Large three-dimensional cell constructs for tissue engineering.
Much research has been conducted on fabricating biomimetic biomaterials in vitro. Tissue engineering approaches are often conducted by combining cells, scaffolds, and growth factors. However, the degradation rate of scaffolds is difficult to control and the degradation byproducts occasionally limit tissue regeneration. To overcome these issues, we have developed a novel system using a thermo-responsive hydrogel that forms scaffold-free, three-dimensional (3D) cell constructs with arbitrary size and morphology. 3D cell constructs prepared using bone marrow-derived stromal stem cells (BMSCs) exhibited self-organizing ability and formed bone-like tissue with endochondral ossification. Endothelial cells were then introduced into the BMSC construct and a vessel-like structure was formed within the constructs. Additionally, the bone formation ability was promoted by endothelial cells and cell constructs could be freeze-dried to improve their clinical application. A pre-treatment with specific protein protectant allowed for the fabrication of novel bone substitutes composed only of cells. This 3D cell construct technology using thermo-responsive hydrogels was then applied to other cell species. Cell constructs composed of dental pulp stem cells were fabricated, and the resulting construct regenerated pulp-like tissue within a human pulpless tooth. In this review, we demonstrate the approaches for the in vitro fabrication of bone and dental pulp-like tissue using thermo-responsive hydrogels and their potential applications.


Stroke after Initiating IV Penicillin for Neurosyphilis: A Possible Jarisch-Herxheimer Reaction.
Introduction. Syphilis incidence has increased in the US in the last decade. Jarisch-Herxheimer reaction (JHR) is a well-documented adverse effect of penicillin treatment in syphilis. Stroke has not been reported as part of its phenomenology. Case Report. A 57-year-old man presented with worsening memory. His minimental status examination score was 14/30. Serum RPR test was positive and VDRL test in the CSF was reactive. Within six hours of first dose of IV crystalline penicillin G, he was found to have hemineglect and difficulty moving the left leg. MRI of the brain showed multiple acute ischemic strokes. Immediate MRA ruled out vascular occlusion. Penicillin treatment was stopped. Four hours later, he was found to be febrile and had two episodes of generalized tonic-clonic seizures. Conclusions. We report a case of confirmed neurosyphilis with no known modifiable stroke risk factors, who developed acute ischemic stroke and other constitutional symptoms consistent with JHR after IV penicillin. This is the first reported case in literature where an acute ischemic stroke can be attributed to Jarisch-Herxheimer reaction. Given an increase in incidence of syphilis in recent years, our case underlies the importance of keeping in mind potential catastrophic drug adverse reactions in neurosyphilis patients.


Ecological momentary assessment of drinking in young adults: An investigation into social context, affect and motives.
Daily assessment studies have examined how day specific factors, such as affect, social context, and drinking motives, alongside dispositional drinking motives, predict young adults' drinking. However, these studies did not examine how the interplay between drinking motives (dispositional and day specific) and multiple features of the drinking situation predict drinking with respect to either the initial decision to drink or the quantity of alcohol consumed. Ecological momentary assessment (EMA) via smartphone technology, enables us to address this gap by evaluating to what extent dispositional drinking motives and day specific factors are associated with: a) the initiation of drinking episodes and; b) the quantity of alcohol consumed. Participants were 83 young adults (63 female) aged 18 to 30 (M = 21.42, SD = 3.09) who resided in Australia and participated in an EMA study for 21 days via their smartphone. On a daily basis, participants received three random-interval prompts that measured momentary affect, drinking motives, social context (e.g., people present in the social context and if these individuals are drinking), and alcohol use. A multilevel hurdle analysis found that young adults were more likely to both initiate a drinking episode and consume a higher quantity of alcohol if they were surrounded by other people who were drinking and were motivated to drink to conform to the reference group. This study is the first of its kind to demonstrate that different drinking behaviors (i.e., initiation and quantity of alcohol consumed) are associated with a similar set of predictors. Drinking-based interventions that address these risk factors could effectively reduce risky drinking as it would intervene on both the decision to initiate alcohol use, and the decision to continue drinking.


Stargardt's disease with neurological involvement: case report.
The diagnosis of Stargardt's disease, in cases of macular degeneration associated with neurological symptomatology, has been questioned. A 23 year old man, suffering from well characterized Stargardt's disease since childhood, presented with progressive spastic tetraparesis and cerebellar involvement since the age of 18. The case is presented and the association between the two clinical pictures is discussed.


Clinical importance of serum HE4 and MMP2 levels in endometrial cancer patients.
Endometrial cancer is the one of the most common cancers of the genital organ. HE4 and MMP2 are both proteins whose serum levels increase in endometrial cancer. To explore the diagnostic potential of the serum levels of HE4 and MMP2 in patients with endometrial cancer and benign endometrial diseases. To assess the relationship between the serum levels of HE4 and MMP2 and the typical prognostic factors in patients with endometrial cancer. Included in the study was a group of 112 patients presenting with bleeding abnormalities at the Pomeranian Medical University in years 2012-2016. Serum HE4 concentrations were measured using the Elecsys Electrochemiluminescence Immunoassay (ECLIA). MMP2 concentrations were quantified in the serum using multiplex immunoassays. We observed statistically significant differences in mean serum levels of HE4 and MMP2 between the group of endometrial cancer patients and the group of patients with no changes in the endometrium (P=0.002/0.003). The diagnostic potential of HE4 and MMP2 in differentiation of high (International Federation of Gynecology and Obstetrics [FIGO] III and IV) vs low (FIGO I and II) clinical stage of tumor and prediction of cellular differentiation grade (G1 vs G3) on the basis of the analysis of the area under the curve is, respectively, 0.86 and 0.82 for HE4 and 0.82 and 0.74 for MMP2. The HE4 marker was significantly more specific than MMP2 in every study group and amounted to 93% vs 86% in all patients included in the analysis, 94% vs 84% in pre-menopausal patients and 84% vs 79% in post-menopausal patients. HE4 and MMP2 are characterized by high specificity and may be useful as biomarkers in the diagnostics of endometrial cancer. When determined preoperatively, HE4 is correlated with the prognostic factors of endometrial cancer and may be helpful in the planning of individual treatment of endometrial cancer patients.


Squamous cell carcinoma arising in a dermoid cyst of the ovary. Clinical and pathological report of a case.
A case of squamous cell carcinoma arising in a dermoid cyst of the ovary of a 38 year-old woman is reported. Its frequency is nearly 1%; nevertheless it is opinion that the real frequency could be higher if the hystological examination of the cyst would be made more accurately. Clinical and pathological findings of this case are presented in this brief note. The importance to always make an intraoperatively frozen-section examination is emphasized. The patient is alive and well after six months after surgery.


Cationic shell-crosslinked knedel-like nanoparticles for highly efficient gene and oligonucleotide transfection of mammalian cells.
In this work, a robust synthetic nanostructure was designed for the effective packaging of DNA and it was shown to be an efficient agent for cell transfection. An amphiphilic block copolymer, poly(acrylamidoethylamine)(128)-b-polystyrene(40) (PAEA(128)-b-PS(40)), was synthesized, micellized in water and shell-crosslinked using a diacid-derivatized crosslinker, to give cationic shell-crosslinked nanoparticles (cSCKs) with a mean hydrodynamic diameter of 14 +/- 2 nm. A series of discrete complexes of the cSCKs with plasmid DNA (pDNA) was able to be formed over a broad range of polymer amine:pDNA phosphate ratios (N/P ratio), 2:1-20:1. The sizes of the complexes and their ability to fully bind the pDNA were dependent upon the N/P ratio, as characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM) and gel retardation assay. A luciferase activity assay and EGFP expression were used to evaluate intracellular delivery of a splice-correcting phosphorothioate and genetic material, respectively, by the cSCKs, which indicated that an N/P ratio of 6:1 gave the highest transfection. It was shown by both luciferase activity assay (48 h) and EGFP transfection data that high transfection efficiencies were achieved for HeLa cells transfected by cSCK/CCUCUUACCUCAGUUACA and cSCK/pEGFP-N1 plasmid, respectively. The cSCK/pEGFP-N1 plasmid transfection efficiency of 27% far exceeded the performance of Polyfect (PAMAM dendrimers), which achieved only 12% transfection efficiency, under the same conditions. Cytotoxicities for the cSCKs were evaluated for HeLa and CHO cells.


CD6: expression during development, apoptosis and selection of human and mouse thymocytes.
CD6, a 130-kDa surface glycoprotein, is expressed primarily on cells of T lineage. A co-stimulatory role for CD6 in mature T cells has been shown, but the function of CD6 during thymocyte development is unknown. Since CD6 ligands are expressed on thymic epithelium, their interactions with CD6 could be important in thymic selection. In this report we show that CD6 is developmentally regulated in human and mouse thymocytes, and further demonstrate that increase in the level of CD6 expression correlates with expression of the selection marker CD69. We also show that activation via CD2 induces CD6 expression on mature human thymocytes and on a subset of immature human thymocytes that are resistant to apoptosis. In human and mouse thymocytes that express heterogeneous TCR, CD6 increases occur as double-positive thymocytes are selected to a single-positive stage. In contrast, in thymocytes from TCR transgenic mice, CD6 is barely increased following selection, suggesting that as functional avidity increases, requirements for CD6-dependent co-stimulation decrease. Taken together, these results indicate that during thymic development CD6-dependent signals may contribute both to thymocyte survival, and to the overall functional avidity of selection in both man and mouse.


The NLR immune receptor ADR1 and lipase-like proteins EDS1 and PAD4 mediate stomatal immunity in Nicotiana benthamiana and Arabidopsis.
In the presence of pathogenic bacteria, plants close their stomata to prevent pathogen entry. Intracellular nucleotide-binding leucine-rich repeat (NLR) immune receptors recognize pathogenic effectors and activate effector-triggered immune responses. However, the regulatory and molecular mechanisms of stomatal immunity involving NLR immune receptors are unknown. Here, we show that the Nicotiana benthamiana RPW8-NLR central immune receptor ACTIVATED DISEASE RESISTANCE 1 (NbADR1), together with the key immune proteins ENHANCED DISEASE SUSCEPTIBILITY 1 (NbEDS1) and PHYTOALEXIN DEFICIENT 4 (NbPAD4), plays an essential role in bacterial pathogen- and flg22-induced stomatal immunity by regulating the expression of salicylic acid (SA) and abscisic acid (ABA) biosynthesis or response-related genes. NbADR1 recruits NbEDS1 and NbPAD4 in stomata to form a stomatal immune response complex. The transcription factor NbWRKY40e, in association with NbEDS1 and NbPAD4, modulates the expression of SA and ABA biosynthesis or response-related genes to influence stomatal immunity. NbADR1, NbEDS1, and NbPAD4 are required for the pathogen infection-enhanced binding of NbWRKY40e to the ISOCHORISMATE SYNTHASE 1 promoter. Moreover, the ADR1-EDS1-PAD4 module regulates stomatal immunity in Arabidopsis (Arabidopsis thaliana). Collectively, our findings show the pivotal role of the core intracellular immune receptor module ADR1-EDS1-PAD4 in stomatal immunity, which enables plants to limit pathogen entry.


Ultrastructural immunolabeling: a unique diagnostic tool in monoclonal light chain-related renal diseases.
Ultrastructural labeling can play a key role in the evaluation of morphologic expressions of monoclonal light chain-related renal diseases in cases where light microscopy, electron microscopy, and immunofluorescence data, even when combined, are not definitive in conveying a diagnosis and, in other cases, in clarifying the findings by providing immunomorphologic correlation. The important role of ultrastructural labeling is highlighted by the fact that in some of these cases bone marrow aspirates and biopsy specimens obtained at the time of the evaluation of the renal specimens are often unable to establish unequivocally a diagnosis of plasma cell dyscrasia. This is in part because renal manifestations commonly precede overt diagnostic bone marrow alterations. Overt bone marrow findings and clinical manifestations may be preceded for as long as 16 years by the renal manifestations. Determination or confirmation of monoclonality and detection of early deposition of monotypical light chains before the finding of ultrastructural morphologic correlates (ie, subendothelial, punctate, granular, electron-dense material) represent unique attributes of this technique. The increased sensitivity of ultrastructural immunolabeling compared with other available diagnostic techniques and its exquisite immunomorphologic correlative capabilities result in a comprehensive evaluation. Sixteen monoclonal light chain-related renal disease cases with early, unusual, or equivocal immunomorphologic manifestations that may have not been characterized properly if ultrastructural labeling had not been performed are presented. The crucial role played by ultrastructural labeling in evaluating these cases and establishing an accurate diagnosis is illustrated and emphasized.


Fully 3D printed integrated reactor array for point-of-care molecular diagnostics.
Molecular diagnostics that involve nucleic acid amplification tests (NAATs) are crucial for prevention and treatment of infectious diseases. In this study, we developed a simple, inexpensive, disposable, fully 3D printed microfluidic reactor array that is capable of carrying out extraction, concentration and isothermal amplification of nucleic acids in variety of body fluids. The method allows rapid molecular diagnostic tests for infectious diseases at point of care. A simple leak-proof polymerization strategy was developed to integrate flow-through nucleic acid isolation membranes into microfluidic devices, yielding a multifunctional diagnostic platform. Static coating technology was adopted to improve the biocompatibility of our 3D printed device. We demonstrated the suitability of our device for both end-point colorimetric qualitative detection and real-time fluorescence quantitative detection. We applied our diagnostic device to detection of Plasmodium falciparum in plasma samples and Neisseria meningitides in cerebrospinal fluid (CSF) samples by loop-mediated, isothermal amplification (LAMP) within 50 min. The detection limits were 100 fg for P. falciparum and 50 colony-forming unit (CFU) for N. meningitidis per reaction, which are comparable to that of benchtop instruments. This rapid and inexpensive 3D printed device has great potential for point-of-care molecular diagnosis of infectious disease in resource-limited settings.


Evidence for an articulatory awareness deficit in adult dyslexics.
Dyslexia is widely considered to be associated with impaired performance on phonological awareness tasks. However, it is likely that orthographic knowledge also influences performance on these tasks. In this study, adult dyslexics, for whom reading is no longer a major problem, were compared to a control group on a measure of articulatory awareness, a task which is not confounded with orthography. The dyslexic group showed deficits on the task in comparison to the control group. We hypothesize that information about articulatory movements for specific phonemes is less accessible to dyslexics because of a deficient phonological processing system.


Quantum Tunnelling Driven H2 Formation on Graphene.
It is commonly believed that it is unfavorable for adsorbed H atoms on carbonaceous surfaces to form H2 without the help of incident H atoms. Using ring-polymer instanton theory to describe multidimensional tunnelling effects, combined with ab initio electronic structure calculations, we find that these quantum-mechanical simulations reveal a qualitatively different picture. Recombination of adsorbed H atoms, which was believed to be irrelevant at low temperature due to high barriers, is enabled by deep tunnelling, with reaction rates enhanced by tens of orders of magnitude. Furthermore, we identify a new path for H recombination that proceeds via multidimensional tunnelling but would have been predicted to be unfeasible by a simple one-dimensional description of the reaction. The results suggest that hydrogen molecule formation at low temperatures are rather fast processes that should not be ignored in experimental settings and natural environments with graphene, graphite, and other planar carbon segments.


Long PCR-based genotyping for a deleted CYP2D6 gene without DNA extraction.
In the post-genome era, a simple and inexpensive method for diagnostic analysis is in high demand. Cytochrome P450 (CYP) 2D6 is one of the most widely investigated CYPs in relation to genetic polymorphism. Detection of CYP2D6*5 is difficult since long PCR is used. Especially for samples without DNA extraction, the detection is not sensitive enough for population analysis. Therefore, we developed a CYP2D6*5 genotyping method that involves nested long PCR, directly using human whole saliva as a template without DNA extraction. This method will be very useful for genetic diagnoses and can be an efficient tool for individualization of drug therapy in clinical studies.


Molecular microbial biodiversity assessment: a biological indicator of soil health.
The soil performs a variety of key functions: (i) provides the food, fuel, and fiber needs of the world's population, (ii) regulates the quality of the air and water, (iii) decomposes organic wastes, (iv) recycles nutrients, and (v) acts as a sink for pollutants (including global gases). Soil degradation is increasingly recognized as an urgent environmental issue and a crucial need exists for the capacity to evaluate soil health. After all, soil health is a key component in sustaining the different world's ecosystems and the myriad of natural and socioeconomic systems they support. In this respect, soil health has been defined as the "continued capacity of a specific kind of soil to function as a vital living system, within natural or managed ecosystem boundaries, to sustain plant and animal productivity, to maintain or enhance the quality of air and water environments, and to support human health and habitation". A biological indicator is an organism, a part of an organism, or a community of organisms, used to obtain information about environmental quality. The assessment of microbial biodiversity has the potential to provide useful insight into the health and functioning of soil. Our inability to culture most microorganisms that are present in soils has, until recently, impaired studying the relationships between the structure and the function of soil microbial communities. This shortcoming has been recently overcome by using several molecular techniques that allow the detection, enumeration, and characterization of soil microorganisms without cultivation.


Disentangling leaf-microbiome interactions in Arabidopsis thaliana by network mapping.
The leaf microbiota plays a key role in plant development, but a detailed mechanism of microbe-plant relationships remains elusive. Many genome-wide association studies (GWAS) have begun to map leaf microbes, but few have systematically characterized the genetics of how microbes act and interact. Previously, we integrated behavioral ecology and game theory to define four types of microbial interactions - mutualism, antagonism, aggression, and altruism, in a microbial community assembly. Here, we apply network mapping to identify specific plant genes that mediate the topological architecture of microbial networks. Analyzing leaf microbiome data from an Arabidopsis GWAS, we identify several heritable hub microbes for leaf microbial communities and detect 140-728 SNPs (Single nucleotide polymorphisms) responsible for emergent properties of microbial network. We reconstruct Bayesian genetic networks from which to identify 22-43 hub genes found to code molecular pathways related to leaf growth, abiotic stress responses, disease resistance and nutrition uptake. A further path analysis visualizes how genetic variants of Arabidopsis affect its fecundity through the internal workings of the leaf microbiome. We find that microbial networks and their genetic control vary along spatiotemporal gradients. Our study provides a new avenue to reveal the "endophenotype" role of microbial networks in linking genotype to end-point phenotypes in plants. Our integrative theory model provides a powerful tool to understand the mechanistic basis of structural-functional relationships within the leaf microbiome and supports the need for future research on plant breeding and synthetic microbial consortia with a specific function.


Human opioid peptide Met-enkephalin binds to anionic phosphatidylserine in high preference to zwitterionic phosphatidylcholine: natural-abundance 13C NMR study on the binding state in large unilamellar vesicles.
A human opioid neuropeptide, Met-enkephalin (M-Enk: Tyr1-Gly2-Gly3-Phe4-Met5), having no net charge binds to anionic phosphatidylserine (PS) in high preference to zwitterionic phosphatidylcholine (PC). The binding mechanism in the PS and PC bilayers was studied on the basis of the inter- and intramolecular interaction data obtained by natural-abundance 13C nuclear magnetic resonance (NMR) of the peptide. Prominent upfield changes of the 13C resonance were observed in the C-terminal residue upon binding to PS, whereas no such marked change was observed upon binding to PC. The upfield chemical shift changes with their characteristic carbon site dependence are ascribed to the electrostatic binding between the peptide C-terminal CO2- and the PS headgroup NH3+. Despite the net negative charge of the PS bilayer surface, M-Enk thus anchors the negatively charged C-terminus. In the N-terminal residue, on the other hand, marked downfield chemical shift changes are observed upon binding to both the PS and PC bilayers, the magnitude of the changes being much larger in the PS system. The downfield changes with their characteristic carbon site dependence are ascribed to the electrostatic binding between the peptide N-terminal NH3+ and the lipid headgroup negative charge(s) (CO2- or PO4- in PS, PO4- in PC). Perturbation on the signal half-widths due to membrane binding also indicates the preferential and deeper binding of M-Enk on the PS membrane surface than on the PC membrane surface. Local charge cancellation takes place efficiently between M-Enk termini and the PS headgroups and compensates for the strong electrostatic hydration of the ionic groups. Distribution of the charged (positive and negative) and uncharged sites in the headgroups along the bilayer normal is responsible for the marked difference between PS and PC headgroups in controlling the binding state of the zwitterionic M-Enk.


[The century of Nencki Institute of Experimental Biology, PAS - the idea and will power].
The Institute was founded in 1918, shortly after the re-establishment of Poland as an independent country. It was based on three pre-existing laboratories affiliated with the Scientific Society of Warsaw. The formation and development of the Institute was supported in part by a donation of Nadine Sieber-Shumova, a close co-worker of Marceli Nencki from Berne and St. Petersburg. Over the next two decades the Institute grew to become the leading biological research centre in Poland. The outbreak of World War II interrupted a period of its intensive expansion and achievement of scientific excellence in the field of experimental biology. After the turmoil of the war, during which over a dozen of the Institute's staff lost their lives, and its premises (including most of its 30.000 volume library)were destroyed, the surviving staff members (Jan Dembowski, Stefania Dembowska, Jerzy Konorski, Liliana Lubińska, Włodzimierz Niemierko and Stella Niemierko) re-established the Nencki Institute. In 1952, the Institute was incorporated into the newly founded Polish Academy of Sciences, and the Institute's director, Prof. Dembowski, became the first President of the Academy. During the period of 1953-55, a newly erected building at 3 Pasteur Street in Warsaw became the final home of the Nencki Institute. The research goals of the Nencki Institute are to arrive at molecular, cellular and systematic explanations of excitability, movement, development, memory, learning, behaviour, ageing and disease. All those tasks need to be both intellectually satisfying and relevant to problems of human health. Institute edit journal Acta Neurobiologiae Experimentalis. Directors from 1947: Jan Dembowski, Włodzimierz Niemierko, Jerzy Konorski, Kazimierz Zieliński, Maciej Jan Nałęcz, Jerzy Duszyński, Adam Szewczyk. At present Agnieszka Dobrzyń.


Relation between in vivo effects and in vitro effects of serine and thiol proteinase inhibitors.
In the present study we investigated the relationship between the in vivo effects and the in vitro effects of serine and thiol proteinase inhibitors. Each one of the inhibitors, leupeptin, dansyl-L-leucyl-L-argininal (Dan-Leu-Argal), pyroglutamyl-L-leucyl-L-argininal (Pyr-Leu-Argal), E-64C or EP-459 was given intraperitoneally to mice for 8 consecutive days and various enzymatic activities were tested in 6 organs: forelimb muscle, hindlimb muscle, heart, spleen, liver and kidney. A multivariate analysis clearly differentiated the effects of leupeptin from those of its two analogues (Dan-Leu-Argal and Pyr-Leu-Argal), while the two analogues showed close relations with each other as to their in vivo effects. E-64C and EP-459 also showed resemblance of in vivo effects between them, but the effects of these two agents were clearly differentiated from those of leupeptin and its two analogues. This kind of study seems important to assess the in vivo effects of physiologically active substances including enzyme inhibitors.


[Prevention of cystoid macular edema following cataract extraction using local indomethacin application].
In a randomized, controlled study drops of 1% aqueous solution of indomethacin were compared to Placebo to assess efficacy in the prevention of cystoid macular edema. The study involved 124 patients who had undergone intracapsular cataract extraction and 40 patients with implanted lenses. The study parameters--visual acuity, biomicroscopy observations of the macula and fluorescein angiography--were assessed for three months postoperatively. The long-term course was reviewed by regular outpatient follow-up visits or by questionnaires completed by the patients' personal ophthalmologists. Fluorescein angiography could only be carried out in 13 of the 40 patients who had intraocular lenses. The relatively high incidence of secondary cataract or pupillary synechiae made technically perfect exposures difficult. In 73 patients with intracapsular extraction, fluorescein angiography showed a distinct diminution in the incidence of edema in the indomethacin group at all three observation periods (3, 6 and 12 weeks postoperatively). The difference compared with placebo was significant at weeks 6 and 12. Ophthalmoscopically visible macular changes were rare. No evidence of edema was noted in any of the 51 patients who could not have fluorescein angiography for one reason or another. The visual acuity of patients who received indomethacin was not significantly different from that of those who received placebo. Furthermore, and this is important for the patients, in the group with proven edema no conclusion could be made about the effect of therapy on the severity and persistence of visual impairment.(ABSTRACT TRUNCATED AT 250 WORDS)


The effects of topical hypothermia and steroids on ATP levels in an in vivo liver ischemia model.
Complex hepatic surgery often requires occlusion of the portal triad in order to decrease parenchymal bleeding. This study was undertaken to evaluate the effects of topical hypothermia and intravenous steroids on liver ischemia by measuring adenosine triphosphate (ATP) levels within the hepatic parenchyma. Forty New Zealand white rabbits were divided into four experimental and four control groups. All experimental animals underwent laparotomy and ligation of the porta hepatis. Serial liver biopsy specimens were obtained at predetermined time intervals. Group I received no further intervention. Group II were topically cooled until intrahepatic temperature reached 30 degrees C. Group III received preligation intravenous methylprednisolone (30 mg/kg). Group IV received both steroids and topical hypothermia. The corresponding control groups underwent laparotomy and isolation of the porta without ligation. Adenosine triphosphate was extracted from the liver parenchyma and quantified by high-performance liquid chromatography (HPLC). The data were analyzed using a three-factor mixed analysis of variance (ANOVA). There was a statistically significant protective effect on ATP levels provided by topical hypothermia at 15 and 30 minutes of ischemia (p < 0.01), but not at 60 minutes (p > 0.05). Steroids were not found to have any protective effect on ATP levels at any time point. The combination of steroids and topical hypothermia provided significant preservation of hepatic parenchymal ATP levels, although less than that of hypothermia alone, at 15 and 30 minutes of ischemia (p < 0.01).


Perspectives and factors associated with informal human milk sharing: a mixed-methods systematic review protocol.
The practice of informal human milk sharing is a relatively new phenomenon and poses significant questions in the domain of infant feeding. Informal human milk sharing is a means of donating human milk from another lactating individual who is not the child's biological parent, in a casual manner, that is, without the involvement of health professionals or agencies. The advent of digital technology has facilitated the donation and receipt of human milk through digital online platforms and thus has amplified this modern practice. This research aims to comprehensively examine and synthesize evidence about the motivations, barriers, facilitators and experiences of individuals who both donate (donors) and the primary care givers of the infants who receive (recipients) human milk informally to provide to infants. A mixed-methods systematic review will be undertaken. This review will consider qualitative, quantitative and primary mixed-methods studies which report on the factors associated with informal milk sharing, and on donors' and recipients' experiences of the practice. Primary mixed-method studies will be included if the individual qualitative and quantitative components can be extracted. Five databases will be searched for studies on informal human milk sharing published from inception of the database. Study quality will be evaluated using the standardized JBI critical appraisal tools, selected based on the methodology in each individual study. Data extraction will be conducted using the JBI mixed methods data extraction form followed by data transformation, synthesis and integration. This mixed-methods systematic review will follow a convergent integrated approach in accordance with JBI guidance. Informal human milk sharing is a novel practice in the domain of infant feeding. This review will enable a thorough understanding of this practice from both the donors and recipients' perspective and will have implications for healthcare professionals, policy and future clinical decision-making. PROSPERO CRD42023405653.


Comparison of phenotype, comorbidities, therapy and adverse events between psoriatic patients with and without psoriatic arthritis. Biobadaderm registry.
There are a limited number of studies comparing psoriasis patients without psoriatic arthritis (PsA) to those with arthritis. Previous results are controversial. To perform a comparative analysis of the phenotype, baseline comorbidities, therapeutic profile and incidence of adverse events (particularly overall adverse events, infections and infestations, malignancies and psychiatric disorders) among psoriatic patients with/without PsA. All the patients on the Biobadaderm registry, a prospective inception cohort of psoriasis patients on systemic therapy, were included. Patients were divided into two groups: those with psoriasis without arthritis at the time of entry into the cohort (Pso group) and those with psoriasis and psoriatic arthritis (PsA group) at entry. Patients were followed until the censorship date (last visit in a lost-to-follow-up patient, or 10 November 2015, whichever occurred first). We excluded all the patients who developed any kind of signs and/or symptoms of joint involvement during the follow-up. A descriptive analysis was performed. We estimated incidence ratios (IRR) of adverse events during systemic treatment using a mixed-effects Poisson regression. We included 2120 patients: 1871 (88%) patients with psoriasis without arthritis and 249 (12%) with psoriasis and PsA. The follow-up time was 5020 patients-year in the Pso group and 762 patients-year in the PsA group. Patients with PsA had more comorbidities, particularly hypertension and liver disease; used a higher number of systemic therapies, particularly anti-TNFα drugs and combination therapy; and presented more adverse events (IRR adjusted = 1.29; 95% CI: [1.05-1.58]), particularly serious adverse events (IRR adjusted = 1.51; 95% CI: [1.01-2.26]) and infections/infestations (IRR adjusted = 1.88; 95% CI: [1.27-2.79]), independently of the associated comorbidities and present/past therapies. Given the differences between patients with psoriasis alone or with psoriasis associated with PsA, patients with psoriasis and PsA should be followed and managed more closely and with specific attention.


Implementation and evaluation of a peer review process for advanced practice nurses in a university hospital setting.
Over the past decade, implementation of the peer review process for the development of the advanced practice nurse (APN) has been emphasized. However, little exists in the literature regarding APN peer review. The peer review process is intended to help demonstrate competency of care, enhance quality improvement measures, and foster the professional growth of the APN. APNs serving on a professional governance council within a university teaching hospital developed a model of peer review for APNs. Nine months after the tool was implemented, an anonymous follow-up survey was conducted. A follow-up request was sent 4 weeks later to increase the number of respondents. Likert scales were used to elicit subjective data regarding the process. Of 81 APNs who participated in the survey, more than half (52%) felt that the process would directly improve their professional practice. Survey results show that the peer review process affected APN professional practice positively. Additional research might include pathways for remediation and education of staff, evaluation of alternate methods to improve application to clinical practice, and collection of outcome data. The models presented provide a foundation for future refinement to accommodate different APN practice settings.


Fixed, object-specific intensity compensation for cone beam optical CT radiation dosimetry.
Optical cone beam computed tomography (CT) scanning of radiochromic gel dosimeters, using a CCD camera and a low stray light convergent source, provides fast, truly 3D radiation dosimetry with high accuracy. However, a key limiting factor in radiochromic gel dosimetry at large (⩾10 cm diameter) volumes is the initial attenuation of the dosimeters. It is not unusual to observe a 5-10× difference in signal intensity through the dosimeter center versus through the surrounding medium in pre-irradiation images. Thus, all dosimetric information in a typical experiment is measured within the lower 10%-20% of the camera sensor's range, and re-use of gels is often not possible due to a lack of transmission. To counteract this, in this note we describe a simple method to create source compensators by printing on transparent films. This technique, which is easily implemented and inexpensive, is an optical analogue to the bowtie filter in x-ray CT. We present transmission images and solution phantom reconstructions to demonstrate that (1) placing compensators beyond the focal zone of the imaging lens prevents high spatial frequency features of the printed films from generating reconstruction artifacts, and (2) object-specific compensation considerably reduces the range of intensities measured in projection images. This will improve the measurable dose range in optical CT dosimetry, and will enable imaging of larger gel volumes (∼15 cm diameter). Additionally, it should enable re-use of dosimeters by printing a new compensator for a second experiment.


Circadian Clock and OxInflammation: Functional Crosstalk in Cutaneous Homeostasis.
Circadian rhythms are biological oscillations that occur with an approximately 24 h period and optimize cellular homeostasis and responses to environmental stimuli. A growing collection of data suggests that chronic circadian disruption caused by novel lifestyle risk factors such as shift work, travel across time zones, or irregular sleep-wake cycles has long-term consequences for human health. Among the multiplicity of physiological systems hypothesized to have a role in the onset of pathologies in case of circadian disruption, there are redox-sensitive defensive pathways and inflammatory machinery. Due to its location and barrier physiological role, the skin is a prototypical tissue to study the influence of environmental insults induced OxInflammation disturbance and circadian system alteration. To better investigate the link among outdoor stressors, OxInflammation, and circadian system, we tested the differential responses of keratinocytes clock synchronized or desynchronized, in an in vitro inflammatory model exposed to O3. Being both NRF2 and NF-κB two key redox-sensitive transcription factors involved in cellular redox homeostasis and inflammation, we analyzed their activation and expression in challenged keratinocytes by O3. Our results suggest that a synchronized circadian clock not only facilitates the protective role of NRF2 in terms of a faster and more efficient defensive response against environmental insults but also moderates the cellular damage resulting from a condition of chronic inflammation. Our results bring new insights on the role of circadian clock in regulating the redox-inflammatory crosstalk influenced by O3 and possibly can be extrapolated to other pollutants able to affect the oxinflammatory cellular processes.


Dual Roles of Coconut Oil and Its Major Component Lauric Acid on Redox Nexus: Focus on Cytoprotection and Cancer Cell Death.
It has been reported that coconut oil supplementation can reduce neuroinflammation. However, coconut oils are available as virgin coconut oil (VCO), crude coconut oil (ECO), and refined coconut oil (RCO). The impact of coconut oil extraction process (and its major fatty acid component lauric acid) at cellular antioxidant level, redox homeostasis and inflammation in neural cells is hitherto unexplained. Herein, we have shown the antioxidant levels and cellular effect of coconut oil extracted by various processes in human neuroblastoma cells (SH-SY5Y) cultured in vitro. Results indicate VCO and ECO treated cells displayed better mitochondrial health when compared to RCO. Similar trend was observed for the release of reactive oxygen species (ROS), key oxidative stress response genes (GCLC, HO-1, and Nqo1) and inflammatory genes (IL6, TNFα, and iNOS) in SH-SY5Y cells. Our results signified that both VCO and ECO offer better neural health primarily by maintaining the cellular redox balance. Further, RCO prepared by solvent extraction and chemical refining process lacks appreciable beneficial effect. Then, we extended our study to find out the reasons behind maintaining the cellular redox balance in neuroblastoma cells by VCO and ECO. Our GC-MS results showed that lauric acid (C14:0) (LA) content was the major difference in the fatty acid composition extracted by various processes. Therefore, we evaluated the efficacy of LA in SH-SY5Y cells. The LA showed dose-dependent effect. At IC50 concentration (11.8 μM), LA down regulated the oxidative stress response genes and inflammatory genes. The results clearly indicate that the LA inhibited the neuroinflammation and provided an efficient cellular antioxidant activity, which protects the cells. The efficiency was also evaluated in normal cell line such as fibroblasts (L929) to cross-validate that the results were not false positive. Different concentration of LA on L929 cells showed high compatibility. From our observation, we conclude that VCO and ECO offers better cellular protection owing to their powerful antioxidant system. Therefore, we advocate the inclusion of either VCO and/or ECO in the diet for a healthy lifestyle.


A new method for inducing copious drinking and the accompanying stimulation on the pars intermedia of the mouse pituitary gland.
A new method for causing excessive liquid intake was developed as the condition opposite to dehydration, and the effect of this copious drinking for 2, 4 and 6 days on the pars intermedia of the hypophysis was evaluated by quantitative electron microscopy in mice. Free access to 5% glucose solution and concurrent food deprivation resulted in the development of polydipsic mice. Drinking volume was 18.9 +/- 1.1 ml/10 g BW on day 2, 20.8 +/- 1.2 on day 4 and 24.1 +/- 1.3 on day 6, respectively, while remaining at 2.2 +/- 0.2 ml/10g BW in the control. Copious drinking was found to elicit secretory activity of the pars intermedia cells. The cytoplasmic volume percentage of rough endoplasmic reticulum (rER) and the numerical density of Golgi granules increased significantly on day 2 reaching a peak on day 4. Contrarily, secretory granules decreased in number, indicating that the granule release exceeded the activated granule formation. The rise in the activity of the gland was followed by a slight fall after 6 days, which was probably due to a malnutritional condition by food deprivation. Neither 20% glucose drinking nor food deprivation for 4 days enhanced the pars intermedia cells. Thus, excessive water intake seems to be a plausible reason for stimulating secretory activity in cells of the pars intermedia of the hypophysis in mice.


Actions of "antioxidants" in the protection against atherosclerosis.
This review addresses the role of oxidative processes in atherosclerosis and its resulting cardiovascular disease by focusing on the outcome of antioxidant interventions. Although there is unambiguous evidence for the presence of heightened oxidative stress and resulting damage in atherosclerosis, it remains to be established whether this represents a cause or a consequence of the disease. This critical question is complicated further by the increasing realization that oxidative processes, including those related to signaling, are part of normal cell function. Overall, the results from animal interventions suggest that antioxidants provide benefit neither generally nor consistently. Where benefit is observed, it appears to be achieved at least in part via modulation of biological processes such as increase in nitric oxide bioavailability and induction of protective enzymes such as heme oxygenase-1, rather than via inhibition of oxidative processes and lipid oxidation in the arterial wall. Exceptions to this may be situations of multiple/excessive stress, the relevance of which for humans is not clear. This interpretation is consistent with the overall disappointing outcome of antioxidant interventions in humans and can be rationalized by the spatial compartmentalization of cellular oxidative signaling and/or damage, complex roles of oxidant-producing enzymes, and the multifactorial nature of atherosclerosis.


Nonaqueous Interfacial Polymerization-Derived Polyphosphazene Films for Sieving or Blocking Hydrogen Gas.
A series of cyclomatrix polyphosphazene films have been prepared by nonaqueous interfacial polymerization (IP) of small aromatic hydroxyl compounds in a potassium hydroxide dimethylsulfoxide solution and hexachlorocyclotriphosphazene in cyclohexane on top of ceramic supports. Via the amount of dissolved potassium hydroxide, the extent of deprotonation of the aromatic hydroxyl compounds can be changed, in turn affecting the molecular structure and permselective properties of the thin polymer networks ranging from hydrogen/oxygen barriers to membranes with persisting hydrogen permselectivities at high temperatures. Barrier films are obtained with a high potassium hydroxide concentration, revealing permeabilities as low as 9.4 × 10-17 cm3 cm cm-2 s-1 Pa-1 for hydrogen and 1.1 × 10-16 cm3 cm cm-2 s-1 Pa-1 for oxygen. For films obtained with a lower concentration of potassium hydroxide, single gas permeation experiments reveal a molecular sieving behavior, with a hydrogen permeance of around 10-8 mol m-2 s-1 Pa-1 and permselectivities of H2/N2 (52.8), H2/CH4 (100), and H2/CO2 (10.1) at 200 °C.


The clinical analysis of small supernumerary marker chromosomes in 17 children with mos 45,X/46,X,+mar karyotype.
Small supernumerary maker chromosome (sSMC) is a type of structurally abnormal chromosome. In order to identify the origin, morphology and other characteristics of sSMCs in children with mos 45,X/46,X,+mar karyotype, 17 patients (16 females and 1 male) were analyzed. All patients underwent general physical examination, gonadal imaging and molecular cytogenetic analyses, including Giemsa banding, dual-color fluorescence in situ hybridization and detection of the sex-determining region Y gene by polymerase chain reaction. Cytogenetic analyses indicated sSMCs in 14/17 cases were derived from the X chromosome, of which 8 individuals presented with ring-shaped sSMCs and 6 with centric minute-shaped sSMCs. The remaining 3 cases were derived from the Y chromosome, and all presented with minute-shaped sSMCs. All female patients exhibited short stature, gonadal dysgenesis and other typical features of Turner syndrome. The male patient exhibited short stature, hypospadias and bilateral cryptorchidism. In conclusion, the majority of the sSMCs in patients with a mos 45,X/46,X,+mar karyotype were derived from the sex chromosomes. The molecular cytogenetic features of sSMCs may provide useful information for genetic counseling, prenatal diagnosis and individualized treatment.


The rate of loss of eyelid reflex following thiopental administration in hypo- and hypergonadism in rabbits.
The aim of this research was to explain whether different hormonal conditions caused by disturbed concentrations of estrogens and progesterone might lead to alteration of CNS reaction following administration of the hypnotic agent thiopental. The investigated factor was the rate of loss of the eyelid reflex after intravenous thiopental administration, since this corresponds with loss of consciousness. The investigation was performed in 24 sexually mature female Chinchilla rabbits divided into 4 groups of 6 rabbits each. The animals were oophorectomized (hypoprogesterone/hypoestrogen), hyperestrogen (sham surgery plus estradiol injections), hyperprogesterone (sham surgery plus 17 alpha-hydroxyprogesterone injections), or normal (sham surgery). Twelve weeks later, thiopental (40 mg/ml) was infused through the intravenous cannula at a constant rate (90 ml/hour) until loss of the eyelid reflex, at which time blood was sampled for determination of thiopental, b-estradiol, progesterone and 17 alpha-hydroxyprogesterone concentrations. The ANOVA and Tukey tests were applied in statistical analysis (p=0.05). Hyperprogesterone rabbits lost consciousness faster (138 +/- 34.6 sec), at a lower plasma thiopental concentration (46.3 +/- 6.6 microg/ml), and required less thiopental (24.63 +/- 6.44 mg/kg) than controls; hypoprogesterone rabbits lost consciousness slower (207.5 +/- 30.9 sec), at a higher plasma thiopental concentration (129.2 +/- 24.9 microg/ml), and required more thiopental (38.51 +/- 2.33 mg/kg) than controls. The time of sleep induction in the control group was 190 +/- 25.7 sec.; the serum thiopental concentration was 77.8 +/- 13.9 microg/ml, and the total thiopental consumption was 35.8 +/- 3.51 mg/kg. Estrogen status has no effect. Various hormonal states are accompanied by different CNS reactions to thiopental. Among the studied groups of sex steroids, only progestins significantly modify the CNS response to barbiturate infusion.


Ginsenoside Rg1 attenuates invasion and migration by inhibiting transforming growth factor-β1-induced epithelial to mesenchymal transition in HepG2 cells.
Ginseng has become one of the most commonly used alternative herbal medicines and its active component, ginsenoside Rg1, possesses known pharmacological effects, including anticancer properties. However, the effects of ginsenoside Rg1 on metastasis have not been investigated. The present study demonstrated that ginsenoside Rg1 was able to suppress transforming growth factor‑β1 (TGF‑β1)‑induced invasion and migration in HepG2 liver cancer cells. This suppression was associated with the inhibition of TGF‑β1‑induced epithelial to mesenchymal transition (EMT). Furthermore, TGF‑β1 induced HepG2 cells to undergo EMT and significantly promoted cell invasion and migration. When cells were pretreated with ginsenoside Rg1 for 24 h and subsequently exposed to TGF‑β1 for 24 h, the results demonstrated that ginsenoside Rg1 inhibited the initiation of TGF‑β1‑induced EMT. In addition, HepG2 cells exhibited a mesenchymal phenotype when exposed to TGF‑β1, but when exposed to ginsenoside Rg1 this effect was reversed and the cells exhibited a classical epithelial morphology. Ginsenoside Rg1 also increased the expression of the epithelial phenotype marker E‑cadherin and repressed the expression of the mesenchymal phenotype marker vimentin. In conclusion, the results of the present study suggest that ginsenoside Rg1 may suppress liver cancer invasion and migration in vitro through inhibiting TGF‑β1‑induced EMT.


Hospital and hospice resource use of HIV-positive patients in Edinburgh.
A prospective study of health service and hospice resource utilization (average length of stay or ALOS, discharge rate, bed day use, outpatient consultation) analysed with reference to gender, risk activity, immunological and clinical staging (1987 definition of AIDS) for the financial year 1992-93 was undertaken at the Regional Infectious Disease Unit (RIDU), City Hospital, Edinburgh, Scotland where 72% of 513 patients were infected via injection drug use. Not surprising therefore overall, drug users were the heaviest users of the inpatient facilities (74% of the discharges and 65% of the bed days) although homosexuals had the highest discharge rate (114 per 100 person years) and rate of bed day use (1654 days per 100 person years). Immunodeficiency (CD4 count < 200 cells/ul) and a clinical diagnosis of AIDS were both associated with greater inpatient and outpatient resource use compared to those without immunodeficiency (CD4 count > or = 200 CD4 cells/ul) or AIDS. Gender effects were complex; the ALOS for women was increased for all risk groups whatever the CD4 count whilst there was no consistent trend of more resource use for women by risk group. Drug users were the heaviest overall users of the local hospice (84% of all admissions, 83% of the bed days and a discharge rate of 76.4 per 100 person years), more than double the rates experienced by the other risk groups. Thus both clinical and immunological staging (AIDS or a CD4 count < 200 cells/ul) were associated with increased resource use in HIV infection and estimates of resource use for AIDS need to be increased by around one-third to take into account hospice use. Despite the preponderance of drug users in Edinburgh, comparisons with other centres did not reveal increased resource use.


[Use of cryoprecipitate in combined therapy of acute purulent pyelonephritis].
The treatment of 120 patients with acute purulent pyelonephritis (APP) brought about the following results. 86% patients given standard therapy showed deterioration of their condition, DIC syndrome and inflammatory process were not cured, pyoinflammatory process in the kidney continued. On the contrary, 84.1% patients on combined conservative treatment improved: DIC syndrome and active inflammation relieved, fibronectin content, IgA, IgM and IgG levels rose. Ultrasonic examination and dopplerography indicated that combined conservative therapy including cryoprecipitate arrested development of a purulent focus in the kidney, reestablished intraorganic circulation. Mean stay in hospital was 13.9 days.


Sickle cell disease, sickle trait and the risk for venous thromboembolism: a systematic review and meta-analysis.
Globally, sickle cell disease (SCD) is one of the most common haemoglobinopathy. Considered a public health problem, it leads to vessel occlusion, blood stasis and chronic activation of the coagulation system responsible for vaso-occlussive crises and venous thromboembolism (VTE) which may be fatal. Although contemporary observational studies suggest a relationship between SCD or sickle trait (SCT) and VTE, there is lack of a summary or meta-analysis data on this possible correlation. Hence, we propose to summarize the available evidence on the association between SCD, SCT and VTE including deep vein thrombosis (DVT) and pulmonary embolism (PE). We searched PubMed and Scopus to identify all cross-sectional, cohort and case-control studies reporting on the association between SCD or SCT and VTE, DVT or PE in adults or children from inception to April 25, 2017. For measuring association between SCD or SCT and VTE, DVT, or PE, a meta-analysis using the random-effects method was performed to pool weighted odds ratios (OR) of risk estimates. From 313 records initially identified from bibliographic databases, 10 studies were eligible and therefore included the meta-analysis. SCD patients had significantly higher risk for VTE (pooled OR 4.4, 95%CI 2.6-7.5, p < 0.001), DVT (OR 1.1, 95% CI 1.1-1.2, p < 0.001) and PE (pooled OR 3.7, 95% CI 3.6-3.8, p < 0.001) as compared to non SCD-adults. A higher risk of VTE (OR 33.2, 95% CI 9.7-113.4, p < 0.001) and DVT (OR 30.7, 95% CI 1.6-578.2, p = 0.02) was found in pregnant or postpartum women with SCD as compared to their counterparts without SCD. Compared to adults with SCT, the risk of VTE was higher in adults with SCD (pooled OR 3.1, 95% CI 1.8-5.3, p < 0.001), and specifically in SCD pregnant or postpartum women (OR 20.3, 95% CI 4.1-102, p = 0.0003). The risk of PE was also higher in adults with SCD (OR 3.1, 95% CCI 1.7-5.9, p = 0.0004) as compared to those with SCT. The risk of VTE was higher in individuals with SCT compared to controls (pooled OR 1.7, 95% CI 1.3-2.2, p < 0.0001), but not in pregnant or postpartum women (OR 0.9, 95% CI 0.3-2.9, p = 0.863). Compared to controls, SCT was associated with a higher risk of PE (pooled OR 2.1, 95% CI 1.2-3.8, p = 0.012) but not of DVT (pooled OR 1.2, 95% CI 0.9-1.7, p = 0.157). Individuals with SCD, especially pregnant or postpartum women, might have a higher risk of VTE compared to the general population. SCT might also increases the risk of VTE. However, currently available data are not sufficient to allow a definite conclusion. Further larger studies are needed to provide a definitive conclusion on the association between SCD, SCT and VTE.


Bone regeneration in 3D printing bioactive ceramic scaffolds with improved tissue/material interface pore architecture in thin-wall bone defect.
Three-dimensional (3D) printing bioactive ceramics have demonstrated alternative approaches to bone tissue repair, but an optimized materials system for improving the recruitment of host osteogenic cells into the bone defect and enhancing targeted repair of the thin-wall craniomaxillofacial defects remains elusive. Herein we systematically evaluated the role of side-wall pore architecture in the direct-ink-writing bioceramic scaffolds on mechanical properties and osteogenic capacity in rabbit calvarial defects. The pure calcium silicate (CSi) and dilute Mg-doped CSi (CSi-Mg6) scaffolds with different layer thickness and macropore sizes were prepared by varying the layer deposition mode from single-layer printing (SLP) to double-layer printing (DLP) and then by undergoing one-, or two-step sintering. It was found that the dilute Mg doping and/or two-step sintering schedule was especially beneficial for improving the compressive strength (∼25-104 MPa) and flexural strength (∼6-18 MPa) of the Ca-silicate scaffolds. The histological analysis for the calvarial bone specimens in vivo revealed that the SLP scaffolds had a high osteoconduction at the early stage (4 weeks) but the DLP scaffolds displayed a higher osteogenic capacity for a long time stage (8-12 weeks). Although the DLP CSi scaffolds displayed somewhat higher osteogenic capacity at 8 and 12 weeks, the DLP CSi-Mg6 scaffolds with excellent fracture resistance also showed appreciable new bone tissue ingrowth. These findings demonstrate that the side-wall pore architecture in 3D printed bioceramic scaffolds is required to optimize for bone repair in calvarial bone defects, and especially the Mg doping wollastontie is promising for 3D printing thin-wall porous scaffolds for craniomaxillofacial bone defect treatment.


[Learning from experience. Applications of control charts to the monitoring of performance in cardiology].
Health services and teams of professionals should be able to monitor the process and outcomes of the care provided, in order to assess and maintain quality. In this paper the application of the control chart technique as a tool for the evaluation of in-hospital mortality of patients with acute myocardial infarction is presented and discussed. The control chart technique, based on the comparison between the outcome observed and the one expected in each individual patient, taking into account prognostic factors, has been applied in the evaluation of in-hospital mortality of 193 patients cared for at four centers of one of the AUSL of Bologna, during the year 2000. Yearly case volume for each of the four centers ranged from 14 to 68 patients. Overall mortality was 8.8, 18.0, 18.0, and 28.6% in the four centers. Use of control charts made it possible to identify relevant differences in outcomes among the four study centers, which would have been otherwise overlooked with usual statistical approaches. In particular, over the whole study period two centers showed relevant variations in their ability to achieve the desired clinical outcome. Clinical teams may use control charts as a tool to monitor their performance and for a timely identification of aspects of care deserving further investigation and analysis.


Enhanced co-stimulatory ability of synovial fluid accessory cells in rheumatoid arthritis.
We have established in vitro assays that allow the examination of co-stimulatory function of rheumatoid arthritis (RA) antigen-presenting cells (APC). Synovial fluid (SF) and peripheral blood (PB) APC co-stimulatory ability was compared in the activation of peptide-specific human T-cell clones. T-cell receptor (TCR) stimulation by peptide or anti-CD3 antibody allowed the direct comparison of SF and PB APC co-stimulatory activity, separately from their ability to process antigen. SF APC from 15 RA patients consistently enhanced T-cell proliferation when compared to their PB counterparts. Moreover, increasing the numbers of PB APC present resulted in only a minor increase in T-cell proliferation, failing to achieve levels stimulated by SF APC. We propose that the enhanced co-stimulatory function of synovial APC may be a significant factor in the persistence of local immune responses in RA.


The transformics assay: first steps for the development of an integrated approach to investigate the malignant cell transformation in vitro.
The development of alternative methods to animal testing is a priority in the context of regulatory toxicology. Carcinogenesis is a field where the demand for alternative methods is particularly high. The standard rodent carcinogenicity bioassay requires a large use of animals, high costs, prolonged duration and shows several limitations, which can affect the comprehension of the human relevance of animal carcinogenesis. The cell transformation assay (CTA) has long been debated as a possible in vitro test to study carcinogenesis. This assay provides an easily detectable endpoint of oncotransformation, which can be used to anchor the exposure to the acquisition of the malignant phenotype. However, the current protocols do not provide information on either molecular key events supporting the carcinogenesis process, nor the mechanism of action of the test chemicals. In order to improve the use of this assay in the integrated testing strategy for carcinogenesis, we developed the transformics method, which combines the CTA and transcriptomics, to highlight the molecular steps leading to in vitro malignant transformation. We studied 3-methylcholanthrene (3-MCA), a genotoxic chemical able to induce in vitro cell transformation, at both transforming and subtransforming concentrations in BALB/c 3T3 cells and evaluated the gene modulation at critical steps of the experimental protocol. The results gave evidence for the potential key role of the immune system and the possible involvement of the aryl hydrocarbon receptor (AhR) pathway as the initial steps of the in vitro transformation process induced by 3-MCA, suggesting that the initiating events are related to non-genotoxic mechanisms.


Linear ablation of left atrium for the treatment of atrial fibrillation guided by double Lasso catheters and three dimensional electroanatomical mapping.
Linear ablation of left atrium (LA) guided by three dimensional (3-D) electroanatomical mapping (Carto) has been used in many centres worldwide for the treatment of atrial fibrillation (AF) instead of pure anatomical approaches. There were little data about linear ablation of LA guided by Carto and double Lasso catheters in China. We report the results of linear ablation of LA guided by both Carto and double Lasso catheters. After the anatomical model of LA and all pulmonary veins (PVs) had been established, circumferential ablations of the left pulmonary vein antrum and the right pulmonary vein antrum were performed with 2 circumferential mapping catheters (Lasso) placed within the ipsilateral superior and inferior PVs. The endpoint of ablation was abolishment or dissociation of the pulmonary vein potentials (PVPs). Oral amiodarone or propafenone was taken for at least 3 months by patients with persistent AF, permanent AF or those whose PVPs had not been isolated completely. The recurrence of atrial tachyarrhythmias was observed 3 months after the procedure. There were 106 patients (mean age, 51.4 +/- 9.9 years). Seventy-eight patients had paroxysmal AF, 12 persistent AF and 16 permanent AF. Onset of atrial fibrillation occurred in 52 patients during ablation procedure. Thirty-two patients restored to sinus rhythm eventually after the procedure. Abolishment or dissociation of PVPs was accomplished during the procedure in 94 patients (88.7%). The duration of procedure and exposure to X-ray were (213 +/- 45) minutes and (32.5 +/- 12.8) minutes, respectively. Among the 87 patients followed up for over 3 months, 62 were free of atrial tachyarrhythmias (including 8 patients who were still taking oral amiodarone). The success rate was 71.3% in the first procedure. Two patients had pericardial effusion treated by pericardial puncture and effusion drainage. No pulmonary vein stenosis, atrioesophageal fistula, stroke or procedural death occurred. Combination of double Lasso catheters with 3-D electroanatomical mapping to guide the linear ablation of left atrium procedure can confirm the isolation of PVPs.


Changes in blood cell membrane properties in HIV type-1-infected patients.
To evaluate the possible HIV-1 infection-induced changes in cell membrane properties and in calcium signaling, membrane fluidity, acetylcholinesterase (AChE, a glycosylphosphatidylinositol-anchored protein) activity, and intracellular calcium concentration ([Ca2(+)](int)) were evaluated in lymphocytes and erythrocytes of infected individuals, previous to their engagement in antiretroviral therapy. Membrane fluidity was assessed by fluorescence spectroscopy measurements, using the fluorescence probes 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-[4-(trimethylamino)-phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). AChE activity was determined by the colorimetric Ellman's method and [Ca2(+)](int) using the fluorescent fura-2 acetoxymethyl ester. When compared with the control group, lymphocytes of infected patients presented significantly decreased membrane fluidity, decreased AChE activity, and increased [Ca2(+)](int). Erythrocytes from HIV-infected patients presented decreased [Ca2(+)](int) when compared with the control group and decreased membrane fluidity near the lipid/water interface. Our data show that HIV-1 infection leads to biochemical and biophysical changes in the membrane itself and in membrane protein activity in lymphocytes (average of infected and noninfected subpopulations) and even in erythrocytes. The present observations are in agreement with a process of facilitated propagation of the infection to new cells, stimulation of virion production, and maintenance of a reservoir of erythrocyte-bound infectious virus.


Human Parechovirus Central Nervous System Infection in a Young Infant Cohort.
In 2022, a surge in cases of pediatric human parechovirus (HPeV) central nervous system infections in young infants was seen at our institution. Despite the dramatic increase in the number of cases seen that year, the clinical features of the illness were similar to prior years. The recent pediatric HPeV surge highlights the need to evaluate treatment options and standardize follow-up to better understand the long-term prognosis of infants with HPeV infection.


An iterative transmission algorithm incorporating cross-talk correction for SPECT.
Simultaneous emission/transmission acquisitions in cardiac SPECT with a Tc99m/Gd153 source combination offer the capability for nonuniform attenuation correction. However, cross-talk of Tc99m photons downscattered into the Gd153 energy window contaminates the reconstructed transmission map used for attenuation correction. The estimated cross-talk contribution can be subtracted prior to transmission reconstruction or incorporated in the reconstruction algorithm itself. In this work, we propose an iterative transmission algorithm (MLTG-S) based on the maximum-likelihood gradient algorithm (MLTG) that explicitly accounts for this cross-talk estimate. Clinical images were acquired on a three-headed SPECT camera, acquiring Tc99m emission and Gd153 transmission images simultaneously. Subtracting the cross-talk estimate prior to transmission reconstruction can result in negative and zero values if the estimate is larger than or equal to the count in the transmission projection bin, especially with increased attenuator size or amount of cross-talk. This results in inaccurate attenuation coefficients for MLTG reconstructions with cross-talk subtraction. MLTG-S reconstructions on the other hand, yield better estimates of attenuation maps, by avoiding the subtraction of the cross-talk estimate. Comparison of emission slices corrected for nonuniform attenuation reveals that inaccuracies in the reconstructed attenuation map caused by cross-talk can artificially enhance the extra-cardiac activity, confounding the ability to visualize the left-ventricular walls.


Breast cancer metastasizing to the upper gastrointestinal tract (the esophagus and the stomach): A comprehensive review of the literature.
Breast cancer can infrequently metastasize to the upper gastrointestinal (GI) tract but the exact incidence is not well established-there is considerable variation between incidence reported from clinical studies and incidence noted in autopsy series. Clinical presentation can be very non-specific and often mimics primary gastrointestinal conditions. Endoscopy alone may not be sufficient to make a diagnosis and misdiagnosis is also common. A high degree of awareness and clinical suspicion is required to establish metastases to the upper GI tract. We undertook a comprehensive review of the available literature on breast cancer metastases to the esophagus and stomach including the clinical symptoms and presentation, endoscopic features, additional diagnostic imaging modalities, treatment and outcomes.


Controlled molecular organization of surface macromolecular assemblies based on stimuli-responsive polypeptide brushes.
End-tethered cationic polypeptide brushes of poly(L-lysine) (t-PLL) were combined with three anionic polymers, poly(acrylic acid) (PAA), poly(L-glutamic acid) (PLGA), and poly(L-aspartic acid) (PLAA), to form reversible polyelectrolyte complex films at surfaces at neutral pH. The polyelectrolyte complex formation was confirmed by an in situ zeta-potential study and by positive fluorescent images after adding prelabeled anionic polymers. The secondary conformations of the t-PLL complex films depend upon the specific polyelectrolyte with which t-PLL was coupled as studied by circular dichroism and FTIR. Specifically, the random coil chain configuration of the t-PLL film was converted to an alpha-helical, beta-sheet, or random coil structure after forming complexes with PAA, PLGA, or PLAA, respectively. Each of these complexes could be returned to the original random coil t-PLL structure by a dilute acid rinse. Additional thickness and morphological studies from ellipsometry and atomic force microscopy have further shown that the corresponding film thicknesses of the individual solvated films were affected more by the secondary structures in films than by the adsorbed mass or surface net charges. The solvated thickness was reduced significantly after the random coil t-PLL film was coupled with polyanions in forming compact regulated structures in films. This biomimetic approach provides a new opportunity for controlling the molecular organization in surface macromolecular assemblies and may provide a model for structural study of protein complexes on a chip.


Optimization of Dental Devices and Tools Used on Teeth.
Taking decisions is important in every aspect of life. Decision-making has become a difficult problem in any situation where there are multiple criteria. The application of multicriteria decision-making methods that can bring mathematical and logical solutions to the problem from an analytical perspective has experienced considerable growth recently. It provides great benefits in solution and subsequent stages. Medical equipment selection is also a challenging, complex, and difficult problem for the decision-maker, due to the requirements of conflicting criteria, which must be taken into account simultaneously. In this context, the aim of this study implicates the principle of multicriteria decision-making theories on various types of instruments used in dentistry. Since the data used in this study are not numeric but linguistic, the Fuzzy PROMETHEE decision-making method is used. In this research, six dental tools most commonly used by professionals to perform operations on patients are compared and evaluated. Fuzzy PROMETHEE decision-making method investigations show that the dental mirror is the most effective tool among all compared tools, followed by dental suction, dental air abrasion, dental handpiece, dental laser, and dental X-ray, consequently, basing on the selected criteria and the importance weight given to each criterion. Using this technique, one can obtain more specific ranking results based on a specific preference level.


Resource-based adaptive divergence in the freshwater fish Telmatherina from Lake Matano, Indonesia.
Adaptive radiations are an important source of biodiversity, but resolving which ecological pressures seed these processes in natural systems remains difficult. Here the adaptive radiation among Telmatherina, a genus of freshwater fish endemic to an ancient lake in central Sulawesi, Indonesia, was examined to determine its causal root. We demonstrate that all Telmatherina in this lake can be categorized into three lineages each possessing specialized skull shapes and pharyngeal jaw bones allowing them to exploit different resources. These data demonstrate a natural example of how resource partitioning has likely initiated adaptive radiation in a resource limited environment.


The Magnetic Levator Prosthesis for Temporary Management of Severe Blepharoptosis: Initial Safety and Efficacy.
We further optimized and evaluated the safety of the magnetic levator prosthesis (MLP) for temporary management of severe blepharoptosis, and compared efficacy and comfort against the ptosis crutch. The interpalpebral fissure (IPF) of participants (n = 12) with ptosis was measured during attempted eyelid opening, volitional closing, and spontaneous closing with no device, ptosis crutch, or the MLP. A 10-point scale documented comfort. Additionally, a 20 minute and then 1 week trial of the MLP was offered. Safety measures were skin erythema rating, change in visual acuity, and change in corneal staining. The MLP and crutch opened the eye (IPF 11.2 and 9.3 mm), but the MLP allowed better volitional closure (IPF 1.0 vs. 4.9 mm, P = 0.009), but was no better in allowing spontaneous blink (IPF 7.5 vs. 7.7 mm, P = 0.722). Both devices were equally comfortable (both median 8/10 comfort, P = 0.46). With extended use, opening with the MLP showed IPF 9.24 mm at 20 minutes and 9.46 mm at 1 week, and volitional closure was IPF 0.95 and 0.52 mm, respectively. Closure on spontaneous blink improved with extended wear to IPF 5.14 and 5.18 mm, respectively (P = 0.002). Two participants exhibited moderate skin erythema and one had increased corneal staining without change in acuity. The MLP is safe and feasible for temporary correction of severe ptosis. First group data in patients showing successful reanimation of the eyelid with magnetic force.


Impact of Neuroeffector Adrenergic Receptor Polymorphisms on Incident Ventricular Fibrillation During Acute Myocardial Ischemia.
Background Cardiac adrenergic receptor gene polymorphisms have the potential to influence risk of developing ventricular fibrillation (VF) during ST-segment-elevation myocardial infarction, but no previous study has comprehensively investigated those most likely to alter norepinephrine release, signal transduction, or biased signaling. Methods and Results In a case-control study, we recruited 953 patients with ST-segment-elevation myocardial infarction without previous cardiac history, 477 with primary VF, and 476 controls without VF, and genotyped them for ADRB1 Arg389Gly and Ser49Gly, ADRB2 Gln27Glu and Gly16Arg, and ADRA2C Ins322-325Del. Within each minor allele-containing genotype, haplotype, or 2-genotype combination, patients with incident VF were compared with non-VF controls by odds ratios (OR) of variant frequencies referenced against major allele homozygotes. Of 156 investigated genetic constructs, 19 (12.2%) exhibited significantly (P<0.05) reduced association with incident VF, and none was associated with increased VF risk except for ADRB1 Gly389 homozygotes in the subset of patients not receiving β-blockers. ADRB1 Gly49 carriers (prevalence 23.0%) had an OR (95% CI) of 0.70 (0.49-0.98), and the ADRA2C 322-325 deletion (Del) carriers (prevalence 13.5%) had an OR of 0.61 (0.39-0.94). When present in genotype combinations (8 each), both ADRB1 Gly49 carriers (OR, 0.67 [0.56-0.80]) and ADRA2C Del carriers (OR, 0.57 [0.45- 0.71]) were associated with reduced VF risk. Conclusions In ST-segment-elevation myocardial infarction, the adrenergic receptor minor alleles ADRB1 Gly49, whose encoded receptor undergoes enhanced agonist-mediated internalization and β-arrestin interactions leading to cardioprotective biased signaling, and ADRA2C Del322-325, whose receptor causes disinhibition of norepinephrine release, are associated with a lower incidence of VF. Registration URL: https://clinicaltrials.gov; Unique identifier: NCT00859300.


[Anesthetic management of a patient with mental disabilities as well as the past history of heat stroke and neuroleptic malignant syndrome].
A 22-year-old man with mental disabilities with the history of neuroleptic malignant syndrome and heat stroke was scheduled for dental treatment under general anesthesia. Heat stroke and neuroleptic malignant syndrome are related to malignant hyperthermia. We suggested the patient and family to undergo preoperative screening tests for malignant hyperthermia susceptibility, but they rejected. We selected slow induction using nitrous oxide, oxygen and sevoflurane to prevent excitement and anxiety for placing a catheter in a peripheral vein. We were very cautious in primary symptoms of malignant hyperthermia, i.e., tachycardia, increased end-tidal carbon dioxide, and rigidity of masseter. In the perioperative period, no complications occurred.


Intracytoplasmic sperm injection (ICSI) and chromosomally abnormal spermatozoa.
An infertile couple was referred for intracytoplasmic sperm injection (ICSI) because of primary infertility and oligoasthenoteratozoospermia (OAT) in the male. It was observed that although the sperm cells presented with an unusual head size and multiple tails they were able to fertilize the oocytes after ICSI. Subsequent molecular cytogenetic analysis demonstrated de-novo chromosome abnormalities in virtually all sperm cells with 40% diploidy and 24% triploidy in addition to aneuploidy for the sex chromosomes.


RNA sequencing reveals niche gene expression effects of beta-hydroxybutyrate in primary myotubes.
Various forms of fasting and ketogenic diet have shown promise in (pre-)clinical studies to normalize body weight, improve metabolic health, and protect against disease. Recent studies suggest that β-hydroxybutyrate (βOHB), a fasting-characteristic ketone body, potentially acts as a signaling molecule mediating its beneficial effects via histone deacetylase inhibition. Here, we have investigated whether βOHB, in comparison to the well-established histone deacetylase inhibitor butyrate, influences cellular differentiation and gene expression. In various cell lines and primary cell types, millimolar concentrations of βOHB did not alter differentiation in vitro, as determined by gene expression and histological assessment, whereas equimolar concentrations of butyrate consistently impaired differentiation. RNA sequencing revealed that unlike butyrate, βOHB minimally impacted gene expression in primary adipocytes, macrophages, and hepatocytes. However, in myocytes, βOHB up-regulated genes involved in the TCA cycle and oxidative phosphorylation, while down-regulating genes belonging to cytokine and chemokine signal transduction. Overall, our data do not support the notion that βOHB serves as a powerful signaling molecule regulating gene expression but suggest that βOHB may act as a niche signaling molecule in myocytes.


Ilheus virus isolation in the Pantanal, west-central Brazil.
The wetlands of the Brazilian Pantanal host large concentrations of diverse wildlife species and hematophagous arthropods, conditions that favor the circulation of zoonotic arboviruses. A recent study from the Nhecolândia sub-region of Pantanal reported serological evidence of various flaviviruses, including West Nile virus and Ilheus virus (ILHV). According to the age of seropositive horses, at least three flaviviruses, including ILHV, circulated in the Brazilian Pantanal between 2005 and 2009. To extend this study, we collected 3,234 adult mosquitoes of 16 species during 2009 and 2010 in the same sub-region. Mosquito pool homogenates were assayed for infectious virus on C6/36 and Vero cell monolayers and also tested for flaviviral RNA by a group-specific real-time RT-PCR. One pool containing 50 non-engorged female specimens of Aedes scapularis tested positive for ILHV by culture and for ILHV RNA by real-time RT-PCR, indicating a minimum infection rate of 2.5 per 1000. Full-length genomic sequence exhibited 95% identity to the only full genome sequence available for ILHV. The present data confirm the circulation of ILHV in the Brazilian Pantanal.


Basal cell carcinoma: what new can be learned about the most common human cancer? A cross-sectional prospective study of 180 cases in a single centre.
Basal cell carcinoma (BCC) is the most common cancer in humans, but the reporting of patients with BCC is still not complete. There are a limited number of analyses in the literature on BCC epidemiology. To study the epidemiological as well as clinical aspects of BCC by analysing a single centre's experience in the Lower Silesia region of Poland. We investigated 180 patients with BCC treated in the Unit of Dermatosurgery, Department of Dermatology, Venereology, and Allergology of Wroclaw Medical University between September 2017 and December 2019. The mean age of patients with BCC was 70.5 ±11.9 years. The most frequently diagnosed type of BCC was nodular type, at 72% of all patients. BCC occurred most commonly on the face and neck area, at 132 (73.3%), without a statistical difference between males and females. The vast majority of cancers were located on the nose. There was a personal history of skin cancer in 29% of our patients. In 127 (70.6%) subjects, the diameter of the BCC on the day of excision was less than 2 cm. The clinical characteristics of our cohort of BCC patients has some similarities with that reported from central Poland and other European countries. However, we found an increase in the incidence of BCC among females.


An Efficient, Robust Method for the Determination of Cannabinoids in Whole Blood by LC-MS-MS.
Due to the high prevalence of cannabinoids in forensic toxicology casework, it is desirable to have an efficient method that uses a small volume of blood and requires a minimal sample preparation. Although many methods have been reported, they are often labor intensive, require special sample preparation materials, use 1 mL or more of specimen or are difficult to replicate. The liquid chromatography with tandem mass spectrometry (LC-MS-MS) method presented herein employs a rapid and simple liquid-liquid extraction, has been successfully applied in two different laboratories, uses 0.5 mL of specimen and was extensively validated. The validated limit of detection and limit of quantitation were 1 ng/mL for delta-9-tetrahydrocannabinol (THC) and 11-hydroxy-delta-9-tetrahydrocannabinol (OH-THC) and 5 ng/mL for 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THCA). Each analyte demonstrated a zero-order linear range (r2 > 0.990) with 1/x weighting of 1-40 ng/mL for THC and OH-THC and 5-200 ng/mL for THCA. The coefficient of variation of replicate analyses was within 14%. Bias was within ±13% of the prepared concentration. The validated method provides a sensitive, efficient and robust procedure for the quantitation of cannabinoids in blood using LC-MS-MS and a sample volume of 0.5 mL.


Organization of Presynaptic Autophagy-Related Processes.
Brain synapses pose special challenges on the quality control of their protein machineries as they are far away from the neuronal soma, display a high potential for plastic adaptation and have a high energy demand to fulfill their physiological tasks. This applies in particular to the presynaptic part where neurotransmitter is released from synaptic vesicles, which in turn have to be recycled and refilled in a complex membrane trafficking cycle. Pathways to remove outdated and damaged proteins include the ubiquitin-proteasome system acting in the cytoplasm as well as membrane-associated endolysosomal and the autophagy systems. Here we focus on the latter systems and review what is known about the spatial organization of autophagy and endolysomal processes within the presynapse. We provide an inventory of which components of these degradative systems were found to be present in presynaptic boutons and where they might be anchored to the presynaptic apparatus. We identify three presynaptic structures reported to interact with known constituents of membrane-based protein-degradation pathways and therefore may serve as docking stations. These are (i) scaffolding proteins of the cytomatrix at the active zone, such as Bassoon or Clarinet, (ii) the endocytic machinery localized mainly at the peri-active zone, and (iii) synaptic vesicles. Finally, we sketch scenarios, how presynaptic autophagic cargos are tagged and recruited and which cellular mechanisms may govern membrane-associated protein turnover in the presynapse.


Tuberculosis Cervical Adenitis: Management Dilemmas.
Peripheral lymphadenopathy (scrofula) is the second most common site for childhood tuberculosis. Paradoxical reactions are commonly seen even in immunocompetent children after therapy is initiated, and this can lead to draining sinus tracts. We describe a multimodal therapeutic option of antituberculosis therapy, corticosteroids and therapeutic ultrasound-guided nodal aspiration in an adolescent with massive cervical adenopathy.


Subretinal semiconductor microphotodiode array.
To examine the function of a semiconductor microphotodiode array (SMA) surgically implanted in the subretinal space. Positive-intrinsic layer-negative (PiN) or negative-intrinsic layer-positive (NiP) SMAs were surgically placed into the subretinal space of rabbits through a pars plana incision and a posterior retinotomy. The implants required no external connections for power and were sensitive to light over the visible and infrared (IR) spectrum; IR stimuli were used to isolate implant-mediated responses from the activity of native photoreceptors. A stimulator ophthalmoscope was used to superimpose IR stimuli on the implant and adjacent retinal areas, and responses were recorded during the postoperative recovery period. SMA responses were also evaluated in vitro. The animals were given lethal anesthetic overdoses, and the retinas were examined histologically. The in vitro implant response consisted of an electrical spike, followed by a small-amplitude DC offset that followed the time course of the IR stimulation, and an overshoot at the stimulus offset. The SMAs placed in the subretinal space retained a stable position and continued to function throughout the postoperative period. The SMA responses recorded in vivo included additional slow-wave components that were absent from the in vitro recordings. These responses reverted to the in vitro configuration following the death of the animal. There was a significant loss of retinal cells in areas overlying the implant, and the retina appeared normal away from the implant and surgical site. SMAs can be successfully implanted into the subretinal space and will generate current in response to light stimulation during an extended period of time.


Identification of a Region of the Polypeptide Chain of Na,K-ATPase α-Subunit Interacting with 67-kDa Melittin-Like Protein.
It was shown earlier that a 67-kDa protein purified from mouse kidney using polyclonal antibodies against melittin (a peptide from bee venom) interacted with Na,K-ATPase from rabbit kidney. In this study, a 43-kDa proteolytic fragment of Na,K-ATPase α-subunit interacting with the 67-kDa melittin-like protein was found. The α-subunit was hydrolyzed by trypsin in the presence of 0.5 mM ouabain (E2-conformation of Na,K-ATPase). A proteolytic fragment interacting with the 67-kDa melittin-like protein that was identified by mass-spectrometry is a region of the cytoplasmic domain of Na,K-ATPase α-subunit located between amino acid residues 591 and 775. The fragment includes a conservative DPPRA motif that occurs in many P-type ATPases. It was shown earlier that this motif of H,K-ATPase from gastric mucosa binds to melittin. We suggest that namely this motif of P-type ATPases is able to interact with proteins containing melittin-like modules.


NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP).
NFATc1 has been characterized as a master regulator of nuclear factor kappaB ligand-induced osteoclast differentiation. Herein, we demonstrate a novel role for NFATc1 as a positive regulator of nuclear factor kappaB ligand-mediated osteoclast fusion as well as other fusion-inducing factors such as TNF-alpha. Exogenous overexpression of a constitutively active form of NFATc1 in bone marrow-derived monocyte/macrophage cells (BMMs) induces formation of multinucleated osteoclasts as well as the expression of fusion-mediating molecules such as the d2 isoform of vacuolar ATPase V(o) domain (Atp6v0d2) and the dendritic cell-specific transmembrane protein (DC-STAMP). Moreover, inactivation of NFATc1 by cyclosporin A treatment attenuates expression of Atp6v0d2 and DC-STAMP and subsequent fusion process of osteoclasts. We show that NFATc1 binds to the promoter regions of Atp6v0d2 and DC-STAMP in osteoclasts and directly induces their expression. Furthermore, overexpression of Atp6v0d2 and DC-STAMP rescues cell-cell fusion of preosteoclasts despite reduced NFATc1 activity. Our data indicate for the first time that the NFATc1/Atp6v0d2 and DC-STAMP signaling axis plays a key role in the osteoclast multinucleation process, which is essential for efficient bone resorption.


The process of engaging members from two underserved populations in the development of interventions to promote the uptake of the HPV vaccine.
We describe a community-engaged research process used to develop multilevel interventions (caregivers, providers, system) to improve the uptake of human papillomavirus vaccine among adolescents by partnering with members from two underserved populations in Ohio. We began by conducting focus groups to better understand the knowledge and attitudes of caregivers and providers about the human papillomavirus vaccine and to develop teams of community members to assist with development of the interventions. The process continued with conducting writing sessions to determine the content and format of the interventions, and initial feedback was obtained during review sessions to refine the interventions prior to implementation. Using this approach, we were able to consider contextual factors that made the interventions more acceptable and relevant to members of the priority populations. Challenges included development and maintenance of a team of community members to participate in the entire intervention development process, rejection of ideas presented by academic researchers, the need to balance community members' suggestions with what was known from evidence-based research, and the time, cost, and effort associated with partnering with community members. The benefits, however, outweigh the challenges associated with using a community-engaged research process to develop interventions aimed at reducing cancer disparities among underserved populations.


Scalable Assembly of High-Quality Graphene Films via Electrostatic-Repulsion Aligning.
Assembling pristine graphene into freestanding films featuring high electrical conductivity, superior flexibility, and robust mechanical strength aims at meeting the all-around high criteria of new-generation electronics. However, voids and defects produced in the macroscopic assembly process of graphene nanosheets severely degrade the performance of graphene films, and mechanical brittleness often limits their applications in wide scenarios. To address such challenges, an electrostatic-repulsion aligning strategy is demonstrated to produce highly conductive, ultraflexible, and multifunctional graphene films. Typically, the high electronegativity of titania nanosheets (TiNS) induces the aligning of negatively charged graphene nanosheets via electrostatic repulsion in the film assembly. The resultant graphene films show fine microstructure, enhanced mechanical properties, and improved electrical conductivity up to 1.285 × 105 S m-1 . Moreover, the graphene films can withstand 5000 repeated folding without structural damage and electrical resistance fluctuation. These comprehensive improved properties, combined with the facile synthesis method and scalable production, make these graphene films a promising platform for electromagnetic interference (EMI) shielding and thermal-management applications in smart and wearable electronics.


[Anesthetic Management for a Patient with Fibromyalgia].
A 22-year-old female with fibromyalgia (FM) was scheduled for tonsillectomy under general anesthesia. Her medication included pregabalin 300 mg x day(-1) and dantrolene 50 mg x day(-1). Anesthesia was maintained with sevoflurane-remifentanil-fentanyl. Intravenous injection of fentanyl 20 μg x hr(-1) and droperidol 100 μg x hr(-1) was continued for 24 hours. On the first postoperative day, she reported that she had slept well and had no pain. There are some perioperative problems in a patient with FM. Therefore, anesthetic managements for a patient with FM is worth reporting.


Schistosoma mansoni associated glomerulopathy with IgA mesangial deposits: case report.
Renal involvement is a severe form of schistosomiasis and occurs in 10% to 15% of patients with the hepatosplenic form of the disease. Nephrotic syndrome is the most common clinical presentation. It is a complication caused by immune complexes (IC), it is rare to appear in the Brazilian context with a immunoglobulin A (IgA) deposits. When installed the renal injury by Schistosoma mansoni, classically presents as membranoproliferative glomerulonephritis (mesangiocapillary) with lobular accentuation. To report a case of schistosomiasis nephropathy that appeared 7 years after treatment of hepatosplenic schistosomiasis with histologic pattern of mesangial proliferative glomerulonephritis with IgA deposits in mesangium. Clinically developed with progressive decrease of proteinuria with angiotensin receptor blocker (ARB). It was reported a case of a 36 years old patient, brown, with classical sintoms of nephrotic syndrome (proteinuria > 3.5 g/24h, hypoalbuminemia and hypercholesterolemia), however with hepatosplenic schistosomiasis history 7 years ago and portal hypertension. Patient underwent renal biopsy which showed IgA deposits in mesangial, being more intense than immunoglobulin G (IgG), accompanied by C1q and C3, with 4/13 glomeruli sclerotic, standard light mesangial glomerulonephritis renal injury with IgA deposits. Patient began taking ARB with progressive improvement in proteinuria. Patients with glomerulonephritis by schistosoma don't show improvement of disease progression with antiparasitic treatment. However the anti-proteinuric treatment can slow the progression of end stage kidney disease.


Tattoos and human identification: investigation into the use of X-ray and infrared radiation in the visualization of tattoos.
Any person with a tattoo known to their family or friends could potentially be identified from the presence of such personal identifying markers. Problems in identification utilizing tattoos may arise when these markers are removed or defaced in some way. This paper uses infrared wavelengths at 760, 850, and 950 nm to improve the visualization of laser-removed or covered up tattoos and also to establish whether the ink pigments used can be observed on radiographs from any metal that may be present. The results obtained indicate that some older inks have a high enough metallic content to allow them to be viewed on a radiograph, while infrared light can demonstrate latent ink still present in the skin after laser removal and can also be utilized to distinguish an original tattoo through a secondary "cover-up" tattoo. Infrared photography and radiography have been shown to improve tattoo visualization in a forensic context.


A new inflammation model.
The UV-edema in the hairless mouse is a suitable inflammation model as the degree of edema can be quantified - the degree of edema being a function of the irradiation time. The UV-edema may also be employed in the testing of antiphlogistics using oral administration as well as cutaneous application.


Learning to fly: building de novo medical home practices to improve experience, outcomes, and affordability.
While most approaches to improving primary care delivery involve making incremental improvements to existing practices, for the past decade we have been engaged in a different approach-to build new practices from the ground up to demonstrate a vision of what is possible. Our Iora model of care combines a new payment model which does away with all fee for service billing, a new delivery model focused on robust team-based population management, and a new IT system to drive this very different care. We have demonstrated in several pilots improved experience, outcomes, and lower total cost of care.