Some aspects of the regulation of pyruvate kinase levels in Neurospora crassa.
Pyruvate kinase levels were monitored in Neurospora crassa mycelium (grown on different carbon sources for varying time intervals) by immunoprecipitation using polyclonal antibodies raised against a purified enzyme preparation. Pyruvate kinase specific mRNA was demonstrated by hybridization of Northern and dot blots of total RNA with a N. crassa pyruvate kinase gene fragment. Two pyruvate kinase specific mRNA species were detected in mycelia of all ages examined. An age-dependent and carbon source dependent variation in the pyruvate kinase protein and mRNA levels was encountered: both registered an increase for up to about 20 h and a subsequent decline; growth on acetate and sucrose resulted in significantly higher yields of both, relative to that on medium containing ethanol and alanine. Stress caused by heat shock depressed the pyruvate kinase mRNA levels.


Endocrine function after bone marrow transplantation without the use of preparative total body irradiation.
Ten children who underwent allogeneic (n = 5) or autologous (n = 5) bone marrow transplantation (BMT) for chronic myelogenous leukaemia (n = 2), acute lymphoblastic leukaemia (n = 1), acute myelogenous leukaemia (n = 2), severe aplastic anaemia (n = 2), malignant histiocytosis (n = 1), neuroblastoma (n = 1) and teratoma (n = 1) were assessed for endocrinological function. Transplant preparative regimens consisted of high-dose cyclophosphamide, high-dose cyclophosphamide in combination with high-dose busulphan, high-dose melphalan as well as BACT (BCNU, cytarabine, cyclophosphamide and 6-thioguanine) chemotherapy. None of the patients received total body irradiation (TBI). Median survival following BMT was 37 months (range 7-115). Growth hormone deficiency was present in only one patient; none of the patients had abnormal thyroid or adrenocortical function. This is in contrast to previous reports in which growth hormone deficiency and abnormal thyroid and adrenocortical function occurred in a much higher percentage of patients after BMT conditioned with TBI.


[Embolism in left-atrial thrombi (ELAT Study): are spontaneous echo contrast, thrombi in the left atrium/appendage and size of the left atrial appendage predictors of possible embolisms?].
Transesophageal echocardiography visualizes the left atrium, the left atrial appendage, thrombi and spontaneous echo contrast within them. The role of these findings as predictors for embolism in atrial fibrillation is unknown. We performed transesophageal echocardiography in 409 non-rheumatic atrial fibrillation outpatients (62 +/- 12 years, 36% female) with no recent (< 1 year) history of embolism. Patients with left atrial/appendage thrombi received oral anticoagulation, those without thrombi Aspirin. The patients were followed up over 2 years. Primary events were stroke, embolism and non stroke/embolism related deaths. Secondary events were initiation of anticoagulation in patients primarily assigned to Aspirin. Left atrial/appendage thrombi were diagnosed in 2.5%. They were associated with diabetes, heart failure and decreased left ventricular fractional shortening (p < 0.05 for each variable). Spontaneous echo contrast was diagnosed in 12%. It was associated with increased age, constant atrial fibrillation, hypertension, heart failure, valvular abnormalities and increased left atrial diameter (p < 0.05 for each variable). Increased left atrial appendage size was associated with constant atrial fibrillation, etiology of atrial fibrillation and valvular abnormalities (p < 0.05 for each variable). Follow-up was 25 +/- 7 months. 29 patients suffered a stroke, 33 further patients died of non stroke/embolism related causes. Secondary events occurred in 19 patients. Neither left atrial/appendage thrombi nor left atrial appendage size were predictors for embolism. Predictors for embolism were increased age (p = 0.003), hypertension (p = 0.01) and increased diastolic blood pressure (p = 0.04). In non-rheumatic atrial fibrillation outpatients with no recent history of embolism, transesophageal echocardiography is of limited value to assess embolic risk. Hypertension and increased diastolic blood pressure have been confirmed in their significance as clinical predictors for embolism.


Experimental rickets in broilers: gross, microscopic, and radiographic lesions. I. Phosphorus deficiency and calcium excess.
Day-old broiler chicks were fed diets high in calcium or low in phosphorus for 30 days. Chicks in both groups had similar gross, microscopic, and radiographic lesions. At 2 weeks, the growth-plate degenerating hypertrophied zone (DHZ) and metaphyseal primary spongiosa were lengthened and decreased in radiopacity. Metaphyseal vessels extended to the growth-plate hypertrophied zone (HZ), and osteoid seams were widened. At 3 and 4 weeks, lesions were similar; however, folding fractures were common and cartilage columns of the metaphysis were thinned and replaced with increased woven bone.


Dynamic Mechanism of a Fluorinated Oxime Reactivator Unbinding from AChE Gorge in Polarizable Water.
A well-tempered metadynamics simulation is performed to study the unbinding process of a fluorinated oxime (FHI-6) drug from the active-site gorge of acetylcholinesterase enzyme in a polarizable water medium. Cation-π interactions and water bridge and hydrogen bridge formations between the protein and the drug molecule are found to strongly influence the unbinding process, forming basins and barriers along the gorge pathway. Distinct unbinding pathways are found when FHI-6 was compared with its recently reported nonfluorinated analogue, HI-6. For example, because of permanent positive charges on both the pyridinium rings of HI-6, it exhibits the minimum in the potential of mean force of the unbinding process in the gorge mouth (where the peripheral anion site, PAS, of the enzyme is located), which is largely caused by cation-π interactions. However, the same interaction, both in the catalytic active-site (CAS) and PAS regions, is found to be greatly enhanced in its lipophilic fluorinated analogue, FHI-6, causing a deep potential energy minimum in the bound state. This may render FHI-6 to be held more firmly in the CAS region of the gorge, as is also evidenced from the microkinetics of unbinding transitions, measured through a combination of metadynamics and hyperdynamics simulations.


Inactivation of AIDS-causing retroviruses by the manufacturing procedures for human plasma proteins.
Human retroviruses causing AIDS (HIV) may occur in human plasma. Since HIV contaminated plasma cannot be completely excluded by testing for anti-HIV, AIDS safety of human plasma products can only be achieved by introducing HIV inactivating and/or eliminating methods into the manufacturing procedure. Here we review a number of methods used when manufacturing plasma derivatives at Behringwerke. These methods were previously developed either to produce a protein of required purity or to manufacture hepatitis safe products. Methods used to produce purer proteins are ethanol fractionation, pepsin treatment, affinity chromatography or various protein precipitation procedures. The method developed at Behringwerke for inactivating infectious viruses in plasma protein preparations not destroying the biological activities of the human protein is pasteurization, i.e. 10 h heat treatment of the aqueous protein solution at 60 degrees C. To investigate the HIV inactivating efficiency of the methods mentioned above, aliquots of an infectious HIV type 1 concentrate were added to a protein preparation, the resulting HIV spiked preparation treated according to the method to be studied and the amount of infectious HIV in this preparation determined before and after treatment. By all methods reported on here the HIV type 1 isolate was completely inactivated resulting in high inactivation factors. In addition, the heat stability of HIV type 2 was tested in aqueous solution at 60 degrees C proving that both HIV-1 and HIV-2 isolates are of comparably low heat stability under these conditions. From the results discussed here it can be concluded that all commercial human protein products of Behringwerke derived from either human plasma or placenta do not contain any infectious retrovirus causing AIDS and thus have a high margin of safety regarding the transmission of AIDS.


Thermodynamic integration network approach to ion transport through protein channels: Perspectives and limits.
We present a molecular dynamics simulation study of alkali metal cation transport through the double-helical and the head-to-head conformers of the gramicidin ion channel. Our approach is based on a thermodynamic integration network, which consists of a sequence of transport reactions, absolute free energies of solvation and cycles of alchemical transmutations of the ions. In this manner, we can reliably estimate free energies and their statistical errors via a least-squares method without imposing external forces on the system. Within the double helical channel, we find a free energy surface typical for hopping transport between isoenergetic sites of ion localization, separated by comparatively large activation barriers. For fast transport through the head-to-head conformation, the thermodynamic network scheme starts to break down. © 2018 Wiley Periodicals, Inc.


Treatment of Sinonasal Aspergillosis by Debridement and Sinonasal Deposition Therapy with Clotrimazole Under Rhinoscopic Guidance.
Rhinoscopy was performed on 10 dogs with sinonasal aspergillosis (SNA). Direct access to the sinus via the nasal ostium was possible with a flexible endoscope to allow sinuscopy. Debridement of fungal plaques in the frontal sinus and the nasal cavity was performed, and a sinus and nasal deposition therapy with clotrimazole (1%) cream was made under rhinoscopic guidance. No oral medication was administered following the procedure. A rhinoscopic follow-up was performed monthly until cure. Six of ten (60%) dogs presented fungal plaques in the nasal cavity and in the frontal sinus and 4/10 (40%) dogs presented fungal plaques only in the frontal sinus. Five of ten (50%) dogs were considered to be cured at the first follow-up rhinoscopy, 4/10 (40%) after the second follow-up, and 1/10 (10%) after the third. Two dogs had delayed recurrence of SNA rhinoscopically assessed 12 and 21 mo, respectively, after the last clotrimazole treatment. Endoscopic debridement of fungal plaques and clotrimazole (1%) cream deposition therapy seems to be a valuable minimally invasive technique for SNA treatment in dogs without the use of complementary oral medication. Delayed recurrence is a potential finding following treatment of SNA.


[Cyclic thrombocytopenia associated with erythroid hypoplasia--a case].
We report the case of a 75-year-old woman with cyclic thrombocytopenia associated with erythroid hypoplasia. One platelet cycle lasted for about 28-30 days, with the platelet count fluctuating from 1.0 x 10(4)/microliter to 56.0 x 10(4)/microliter. Megakaryocyte count increased in the phase during which platelet count increased, and decreased in the phase during which platelet count decreased. Bone marrow colony formation was observed in serum-free agar, and megakaryocyte colony count was correlated with the platelet cycle. Platelet-associated immunoglobulin was in the normal range when platelet count increased, but increased when platelet count decreased. These findings suggest that the observed platelet count fluctuation was related to the production and destruction of platelet. Our patient also had erythroid hypoplasia, but her erythrocyte count did not fluctuate. This is the first reported case of cyclic thrombocytopenia and erythroid hypoplasia.


Parental Management of Discharge Instructions: A Systematic Review.
Parents often manage complex instructions when their children are discharged from the inpatient setting or emergency department (ED); misunderstanding instructions can put children at risk for adverse outcomes. Parents' ability to manage discharge instructions has not been examined before in a systematic review. To perform a systematic review of the literature related to parental management (knowledge and execution) of inpatient and ED discharge instructions. We consulted PubMed/Medline, Embase, Cumulative Index to Nursing and Allied Health Literature, and Cochrane CENTRAL (from database inception to January 1, 2017). We selected experimental or observational studies in the inpatient or ED settings in which parental knowledge or execution of discharge instructions were evaluated. Two authors independently screened potential studies for inclusion and extracted data from eligible articles by using a structured form. Sixty-four studies met inclusion criteria; most (n = 48) were ED studies. Medication dosing and adherence errors were common; knowledge of medication side effects was understudied (n = 1). Parents frequently missed follow-up appointments and misunderstood return precaution instructions. Few researchers conducted studies that assessed management of instructions related to diagnosis (n = 3), restrictions (n = 2), or equipment (n = 1). Complex discharge plans (eg, multiple medicines or appointments), limited English proficiency, and public or no insurance were associated with errors. Few researchers conducted studies that evaluated the role of parent health literacy (ED, n = 5; inpatient, n = 0). The studies were primarily observational in nature. Parents frequently make errors related to knowledge and execution of inpatient and ED discharge instructions. Researchers in the future should assess parental management of instructions for domains that are less well studied and focus on the design of interventions to improve discharge plan management.


Understanding the role of microRNAs in the interaction of Aedes aegypti mosquitoes with an insect-specific flavivirus.
The Flavivirus genus contains some of the most prevalent vector-borne viruses, such as the dengue, Zika and yellow fever viruses that cause devastating diseases in humans. However, the insect-specific clade of flaviviruses is restricted to mosquito hosts, albeit they have retained the general features of the genus, such as genome structure and replication. The interactions between insect-specific flaviviruses (ISFs) and their mosquito hosts are largely unknown. Pathogenic flaviviruses are known to modulate host-derived microRNAs (miRNAs), a class of non-coding RNAs that are important in controlling gene expression. Alterations in miRNAs may represent changes in host gene expression and promote understanding of virus-host interactions. The role of miRNAs in ISF-mosquito interactions is largely unknown. A recently discovered Australian ISF, Palm Creek virus (PCV), has the ability to suppress medically relevant flaviviruses. Here, we investigated the potential involvement of miRNAs in PCV infection using the model mosquito Aedes aegypti. By combining small-RNA sequencing and bioinformatics analysis, differentially expressed miRNAs were determined. Our results indicated that PCV infection hardly affects host miRNAs. Out of 101 reported miRNAs of Ae. aegypti, only aae-miR-2940-5p had a significantly altered expression over the course of infection. However, further analysis of aae-miR-2940-5p revealed that this miRNA does not have any direct impact on PCV replication in vitro. Thus, overall the results suggest that PCV infection has a limited effect on the mosquito miRNA profile and therefore miRNAs may not play a significant role in the PCV-Ae. aegypti interaction.


Importance of Resection Margins in the Treatment of Rectal Adenomas by Transanal Endoscopic Surgery.
Polypectomy is the gold standard for treating colorectal adenomas up to 2 cm in size. For larger lesions, various procedures ranging from endoscopy to transanal surgery can be performed and achieve varying results for en bloc resection and recurrence. There are no clear guidelines for dealing with involved resection margins. We assess the recurrence of rectal adenomas operated using TEM with full-thickness wall excision with or without free resection margins and define optimal endoscopic follow-up. Observational study with prospective data collection, including patients undergoing TEM between 6/2004 and 11/2017, with definitive diagnosis of rectal adenoma. Data on epidemiological, preoperative, surgical, postoperative, pathological, and follow-up variables were recorded. Univariate analysis, follow-up risk function, and multivariate logistic regression analysis were performed to detect risk factors for recurrence. TEM was indicated in 736 patients; 481 adenomas were identified in the preoperative biopsy, of which 95 were infiltrating adenocarcinomas (19.8%) in the definitive pathology study. With a minimum follow-up of 1 year, 372 patients were included. Pathology study showed free margins in 324 (87%). Recurrences were recorded in 15 patients (4%), up to 18 months in the free margins group and up to 24 months in the involved margins group. Thirteen patients with recurrence (86.6%) were treated with TEM. No predictors of recurrence were found in the multivariate analysis. TEM is the technique of choice for treating rectal adenomas and recurrences, achieving a low relapse rate. Follow-up must be adapted to resection margins and should be extended to 24 months.


Activity of divicine in Plasmodium vinckei-infected mice has implications for treatment of favism and epidemiology of G-6-PD deficiency.
Intravenous injection of divicine into mice infected with Plasmodium vinckei rapidly killed the parasites and caused haemolysis. Degenerating parasites were observed frequently inside intact circulating erythrocytes, implying that parasite death was not a passive consequence of haemolysis. Both parasite death and haemolysis were prevented by the iron chelator desferrioxamine. In vitro, divicine caused the accumulation of malonyldialdehyde and the depletion of reduced glutathione in normal mouse erythrocytes. Desferrioxamine inhibited the former event, but not the latter. These observations support the hypothesis advanced by Huheey & Martin (Experientia, 31, 1145, 1975) to explain the patchy geographical distribution of glucose-6-phosphate dehydrogenase deficiency in historic malarial areas and also suggest that desferrioxamine, a drug already in clinical use, is a potential treatment for favism and other examples of oxidative haemolysis.


Influence of particle size and reactive oxygen species on cobalt chrome nanoparticle-mediated genotoxicity.
Patients with cobalt chrome (CoCr) metal-on-metal (MOM) implants may be exposed to a wide size range of metallic nanoparticles as a result of wear. In this study we have characterised the biological responses of human fibroblasts to two types of synthetically derived CoCr particles [(a) from a tribometer (30 nm) and (b) thermal plasma technology (20, 35, and 80 nm)] in vitro, testing their dependence on nanoparticle size or the generation of oxygen free radicals, or both. Metal ions were released from the surface of nanoparticles, particularly from larger (80 nm) particles generated by thermal plasma technology. Exposure of fibroblasts to these nanoparticles triggered rapid (2 h) generation of reactive oxygen species (ROS) that could be eliminated by inhibition of NADPH oxidase, suggesting that it was mediated by phagocytosis of the particles. The exposure also caused a more prolonged, MitoQ sensitive production of ROS (24 h), suggesting involvement of mitochondria. Consequently, we recorded elevated levels of aneuploidy, chromosome clumping, fragmentation of mitochondria and damage to the cytoskeleton particularly to the microtubule network. Exposure to the nanoparticles resulted in misshapen nuclei, disruption of mature lamin B1 and increased nucleoplasmic bridges, which could be prevented by MitoQ. In addition, increased numbers of micronuclei were observed and these were only partly prevented by MitoQ, and the incidence of micronuclei and ion release from the nanoparticles were positively correlated with nanoparticle size, although the cytogenetic changes, modifications in nuclear shape and the amount of ROS were not. These results suggest that cells exhibit diverse mitochondrial ROS-dependent and independent responses to CoCr particles, and that nanoparticle size and the amount of metal ion released are influential.


A novel antihypertension agent, sargachromenol D from marine brown algae, Sargassum siliquastrum, exerts dual action as an L-type Ca2+ channel blocker and endothelin A/B2 receptor antagonist.
We isolated the novel vasoactive marine natural products, (5E,10E)-14-hydroxy-2,6,10-trimethylpentadeca-5,10-dien-4-one (4) and sargachromenol D (5), from Sargassum siliquastrum collected from the coast of the East Sea in South Korea by using activity-guided HPLC purification. The compounds effectively dilated depolarization (50mMK+)-induced basilar artery contraction with EC50 values of 3.52±0.42 and 1.62±0.63μM, respectively, but only sargachromenol D (5) showed a vasodilatory effect on endothelin-1 (ET-1)-induced basilar artery contraction (EC50=9.8±0.6μM). These results indicated that sargachromenol D (5) could act as a dual antagonist of l-type Ca2+ channel and endothelin A/B2 receptors. Moreover, sargachromenol D (5) lowered blood pressure in spontaneous hypertensive rats (SHRs) 2h after oral treatment at a dose of 80mg/kg dose and the effect was maintained for 24h. Based on our ex vivo and in vivo experiments, we propose that sargachromenol D (5) is a strong candidate for the treatment of hypertension that is not controlled by conventional drugs, in particular, severe-, type II diabetes-, salt-sensitive, and metabolic disease-induced hypertension.


'Hibernating' accessory pathway.
A 13-year-old boy presented with recurrent narrow QRS tachycardia from which he had been suffering for one year. An electrophysiology study revealed a concealed left lateral accessory pathway with inducible orthodromic atrioventricular re-entry tachycardia using the same pathway. The accessory pathway was ablated in January 2004. After the ablation, atrioventricular block was demonstrated (with adenosine), as was ventriculo-atrial block, and the patient had paroxysmal tachycardia one month later. This time, however, the electrocardiogram in sinus rhythm showed a negative delta wave in V1, suggestive of a right-sided accessory pathway. Another electrophysiology study showed no conduction across the left accessory pathway found earlier. The right accessory pathway, which had 'emerged' after the ablation, had an antegrade effective refractory period of 440 msec and a retrograde effective refractory period of <250 msec. Orthodromic atrioventricular re-entry tachycardia using this accessory pathway was easily inducible. The tricuspid annulus was mapped and the second accessory pathway was localized to the right anterolateral region. This was ablated successfully. This is a unique instance of a symptomatic 'hibernating' accessory pathway emerging in adolescence.


Interaction pathways of implant metal localized corrosion and macrophage inflammatory reactions.
Macrophages play a central role in immunological responses to metallic species associated with the localized corrosion of metallic implants, and mediating in peri-implant inflammations. Herein, the pathways of localized corrosion-macrophage interactions were systematically investigated on 316L stainless steel (SS) implant metals. Electrochemical monitoring under macrophage-mediated inflammatory conditions showed a decreased pitting corrosion resistance of 316L SSs in the presence of RAW264.7 cells as the cells would disrupt biomolecule adsorbed layer on the metal surface. The pitting potentials were furtherly decreased when the RAW264.7 cells were induced to the M1 pro-inflammatory phenotype by the addition of lipopolysaccharide (LPS), and pitting corrosion preferentially initiated at the peripheries of macrophages. The overproduction of aggressive ROS under inflammatory conditions would accelerate the localized corrosion of 316L SS around macrophages. Under pitting corrosion condition, the viability and pro-inflammatory polarization of RAW264.7 cells were region-dependent, lower viability and more remarkable morphology transformation of macrophages in the pitting corrosion region than the pitting-free region. The pitting corrosion of 316L SS induced high expression of CD86, TNF-α, IL-6 and high level of intracellular ROS in macrophages. Uneven release of metallic species (Fe2+, Cr3+, Ni2+, etc) and uneven distribution of surface overpotential stimulated macrophage inflammatory responses near the corrosion pits. A synergetic effect of localized corrosion and macrophages was revealed, which could furtherly promote localized corrosion of 316L SS and macrophage inflammatory reactions. Our results provided direct evidence of corrosion-macrophage interaction in metallic implants and disclosed the pathways of this mutual stimulation effect.


[Immunological and oxidase measurement of ceruloplasmin in pregnant women and newborn (author's transl)].
Ceruloplasmin measurements were made by oxidase and immunochemical methods in serum from three groups of people, normal, pregnant women and their newborns. Whatever the method ceruloplasmin showed a decrease in the newborns and an increase in pregnant women as compared with the normal controls. The three groups showed a significant correlation between both measurement. The correlation ratio between the two methods of measurement in the newborns is significantly different from the pregnant and the normal groups. It appears that ceruloplasmin in the newborns' serum has a lower oxidase activity per molecule than that from adult's serum.


Evaluation of intracellular anion superoxide level, heat shock protein A2 and protamine positive spermatozoa percentages in teratoasthenozoospermia.
Teratoasthenozoospermia (TA) is a severe form of male infertility with no clear etiology. To compare the level of intracellular anion superoxide (O2-), heat shock protein A2 (HSPA2) and protamine deficiencies in ejaculated spermatozoa between teratoasthenozoospermic and normozoospermic men. In this case- control study, semen samples of 20 infertile men, with TA (with normal morphology lower than 4%_ and total motility lower than 40% ) as the case group and 20 normozoospermic fertile men as the control group were evaluated for intracellular O2- and HSPA2 by flow cytometry and protamine deficiency by Chromomycin A3 (CMA3) test. The rate of CMA3+ spermatozoa in the case group was higher than controls (p=0.001). The percentages of HSPA2+ spermatozoa in the cases were significantly lower than controls (p=0.001). Also, intracellular O2- levels in the case group were significantly higher than controls (p=0.001) and had positive correlations with sperm apoptosis (r=0.79, p=0.01) and CMA3 positive sperm (r=0.76, p=0.01), but negative correlations with normal morphology (r=-0.81, p=0.01) and motility (r=-0.81, p=0.01). There was no significant correlation between intracellular O2- and HSPA2 in the case group (r=0.041, p=0.79). We suggest that the increase in intracellular O2-, decrease in spermatozoa HSPA2+, and high percentages of spermatozoa with immature chromatin might be considered as etiologies of infertility in TA patients.


Electrical neurostimulation in glaucoma with progressive vision loss.
The retrospective study provides real-world evidence for long-term clinical efficacy of electrical optic nerve stimulation (ONS) in glaucoma with progressive vision loss. Seventy glaucoma patients (45 to 86 y) with progressive vision loss despite therapeutic reduction of intraocular pressure (IOP) underwent electrical ONS. Closed eyes were separately stimulated by bipolar rectangular pulses with stimulus intensities up to 1.2 mA sufficient to provoke phosphenes. Ten daily stimulation sessions within 2 weeks lasted about 80 min each. Right before ONS at baseline (PRE), vision loss was documented by static threshold perimetry and compared to the same assessment approximately 1 year afterwards (POST). Mean defect (MD) was defined as primary outcome parameter. Perimetries with a reliability factor (RF) of max. 20% were considered. Perimetry follow-up of 101 eyes in 70 patients fulfilled the criterion of a max. 20% RF. Follow-up was performed on average 362.2 days after ONS. MD significantly decreased from PRE 14.0 dB (median) to POST 13.4 dB (p < 0.01). 64 eyes in 49 patients showed constant or reduced MD as compared to baseline (PRE 13.4 dB vs. POST 11.2 dB). In 37 eyes of 30 patients, MD increased from PRE 14.9 dB to POST 15.6 dB. Innovative treatments that preserve visual function through mechanisms other than lowering IOP are required for glaucoma with progressive vision loss. The present long-term data document progression halt in more than 63% of affected eyes after ONS and, thus, extend existing evidence from clinical trials.


Review of patents on microneedle applicators.
Transdermal drug delivery offers certain advantages over conventional oral or parenteral administration. However, transdermal delivery is not available to many promising therapeutic agents, especially high molecular weight hydrophilic compounds. This is due to the excellent barrier property of the superficial skin layer, the stratum corneum (SC). Only drugs with very specific physicochemical properties (molecular weight < 500 Da, adequate lipophilicity, and low melting point) can be successfully administered transdermally. Of the several active approaches used to enhance the transport of drugs through the SC, the use of microneedles (MNs) has recently been shown to be very promising and has attracted considerable attention by researchers from both industry and academia. MNs, when used to puncture skin, will by-pass the SC and create transient aqueous transport pathways of micron dimensions and enhance the transdermal permeability. However, for effective performance of these MNs in drug delivery applications, irrespective of the type, material, height and density, it is imperative that they penetrate into the skin with the greatest possible accuracy and reproducibility. Due to the inherent elasticity and irregular surface topography of the skin, it remains a major challenge to the reproducibility of MN penetration. Therefore, in order to achieve uniform and reproducible MN penetration into skin, an external source of assistance could be very useful. Accordingly, this review deals with various innovative applicator designs developed by industry and research centres in the context of effective application of MN arrays for transdermal drug delivery, as disclosed in the recent patent literature.


[Exogenous harmful substances in pregnancy and their effects on the embryo and fetus].
When using drugs during pregnancy the advantages and risks of a therapy have to be considered carefully. In order to assess the risk of possible teratogenic effects under the influence of external harmful substances the following factors have to be taken into account: the type of the drug administered, kind, dose and duration of the medication, impaired metabolism of drugs owing to diseases of te mother, placenta transfer, time of the exposition during pregnancy, genetic disposition. As in other cases in pharmacology and toxicology there are clear relations of dose and effect as a result of an exposition. High doses, administered for a short time, are extremely dangerous as they may affect the fetus. A deleterious effect on the fetus in the first trimester results in abortions and malformations, in the second and third trimester in general retardations of the fetal development, postnatal impairments of the functional development, increasing perinatal mortality, premature deliveries, stillbirths, transplacental carcinogesis. A long-term exposition or the administration of high doses of different substances at the end of pregnancy may produce syndromes which may possibly endanger the newborn. Following drugs are among these pharmacons: Benzodiazepines, Neuroleptics, tricyclic antidepressants, Lithium, Beta-receptor-blocking agents, Beta-sympathomimetics using as tocolytics, Captopril. Withdrawal syndromes of the newborns have been described after a long-term therapy with certain drugs or due to drug addiction of the mother. In every pregnancy there is a certain risk of giving birth to an abnormal child. Malformations observed at birth or immediately afterwards happen according to the definition with a frequency of 2 to 4% (FRG), according to sources from other countries with a frequency of 2.5 to 6.5%. If you add anomalies diagnosed within first year of life a deformity rate of 10% can be worked out, approximately 4 to 5% of which can be traced back to the exposition to drugs and environmental chemicals. A teratogenic influence would always have to work together with other factors to cause a faulty development of the fetus.


Treatment of idiopathic granulomatous mastitis using ultrasound-guided microwave ablation: a report of 50 cases.
This study aimed to evaluate the efficacy and safety of ultrasound-guided microwave ablation combined with glucocorticoid therapy for treating idiopathic granulomatous mastitis (IGM). From June 2017 to March 2020, 50 consecutive patients diagnosed with IGM using puncture histology were included. All patients received prednisone and ultrasound-guided microwave ablation and were closely monitored for 12-15 months. A total of 222 lesions in 50 patients were ablated. The results indicated that 78% of cases were cured within 12 months and an additional 20% were cured within 15 months; the recurrence rate was 2%. The clinical and pathological remission rate of the entire group was 98%. The main postoperative complications were local pain, skin ulcerations and sinus formation, skin and areola heat damage, subcutaneous congestion, and fat liquefaction, all of which were conservatively treated. Microwave ablation combined with glucocorticoid therapy was safe and effective for the treatment of IGM, with a low recurrence rate. In addition, the cosmetic appearance of the affected breast was preserved with little trauma. Therefore, microwave ablation is a viable method that can be successfully applied in clinical practice.


Assessment of baseline characteristics and risk factors among Emergency Department patients presenting with recent onset atrial fibrillation: a retrospective cohort study.
The Cardioversion of Atrial Fibrillation in Emergency (CAFE) study was an observational, retrospective, multicenter study focusing on patients with recent onset atrial fibrillation (AF) seen in six different Emergency Departments (ED) of Rome, Italy. The aim of this study was to present the baseline characteristics and risk factors of the patients enrolled to the CAFE study. We retrospectively reviewed 3085 eligible patients diagnosed with recent onset AF in any of the EDs between January 2008 and December 2009. Inclusion criteria required documented ICD-9 primary discharge/admission diagnosis of AF in the ED and stable hemodynamic conditions at presentation (systolic blood pressure > 90 mmHg). Exclusion criteria were permanent AF or an ongoing acute coronary syndrome. Median age was 71 years (interquartile ranges, 62-78 years) and 50.8% were men. Palpitations was the most common symptom at ED presentation and was present in 73.5% of the study subjects. Hypertension was the most prevalent comorbidity, affecting 59.3% of the patients evaluated, and the presence of previous episode(s) of AF was also common (52.3%). Regarding home treatment, the drugs most prescribed were antiplatelets (31.2%) and diuretics (25.2%). A CHADS2 score of 0 was found in 814 patients (26.4%), while a CHADS2 score of 1 was reported in 1114 patients (36.1%). Finally, a CHADS2 score ≥ 2 was reported in 1157 patients (37.5%). The present study represents an important snapshot of demographics, comorbidities, risk factors and anticoagulation management about patients with recent onset AF. Disparities were noted in anticoagulation management, suggesting that this is still a main problem among patients with AF.


An anomalous flexor carpi ulnaris.
We report a muscle with features suggesting an anomalous conjoined palmaris longus and flexor carpi ulnaris. To our knowledge this has not been described previously. In light of improved imaging techniques we question the previously published view that surgery is indicated for swellings suspected as forearm muscle anomalies that are clinically benign and otherwise asymptomatic.


Spatial and Temporal Dynamics in Air Pollution Exposure Assessment.
Analyzing individual exposure in urban areas offers several challenges where both the individual's activities and air pollution levels demonstrate a large degree of spatial and temporal dynamics. This review article discusses the concepts, key elements, current developments in assessing personal exposure to urban air pollution (seventy-two studies reviewed) and respective advantages and disadvantages. A new conceptual structure to organize personal exposure assessment methods is proposed according to two classification criteria: (i) spatial-temporal variations of individuals' activities (point-fixed or trajectory based) and (ii) characterization of air quality (variable or uniform). This review suggests that the spatial and temporal variability of urban air pollution levels in combination with indoor exposures and individual's time-activity patterns are key elements of personal exposure assessment. In the literature review, the majority of revised studies (44 studies) indicate that the trajectory based with variable air quality approach provides a promising framework for tackling the important question of inter- and intra-variability of individual exposure. However, future quantitative comparison between the different approaches should be performed, and the selection of the most appropriate approach for exposure quantification should take into account the purpose of the health study. This review provides a structured basis for the intercomparing of different methodologies and to make their advantages and limitations more transparent in addressing specific research objectives.


Three-dimensional supramolecular architecture in imidazolium hydrogen 2,3,5,6-tetrafluoroterephthalate.
The asymmetric unit of the title salt formed between 2,3,5,6-tetrafluoroterephthalic acid (H(2)tfbdc) and imidazolium (ImH), C(3)H(5)N(2)(+).C(8)HF(4)O(4)(-), contains one Htfbdc(-) anion and one ImH(2)(+) cation, joined by a classical N-H...O hydrogen bond. The acid and base subunits are further linked by N-H...O and O-H...O hydrogen bonds into infinite two-dimensional layers with R(6)(5)(32) hydrogen-bond motifs. The resulting (4,4) network layers interpenetrate to produce an interlocked three-dimensional structure. The final three-dimensional supramolecular architecture is further stabilized by the linkages of two C-H...O interactions.


A controlled study of sacroiliitis in Behçet's disease.
Radiographs of the sacroiliac joints of 34 patients with Behçet's disease were read blind by a senior radiologist. They were compared with those from 25 controls, 14 of whom were healthy. The remainder (11) had other rheumatological disorders including six who had AS. One patient with Behçet's disease had radiologically proven sacroiliitis only but her history was atypical and she was B27 positive. All patients with AS had definite sacroiliitis. Five out of 34 patients with Behçet's disease and four of 25 controls (three healthy, one with SLE) had equivocal sacroiliitis. Whilst appreciating that the numbers in this study are not large, we find no evidence of an increased prevalence of sacroiliitis in Behçet's disease.


[Role of NO in the coronary vessel responses to agonists of various adrenoceptor subtypes].
We have studied the role of NO in coronary flow responses to agonists of alfa-2, beta-1, beta-2 and beta-3 adrenoceptors in the isolated mouse heart perfused according to the Langendorff method. The selective alfa-2 adrenoceptor agonist clonidine (10(-8) - 10(-6) M) displayed coronary vasoconstrictor properties as it induced dose-dependent decrease in the coronary flow. On the contrary, the nonselective beta-1/beta2/beta-3 adrenoceptor agonist isoprenaline (10(-8) - 10(-7) M) was a coronary vasodilator and caused dose-dependent increase in the coronary flow. The response to clonidine or isoprenaline was not changed by the presence of the NO-synthase inhibitor L-NO-nitroarginine methyl ester (L-NAME, 5 x 10(-4) M). The selective beta-3 adrenoceptor agonist BRL 37344 at high concentrations (10(-6) - 10(-5) M) produced a coronary vasodilator effect, but this effect was completely blocked by the beta-1/beta-2 adrenoceptor antagonist nadolol (10(-5) M). It is concluded that NO does not play any significant role in the coronary vascular responces mediated by alfa-2, beta-1, and beta-2 adrenoceptors in the isolated mouse heart. The functionally active beta-3 adrenoceptors seem to be not active in the coronary circulation in the isolated mouse heart.


Epidermal stem cells participate in the repair of scalds via Nanog and Myc regulation.
Epidermal stem cells (EpSCs) with high expression of regulatory factor Nanog can promote wound healing. The aim of the present study was to investigate the effectiveness and mechanism of epidermal stem cells (EpSCs) in healing scalds and the underlying molecular mechanism. Mouse EpSCs were isolated from skin tissues and cultured in vitro. First, the proliferative ability of EpSCs was determined via the upregulation and downregulation of Nanog expression levels in EpSCs using the MTS‑assay. Second, a wound healing assay of the EpSCs with different Nanog expression levels was performed to investigate cell migratory capacities. Third, the protein expression levels of various proteins in EpSCs with Nanog overexpression or knockdown, were determined. Finally, the transfected EpSCs were applied to the rat scald model to observe their effect on scald healing. Subsequently, wound scores, re‑epithelialization and capillary density were determined histologically. The results demonstrated that Nanog overexpression enhanced the proliferative ability of EpSCs via cellular (c)‑Myc. Moreover, the LV‑Nanog group of EpSCs with increased Nanog expression levels exhibited improved healing abilities in the wound healing test than control group. Using western blotting, it was demonstrated that EpSCs that were transfected with a Nanog‑overexpression vector expressed high Nanog protein expression levels, whereas small interfering RNA‑Nanog‑transfected EpSCs exhibited low Nanog protein expression levels. Furthermore, c‑Myc expression was synchronized with Nanog expression. It was also revealed that as the expression levels of c‑Myc increased, p53 expression levels also increased. In the rat scald model, Nanog‑overexpressing EpSCs enhanced wound closure and re‑epithelialization. The EpSCs with Nanog knockdown exhibited the opposite effects. The present study therefore indicated that Nanog may have a positive effect on scald healing in rats, which supports its use in EpSC‑based treatments against scalds. Furthermore, it was suggested that c‑Myc potentially serves a key role in this process and that this process avoids cancerization by relying on the supervision of p53.


Acetylated and propionated derivatives of swertiamarin have anti-adipogenic effects.
To investigate whether the acetylated and propionated derivatives (LMP-09-1 and -2) of swertiamarin have anti-adipogenic effects. 3T3-L1 pre-adipocytes were grown in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% calf serum; fully confluent cells were differentiated with insulin, dexamethasone, and 3-isobutylmethylxanthine in the presence and absence of LMP-09-1 and -2 (100 μg/mL) for 10 days. Control cells received same amount of dimethylsulfoxide (DMSO). On day ten, cells were analyzed for triglycerides accumulation and the expression of genes involved in adipogenesis, lipogenesis, and lipolysis. In another set of experiment, effects of LMP-09-1 and 2 were studied for isoproterenol induced lipolysis using fully mature adipocytes. LMP-09-1 and -2 caused a significant (P < 0.001) reduction in intracellular triglycerides accumulation. Both LMP-09-1 and -2 significantly (P < 0.001) decreased the mRNA expression of peroxisome proliferator activated receptor-γ and acetyl-CoA carboxylase-1, and increased isoproterenol induced lipolysis in adipocytes. LMP-09-1 induced lipolysis even in the absence of isoproterenol, and also showed a significant up-regulation of carnitine palmitoyl transferase-1α and hormone sensitive lipase (HSL) gene expression. These findings show that swertiamarin derivatives, LMP-09-1 and -2 have a potent anti-adipogenic effect.


APP processing is regulated by cytoplasmic phosphorylation.
Amyloid-beta peptide (Abeta) aggregate in senile plaque is a key characteristic of Alzheimer's disease (AD). Here, we show that phosphorylation of amyloid precursor protein (APP) on threonine 668 (P-APP) may play a role in APP metabolism. In AD brains, P-APP accumulates in large vesicular structures in afflicted hippocampal pyramidal neurons that costain with antibodies against endosome markers and the beta-secretase, BACE1. Western blot analysis reveals increased levels of T668-phosphorylated APP COOH-terminal fragments in hippocampal lysates from many AD but not control subjects. Importantly, P-APP cofractionates with endosome markers and BACE1 in an iodixanol gradient and displays extensive colocalization with BACE1 in rat primary cortical neurons. Furthermore, APP COOH-terminal fragments generated by BACE1 are preferentially phosphorylated on T668 verses those produced by alpha-secretase. The production of Abeta is significantly reduced when phosphorylation of T668 is either abolished by mutation or inhibited by T668 kinase inhibitors. Together, these results suggest that T668 phosphorylation may facilitate the BACE1 cleavage of APP to increase Abeta generation.


Decreasing Stress and Burnout in Nurses: Efficacy of Blended Learning With Stress Management and Resilience Training Program.
The study's purpose was to assess efficacy of blended learning to decrease stress and burnout among nurses through use of the Stress Management and Resiliency Training (SMART) program. Job-related stress in nurses leads to high rates of burnout, compromises patient care, and costs US healthcare organizations billions of dollars annually. Many mindfulness and resiliency programs are taught in a format that limits nurses' attendance. Consistent with blended learning, participants chose the format that met their learning styles and goals; Web-based, independent reading, facilitated discussions. The end points of mindfulness, resilience, anxiety, stress, happiness, and burnout were measured at baseline, postintervention, and 3-month follow-up to examine within-group differences. Findings showed statistically significant, clinically meaningful decreases in anxiety, stress, and burnout and increases in resilience, happiness, and mindfulness. Results support blended learning using SMART as a strategy to increase access to resiliency training for nursing staff.


Detection of a new heparin-dependent inhibitor of thrombin in human plasma.
We have demonstrated that human plasma contains a heparin-dependent inhibitor of thrombin that is distinguishable from antithrombin III (AT III). When a 1:50 dilution of plasma was incubated with greater than or equal to 0.01 U/ml heparin and 1 U/ml 125I-thrombin, the labeled thrombin B-chains became incorporated into two complexes of Mr-96,000 and Mr-85,000 that were separated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and beta-mercaptoethanol. Neither complex was detectable at heparin concentrations less than 0.01 U/ml. When a limiting amount of 125I-thrombin was present, the proportion of radioactivity incorporated into each of the two complexes varied with the heparin concentration. Thus, the Mr-85,000 complex predominated at 0.01-5 U/ml heparin, whereas the Mr-96,000 complex predominated at 5-100 U/ml heparin. The Mr-85,000 complex reacted with antibodies to human AT III and comigrated with the purified thrombin-AT III complex. The Mr-96,000 complex did not react with antibodies to AT III or to alpha 1-antitrypsin, and it was detected in normal quantities after incubating 125I-thrombin with plasma immunodepleted of AT III, alpha 2-antiplasmin, alpha 2-macroglobulin, C1 inactivator, alpha 1-antichymotrypsin, or inter-alpha-trypsin inhibitor. The protein that combines with thrombin to form the Mr-96,000 complex was estimated to be present at a minimum concentration of 90 +/- 26 micrograms/ml (mean +/- SD) in identical to any of the known plasma protease inhibitors and that at relatively high heparin concentrations in vitro it reacts with thrombin more rapidly than does AT III.


Suspensions as a Valuable Alternative to Extemporaneously Compounded Capsules.
The objective of this study was to determine the variation in content of 74 different active pharmaceutical ingredients (APIs) and compare it with what is known in the literature for the content uniformity of extemporaneous prepared capsules. Active pharmaceutical ingredients quantification was performed by high-performance liquid chromatography, via a stability-indicating method. Samples for all active pharmaceutical ingredients were taken throughout a 90-day period and the content was determined. In total, 5,190 different samples were analyzed for 74 different active pharmaceutical ingredients at room (15°C to 25°C) or controlled refrigerated temperature (2°C to 8°C). Each of these datasets was analyzed according to the United States Pharmacopeia Content Uniformity monograph, corrected for the sample number. The mean acceptance values were well within specifications. In addition, all suspensions complied with the criteria defined by the British Pharmacopoeia monograph for Content Uniformity of Liquid Dispersions for both room and controlled refrigerated temperature. In previous studies, it was found that a routine weight variation check is often not sufficient for quality assurance of extemporaneous prepared capsules. Compounded oral liquids show little variation in content for 74 different active pharmaceutical ingredients; therefore, compounded oral liquids are a suitable alternative when compounding individualized medications for patients.


[Degradation of prolactin 125-I in the mammary gland of lactating rats].
Prolactin-125I metabolism in the mammary gland of lactating rats was studied; the hormone was injected intraperitoneally. Radioactive products accumulated by the mammary gland tissue were extracted with isotonic medium. Tissue extracts, blood serum and milk were analyzed by gel filtration on Sephadex G-200. The Blood displayed a gradual reduction of prolactin-125I content as a result of its splitting in the organs and binding with blood proteins; as to the mammary gland--there occurred accumulation of the products of prolactin-125I degradation. Some hormone was inactivated losing immunological properties without any significant changes in the molecular weight. Besides, the mammary gland displayed an intensive accumulation of the products of prolactin-125I splitting in the other organs and in the gland proper. Radioactivity accumulated in the milk was mainly referred to the products of prolactin-125I degradation. There was also shown the presence of immunologically active prolactin-125I in the milk.


Immunocytochemical localization of the Menkes copper transport protein (ATP7A) to the trans-Golgi network.
We have generated polyclonal antibodies against the amino-terminal third of the Menkes protein (ATP7A; MNK) by immunizing rabbits with a histidine-tagged MNK fusion construct containing metal-binding domains 1-4. The purified antibodies were used in Western analysis of cell lysates and in indirect immunofluorescence experiments on cultured cells. On Western blots, the antibodies recognized the approximately 165 kDa MNK protein in CHO cells and human fibroblasts. No MNK signal could be detected in fibroblasts from a patient with Menkes disease or in Hep3B hepatocellular carcinoma cells, confirming the specificity of the antibodies. Immunocytochemical analysis of CHO cells and human fibroblasts showed a distinct perinuclear signal corresponding to the pattern of the Golgi complex. This staining pattern was similar to that of alpha-mannosidase II which is a known resident enzyme of the Golgi complex. Using brefeldin A, a fungal inhibitor of protein secretion, we further demonstrated that the MNK protein is localized to the trans-Golgi network. This data provides direct evidence for a subcellular localization of the MNK protein which is similar to the proposed vacuolar localization of Ccc2p, the yeast homolog of MNK and WND (ATP7B), the Wilson disease gene product. In light of the proposed role of MNK both in subcellular copper trafficking and in copper efflux, these data suggest a model for how these two processes are linked and represent an important step in the functional analysis of the MNK protein.


Biliary and pancreatic stents.
Endoscopic retrograde cholangiopancreatography is one of the most challenging and demanding procedures performed in endoscopy units. Many endoscopy staff nurses become overwhelmed by the complexity of patient management and the realm of equipment necessary in providing endoscopic therapy for patients with biliary and pancreatic disease. The case of Ray, a 45-year-old patient, is used in this article to demonstrate the current value of biliary stenting and its potential impact on a patient's life. An understanding of the indications for and uses of stents in the treatment of biliary occlusion and pancreatic disease will provide a foundation for those assisting in endoscopic retrograde cholangiopancreatography.


Enhance your technology and enrich your bottom line.
Electronic systems can assist medical practice executives with their biggest practice management challenges, from finding uncollected revenue to tracking patients and supplies, and helping manage documentation and reports. This article explores a few technology opportunities that will boost your bottom line. You can start small and work up to the big purchases. Short-term successes in smoothing work flow or saving money will encourage bigger steps.


The 1942 Massive Contamination of Yellow Fever Vaccine: A Public Health Consequence of Scientific Arrogance.
In the late 1930s, the 17D vaccine against yellow fever was produced in record time. 17D was and is an excellent vaccine. Its rapid diffusion led, however, to several problems, the most important among them being the 1942 massive contamination of the vaccine distributed to the US Army by the hepatitis B virus. The US part of this story is relatively well-known, but its Brazilian part much less so. In 1940, scientists who were producing the 17D vaccine in Rio de Janeiro found that it was contaminated by an "icterus virus" that originated in normal human serum. They solved this problem through the exclusion of human serum from vaccine production, but failed to persuade their US colleagues to do the same. The Rio experts, aware of the potential pitfalls of a new technology, carefully supervised the consequences of their vaccination campaigns. They were thus able to rapidly spot problems and eliminate them. By contrast, US scientists, persuaded of their technical superiority and distrustful of warnings that originated from a "less developed" country, neglected to implement basic public health rules. A major disaster followed. (Am J Public Health. 2021;111(9): 1654-1660. https://doi.org/10.2105/AJPH.2021.306313).


Right brain-left brain: the nurse consultant and behavior change following stroke.
In summary, I have made suggestions for working with staff to promote empathy for the stroke patient presenting with behavior problems. All of us take for granted our "two ways of knowing," the richness of our experience being promoted by the two sides of our brain. When brain damage occurs, the results are sometimes dramatic as with paralysis or expressive aphasia, and sometimes subtle as with receptive aphasia and certain visual-spatial problems. Nursing staff provide the care which otherwise would have been self care. When expectations are low, patients withdraw and staff "care ... less." When expectations are high, patients become symptomatic, staff burned out. The nurse consultant can intervene most effectively by addressing the total milieu, enriching the perspective of staff and family members who are seeing only one side of the whole.


Central and peripheral mechanisms in the stimulation of adrenocortical secretion by the 5-hydroxytryptamine2 agonist, (+-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane.
The mechanisms by which the serotonin2A/2C (5-HT2A/2C) receptor agonist, (+-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) increase adrenocortical secretion were examined. Intraperitoneal (i.p., 2 mumol/kg) and intracerebroventricular (i.c.v., 0.1 pmol -40 nmol) administration of DOI increased plasma corticosterone (CS) concentrations. Administration of the 5-HT2A/2C antagonist, ketanserin (i.c.v.) also increased adrenocortical secretion, although the more selective 5-HT2A/2C antagonist, 6-methyl-1-(1-methylethyl)-ergoline-8 beta-carboxylic acid 2-hydroxy-1-methylpropyl ester (LY 53857) did not. In pentobarbital-anesthetized rats, DOI (i.c.v.) decreased adrenocortical secretion, whereas i.p. administration increased adrenocortical secretion. Ketanserin and LY 53857 (i.c.v.) failed to block CS responses after i.p. DOI, indicating that these increases were not principally due to central 5-HT2 receptor activation; only i.p. administration of ketanserin was able to block responses to DOI. Adrenocortical responses to DOI (i.p.) were preserved after lesions of the hypothalamic paraventricular nucleus or posterior hypothalamic deafferentations. However, responses to DOI were almost abolished in hypophysectomized animals, whereas neurohypophysectomy and treatment with dexamethasone were without effect. Adrenocortical responses after DOI (i.p.) were attenuated in medullectomized and in combined 6-hydroxydopamine-sympathectomized/medullectomized animals, but responses to i.c.v. DOI were not affected by such treatment. 6-Hydroxydopamine lesions of the ventral noradrenergic ascending bundle abolished the response to i.p. DOI, indicating that activation of central noradrenergic systems mediates the increases in adrenocortical secretion. These results demonstrate that central and peripheral 5-HT2 receptors differentially regulate adrenocortical secretion, the central component being sensitive to pentobarbital anesthesia and the peripheral component being dependent upon central and peripheral noradrenergic systems.


Auditory early- and middle-latency evoked potentials in patients with quadrigeminal plate tumors.
Both early auditory evoked potentials (BAEPs) and middle-latency auditory evoked potentials (MLAEPs), were recorded in nine patients suffering from a quadrigeminal plate tumor. These recordings were performed before surgery in six cases and after surgery in three cases. The results of these examinations were correlated with impairments of the midbrain auditory pathways as shown by magnetic resonance imaging. BAEPs and MLAEPs were abnormal in five of nine cases and eight of nine cases, respectively. The two examinations yielded normal results in only one case. These data show that the functional evaluation of the midbrain should not be limited to the recording of BAEPs, routinely performed for brain stem functional evaluation, but should also include recording of MLAEPs, although the technique is a little more delicate. In the five patients with abnormal BAEPs, I-V conduction time was increased unilaterally (three patients) and bilaterally (two patients), and the I/V amplitude ratio was abnormal in two patients. In one of these two patients, isolated destruction of the right inferior colliculus was responsible for an abnormality affecting Wave V of the BAEP that was visible only after left ear stimulation. The most frequently observed MLAEP abnormality was a delay in the peaking of the Pa component, assumed to be of cortical origin. Therefore, a limited impairment of the midbrain may delay the peaking latency of Pa. Unilateral hypovoltage of Na-Pa was also observed. In only one case were Na and Pa components unilaterally abolished, in a patient suffering from a postoperative lesion extending from the right inferior colliculus to the right medial geniculate body.(ABSTRACT TRUNCATED AT 250 WORDS)


Screening the optimal modified biochar for nitrogen retention in black soil.
Reducing the environmental problems caused by nitrogen loss and nitrogen pollution is of great significance. The addition of biochar to soil is a new method for increasing nitrogen interception due to the special structural and physicochemical properties of biochar. The optimal modified biochar was screened out after acid-base modification and batch adsorption test in this paper. And then the effects of different soil and biochar mixing methods on soil physicochemical properties and nitrogen adsorption and retention were explored through soil column leaching test. The results showed that the biochar with a pyrolysis temperature of 700 °C had the best adsorption effect on nitrogen after being modified by 0.1 mol/L HCI, and the adsorption capacity of nitrate nitrogen reached 121.46 mg/g. The adsorption process of ammonia nitrogen and nitrate nitrogen conformed to the Langmuir model and was mainly homogeneous monolayer. After mixing the selected modified biochar with black soil, the pH increased by 4.77%, the content of ammonia nitrogen increased by 4.89%, and the nitrate content increased by 16.62%. In this study, the adsorption effect of biochar on nitrogen in black soil was discussed, so as to explore the optimal use of biochar in soil, which provided some reference basis for the relevant research.


Thermal transitions in dry and hydrated layer-by-layer assemblies exhibiting linear and exponential growth.
Layer-by-layer (LbL) assemblies are remarkable materials, known for their tunable mechanical, optical, and surface properties in nanoscale films. However, questions related to their thermal properties still remain unclear. Here, the thermal properties of a model LbL assembly of strong polyelectrolytes, poly(diallyldimethylammonium chloride)/poly(styrene sulfonate) (PDAC/PSS), assembled from solutions of varying ionic strength (0-1.25 M NaCl) are investigated using quartz crystal microbalance with dissipation (QCM-D) and modulated differential scanning calorimetry. Hydrated exponentially growing films (assembled from 0.25 to 1.25 M NaCl) exhibited distinct thermal transitions akin to a glass transition at 49-56 °C; linearly growing films (assembled without added salt) did not exhibit a transition in the temperature range investigated and were glassy. Results support the idea that exponentially growing films have greater segmental mobility than that of linearly growing films. On the other hand, all dry LbL assemblies investigated were glassy at room temperature and did not exhibit a T(g) up to 250 °C, independent of ionic strength. For the first time, thermal transitions such as T(g) values can be measured for LbL assemblies using QCM-D by monitoring fluctuations in changes in dissipation, allowing us to probe the film's internal structure as a function of film depth.


New Insights into the Role of Matrix Metalloproteinases in Preeclampsia.
Preeclampsia is a severe pregnancy complication globally, characterized by poor placentation triggering vascular dysfunction. Matrix metalloproteinases (MMPs) exhibit proteolytic activity implicated in the efficiency of trophoblast invasion to the uterine wall, and a dysregulation of these enzymes has been linked to preeclampsia. A decrease in MMP-2 and MMP-9 interferes with the normal remodeling of spiral arteries at early pregnancy stages, leading to the initial pathophysiological changes observed in preeclampsia. Later in pregnancy, an elevation in MMP-2 and MMP-9 induces abnormal release of vasoactive factors conditioning hypertension. Although these two enzymes lead the scene, other MMPs like MMP-1 and MMP-14 seem to have a role in this pathology. This review gathers published recent evidence about the implications of different MMPs in preeclampsia, and the potential use of these enzymes as emergent biomarkers and biological therapeutic targets, focusing on studies involving human subjects.


Sensitive detection of electric field-induced second harmonic signals.
We demonstrate sensitive electric field measurements by coherent homodyne amplification of the electric field induced second harmonic generation (E-FISH) technique. In the process of E-FISH, an applied electric field breaks the centrosymmetry of an otherwise homogeneous medium, in turn promoting the generation of the second harmonic frequency of an incident field. Due to weak third-order hyperpolarizability and the requirement of an applied field to break the symmetry, the E-FISH technique has been mainly used to study high fields, also requiring a strong optical field and sensitive detection. Here we superimpose the E-FISH signal with an auxiliary beam, also termed a local oscillator (LO), at double the incident frequency. Coherent superposition of the LO and the E-FISH output (LOE-FISH) allows for a homodyne amplification of the otherwise weak nonlinear signal. We have demonstrated an increase of signal-to-noise by a factor of seven, which results in a measurement time reduction of a factor of 49. This technique, LOE-FISH, has a number of advantages: detection with intensified detectors is not required. Furthermore, instead of millijoule pulsed lasers, we can work with microjoule pulsed lasers, which allows measuring at repetition rates of megahertz and opens single shot and real-time capability. The LOE-FISH technique increases in sensitivity at lower electric field values. Our work is a demonstration of the principle. Already with our first results from the demonstration, one can see the high potential of LOE-FISH.


Syndactyly and intracranial arteriovenous malformation: case report.
A case of a 14-year-old boy with syndactyly of all limbs and intracranial dural arteriovenous malformation (AVM) is presented. A failure of differentiation of cerebral vessels, possibly at 3 weeks gestational age, is discussed along with several alternative theories relating to the pathogenesis of dural AVMs. The embryogenesis of syndactyly, occurring as an arrest of normal interdigital tissue regression at 5-7 weeks gestational age, is also reviewed. It is postulated that a single common intrauterine environmental insult of vascular origin occurring during the second month of gestation, is the reason for their simultaneous manifestation.


A Technical Look at Fetoscopic Laser Ablation for Fetal Laryngeal Surgical Recanalization in Congenital High Airway Obstruction Syndrome.
Background: Congenital high airway obstruction syndrome (CHAOS) is a rare condition characterized by complete obstruction of the upper fetal airways. Left untreated, it is uniformly fatal. Ex utero intrapartum treatment (EXIT) has been used to establish a surgical airway in affected fetuses during delivery. While this procedure benefits those fetuses that survive to delivery, high mortality in the prenatal period necessitates earlier innovative strategies. Herein, we report a novel technique for in utero intervention. Methods: A fetoscopic intervention was performed at 28 weeks on a 35-year-old G1P0 woman with fetal CHAOS from a laryngeal obstruction measuring 11 mm in length on prenatal imaging. Under ultrasound guidance, a 3.3-mm curved fetoscope was used to access the uterine cavity through a single subcentimeter maternal skin incision. The scope was driven through the fetal oral cavity and manipulated to attain a view of the vocal cords. A subglottic obstruction was observed. A 600-micron laser fiber was passed through the working channel of the scope and used to ablate the obstructed airway. Using the laser fiber and a guidewire, the ablated opening was traversed with the fetoscope to the level of the carina. Results: Postoperatively, the lungs became less hyperinflated. There was improvement in ascites and diaphragmatic eversion. At 31 1/7 weeks' gestation, the mother experienced preterm premature rupture of membranes with active labor and the fetus was delivered through EXIT to tracheostomy. The infant was managed on mechanical ventilation and is currently thriving at home with a tracheostomy at 2 years of age. Conclusion: Fetoscopy with laser ablation of the airway obstruction is an effective prenatal management strategy that offers the potential to alter the devastating natural course of CHAOS.


The daily spiritual experience scale: development, theoretical description, reliability, exploratory factor analysis, and preliminary construct validity using health-related data.
Spirituality and religiousness are gaining increasing attention as health research variables. However, the particular aspects examined vary from study to study, ranging from church attendance to religious coping to meaning in life. This frequently results in a lack of clarity regarding what is being measured, the meaning of the relationships between health variables and spirituality, and implications for action. This article describes the Daily Spiritual Experience Scale (DSES) and its development, reliability, exploratory factor analyses, and preliminary construct validity. Normative data from random samples and preliminary relationships of health-related data with the DSES also are included. Detailed data for the 16-item DSES are provided from two studies; a third study provided data on a subset of 6 items, and afourth study was done on the interrater reliability of the item subset. A 6-item version was used in the General Social Survey because of the need to shorten the measure for the survey. A rationale for the conceptual underpinnings and item selection is provided, as are suggested pathways for linkages to health and well-being. This scale addresses reported ordinary experiences of spirituality such as awe, joy that lifts one out of the mundane, and a sense of deep inner peace. Studies using the DSES may identify ways in which this element of life may influence emotion, cognition and behavior, and health or ways in which this element may be treated as an outcome in itself a particular component of well-being. The DSES evidenced good reliability across several studies with internal consistency estimates in the .90s. Preliminary evidence showed that daily spiritual experience is related to decreased total alcohol intake, improved quality of life, and positive psychosocial status.


Immediate consequences of spinal cord injury: possible role of potassium in axonal conduction block.
Compression injury of the spinal cord of cats produced increased outflux of potassium from the damaged site. Conduction by large axons in sensory pathways of the spinal cord was blocked partially by bathing the cord in solutions containing potassium concentrations of the order of 50-100 mEq/liter. It is suggested that block by increased K+ concentration in the extracellular space could interfere with long tract function immediately after cord injury before the onset of gross structural changes, and that increased extracellular potassium may be a link in the mechanisms producing thses later changes.


Socioeconomic and psychosocial correlates of oral health.
It has been proposed that psychosocial variables are important determinants of oral health outcomes. In addition, the effect of socioeconomic factors in oral health has been argued to work through the shaping of psychosocial stressors and resources. This study therefore aimed to examine the role of psychosocial factors in oral health after controlling for selected socioeconomic and behavioural factors. Logistic and generalised linear regression analyses were conducted on self-rated oral health, untreated decayed teeth and number of decayed, missing and filled teeth (DMFT) from dentate participants in a national survey of adult oral health (n = 5364) conducted in 2004-2006 in Australia. After controlling for all other variables, more frequent dental visiting and toothbrushing were associated with poorer self-rated oral health, more untreated decay and higher DMFT. Pervasive socioeconomic inequalities were demonstrated, with higher income, having a tertiary degree, higher self-perceived social standing and not being employed all significantly associated with oral health after controlling for the other variables. The only psychosocial variables related to self-rated oral health were the stressors perceived stress and perceived constraints. Psychosocial resources were not statistically associated with self-rated oral health and no psychosocial variables were significantly associated with either untreated decayed teeth or DMFT after controlling for the other variables. Although the role of behavioural and socioeconomic variables as determinants of oral health was supported, the role of psychosocial variables in oral health outcomes received mixed support.


Isolation and functional characterization of murine prostate stem cells.
The ability to isolate prostate stem cells is essential to explore their role in prostate development and disease. In vitro prostate colony- and sphere-forming assays were used to quantitatively measure murine prostate stem/progenitor cell enrichment and self-renewal. Cell surface markers were screened for their ability to positively or negatively enrich for cells with enhanced growth potential in these assays. Immunohistochemical and FACS analyses demonstrate that specific cell surface markers can be used to discriminate prostate stromal (CD34(+)), luminal epithelial (CD24(+)CD49f(-)), basal epithelial (CD24(+)CD49f(+)), hematopoietic (CD45(+), Ter119(+)), and endothelial (CD31(+)) lineages. Sorting for cells with a CD45(-)CD31(-)Ter119(-)Sca-1(+)CD49f(+) antigenic profile results in a 60-fold enrichment for colony- and sphere-forming cells. These cells can self-renew and expand to form spheres for many generations and can differentiate to produce prostatic tubule structures containing both basal and luminal cells in vivo. These cells also localize to the basal cell layer within the region of the gland that is proximal to the urethra, which has been identified as the prostate stem cell niche. Prostate stem cells can be isolated to a purity of up to 1 in 35 by using this antigenic profile. The remarkable similarity in cell surface profile between prostate and mammary gland stem cells suggests these markers may be conserved among epithelial stem cell populations.


The Prevalence and Characteristic Differences in Prostatic Calcification between Health Promotion Center and Urology Department Outpatients.
We evaluated the differences in calculi characteristics and their prevalence according to the presence of lower urinary tract symptoms between adult patients examined at the Urology Department and those examined at the Health Promotion Center (HPC). The prevalence of prostatic calcification, characteristics of calculi (number, size, and location), and differences in lower urinary tract symptoms were compared and analyzed for 479 subjects who underwent transrectal ultrasonography at the HPC and the Urology Outpatients Department at our hospital from October 2009 to October 2010. Of 479 subjects, 268 patients were examined at the HPC, and 211 were examined at the Urology Outpatients Department. Between the two groups, age, prostate-specific antigen levels, prostate volume transrectal ultrasonography, International Prostate Symptom Score (total, voiding, and storage), quality of life, and the prostatic calcification rate were significantly higher in the patients who visited the Urology Outpatients Department. The prevalence of prostatic calcification was 41.5% (199/479), with 36.1% (97/268) from the HPC and 48.3% (102/211) from the Urology Outpatients Department. When the characteristics of prostatic calcification were compared, there were no significant differences in the appearance, size, or location of the calculi between the two groups. The prevalence of prostatic calcification was high in patients complaining of lower urinary tract symptoms; however, there were no significant differences in the characteristics of the calculi. This finding leads us to believe that prostatic calcification can aggravate lower urinary tract symptoms but does not result in differences according to the number, size, or appearance of the calculi.


Charcot-Marie-Tooth disease: genetic subtypes in the Sardinian population.
Charcot-Marie-Tooth disease (CMT) is characterised by great variability of genetic subtypes. This study aimed to assess the genetic subtypes of CMT disease in the Sardinian population. Genetic screening was performed for CMT cases (CMT1, CMT2, and hereditary neuropathy with susceptibility to pressure palsies [HNPP]). A total of 1,043 subjects (119 index cases) were evaluated. In CMT1 index cases (69/119; 58%), PMP22 duplication at 17p11.2 was the most frequent genetic diagnosis (60/69; 87%), followed by mutations in the GJB1 gene (5/69; 7.2%), in the SH3TC2 gene (3/69; 4.4%) and PMP22 Gly107Val point mutation (1/69; 1.4%). The CMT2 group (24/119; 20.1%) comprised 10/24 (41.6%) patients carrying MPZ gene Ser44Phe mutation, 6/24 (25%) with mutations in MFN2 and HSPB1, and 1/24 (4.2%) in GJB1 and LRSAM1. In the HNPP group (26/119; 21.9%), the majority of patients reported the PMP22 deletion (25/26; 96.2%). Further studies are needed to comprehend the overall picture of the disease in Mediterranean area.


Comparison of glucosuria in the first- and second-voided specimens and the time of day as tested by four commercial testing products.
The questions addressed in this study were whether testing first- and second-voided urine specimens yielded different results and whether selected commercial testing product (Clinitest 2 drop, Clinitest 5 drop, Tes-Tape, and Keto-Diastix) yielded similar estimates of glucosuria. Twenty adult diabetic experiencing glucosuria served as subjects. First- and second-voided specimens were collected before meals and before retiring. Data were analyzed using a three-way ANOVA with repeated measures on all factors. The findings indicated significant main effects for product type, F (3,57) = 7.61, p = .0004, and time of day, F (3,57) = 8.60, p = .0002. The main effect for first- and second-voided specimens did not approach significance.


Efficacy and safety of combined targeted therapy and immunotherapy versus targeted monotherapy in unresectable hepatocellular carcinoma: a systematic review and meta-analysis.
Cancer therapy has evolved from non-specific cytotoxic agents to a selective, mechanism-based approach that includes targeted agents and immunotherapy. Although the response to targeted therapies for unresectable hepatocellular carcinoma (HCC) is acceptable with the improved survival, the high tumor recurrence rate and drug-related side effects continue to be problematic. Given that immune checkpoint inhibitor alone are not robust enough to improve survival in unresectable HCC, growing evidence supports the combination of targeted therapy and immunotherapy with synergistic effect. Online databases including PubMed, EMBASE, Cochrane Library, and Web of Science were searched for the studies that compared targeted monotherapy with the combination therapy of targeted drug and checkpoint inhibitors in unresectable HCC patients. Eligibility criteria were the presence of at least one measurable lesion as defined by the Response Evaluation Criteria in Solid Tumors (version 1.1) for unresectable HCC patients, an Eastern Cooperative Oncology Group performance status of 0-2, and a Child-Pugh score ≤ 7. Outcome measurements include overall survival (OS), progression-free survival (PFS), and treatment-related adverse event (TRAE). Three phase II/III randomized controlled trials were included in this study. The pooled results showed that combination therapy significantly improved survival than targeted monotherapy, in terms of OS (hazard ratio (HR) = 0.67; 95% confidence interval [CI]: 0.50-0.91) and PFS (HR = 0.58; 95% CI: 0.51-0.67), respectively. In the incidence of grade 3-5 TRAEs, the combination therapy was significantly higher than targeted monotherapy (odds ratio = 1.98; 95% CI: 1.13-3.48). For unresectable HCC, combined targeted drug and immunotherapy significantly improved survival compared with targeted monotherapy. However, the incidences of AEs of combinational therapy were higher than targeted monotherapy.


Efficacy of oral and intravenous dexamethasone in horses with recurrent airway obstruction.
Although the efficacy of dexamethasone for the treatment of recurrent airway obstruction (RAO) has been documented, the speed of onset of effect and duration of action are unknown, as is the efficacy of orally administered dexamethasone with or without fasting. To document the time of onset of effect and duration of action of a dexamethasone solution i.v. or orally with and without fasting. Protocol 1 used 8 RAO-affected horses with airway obstruction in a crossover design experiment that compared the effect of i.v. saline and dexamethasone (0.1 mg/kg bwt) on pulmonary function over 4 h. Protocol 2 used 6 similar horses to compare, in a crossover design, the effects of dexamethasone i.v. (0.1 mg/kg bwt), dexamethasone per os (0.164 mg/kg bwt) with and without prior fasting, and dexamethasone per os (0.082 mg/kg) with fasting. Dexamethasone i.v. caused significant improvement in lung function within 2 h with a peak effect at 4-6 h. Dexamethasone per os was effective within 6 h with peak effect at 24 h at a dose of 0.164 mg/kg bwt prior to feeding. The duration of effect was, for all dexamethasone treatments, statistically significant for 30 h when compared to saline and tended to have a longer duration of effect when used orally. Dexamethasone per os at a dose of 0.164 mg/kg bwt to fed horses had mean effects comparable to dexamethasone at a dose of 0.082 mg/kg bwt per os given to fasted horses, indicating that feeding decreases bioavailability. Dexamethasone administered i.v. has a rapid onset of action in RAO-affected horses. Oral administration of a bioequivalent dose of the same solution to fasted horses is as effective as i.v. administration and tends to have longer duration of action. Fasting horses before oral administration of dexamethasone improves the efficacy of treatment. Oral administration to fasted horses of a dexamethasone solution intended for i.v. use provides an effective treatment for RAO-affected animals.


Drug resistance of Mycobacterium tuberculosis strains isolated from patients with pulmonary tuberculosis in Archangels, Russia.
The Archangels oblast, Russia, 1998-2000. To study Mycobacterium tuberculosis resistance to anti-tuberculosis drugs in the Archangels oblast, and to reveal risk factors for the development of drug-resistant tuberculosis. The drug susceptibility of strains isolated from 119 patients with pulmonary tuberculosis was studied using the BACTEC method. Medical records of the patients were reviewed, retrospectively, to identify factors associated with drug resistance. Sixty-seven strains (56.3%) were resistant to at least one anti-tuberculosis drug. The highest rates of resistance were observed for streptomycin and isoniazid: respectively 40.4% and 66.7% of strains isolated from newly and previously treated patients were resistant to streptomycin, and respectively 37.1% and 73.3% of strains isolated from newly and previously treated patients were resistant to isoniazid. Thirty of the 119 strains (25.2%) were multidrug-resistant. Multidrug resistance was four times higher among previously treated patients than among new patients. A history of previous or interrupted treatment for tuberculosis and being female were significantly associated with drug resistance. Drug-resistant tuberculosis is an important problem in the Archangels oblast, Russia. The spread of drug resistance is attributed to several risk factors. Being female and evidence of previous treatment for tuberculosis are risk factors for the development of drug-resistant tuberculosis in the Archangels oblast. Patients with drug-resistant tuberculosis also showed a higher risk of interrupting their treatment.


Manufacture of magnesium-fortified Chihuahua cheese.
The aim of this study was to manufacture magnesium-fortified Chihuahua cheese and to evaluate the effect of magnesium fortification on quality parameters. Addition of magnesium chloride to milk during pasteurization (5.44, 10.80, 16.40, 22.00, and 25.20 g of MgCl2·6H2O/L of milk) resulted in cheese with increased magnesium content, proportional to the amount of magnesium added (up to 2,957.13 mg of Mg/kg of cheese). As magnesium content increased, coagulation time and moisture content also increased, whereas calcium content decreased. Higher levels of magnesium fortification (16.40 g of MgCl2·6H2O/L of milk or more) induced the development of bitter-acid flavors and softer texture. Addition of 10.80 g of MgCl2·6H2O/L to milk resulted in Chihuahua cheese that meets regulatory standards and possesses physicochemical and sensory characteristics similar to those of nonfortified Chihuahua cheese. Under this milk fortification level, the manufactured cheese is able to provide 148.4 mg of magnesium per day (35% of the recommended daily intake of magnesium for adult males and 46% for adult females) assuming 3 portions (28 g each) are consumed.


Fibre-years, pulmonary asbestos burden and asbestosis.
The relations between cumulative asbestos fibre doses at the work-places and asbestos burden of the lung evaluated by lung dust analyses have been tested on 3 different groups of patients of the German Mesothelioma Register: 1. total collective (n = 366), 2. collective without elevated asbestos burden of the lungs (n = 193), 3. collective with asbestoses/minimal asbestoses (n = 64). The relations between the above mentioned parameters are in general only weak. The limit value of > 25 fibre-years is found in 19.6% of persons without increased pulmonary asbestos burden. In spite of reaching or exceeding the cumulative doses of 25 fibre-years, 24% of the whole collective also show no elevated asbestos-concentrations in their lung tissues. By contrast, 42% of patients with asbestos-associated lung fibroses do not attain 25 fibre-years at their work-places. Considering our data it is doubtful that the postulated limit value of 25 fibre-years can be an adequate parameter for the evaluation of asbestos-associated lung fibroses.


Platelet lysates produced from expired platelet concentrates support growth and osteogenic differentiation of mesenchymal stem cells.
Mesenchymal stem cells are promising candidates in regenerative cell therapy. Conventional culture methods involve the use of animal substances, specifically fetal bovine serum as growth supplement. Since the use of animal-derived products is undesirable for human applications, platelet lysates produced from human platelets are an attractive alternative. This is especially true if platelet lysates from already approved transfusion units at blood banks can be utilized. The purpose of this study was to produce human platelet lysates from expired, blood bank-approved platelet concentrates and evaluate their use as growth supplement in the culture of mesenchymal stem cells. In this study, bone marrow-derived mesenchymal stem cells were cultured with one of three culture supplements; fetal bovine serum, lysates from freshly prepared human platelet concentrates, or lysates from expired human platelet concentrates. The effects of these platelet-derived culture supplements on basic mesenchymal stem cell characteristics were evaluated. All cultures maintained the typical mesenchymal stem cell surface marker expression, trilineage differentiation potential, and the ability to suppress in vitro immune responses. However, mesenchymal stem cells supplemented with platelet lysates proliferated faster than traditionally cultured cells and increased the expression of the osteogenic marker gene RUNX-2; yet no difference between the use of fresh and expired platelet concentrates was observed. Our findings suggest that human platelet lysates produced from expired platelet concentrates can be used as an alternative to fetal bovine serum for mesenchymal stem cell culture to the same extent as lysates from fresh platelets.


Receptor-bound uPA is reversibly protected from inhibition by low molecular weight inhibitors.
Urokinase-type plasminogen activator (uPA) plays a ubiquitous role in cell migration and invasiveness. Amiloride, a competitive inhibitor of uPA, can inhibit endothelial cell (EC) outgrowth during angiogenesis. To address the question of whether amiloride blocked angiogenesis by inhibiting uPA, we undertook a study of uPA expression in sprouting EC in vitro and the effects of amiloride on both enzymatic and morphogenetic activity. As expected, amiloride inhibited soluble uPA (suPA) with an IC(50) of 45-85 microm, however, receptor-bound uPA (rbuPA) from the sprouting EC was insensitive to amiloride. Removal of uPA from its receptors confers sensitivity to inhibition by amiloride suggesting that a reversible conformational change may mediate the insensitivity of rbuPA to amiloride and its analogs. In summary, we found no evidence to support the hypothesis that amiloride blocks capillary outgrowth by inhibition of uPA, but were able to successfully demonstrate a functional difference between two physiological forms of this important matrix-degrading enzyme.


Social Capital in Relation to Mental Health-The Voices of Adolescents in Sweden.
The social environment that adolescents interact in has undoubtedly changed over the past decades. The latent constructs of social capital that have been described in theory may be universal, but it is necessary to reveal sociocultural specific pathways and manifestation in order to validly operationalize social capital for adolescents. There is a call for qualitative data to enhance our understanding of social capital for adolescents today and the specific sociocultural context they live in. The aim of this study was to explore social capital from the perspective of adolescents in relation to mental health. Twenty-three semi-structured interviews were conducted in a school setting with a sample of adolescents aged 11 and 15 years. Qualitative content analysis was applied, and analysis remained on a manifest level. From having adolescents describe their social relations and networks in relation to mental health, three main categories were formed: accessing a safe space, with sub-categories of trusting enough to share, having someone close to you, and being part of an inclusive and honest environment; feeling connected to others, with sub-categories of hanging out and having things in common; and maintaining control, with sub-categories of deciding for yourself, dealing with change, and having social skills. Having access to a safe space is vital for adolescents' mental health, by providing resources such as mutual trust, honesty, and unconditional access. Feeling connected to others is important in close relationships and reveals the glue that holds networks together, but also links to sociability in a wider sense. Predictability in adolescents' social relationships and networks, influenced by internal and external factors, may be a resource of increasing importance in todays' society and an interesting subject for intervention and future research on social capital and adolescent mental health.


Regulation of Extrasynaptic GABAA α4 Receptors by Ethanol-Induced Protein Kinase A, but Not Protein Kinase C Activation in Cultured Rat Cerebral Cortical Neurons.
Ethanol produces changes in GABAA receptor trafficking and function that contribute to ethanol dependence symptomatology. Extrasynaptic γ-aminobutyric acid A receptors (GABAA-R) mediate inhibitory tonic current and are of particular interest because they are potentiated by physiologically relevant doses of ethanol. Here, we isolate GABAA α4δ receptors by western blotting in subsynaptic fractions to investigate protein kinase A (PKA) and protein kinase C (PKC) modulation of ethanol-induced receptor trafficking, while extrasynaptic receptor function is determined by measurement of tonic inhibition and responses evoked by 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP). Rat cerebral cortical neurons were grown for 18 days in vitro and exposed to ethanol and/or PKA/PKC modulators. Ethanol exposure (1 hour) did not alter GABAA α4 receptor abundance, but it increased tonic current amplitude, an effect that was prevented by inhibiting PKA, but not PKC. Direct activation of PKA, but not PKC, increased the abundance and tonic current of extrasynaptic α4δ receptors. In contrast, prolonged ethanol exposure (4 hours) reduced α4δ receptor abundance as well as tonic current, and this effect was also PKA dependent. Finally, PKC activation by ethanol or phorbol-12,13-dibutyrate (PdBu) had no effect on extrasynaptic α4δ subunit abundance or activity. We conclude that ethanol alters extrasynaptic α4δ receptor function and expression in cortical neurons in a PKA-dependent manner, but ethanol activation of PKC does not influence these receptors. These results could have clinical relevance for therapeutic strategies to restore normal GABAergic functioning for the treatment of alcohol use disorders.


Association of single nucleotide polymorphism (SNP) markers in candidate genes and QTL regions with pork quality traits in commercial pigs.
Numerous reports have described genetic markers or genomic regions (QTL) associated with pork quality and/or palatability but few validation studies have been reported. Therefore, 156 SNP markers from 45 candidate genes and eight QTL regions were analyzed for association with pork quality and palatability traits from 888 pork loins. Loins were collected at three slaughter facilities and selected to represent a wide range of pork color, pH and marbling. Phenotypic data recorded included objective and subjective measures of color and marbling, purge loss, shear force, and cooking loss. Data were analyzed with SAS PROC MIXED where loin was fit as a random effect. Results indicated some of the markers tested should be useful in industry, while others are not segregating in all populations or linkage disequilibrium between markers and causative genetic variation fluctuates among populations limiting their universal utility. Genes with the largest effects on pork quality were MC4R, IGF2, CAST and PRKAG3.


[Identification of mutations associated with coronary artery lesion susceptibility in Kawasaki disease by targeted enrichment of genomic region sequencing technique].
Objective: To screen and identify the mutations in Kawasaki disease by targeted enrichment of genomic region sequencing technique and investigate susceptibility genes associated with coronary artery lesion. Method: This was a case-control study.A total of 114 patients diagnosed as Kawasaki disease treated in Shanghai Children's Hospital between December 2015 and November 2016 were studied and another 45 healthy children who were physically examined in outpatient department were enrolled as control group. Patients were divided into two groups based on the results of echocardiogram. Peripheral venous blood was obtained from patients and controls. Genomic DNA was extracted. SeqCap EZ Choice libraries were prepared by targeted enrichment of genomic region technology. Then the libraries were sequenced to identify susceptibility genes associated with coronary artery lesion in patients diagnosed as Kawasaki disease.Susceptible genes were identified by Burden test, Pearson chi-square test or Fisher's exact probability test. Result: There was statistically significant difference in TNFRSF11B(rs2073618)G>C(p.N3K)mutation and GG/GC/CC genotype between Kawasaki disease group and control group(χ(2)=15.52, P=0.00). There was statistically significant difference in TNFRSF13B(rs34562254)C>T(p.P251L)mutation(χ(2)=10.40, P=0.01)and LEFTY1(rs360057)T>G(p.D322A)mutation(χ(2)=8.505, P=0.01)between patients with coronary artery lesions and those without. Conclusion: Targeted enrichment of genomic region sequencing technology can be used to do primary screening for the susceptible genes associated with coronary artery lesions in Chinese Kawasaki patients and may provide theoretical basis for larger sample investigation of risk prediction score standard in Kawasaki disease.


Assessment of the intraobserver and interobserver reliability of a communicating vessels volumeter to measure wrist-hand volume.
Traditional volumetry based on Archimedes' principle is the gold standard for the measurement of limb volume, but the routine use of this technique is discouraged because of several disadvantages. The purpose of this study was to evaluate intraobserver and interobserver reliability of direct measurements of wrist-hand volume using a new communicating vessels volumeter based on Pascal's law. A reliability study was conducted. To evaluate the reliability of the communicating vessels volumeter in generating measurements, 30 hands of 15 participants (9 women, 6 men) were measured 3 times each by 3 observers, totaling 270 volumetric results. Measurement time was short (X =3 minutes 42 seconds). The intraclass correlation coefficient (ICC) was .9977 for observer 1 and .9976 for observers 2 and 3. The interobserver ICC was .9998. The standard error of measurement was about 3 mL for all observers; the interobserver result was 1 mL. The interrater coefficient of variance (CV) was 1.15% for the series of 9 measurements collected for each segment; the intrarater CV was 1.20%. Limitations No swollen hands were measured, and measurements were not compared with the gold standard technique. Thus, accuracy of the new volumeter was not determined in this study. A new device has been developed for plethysmography of the extremities, and the results of its use to measure the volume of the wrist-hand segment were reliable in both intraobserver and interobserver analyses.