stored with all three anticoagulants, the platelet count changed little, but the mean platelet volume (MPV) at first decreased and then increased, whereas the mean platelet component (MPC; an indicator of activation) changed in a reciprocal manner. The changes in MPV and MPC, which reflect platelet sphering and swelling, were greatest between 30 and 60 min in blood stored at ambient temperature, irrespective of which anticoagulant was used, and between 60 and 180 min when blood anticoagulated with E/C was stored at 4 degrees C. In all anticoagulants, the percentages of platelets expressing CD62P and of leukocytes in platelet-leukocyte aggregates increased significantly (P <0.01) over 180 min at ambient temperature. Only minimal (<2%) increases occurred when blood with E/C was stored at 4 degrees C. When determining platelet activation ex vivo on the ADVIA 120, blood should be collected into E/C, stored at 4 degrees C, and analyzed between 60 and 180 min later; these conditions ensure maximum platelet sphering without concurrent artifactual platelet activation.


[Acute low back pain with progressive sensorimotor paralysis. Differential diagnosis and therapy of acute decompression disease].
A 38-year-old man presented with acute low back pain and paraesthesia in dermatome S1. 4 hours before onset of symptoms he had finished the last of three scuba dives with a maximum depth of 30 m and a total diving time of 3 hours. No alcoholic beverages were taken during or before diving. The patient was complaining of local pain in the thoracic and lumbar spine and showed a 3/5 weakness of the big toe and impaired sensitivity in the S1 dermatome. 90 minutes later the patient developed a hemiparesis of the right side including hypaesthesia and additional meningism (stiff neck). The X-rays of the thoracic and lumbar spine in two standard planes, cerebrospinal fluid examination, a cranial spiral-computer tomographie (CCT) and laboratory investigations showed no pathologic values. Acute decompression sickness (DCS) type II. The patient received an intravenous infusion, antiinflammatory prophylaxis with dexamethasone and an immediate submission to hyperbaric oxygenation therapy. Complete recovery of neurological symptoms appeared after 4 decompression sessions in a hyperbaric chamber. In patients presenting low back pain as common symptoms the differential diagnosis needs to be worked out and DCS should be included especially if progressive neurologic deficits are present. Only emergency hyperbaric oxygen therapy can compromise a sufficient therapy of these patients.


Porcine surfactant protein D is N-glycosylated in its carbohydrate recognition domain and is assembled into differently charged oligomers.
Surfactant protein D (SP-D) belongs to a subgroup of mammalian collagenous Ca(2+)-dependent lectins known as the collectins. It is thought to play a significant role in the innate immune response against microorganisms within the lungs and at other mucosal surfaces. This report documents the isolation and characterization of SP-D purified from porcine lung lavage using mannan affinity chromatography and gel filtration. Ultrastructural analysis shows both dodecameric and higher order oligomeric complexes of SP-D. The molecular mass of monomeric porcine SP-D (50 kD) is larger than that of SP-D from humans (43 kD). The difference in mass is due to the presence of an Asparagine-linked glycosylation in the carbohydrate recognition domain of porcine SP-D, which is absent in SP-D of other species investigated so far. Analysis of this carbohydrate moiety indicates that it is a highly heterogeneous, complex type oligosaccharide which is sialylated. The heterogeneity of oligosaccharide sialylation results in the existence of many differently charged porcine SP-D isoforms. The removal of the carbohydrate moiety reduces the inhibitory effect of porcine SP-D on influenza A virus haemagglutination. Therefore, the carbohydrate moiety may influence interactions with pathogens.


Distinctive p53 and mdm2 immunohistochemical expression profiles suggest different pathogenetic pathways in poorly differentiated endometrial carcinoma.
Endometrial serous carcinoma (ESC) and FIGO (International Federation of Gynecology and Obstetrics) grade 3 endometrioid adenocarcinoma (EC) are high-grade endometrial tumors that have different clinical and morphologic attributes. Alteration of p53 tumor suppressor protein function has been implicated in the pathogenesis of both tumors, although the mechanisms may differ. We sought to investigate this difference by comparing immunohistochemical expression of p53 and mdm2. p53 immunoreactivity often correlates with gene mutation, whereas increased mdm2 expression is linked to complex formation with wild-type p53 resulting in its inactivation. Twenty cases of ESC and 21 cases of EC were evaluated and an immunoreactivity score (IRS) was assigned using both the percentage of cells stained and the intensity of staining. The overall IRSs were significantly different in ESCs versus ECs for both p53 and mdm2 (p < 0.001 and p < 0.01, respectively). Strong mean immunoreactivity for p53 was detected in 15 (75%) ESCs as compared to only weak mean immunoreactivity in 17 (81%) ECs. Conversely, for mdm2 expression, 17 (81%) ECs had moderate mean immunoreactivity whereas 9 (45%) ESCs showed only weak mean immunoreactivity. mdm2 expression more closely correlated with p53 expression in ECs than in ESCs. In ECs, mdm2 was detected in 16 of 17 (94%) p53-positive tumors but in only 1 of 3 (33%) p53-negative tumors (p < 0.025). In ESCs, mdm2 was detected in 9 of 15 (60%) p53-positive tumors but in none of the 5 p53-negative tumors (p < 0.10). Overall, our results demonstrate an inverse relationship between the expression of p53 and mdm2 in ESC versus high-grade EC. Specifically, strong p53 immunoreactivity is associated with weak mdm2 expression-in ESC and weak p53 expression is associated with moderate mdm2 expression in EC. These results suggest different pathogenetic pathways resulting in loss of normal p53 function in these two tumors: by p53 gene mutation (strong p53 overexpression) in ESCs, or by mdm2 complex formation and inactivation of p53 in high-grade ECs.


POLYGENIC INHERITANCE OF A BEHAVIORAL PHENOTYPE: INTERSPECIFIC GENETICS OF SONG IN THE HAWAIIAN CRICKET GENUS LAUPALA.
The Hawaiian cricket genus Laupala (family Gryllidae) is one of several native genera of flightless crickets found in rain-forest habitat across the Hawaiian archipelago. Species in this genus are morphologically quite similar, but the songs produced by adult males are acoustically distinct. I examined the nature of song variation found within Laupala paranigra and between Laupala kohalensis and L. paranigra, both endemic to the island of Hawaii. Variation within and among species was most notable in the temporal structure of the song, as quantified by the pulse rate. The variation in pulse rate present in natural populations of L. paranigra bred true through the F1 laboratory generation, suggesting that the intraspecific variation in this species has a genetic basis. Interspecific hybridizations between L. kohalensis and L. paranigra successfully produced F1 , F2 , and backcross generations. Hybrid F1 males from reciprocal crosses sang with significantly different pulse rates, implicating an X chromosomal contribution to the phenotypic difference between these species. Interspecific patterns of inheritance are most consistent with a type-I genetic architecture. Polygenic inheritance of the interspecific pulse-rate variation was observed, and approximately eight genetic factors were estimated to underlie the difference in pulse rate between L. kohalensis and L. paranigra.


Rapid and Sensitive Determination of Timosaponin AIII in Rat Plasma by LC-MS/MS and Its Pharmacokinetic Application.
A rapid sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for determination of timosaponin AIII (TA-III) in rat plasma, using ginsenoside Re as an internal standard (IS). TA-III and the IS were detected in MRM mode with a negative ionization electrospray mass spectrometer. The calibration curves were linear over the concentration ranges from 11.14 to 1114 ng/mL and the lower limit of quantification (LLOQ) was 11.14 ng/mL. Intra-day and inter-day precisions (RSD) were within 10%, and accuracy ranged from 6.4% to 9.1%. The extraction recovery at three concentrations ranged from 92.3% to 95.5%. The validated method was successfully applied to monitor the concentrations of TA-III in rat plasma after intragastric administration. The best fit pharmacokinetic model to estimate the pharmacokinetic parameters was a single compartment model with weight of 1/x2 for oral administration groups of rats for TA-III.


Binary Ni-Co-Based Layered Double Hydroxide Nanoneedle Arrays for High Performance of Oxygen Evolution Reaction.
Low-cost and high-performance electrocatalysts are crucial for water-splitting reactions. Some non-precious metal electrocatalysts are proved to be good replacements for noble metal due to the unique electronic structure features and excellent performance. In this work, binary Ni-Co-based layered double hydroxide nanoneedle arrays electrocatalysts are synthesized on Ni foam (NF) via a hydrothermal process. The microstructure and the catalytic performance of the catalyst changes significantly by regulating the molar ratio of Ni/Co. The theoretical analysis confirmed that the as-prepared NiCo-LDH nanoneedle arrays reveal a potential behavior in oxygen evolution reaction (OER) at a lower overpotential of 305 mV at 10.0 mA cm-2 and a Tafel slope of 110.38 mV dec-1. The double-layer capacitance (Cdl) is 776 mF cm-2, which indicates that there are many active sites that are exposed on the surface for the electrocatalytic reaction. The results provide an obvious reference value to other types of LDH catalysts for the development of water electrolysis.


Bone-Targeted Nanoplatform Combining Zoledronate and Photothermal Therapy To Treat Breast Cancer Bone Metastasis.
Bone metastasis, a clinical complication of patients with advanced breast cancer, seriously reduces the quality of life. To avoid destruction of the bone matrix, current treatments focus on inhibiting the cancer cell growth and the osteoclast activity through combination therapy. Therefore, it could be beneficial to develop a bone-targeted drug delivery system to treat bone metastasis. Here, a bone-targeted nanoplatform was developed using gold nanorods enclosed inside mesoporous silica nanoparticles (Au@MSNs) which were then conjugated with zoledronic acid (ZOL). The nanoparticles (Au@MSNs-ZOL) not only showed bone-targeting ability in vivo but also inhibited the formation of osteoclast-like cells and promoted osteoblast differentiation in vitro. The combination of Au@MSNs-ZOL and photothermal therapy (PTT), triggered by near-infrared irradiation, inhibited tumor growth both in vitro and in vivo and relieved pain and bone resorption in vivo by inducing apoptosis in cancer cells and improving the bone microenvironment. This single nanoplatform combines ZOL and PTT to provide an exciting strategy for treating breast cancer bone metastasis.


Pleomorphic adenoma of the labial gland, characterized by reticular pattern of myoepithelial cells in a dog.
An 11-year-old female golden retriever dog had a mass at the right corner of the upper lip, which gradually increased in size and protruded into the oral cavity. The mass was removed surgically. The cut surface of the mass was smooth, whitish and solid, and covered by the oral mucosal membrane. Histopathologically, the mass consisted mainly of reticular pattern of short spindle cells that stained positively for cytokeratin AE1/AE3, α-smooth muscle actin and p63, suggestive of a myoepithelial cell phenotype. Between the neoplastic cords, there was myxoid or edematous connective tissue. Additionally, neoplastic cells with luminal epithelial and basal cell phenotypes were arranged in ducts and small islands, respectively. Based on the diverse histological and immunohistochemical features, the tumor was diagnosed as pleomorphic adenoma of the labial gland. To our knowledge, the reticular proliferation pattern of myoepithelial cells has not been described in salivary gland tumors of domestic animals.


Asymmetry of anticipatory postural adjustment during gait initiation.
The purpose of this study was to investigate the asymmetry of anticipatory postural adjustment (APA) during gait initiation and to determine whether the process of choosing the initial swing leg affects APA during gait initiation. The participants initiated gait with the leg indicated by a start tone or initiated gait with the leg spontaneously chosen. The dependent variables of APA were not significantly different among the condition of initiating gait with the preferred leg indicated by the start tone, the condition of initiating gait with the non-preferred leg indicated by the start tone, and the condition of initiating gait with the leg spontaneously chosen. These findings fail to support the view that the process of choosing the initial swing leg affects APA during gait initiation. The lateral displacement of the center of pressure in the period in which shifting the center of pressure to the initial swing phase before initiating gait with the left leg indicated by the external cue was significantly larger than that when initiating gait with the right leg indicated by the external cue, and significantly larger than that when initiating gait with the leg spontaneously chosen. Weight shift to the initial swing side during APA during gait initiation was found to be asymmetrical when choosing the leg in response to an external cue.


Novel approaches to the design of bioavailable melanotropins.
The melanocortin system is a primordial and critical system for survival, involved in a wide variety of physiological functions. It includes melanocortin receptors (MCRs) and melanotropin ligands (MCLs). MCRs are important drug targets that can regulate several key physiological processes. Extensive efforts have been made to develop peptide and peptidomimetics targeting melanocortin receptors including MC1R, MC3R, MC4R and MC5R. Most research is focused on developing potent and selective melanotropins. However, developing bioavailable melanotropins remains challenging. Areas covered: Herein, the authors summarize promising strategies for developing bioavailable MCLs by using cyclized N-methylated melanotropins, and using cyclotide and tetrapeptide as templates. They discuss their unique advantages in oral availability and targeting MCRs in the central nervous system or in peripheral tissues. Finally, they discuss the observed differences in thepharmacology of MCRs between in vitro and in vivo tests. Expert opinion: N-methylated cyclized melanotropins have great potential to become bio- available drugs targeting MCRs in the brain, while MCR-grafted cyclotides tend to target MCRs in peripheral tissue. A better understanding of the biased signaling process is a new challenge and opportunity for the future discovery of bioavailable MCLs.


Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy.
The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label- and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.


Interactive histogenesis of axonal strata and proliferative zones in the human fetal cerebral wall.
Development of the cerebral wall is characterized by partially overlapping histogenetic events. However, little is known with regards to when, where, and how growing axonal pathways interact with progenitor cell lineages in the proliferative zones of the human fetal cerebrum. We analyzed the developmental continuity and spatial distribution of the axonal sagittal strata (SS) and their relationship with proliferative zones in a series of human brains (8-40 post-conceptional weeks; PCW) by comparing histological, histochemical, and immunocytochemical data with magnetic resonance imaging (MRI). Between 8.5 and 11 PCW, thalamocortical fibers from the intermediate zone (IZ) were initially dispersed throughout the subventricular zone (SVZ), while sizeable axonal "invasion" occurred between 12.5 and 15 PCW followed by callosal fibers which "delaminated" the ventricular zone-inner SVZ from the outer SVZ (OSVZ). During midgestation, the SS extensively invaded the OSVZ, separating cell bands, and a new multilaminar axonal-cellular compartment (MACC) was formed. Preterm period reveals increased complexity of the MACC in terms of glial architecture and the thinning of proliferative bands. The addition of associative fibers and the formation of the centrum semiovale separated the SS from the subplate. In vivo MRI of the occipital SS indicates a "triplet" structure of alternating hypointense and hyperintense bands. Our results highlighted the developmental continuity of sagittally oriented "corridors" of projection, commissural and associative fibers, and histogenetic interaction with progenitors, neurons, and glia. Histogenetical changes in the MACC, and consequently, delineation of the SS on MRI, may serve as a relevant indicator of white matter microstructural integrity in the developing brain.


Identifying health service barriers in the management of co-morbid diabetes and chronic kidney disease in primary care: a mixed-methods exploration.
Co-morbid diabetes and chronic kidney disease (CKD) are common in primary care but health care can be suboptimal. In this multi-centre mixed-methods study, we investigated GPs' perspectives on health service barriers in managing diabetes and CKD as an initial step towards health care improvement. Four focus groups were conducted among GPs in Australia's two largest cities. Transcripts underwent content analysis to inform development of a survey exploring health service barriers. This survey was then emailed/mailed to 840 GPs. Statistical analyses were performed using STATA v2.1. Responses were received from 13.7% of GPs (n = 115), mean (±SD) age 55.3 (10.1) years and mean duration of practice 26.6 (10.6). The majority (88.4%) reported wanting to manage diabetes and CKD in primary care with specialist assistance. However, 34.8% were unclear about the definition of CKD with 73.2% wanting more education. Access to specialist services was problematic with 39.3% and 28.2% reporting the process of referring patients to diabetes or CKD services, respectively, as hard. Coordination of care was also a problem with 35.6% unclear about each health care provider's role, 50.5% believing patients faced difficulties due to poor coordination across providers and 51.6% reporting duplication of tests. GPs expressed a clear interest in being the principal health care providers for patients with co-morbid diabetes and CKD. Supporting GPs and health care improvement focusing on overcoming reported barriers such as inadequate knowledge about CKD, access to specialist services and coordination of care may improve outcomes for people with co-morbid diabetes and CKD.


A demyelinating disorder associated with cerebrovascular amyloid angiopathy.
Five patients with a demyelinating disorder and associated amyloid angiopathy are presented. The disease affected middle-aged individuals, pursued a fluctuating course, and ended in progressive, fatal deterioration of the central nervous system. Neurologic findings indicated multiple lesions within the neuraxis; profound dementia was prominent in all cases. Pathologically, numerous demyelinated plaques, similar to those in multiple sclerosis, were found in the cerebral white matter, and less consistently in other locations such as optic nerve, brain stem, and spinal cord. Amyloid accumulated massively in and around blood vessels, usually in the immediate vicinity of the plaques. At least one similar case is reported in the literature, but the nosologic status of the condition is uncertain.


Cerebral damage in epilepsy: a population-based longitudinal quantitative MRI study.
Whether cerebral damage results from epileptic seizures remains a contentious issue. We report on the first longitudinal community-based quantitative magnetic resonance imaging (MRI) study to investigate the effect of seizures on the hippocampus, cerebellum, and neocortex. One hundred seventy-nine patients with epilepsy (66 temporal lobe epilepsy, 51 extratemporal partial epilepsy, and 62 generalized epilepsy) and 90 control subjects underwent two MRI brain scans 3.5 years apart. Automated and manual measurement techniques identified changes in global and regional brain volumes and hippocampal T2 relaxation times. Baseline hippocampal volumes were significantly reduced in patients with temporal lobe epilepsy and could be attributed to an antecedent neurologic insult. Rates of hippocampal, cerebral, and cerebellar atrophy were not syndrome specific and were similar in control and patient groups. Global and regional brain atrophy was determined primarily by age. A prior neurologic insult was associated with reduced hippocampal and cerebellar volumes and an increased rate of cerebellar atrophy. Significant atrophy of the hippocampus, neocortex, or cerebellum occurred in 17% of patients compared with 6.7% of control subjects. Patients with and without significant volume reduction were comparable in terms of seizure frequency, antiepileptic drug (AED) use, and epilepsy duration, with no identifiable risk factors for the development of atrophy. Overt structural cerebral damage is not an inevitable consequence of epileptic seizures. In general, brain volume reduction in epilepsy is the cumulative effect of an initial precipitating injury and age-related cerebral atrophy. Significant atrophy developed in individual patients, particularly those with temporal lobe and generalized epilepsy. Longer periods of observation may detect more subtle effects of seizures.


On three-dimensional dose calculation of photon beam with wedge filters.
A wedge filter is one of the most commonly used beam modifying devices. The introduction of a wedge filter alters both the distribution of the primary photon fluence and the first and second scatter patterns. The effect of the wedge filter is normally expressed in terms of an exponential attenuation function that is used to modify the primary photon fluence. This paper describes a new three-dimensional (3-D) wedged field dose calculation algorithm. This algorithm automatically takes into account (1) the scatter distribution pattern correction due to the effect of wedge filters and (2) the fixed primary photon fluence correction, by utilizing a tissue maximum ratio table calculated from the measured percent depth dose of the wedge, a measured in-plane scan and a measured cross-plane scan of the largest rectangular wedge field size. As a result, the calculation accuracy achieved by this algorithm is better than the calculation accuracy achieved by other algorithms. The results show better than 1% accuracy of the wedge central axis percent depth dose calculation and better than 3% accuracy of the off-axis dose calculation when compared with measurements.


A study on the electrical, optical, and physicochemical properties of poly(MMA-co-MAA)/ poly(3,4-ethylenedioxythiophene) hybrid thin films.
Poly(3,4-ethylenedioxythiophene) (PEDOT) has good properties as a conductive polymer such as high conductivity, optical transmittance, and chemical stability, while offering relatively weak physicochemical properties. The main purpose of this paper is to improve physicochemical properties such as solvent resistance and pencil hardness of PEDOT. Carboxyl groups in the poly(MMA-co-MAA) polymer chains can effectively crosslink each other in the presence of aziridine, resulting in physicochemically robust PEDOT/poly(MMA-co-MAA) hybrid conductive films. The electrical conductivity, optical properties, and physicochemical properties of the hybrid conductive film were compared by varying the solid content and poly(MMA-co-MAA) portion in the coating precursor solution. From the results, the transparency and surface resistance of the hybrid film show a tendency to decrease with increasing solid content in the coating precursor. Moreover, solvent resistance and hardness were dramatically enhanced by hybridization of PEDOT and crosslinked poly(MMA-co-MAA) due to curing reactions between carboxyl groups. The chemical composition of 30 wt-% of poly(MMA-co-MAA) (MMA:MAA mole ratio 9:1) and 3 wt-% - 5 wt-% of aziridine yields the best physicochemical properties of poly(MMA-co-MAA)/PEDOT hybrid thin films.


Modification of gastric pH with oral glutamic acid hydrochloride.
The extent to which oral glutamic acid hydrochloride decreases mean gastric pH in fasting persons with and without simulated hypochlorhydria was studied. Healthy nonsmoking men were randomly assigned to one of two drug regimens followed by the other regimen after a one-week washout period. In regimen 1 the fasting subjects received two 680-mg doses of glutamic acid hydrochloride given 10 minutes apart. Regimen 2 was the same, except that an oral dose of ranitidine 300 mg (as the hydrochloride salt) was administered one to two hours before the first dose of glutamic acid hydrochloride to simulate hypochlorhydria. Gastric pH was monitored radiotelemetrically before and after glutamic acid hydrochloride administration by using the Heidelberg capsule technique. Six men 20 to 28 years of age participated in the study. For regimen 1, the gastric pH before glutamic acid hydrochloride was given was not significantly different from that after administration (grand medians, 1.4 and 1.3, respectively). In regimen 2, the median gastric pH increased to greater than 4.0 within two hours after ranitidine treatment. Median gastric pH after the second dose of glutamic acid hydrochloride was significantly lower than before the first dose (grand medians, 1.6 and 6.2, respectively). The time to minimum pH was 2 to 15 minutes, and pH remained less than 3.0 for a mean of 45 minutes. Glutamic acid hydrochloride alone did not decrease fasting gastric pH, but it significantly reduced pH in subjects with simulated hypochlorhydria produced by orally administered ranitidine.


Acute upper gastrointestinal bleeding.
Acute upper gastrointestinal bleeding (AUGIB) is a frequently encountered medical emergency with an incidence of 84-160/100000 and associated with mortality of approximately 10%. Guidelines from the National Institute for Care and Care Excellence outline key features in the management of AUGIB. Patients require prompt resuscitation and risk assessment using validated tools. Upper gastrointestinal endoscopy provides accurate diagnosis, aids in estimating prognosis and allows therapeutic intervention. Endoscopy should be undertaken immediately after resuscitation in unstable patients and within 24 hours in all other patients. Interventional radiology may be required for bleeding unresponsive to endoscopic intervention. Drug therapy depends on the cause of bleeding. Intravenous proton pump inhibitors should be used in patients with high-risk ulcers. Terlipressin and broad-spectrum antibiotics should be used following variceal haemorrhage. Hospitals admitting patients with AUGIB need to provide well organised services and ensure access to relevant services for all patients, and particularly to out of hours endoscopy.


Hospitalizations for ambulatory care sensitive conditions across primary care models in Ontario, Canada.
The study analyzes the relationship between the risk of a hospitalization for an ambulatory care sensitive condition (ACSC), and the primary care payment and the organizational model used by the patient (fee-for-service, enhanced fee-for-service, blended capitation, blended capitation with interdisciplinary teams). The study used linked patient-level health administrative databases and census data housed at the Institute for Clinical Evaluative Sciences in Ontario. Since the province provides universal health care, the data capture all patients in Ontario, Canada's most populous province, with about 13 million inhabitants. All Ontario patients diagnosed with an ACSC prior to April 1, 2012, who had at least one visit with a physician between April 1, 2012, and March 31, 2013, were included in the study (n = 1,710,310). Each patient was assigned to the primary care model of his/her physician. The different models were categorized as Fee-for-Service (FFS), enhanced-FFS, blended capitation, and interdisciplinary team. A logistic regression was used to model the risk of having an ACSC hospitalization during the one-year observation period. Adjustments were made for patient characteristics (age, sex, health status, and socio-economic status) and for the geographic location of the practice. Using patients belonging to FFS models as the reference group, the risk of an ACSC hospitalization was higher for patients belonging to the blended-capitation model using interdisciplinary teams (Adjusted Odds Ratio [AOR] = 1.06, 95% confidence interval [CI] = 1.00-1.12) and lower for enhanced-FFS (AOR = 0.78, CI = 0.74-0.82) and blended capitation patients (AOR = 0.91, CI = 0.86-0.96). Using patients with hypertension as the reference group, the odds of an ACSC hospitalization were much higher for patients with any other ACSC and increased with patients' morbidity. The risk was lower for patients of higher socio-economic status (AOR = 0.63, CI = 0.60-0.67) in the highest neighborhood income quintile.


Breastfeeding and health professional encouragement.
The objective of the study was to ascertain the nature of health professional contacts related to breastfeeding for 341 women. Subjects were prospectively queried by phone or home visit over a 20-week period regarding the number and nature of contacts with health care professionals related to breastfeeding. Lactation consultants gave significantly more positive encouragement (98%) (p = .01) than either nurses (75%) or physicians (68%) did. However, of the 233 health provider contacts that included breastfeeding advice, only 21% involved lactation consultants. Primiparae were likely to decrease their level of breastfeeding if a health professional encouraged supplemental foods (39%) or weaning (86%) during the prior week. Multiparae who were experienced at breastfeeding (3+ weeks breastfeeding the previous infant) decreased their breastfeeding slowly across the weeks with the current infant, and their level of breastfeeding in general appeared independent of health care provider advice. Multiparae who lacked previous sustained breastfeeding experience (0-3 weeks) had the most rapid decrease (65%) in their breastfeeding rates even with health care provider encouragement to continue. Thus, they too appeared to act independently of health care provider advice.


Patient experience of general practice and use of emergency hospital services in England: regression analysis of national cross-sectional time series data.
The UK Government has introduced several national policies to improve access to primary care. We examined associations between patient experience of general practice and rates of visits to accident and emergency (A&E) departments and emergency hospital admissions in England. The study included 8124 general practices between 2011-2012 and 2013-2014. Outcome measures were annual rates of A&E visits and emergency admissions by general practice population, according to administrative hospital records. Explanatory variables included three patient experience measures from the General Practice Patient Survey: practice-level means of experience of making an appointment, satisfaction with opening hours and overall experience (on 0-100 scales). The main analysis used random-effects Poisson regression for cross-sectional time series. Five sensitivity analyses examined changes in model specification. Mean practice-level rates of A&E visits and emergency admissions increased from 2011-2012 to 2013-2014 (310.3-324.4 and 98.8-102.9 per 1000 patients). Each patient experience measure decreased; for example, mean satisfaction with opening hours was 79.4 in 2011-2012 and 76.6 in 2013-2014. In the adjusted regression analysis, an SD increase in experience of making appointments (equal to 9 points) predicted decreases of 1.8% (95% CI -2.4% to -1.2%) in A&E visit rates and 1.4% (95% CI -1.9% to -0.9%) in admission rates. This equalled 301 174 fewer A&E visits and 74 610 fewer admissions nationally per year. Satisfaction with opening hours and overall experience were not consistently associated with either outcome measure across the main and sensitivity analyses. Associations between patient experience of general practice and use of emergency hospital services were small or inconsistent. In England, realistic short-term improvements in patient experience of general practice may only have modest effects on A&E visits and emergency admissions.


Fractionation of human serum lipoproteins by single-spin gradient ultracentrifugation: quantification of apolipoproteins B and A-1 and lipid components.
A sensitive and reproducible method has been developed for separation of the major serum lipoproteins from 1 ml or less of human serum by isopycnic density gradient ultracentrifugation. The serum, applied to a step gradient (total volume 12.8 ml), was spun for 48 hr at 38,000 rpm at 10 degrees C and, in each of the fractions, apolipoproteins B and A-I were quantified by the respective radioimmunoassays. The markers for lipid distribution used were [4-(14)C]cholesterol and [U-(14)C]lecithin, each incubated with an aliquot of serum at 20 degrees C for 75 min prior to ultracentrifugation. In control sera, three main fractions, very low density (VLDL), low density (LDL), and high density (HDL) lipoproteins were clearly separated from a bottom fraction. Their flotational, electrophoretic, and chemical properties were in good agreement with those reported for the corresponding lipoproteins separated by conventional ultracentrifugation. Both apo B and apo A-I were fully recovered. Essentially all of the apo B was found in VLDL (9.3 +/- 3.5%) and LDL (87 +/- 4.6%); of the apo A-I, 81.0 +/- 5.7% was in HDL and the remainder (17.0 +/- 5.8%) was in the bottom fraction. The peak activities of [(14)C]cholesterol coincided with the peak of apo B in both LDL and VLDL, and with the peak of apo A-I in HDL. The results with the radiolabeled cholesterol were in good agreement with those obtained by chemical analyses. Carbon 14-labeled lecithin, although fully recovered, was not an accurate marker of phospholipid distribution because, under our experimental conditions, a significant amount of the lecithin was converted into its lyso derivative. The mechanism of the conversion was not established; it appeared to be unrelated to the activities of either lecithin-cholesterol acyl transferase or a Ca(2+)-dependent phospholipase. Besides its validity in the study of control sera, our method also proved successful in the separation of the serum lipoproteins of the few patients with dyslipoproteinemia (abetalipoproteinemia and familial hypercholesterolemia) who were examined. However, the applicability of the method to all dyslipoproteinemias was not assessed. Taken together, the results indicate that the single-spin method could be useful in clinical studies as a complement to other established techniques.


Fundamentals of interventional pain medicine.
Chronic pain is one of the most common and challenging medical problems facing our society. The specialty of pain medicine has grown steadily in recent years, largely because of the recognition that multiple factors contribute to chronic pain. The practice of pain medicine is multidisciplinary in approach, incorporating modalities from various specialties to ensure the comprehensive evaluation and treatment of the pain patient. The integration of various specialties such as anesthesiology, neurology, neurologic surgery, orthopedic surgery, physical medicine, and psychiatry is essential to treating patients with chronic pain and to establishing continuity of care. Research in the last 30 years has developed a variety of alternatives or adjuncts to opiates for chronic pain, including neuroactive medications, counterstimulation methods, and cognitive-behavioral therapies. Pain medicine specialists have provided leadership in the development of the practice, with the application of a wide verity of central and peripheral nerve blocks, sympathetic and neurolytic blocks, intradiscal procedures, neuromodulation techniques, intrathecal infusion systems, and other technical procedures that are firmly linked to a biomedical model of pain.


Biogenesis of the Gram-negative bacterial outer membrane.
Gram-negative bacteria are bounded by two membranes. The outer membrane consists of phospholipids, lipopolysaccharides, lipoproteins and integral outer membrane proteins, all of which are synthesized in the cytoplasm. Recently, much progress has been made in the elucidation of the mechanisms of transport of these molecules over the inner membrane, through the periplasm and into the outer membrane, in part by exploiting the extraordinary capacity of Neisseria meningitidis to survive without lipopolysaccharide.


Hepatocyte growth factor is a pleiotropic factor protecting epithelial cells from apoptosis.
Hepatocyte Growth Factor (HGF) is a pleiotropic factor, capable of evoking complex biological responses such as mitogenesis, motogenesis and morphogenesis in a variety of epithelial and endothelial cells. Nonetheless, the meaning of the acronym is consistent with the key role of the factor in liver regeneration, in vivo and in liver development during embryogenesis. The receptor for HGF is the tyrosine kinase encoded by the c-MET proto-oncogene. Upon ligand binding, the receptor kinase is activated by tyrosine autophosphorylation and recruits cytoplasmic transducers involved in HGF-triggered signal transduction. We investigated the role of HGF as a survival factor in protecting cells from apoptosis and we show that HGF is able to counteract staurosporin-induced apoptosis of epithelial cells.


Phylogeny, classification and diversity of Choreotrichia and Oligotrichia (Ciliophora, Spirotrichea).
Ciliated protists in the subclasses Choreotrichia and Oligotrichia are major components of marine plankton. Despite their ecological relevance, there are uncertainties in their systematics and diversity. We retrieved and curated all the GenBank ribosomal DNA (rDNA) sequences from these groups, which were analyzed in two ways. The first approach was based on morphologically-identified sequences (including those of two families and six genera newly studied here by single-cell sequencing), and aimed at improving phylogenetic inferences using concatenated sequences of three rDNA loci. Based on phylogenetic and morphological support, we update the taxonomic classification of these subclasses into 23 families, including the re-established Favellidae. We also propose an informal naming system for incertae sedis taxa, namely Tintinnopsis and five related genera that are spread among eleven lineages. The second approach included unidentified environmental sequences, and was used to explore potentially novel diversity in these subclasses. Our results support high proportions of both synonyms in tintinnids and uncharacterized taxa in choreotrichids and oligotrichs. One previously unidentified, environmental clade is here linked to our new Leegaardiellidae sequences. Our curation of almost 4000 rDNA sequences exemplifies known issues of public repositories, and suggests caution in both the use and contribution to these unique resources for evolutionary and diversity studies.


Testing Multiple Conceptualizations of Oppositional Defiant Disorder in Youth.
Recent theories conceptualize oppositional defiant disorder (ODD) as a two-dimensional construct with angry/irritable (i.e., affective) and argumentative/defiant (i.e., behavioral) components. This view has been supported by studies of nonreferred youth but not yet examined in clinic-referred youth. In a reanalysis of data regarding children who received one of two psychosocial ODD treatments, we examined multiple conceptualizations of ODD, whether children showed improvements across these ODD dimensions, and whether main and joint effects of ODD dimension improvement predicted clinical outcome. One hundred thirty-four clinic-referred youth (ages 7-14 years, 38% female, 84% White) who met Diagnostic and Statistical Manual of Mental Disorders (4th ed.) criteria for ODD received 1 of 2 psychosocial treatments. At pretreatment, 1-week follow-up, and 6-month follow-up, mothers reported child aggression and conduct problems, clinicians reported global clinical impairment and clinical improvement, and ODD symptom counts were collected from a semistructured diagnostic interview with mothers. Baseline ODD symptom were used to test previously supported multidimensional models. One- and two-factor conceptualizations were supported; however, the two-factor solution was preferred. With this solution, each dimension significantly and similarly improved across treatment conditions. Improvements across affective and behavioral ODD factors also had significant effects on clinician- and mother-reported clinical outcomes. The current findings provide empirical support for the ongoing study of multidimensional ODD conceptualizations in clinic-referred youth.


Conformation of an RNA pseudoknot.
The structure of the 5' GCGAUUUCUGACCGCUUUUUUGUCAG 3' RNA oligonucleotide was investigated using biochemical and chemical probes and nuclear magnetic resonance spectroscopy. Formation of a pseudoknot is indicated by the imino proton spectrum. Imino protons are observed consistent with formation of two helical stem regions; nuclear Overhauser enhancements between imino protons show that the two stem regions stack to form a continuous helix. In the stem regions, nucleotide conformations (3'-endo, anti) and internucleotide distances, derived from two-dimensional correlated, spectroscopy and two-dimensional nuclear Overhauser effect spectra, are characteristic of A-form geometry. The data suggest minor distortion in helical stacking at the junctions of stems and loops. The model of the pseudoknot is consistent with the structure originally proposed by Pleij et al.


HLA phenotypes and multiple basal cell carcinomas.
Previous investigators noted an association of multiple basal cell carcinomas (BCC) with certain HLA antigens; however, these findings were contradictory, and the associations were only weak. The aim of the study was to objectify the previously found associations. Serologic HLA typing for class I and class II antigens was performed in 49 unrelated patients with 5 or more BCCs. HLA-DR4 showed decreased frequencies in the patient group as compared with healthy controls (n = 716). Cw7 was found to be increased in the total group of patients as well as in a subgroup with multiple BCCs of the face (n = 24), while a subgroup with BCCs mainly on the trunk (n = 25) revealed increased frequencies of HLA-A11, -B17, -B22 and -Cw3. However, none of these deviations appeared significant after correction of p values. We conclude that, if at all, the HLA system plays only a minor role in the development of multiple BCCs.


Diffuse cutaneous bullous mastocytosis with IgM deposits at dermo-epidermal junction.
Cutaneous mastocytosis is a disease characterized by the infiltration and proliferation of mast cells in the skin. In children, the most common form of presentation is urticaria pigmentosa, while the diffuse cutaneous bullous mastocytosis is one of the rarest subtypes seen. The aim of this paper is to present a case of diffuse bullous mastocytosis with detection of IgM deposits at dermo-epidermal junction using direct immunofluorescence (DIF) microscopy. The diagnosis of diffuse bullous mastocytosis is a challenge, and DIF microscopy is necessary in order to exclude an autoimmune bullous disorder. However, IgM deposits at dermo-epidermal junction can be nonspecific, being found in a variety of skin disorders. A 6-month-old girl presented with bullous lesions and erosions on the scalp and the trunk. During hospitalization, further bullous lesions appeared, along with generalized erythrodermia. Skin biopsy revealed aspects of urticaria pigmentosa. Taking into account the clinical findings, the case was enclosed as bullous mastocytosis. Treatment included the avoidance of trigger factors, and administration of antihistamines along with a short-term course of systemic steroids. The evolution was favorable, with remission of the existing lesions and without occurrence of new ones.


Treatment of Halogen Bonding in the OPLS-AA Force Field; Application to Potent Anti-HIV Agents.
The representation of chlorine, bromine, and iodine in aryl halides has been modified in the OPLS-AA and OPLS/CM1A force fields in order to incorporate halogen bonding. The enhanced force fields, OPLS-AAx and OPLS/CM1Ax, have been tested in calculations on gas-phase complexes of halobenzenes with Lewis bases, and for free energies of hydration, densities, and heats of vaporization of halobenzenes. Comparisons with results of MP2/aug-cc-pVDZ(-PP) calculations for the complexes are included. Implementation in the MCPRO software also allowed computation of relative free energies of binding for a series of HIV reverse transcriptase inhibitors via Monte Carlo/free-energy perturbation calculations. The results support the notion that the activity of an unusually potent chloro analog likely benefits from halogen bonding with the carbonyl group of a proline residue.


Development of multiplex PCR assay for simultaneous detection of African swine fever, porcine circo and porcine parvo viral infection from clinical samples.
A diagnostic method for simultaneously detecting and distinguishing African Swine Fever (ASF), porcine circovirus type 2 (PCV2), and porcine parvovirus (PPV) in clinical specimens is critical for differential diagnosis, monitoring, and control in the field. Three primer pairs were designed and used to create a multiplex PCR assay. In addition, 356 porcine post mortem tissue samples from various parts of India's North Eastern region were tested by the developed multiplex PCR assay to demonstrate its accuracy. Using the designed primers, each of the ASF, PCV2 and PPV target genes was amplified, but no other porcine virus genes were detected. The assay's limit of detection was 102 copies/µl of PCV2, PPV, or ASFV. The detection of PCV2, PPV, and ASF in postmortem tissue samples revealed that they are co-circulating in India's North-Eastern region. The percentage positivity (PP) for PCV2, PPV and ASF single infection were 7.02% (25/356), 3.93% (14/356), and 3.37% (12/356), respectively, while the PP for PCV2& PPV co-infection was 2.80% (10/356), ASF & PCV2 co infection was 1.4% (5/356) and the ASF, PPV& PCV2 co-infection was1.40% (5/356). The results also indicate that the ASF can infect pigs alongside PCV and PPV.


Curious fatal intentional poisoning case with organophosphate pesticide.
A case of organophosphorus intoxication with rebounding symptoms is reported. Case report of a 24-year-old man who poisoned himself with organophosphorus pesticide and was hospitalized in a 22-bed adult medical surgical intensive care unit at a tertiary care hospital. The patient had ingested organophosphorus pesticide after an argument and fight with his family and had presented typical clinical and biological manifestations of intoxication by this chemical. He was treated by mechanical ventilation, large fluid infusion, gastric lavage, as well as atropine and pralidoxime. After 48 hours of intensive care, the patient improved considerably, but shortly after this improvement, mental, hemodynamic, and respiratory status altered again. Gastric fibroscopy showed a small plastic bag containing powder in the stomach which was responsible for the rebounding symptoms of the intoxication. Gastric fibroscopy can be helpful in case of organophosphorus intoxication with persistent or rebounding symptoms.


Metalworking fluids: oil mist and beyond.
This article is based upon my own experiences with metalworking fluids and the adverse health effects and medical conditions associated with exposure to metalworking fluids. I have researched and witnessed the benefits that can be achieved when metalworking fluids are properly maintained and managed. My experiences have provided insight into how a shop operates, including comprehension of the equipment used, processes, mist generating points, engineering controls currently being adopted, and procedures that are used to maintain metalworking fluids. I have been able to share my personal experiences with the country's leading experts in the field of metalworking fluids. I have presented my insights on the topic in Washington, D.C., to the Standard Advisory Committee of OSHA, as well as at many other conferences nationwide. I have provided awareness training for a number of union and nonunion workers. Being a part of developing successful metal removal fluid programs, I realize the importance of transferring and sharing information. Many times an organization is not fully aware of certain conditions and how to combat them. My mission and intent is to properly educate those who are exposed to the harm that metalworking fluids can invoke and to inform those involved of the possible methods of reducing long- and short-term risk. One thing that must be kept in mind is the way we view these fluids. Many shops categorize the fluids as a type of "operating expense" when they should actually be seen as a sort of investment. Just as performing a scheduled maintenance on a machine promises the best possible longevity of that machine, the upkeep of metalworking fluid also provides longer "tool life." Monitoring and maintaining the fluids also provides for more effective and efficient productivity. If we fail to consider that proper management of the fluids can cut cost dramatically, then we will miss out on the financial impact they can have on a company. Try looking at the fluids as a liquid tool. Doing so I believe will bring a better understanding of the value of a successful metalworking fluids program. With this new understanding, it can be seen just who must play a role in the management of metalworking fluids. The employees who deal with the daily tasks involving the coolant play a major part. They are on the floor where these metalworking fluids are being used. In many shops, it is assumed that the environmental health & safety departments are responsible for standard operating procedures and management of fluids. The EH&S department should only be responsible for the protection from exposure and the transfer of information regarding policy and procedure to their employees. Not all shops have the resources required to develop and implement the proper standard operating procedure. Therefore, we must understand that what is feasible for one may not be for another. Companies that lack the sufficient resources should not be neglected. It is crucial that awareness of proper standard operating procedure is shared with everyone involved with the fluids in order to provide proper metalworking fluids management. Fluids are as dynamic as the formulations themselves (complex & dynamic). These fluids can quickly become contaminated with foreign materials and chemicals, thereby become aerosolized into mist. With proper education and training, one will be able to control what gets aerosolized.


Can we avoid high levels of dose escalation for high-risk prostate cancer in the setting of androgen deprivation?
Both dose-escalated external beam radiotherapy (DE-EBRT) and androgen deprivation therapy (ADT) improve outcomes in patients with high-risk prostate cancer. However, there is little evidence specifically evaluating DE-EBRT for patients with high-risk prostate cancer receiving ADT, particularly for EBRT doses >74 Gy. We aimed to determine whether DE-EBRT >74 Gy improves outcomes for patients with high-risk prostate cancer receiving long-term ADT. Patients with high-risk prostate cancer were treated on an institutional protocol prescribing 3-6 months neoadjuvant ADT and DE-EBRT, followed by 2 years of adjuvant ADT. Between 2006 and 2012, EBRT doses were escalated from 74 Gy to 76 Gy and then to 78 Gy. We interrogated our electronic medical record to identify these patients and analyzed our results by comparing dose levels. In all, 479 patients were treated with a 68-month median follow-up. The 5-year biochemical disease-free survivals for the 74 Gy, 76 Gy, and 78 Gy groups were 87.8%, 86.9%, and 91.6%, respectively. The metastasis-free survivals were 95.5%, 94.5%, and 93.9%, respectively, and the prostate cancer-specific survivals were 100%, 94.4%, and 98.1%, respectively. Dose escalation had no impact on any outcome in either univariate or multivariate analysis. There was no benefit of DE-EBRT >74 Gy in our cohort of high-risk prostate patients treated with long-term ADT. As dose escalation has higher risks of radiotherapy-induced toxicity, it may be feasible to omit dose escalation beyond 74 Gy in this group of patients. Randomized studies evaluating dose escalation for high-risk patients receiving ADT should be considered.


Lifetime Physical Activity and White Matter Hyperintensities in Cognitively Intact Adults.
White matter hyperintensities (WMHs) observed on magnetic resonance images are associated with depression and increase the risk of stroke, dementia, and death. The association between physical activity and WMHs has been inconsistently reported in the literature, perhaps because studies did not account for a lifetime of physical activity or depression. The aim of this study was to determine the extent to which a lifetime of leisure-time physical activity is associated with less WMHs while accounting for depression. Face-to-face interviews were conducted with the Lifetime Total Physical Activity Questionnaire, where the metabolic equivalent of task hours per week per year was calculated. Cognitively intact participants also underwent magnetic resonance imaging, where WMHs as a percentage of intracranial volume was obtained. Hierarchical multiple linear regression was performed to compare WMHs in a more active group with a group with no psychiatric history (n = 20, mean age = 62.2 years), with a less active group with no psychiatric history (n = 13, mean age = 64.0 years), and a less active group with history of late-onset depression (n = 14, mean age = 62.8 years). There was not a statistically significant difference in WMHlg10 between the more and less active groups without a psychiatric history (b = .09, p > .05) or between the more active group without a psychiatric history and the less active group with a history of depression (b = .01, p > .05). The model was predictive of WMHlg10, explaining an adjusted 15% of the variance in WMHs (p = .041). A lifetime of leisure-time physical activity was not associated with WMHs when accounting for depression.


Plectreurys tristis venome: A proteomic and transcriptomic analysis.
Spider venoms are complex cocktails rich in peptides, proteins and organic molecules that collectively act to immobilize prey. Venoms of the primitive hunting spider, Plectreurys tristis, have numerous neurotoxic peptides called "plectoxins" (PLTX), a unique acylpolyamine called bis(agmatine)oxalamide, and larger unidentified protein components. These spiders also have unconventional multi-lobed venom glands. Inspired by these unusual characteristics and their phylogenetic position as Haplogynes, we have partially characterized the venome of P. tristis using combined transcriptomic and proteomic methods. With these analyses we found known venom neurotoxins U1-PLTX-Pt1a, U3-PLTX-Pt1a, and we discovered new groups of potential neurotoxins, expanding the U1- and ω-PLTX families and adding U4-through U9-PLTX as six new groups. The venom also contains proteins that are homologs of astacin metalloproteases that, combined with venom peptides, make up 94% of components detected in crude venom, while the remaining 6% is a single undescribed protein with unknown function. Other proteins detected in the transcriptome were found to be members of conserved gene families and make up 20% of the transcripts. These include cDNA sequences that match venom proteins from Mesobuthus and Hottentotta scorpions, Loxosceles and Dysdera spiders, and also salivary and secreted peptide sequences from Ixodes, Amblyomma and Rhipicephalus ticks. Finally, we show that crude venom has neurotoxic effects and an effective paralytic dose on crickets of 3.3µg/gm.


Effects of long-term high-altitude exposure on fibrinolytic system.
High altitude (HA), with the main feature of hypobaric hypoxia, is an independent risk factor for thrombosis. However, little is known on the alterations of fibrinolytic system in adaptation to HA. In this study, we investigated changes of fibrinolytic system parameters between individuals permanently living at HA and low altitude (LA) regions, and provided data for further studies on HA-induced thrombotic disease. A total of 226 eligible participants, including 103 LA participants, 100 healthy HA subjects and 23 high altitude polycythemia (HAPC) patients, were recruited in this study. Six fibrinolytic parameters, i.e. fibrinogen (Fbg), D-dimer (DDi), antithrombin III (AT-III), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (tPA) and plasminogen (PLG) were analyzed respectively. PAI-1 and tPA were performed by using bio-immuno-assays and an automated coagulation analyzer was used to conduct Fbg, DDi, AT-III and PLG tests. Plasma levels of Fbg, DDi, PAI-1 and PLG were significantly higher in healthy HA group than in LA group (all p < 0.05), whereas tPA was significantly lower in healthy HA group. No significant difference in AT-III was observed between healthy HA and LA groups (p > 0.05). All these fibrinolytic parameters showed no significant distinctions between healthy HA subjects and HAPC patients (all p > 0.05). HGB showed no relationship with fibrinolytic parameters in HA cohort. This study demonstrates that HA environment has a significant effect on fibrinolytic system and provides a foundation for further studies on HA hypobaric hypoxia-induced thrombotic disease.


Positive epistasis between viral polymerase and the 3' untranslated region of its genome reveals the epidemiologic fitness of dengue virus.
Dengue virus (DENV) is a global health threat, causing repeated epidemics throughout the tropical world. While low herd immunity levels to any one of the four antigenic types of DENV predispose populations to outbreaks, viral genetic determinants that confer greater fitness for epidemic spread is an important but poorly understood contributor of dengue outbreaks. Here we report that positive epistasis between the coding and noncoding regions of the viral genome combined to elicit an epidemiologic fitness phenotype associated with the 1994 DENV2 outbreak in Puerto Rico. We found that five amino acid substitutions in the NS5 protein reduced viral genomic RNA (gRNA) replication rate to achieve a more favorable and relatively more abundant subgenomic flavivirus RNA (sfRNA), a byproduct of host 5'-3' exoribonuclease activity. The resulting increase in sfRNA relative to gRNA levels not only inhibited type I interferon (IFN) expression in infected cells through a previously described mechanism, but also enabled sfRNA to compete with gRNA for packaging into infectious particles. We suggest that delivery of sfRNA to new susceptible cells to inhibit type I IFN induction before gRNA replication and without the need for further de novo sfRNA synthesis could form a "preemptive strike" strategy against DENV.


Neuroepithelial cells and the hypoxia emersion response in the amphibious fish Kryptolebias marmoratus.
Teleost fish have oxygen-sensitive neuroepithelial cells (NECs) in the gills that appear to mediate physiological responses to hypoxia, but little is known about oxygen sensing in amphibious fish. The mangrove rivulus, Kryptolebias marmoratus, is an amphibious fish that respires via the gills and/or the skin. First, we hypothesized that both the skin and gills are sites of oxygen sensing in K. marmoratus. Serotonin-positive NECs were abundant in both gills and skin, as determined by immunohistochemical labelling and fluorescence microscopy. NECs retained synaptic vesicles and were found near nerve fibres labelled with the neuronal marker zn-12. Skin NECs were 42% larger than those of the gill, as estimated by measurement of projection area, and 45% greater in number. Moreover, for both skin and gill NECs, NEC area increased significantly (30-60%) following 7 days of exposure to hypoxia (1.5 mg l(-1) dissolved oxygen). Another population of cells containing vesicular acetylcholine transporter (VAChT) proteins were also observed in the skin and gills. The second hypothesis we tested was that K. marmoratus emerse in order to breathe air cutaneously when challenged with severe aquatic hypoxia, and this response will be modulated by neurochemicals associated chemoreceptor activity. Acute exposure to hypoxia induced fish to emerse at 0.2 mg l(-1). When K. marmoratus were pre-exposed to serotonin or acetylcholine, they emersed at a significantly higher concentration of oxygen than untreated fish. Pre-exposure to receptor antagonists (ketanserin and hexamethonium) predictably resulted in fish emersing at a lower concentration of oxygen. Taken together, these results suggest that oxygen sensing occurs at the branchial and/or cutaneous surfaces in K. marmoratus and that serotonin and acetylcholine mediate, in part, the emersion response.


Dengue surveillance in Metro Manila.
In a one-year surveillance of dengue infection in four hospitals in Metro Manila, Philippines, 143 patients were enrolled. Sixty seven were diagnosed by virus isolation and serology. Majority of patients had secondary antibody response. Only 5 patients developed dengue shock syndrome. Only dengue types 1 and type 2 were isolated. Highest incidence occurred among older children and young adults. Thrombocytopenia and hemoconcentration were observed in 37% and 77% of patients, respectively. There was no fatality. Several problems were identified in the course of the study which need to be corrected in order to have good quality surveillance data in future undertakings.


Cognitive Skills Assessment for the Aging Orthopaedic Surgeon: AOA Critical Issues.
Physician leaders are increasingly confronted with the challenge of evaluating the performance of surgical colleagues during the last phase of their careers. Identification of physicians with declining cognitive and clinical skills must be balanced against awareness of laws protecting colleagues from discrimination. Responding successfully to this challenge requires appropriate policies, reliable data, standardized evaluation tools, consistent documentation, and the development of compassionate and effective solutions that avoid discriminatory practices. Surgical champions are essential for promoting self-regulation. Performance evaluations should comply with medical staff bylaws and institutional wellness policies.


[Anatomical presentation of edentulous sites in the posterior maxillary in cone beam computed tomography].
This study aimed to analyze the bone anatomy of edentulous sites in the posterior maxillary by cone beam computed tomography (CBCT). A total of 100 CBCT radiographs from patients with missing maxillary posterior teeth were obtained, resulting in a sample size of 217 edentulous sites. The width and height of edentulous were assessed by three-dimensional reconstruction. In addition, the angle A and morphology of the maxillary sinus walls were evaluated. The mean bone height was 9.53 mm, and the percentage of sites than 10 mm was 62.67% (136/217). The mean bone width was 9.30 mm, and the percentage of sites more than 6 mm was 91.71% (199/217). The bone height decreased from premolar to molar areas, but the opposite trend was observed in bone width. Regarding the morphology of the sinus floor, 64.52% exhibited an oblique configuration. In angle A, the group of less than 30° was 10.14%, 30°-60° was 42.40%, and greater than 60° was 47.47%. A high percentage of edentulous sites in the posterior maxillary requires sinus floor elevation to allow the placement of dental implants. Thus, the use of CBCT scans is recommended to evaluate the anatomical structure of the maxillary sinus for reasonable implant planning.


Identification of roots from grass swards using PCR-RFLP and FFLP of the plastid trnL (UAA) intron.
The specific associations between plant roots and the soil microbial community are key to understanding nutrient cycling in grasslands, but grass roots can be difficult to identify using morphology alone. A molecular technique to identify plant species from root DNA would greatly facilitate investigations of the root rhizosphere. We show that trnL PCR product length heterogeneity and a maximum of two restriction digests can separate 14 common grassland species. The RFLP key was used to identify root fragments at least to genus level in a field study of upland grassland community diversity. Roots which could not be matched to known types were putatively identified by comparison of the nuclear ribosomal ITS sequence to the GenBank database. Ten taxa were identified among almost 600 root fragments. Additionally, we have employed capillary electrophoresis of fluorescent trnL PCR products (fluorescent fragment length polymorphism, FFLP) to discriminate all taxa identified at the field site. We have developed a molecular database for the identification of some common grassland species based on PCR-RFLP of the plastid transfer RNA leucine (trnL) UAA gene intron. This technique will allow fine-scale studies of the rhizosphere, where root identification by morphology is unrealistic and high throughput is desirable.


Correlation of Fine Needle Aspiration Cytology of Thyroid Gland with Histopathological Results.
Fine needle aspiration cytology (FNAC) is considered as the gold standard diagnostic test for the diagnosis of thyroid nodules. It is a cost-effective procedure that provides specific diagnosis rapidly with minimal complications. It plays an important role in the determination of treatment- patients with suspected malignancy diagnosis can be subjected to surgery. On the other hand it can decrease the rate of unnecessary surgeries. The aim of this study was to evaluate and compare the correlation, accuracy of fine needle aspirational cytology (FNAC) in the diagnosis of thyroid lesions with the final histopathologic diagnosis in the surgical specimens. In our study we have performed a retrospective analysis of a case series of patients who were admitted to the Department of Endocrine, General and Oncological Surgery of Hospital of M. Kopernik in Łodź (Poland) between May 2016 and December 2017 and underwent FNAC with subsequent surgery. Cytological diagnosis was classified into six Bethesda categories. On cytological examination 1070/1262 were reported as benign, 49 malignant and 143 suspicious. On histopathological examination, 956/1070 cases were confirmed as benign but there were 114 discordant cases. Among the other cases histopathology diagnosis of malignancy matched in 45/49 and 128/143 cases.The sensitivity and specificity were 60,28% and 98,05% respectively. False positive rate was 1.95% and false negative rate was 39.72%. The positive predictive value was 90.1% and negative predictive value was 89.35%. Accuracy of FNA in differentiating benign from malignant thyroid lesions was 89,46%. Fine needle aspiration cytology is a simple, cost-effective and popular procedure for the diagnosis of thyroid cancer. It is recommended as the first line investigation for the diagnosis of thyroid lessions.


Bioactive compounds from Aspergillus niger extract enhance the antioxidant activity and prevent the genotoxicity in aflatoxin B1-treated rats.
This study aimed to identify the bioactive compounds of the ethyl acetate extract of Aspergillus niger SH2-EGY using GC-MS and to evaluate their protective role against aflatoxin B1 (AFB1)-induced oxidative stress, genotoxicity and cytotoxicity in rats. Six groups of male Sprague-Dawley rats were treated orally for 4 weeks included the control group, AFB1-treated group (80 μg/kg b.w); fungal extract (FE)-treated groups at low (140) or high dose (280) mg/kg b.w and the groups treated with AFB1 plus FE at the two tested doses. The GC-MS analysis identified 26 compounds. The major compounds found were 1,2,3,4,6-Penta-trimethylsilyl Glucopyranose, Fmoc-L-3-(2-Naphthyl)-alanine, D-(-)-Fructopyranose, pentakis (trimethylsilyl) ether, bis (2-ethylhexyl) phthalate, trimethylsilyl ether-glucitol, and octadecanamide, N-(2- methylpropyl)-N-nitroso. The in vivo results showed that AFB1 significantly increased serum ALT, AST, creatinine, uric acid, urea, cholesterol, triglycerides, LDL, carcinoembryonic antigen, alpha-fetoprotein, interleukin-6, Malondialdehyde, nitric oxide, Bax, caspase-3 and P53 mRNA expression, chromosomal aberrations and DNA fragmentation. It decreased serum TP, albumin, HDL, Bcl-2 mRNA expression, hepatic and renal TAC, SOD and GPx content and induced histological changes in the liver and kidney. FE prevented these disturbances in a dosage-dependent manner. It could be concluded that A. niger SH2-EGY extract is safe a promising agent for pharmaceutical and food industries.


A DECADE OF HEALTH TECHNOLOGY ASSESSMENT IN POLAND.
The objective of this study is to illustrate and provide a better understanding of the role of health technology assessment (HTA) processes in decision making for drug reimbursement in Poland and how this approach could be considered by other countries of limited resources. We analyzed the evolution of the HTA system and processes in Poland over the past decade and current developments based on publicly available information. The role of HTA in drug-reimbursement process in Poland has increased substantially over the recent decade, starting in 2005 with the formation the Agency for Health Technology Assessment and Tariff System (AOTMiT). The key success factors in this development were effective capacity building based on the use of international expertise, the implementation of transparent criteria into the drug reimbursement processes, and the selective approach to the adoption of innovative medicines based on the cost-effectiveness threshold among other criteria. While Poland is regarded as a leader in Central and Eastern Europe, there is room for improvement, especially with regard to the quality of HTA processes and the consistency of HTA guidelines with reimbursement law. In the "pragmatic" HTA model use by AOTMiT, the pharmaceutical company is responsible for the preparation of a reimbursement dossier of good quality in line with HTA guidelines while the assessment team in AOTMiT is responsible for critical review of that dossier. Adoption of this model may be considered by other countries with limited resources to balance differing priorities and ensure transparent and objective access to medicines for patients who need them.


Effects on Surface and Physicochemical Properties of Dielectric Barrier Discharge Plasma-Treated Whey Protein Concentrate/Wheat Cross-Linked Starch Composite Film.
Dielectric barrier discharge (DBD) plasma is a new type of polymer surface modification technology. This study is mainly about the changes in film surface structure and physicochemical properties of whey protein concentrate (WPC)/wheat cross-linked starch (WCS) composite films after DBD plasma treatment with different plasma parameters. The results show that the proper plasma treatment parameters (400 W to 60 s) can increase the surface roughness, tensile strength, barrier properties, and thermal stability of the edible film and decrease elongation at break and the water contact angle. X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction showed that DBD plasma treatment could increase the content of oxygen-containing groups on the WPC/WCS film surfaces instead of damaging the internal crystal structure. The results showed that use of proper DBD plasma treatment technology has a positive effect on the mechanical and barrier properties and thermal stability of WPC/WCS films. PRACTICAL APPLICATION: DBD plasma treatment can improve the mechanical, barrier, and thermal properties of WPC/WCS films without generating any pollution. The DBD plasma can be potentially applied in the enhancement of edible film properties. WPC/WCS films are more environmentally friendly than plastics and can be a replacement for traditional plastics.


Smart particles for noble drug delivery system.
Optically encoded smart particles were prepared for noble drug delivery materials. Distributed Bragg reflector (DBR) porous silicon (PSi) was generated by applying a computer-generated pseudo-square wave current waveform. This DBR PSi film was lifted off from the Si substrate and thermally oxidized to convert PSi to porous silicon dioxide (PSD). DBR PSD film was derivatized with 20(S)-Camptothecin (CPT) and fractured by ultrasono-method to give smart particles. DBR PSD smart particles exhibited a sharp photonic band gap in the optical reflectivity spectrum. Optical characteristic of PSD smart particles retained DBR photonic property in aqueous buffer solution. The release of CPT and change of reflection wavelength were measured by UV-vis and reflectance spectrometer, respectively. The intensity of differential peak from reflection resonances of the smart particles was increased with a drug release. The blue shift of reflection peak resulted in the decrease of refractive index of PSD smart particles during the drug release. The concentration of released drug exhibited an linear relationship with a release time.


New Amorphous Hydrogels with Proliferative Properties as Potential Tools in Wound Healing.
The study and discovery of bioactive compounds and new formulations as potential tools for promoting the repair of dermoepidermal tissue in wound healing is of continuing interest. We have developed a new formulation of amorphous hydrogel based on sodium alginate (NaAlg); type I collagen, isolated by the authors from silver carp tails (COL); glycerol (Gli); Aloe vera gel powder (AV); and silver nanoparticles obtained by green synthesis with aqueous Cinnamomum verum extract (AgNPs@CIN) and vitamin C, respectively. The gel texture of the amorphous hydrogels was achieved by the addition of Aloe vera, demonstrated by a rheological analysis. The evaluations of the cytotoxicity and cell proliferation capacity of the experimental amorphous hydrogels were performed against human foreskin fibroblast Hs27 cells (CRL-1634-ATCC). The developed gel formulations did not show a cytotoxic effect. The hydrogel variant containing AgNPs@CIN in a concentration of 8 µg Ag/gel formulation and hydrogel variant with vitamin C had proliferative activity. In addition, the antibacterial activity of the hydrogels was evaluated against S. aureus ATCC 6538, Ps. aeruginosa ATCC 27853, and E. coli ATCC 25922. The results demonstrated that the gel variant based on AgNPs@CIN in a concentration of 95 µg Ag/gel formulation and the hydrogel based on vitamin C show antibacterial activity. Therefore, the developed hydrogels with AgNPs@CIN and vitamin C could be promising alternatives in wound healing.


Isolation of putative stem cells present in human adult olfactory mucosa.
The olfactory mucosa (OM) has the unique characteristic of performing an almost continuous and lifelong neurogenesis in response to external injuries, due to the presence of olfactory stem cells that guarantee the maintenance of the olfactory function. The easy accessibility of the OM in humans makes these stem cells feasible candidates for the development of regenerative therapies. In this report we present a detailed characterization of a patient-derived OM, together with a description of cell cultures obtained from the OM. In addition, we present a method for the enrichment and isolation of OM stem cells that might be used for future translational studies dealing with neuronal plasticity, neuro-regeneration or disease modeling.


Adaptive walking assistance does not impact long-range stride-to-stride autocorrelations in healthy people.
Many studies have demonstrated in the past that the level of long-range autocorrelations in series of stride durations, characterizing natural gait variability, is impacted by external constraints, such as treadmill or metronome, or by pathologies, such as Parkinson's or Huntington's disease. Nevertheless, no one has analyzed the effects on this metric of a gait constrained by a robot-mediated walking assistance, which intrinsically tends to normalize the gait pattern. This paper focuses on the influence of a wearable active pelvis orthosis on the level of long-range autocorrelations in series of stride durations. Ten healthy participants, aged between 55 and 77 yr, performed four overground walking sessions, wearing this orthosis, and with different assistive parameters. This study showed that the adaptive assistance provided by this device has the potential of improving gait metrics that are typically affected by aging, such as the hip range of motion, walking speed, stride length, and stride duration, without impacting natural gait variability, i.e., the level of long-range autocorrelations in series of stride durations. This combination is virtuous toward the design of an assistive device for people with locomotion disorders resulting in deteriorated levels of long-range autocorrelations, such as patients with Parkinson's disease.NEW & NOTEWORTHY This study is the first that investigates the effects of a wearable active pelvis orthosis using an oscillator-based adaptive assistance on the level of long-range autocorrelations in series of stride durations during overground walking. It is also the first to compare the effects of different assistance settings on spatiotemporal gait metrics.


Machining of a bioactive nanocomposite orthopedic fixation device.
Bioactive ceramics bond to bone and enhance bone formation. However, they have poor mechanical properties which restrict their machinability as well as their application as load bearing implants. The goal of this study was to machine bioactive fixation screws using a silica-calcium phosphate nanocomposite (SCPC50). The effect of compact pressure, holding time, and thermal treatment on the microstructure, machinability, and mechanical properties of SCPC50 cylinders were investigated. Samples prepared by powder metallurgy technique at compact pressure range of 100-300 MPa and treated at 900°C/1 h scored a poor machinability rating of (1/5) due to the significant formation of amorphous silicate phase at the grain boundaries. On the other hand, lowering of compact pressure and sintering temperature to 30 MPa/3 h and 700°C/2 h, respectively, minimized the formation of the amorphous phase and raised the machinability rating to (5/5). The modulus of elasticity and ultimate strength of machinable SCPC50 were 10.8 ± 2.0 GPa and 72.8 ± 22.8 MPa, respectively, which are comparable to the corresponding values for adult human cortical bone. qRT-PCR analyses showed that bone cells attached to SCPC50 significantly upregulated osteocalcin mRNA expression as compared to the cells on Ti-6Al-4V. Moreover, cells attached to SCPC50 produced mineralized bone-like tissue within 8 days. On the other hand, cells attached to Ti-6Al-4V failed to produce bone mineral under the same experimental conditions. Results of the study suggest that machinable SCPC50 has the potential to serve as an attractive new material for orthopedic fixation devices.


The effect of ultrasound on collagen synthesis and fibroblast proliferation in vitro.
Ultrasound has been applied therapeutically to accelerate connective tissue healing although there is little direct scientific evidence to support its use. This investigation was conducted to determine the effects of ultrasound on the rate of collagen synthesis and cell proliferation using cultured fibroblasts derived from Achilles tendons of neonatal rats. Ultrasound (intensity = 0.4 W.cm-2; frequency = 1 MHz) was applied to experimental cells growing as monolayers in culture flasks. Ultrasound had no effect on the rate of collagen synthesis by control fibroblasts over a period of 9 d. The addition of vitamin C to culture media stimulated collagen synthesis to the same extent in both control and ultrasound-treated cultures. Partial digestion of cell matrices with collagenase (used to simulate injury) resulted in an approximately 20% increase in the rate of collagen synthesis. Synthesis was further increased with ultrasound treatment (50-67%). For example, after a single ultrasound treatment, the rate of collagen synthesis was 3.0 +/- 0.4 pg.micrograms-1 DNA.h-1 in cultures treated with collagenase, compared with 1.8 +/- 0.3 pg.micrograms-1 DNA.h-1 in collagenase-treated cultures not treated with ultrasound and 1.4 +/- 0.3 pg.micrograms-1 DNA.h-1 in controls. Ultrasound applied to preconfluent cultures resulted in significant increases in the rate of thymidine incorporation and DNA content. Three daily ultrasound treatments caused a 100% increase in the rate of thymidine incorporation and a 28% increase in DNA content. The results indicate that ultrasound stimulates collagen synthesis in tendon fibroblasts in response to an injury of the connective tissue matrix and that ultrasound stimulates cell division during periods of rapid cell proliferation.


Augmentation of c-fos mRNA expression by activators of protein kinase C in fresh, terminally differentiated resting macrophages.
Expression of c-fos mRNA was investigated in fresh, normal peritoneal macrophages (M phi), which are terminally differentiated, nonproliferating cells. The levels of c-fos mRNA were dramatically increased by stimulation with phorbol myristate acetate (PMA), calcium ionophore, or 1-oleoyl-2-acetoyl glycerol (OAG). Induction of c-fos mRNA by all the above agents followed similar kinetics, with a peak of mRNA 30 min after stimulation. These results demonstrate that c-fos mRNA can be augmented in fresh, terminally differentiated cells. Since the stimuli increasing c-fos mRNA are direct or indirect activators of protein kinase C, our data suggest that in M phi c-fos mRNA is controlled by protein kinase C activation. PMA, calcium ionophore, and OAG were biologically active in M phi. PMA and calcium ionophore induced respiratory burst and tumoricidal activity, respectively, whereas OAG and PMA were chemotactic for M phi. Interferons beta and gamma, potent M phi activators eliciting tumoricidal activity, did not alter the levels of c-fos mRNA. These results indicate that c-fos mRNA augmentation is a stimulus-specific rather than a function-specific response connected to activation of protein kinase C.