Evaluation of a kDNA-Based qPCR Assay for the Detection and Quantification of Old World Leishmania Species.
The parasite protozoan Leishmania, the causative agent of leishmaniasis, includes two subgenera of medical interest: Leishmania (Leishmania) and Leishmania (Viannia). Parasite species detection and characterization is crucial to choose treatment protocols and to monitor the disease evolution. Molecular approaches can speed up and simplify the diagnostic process. In particular, several molecular assays target the mitochondrial DNA minicircle network (kDNA) that characterizes the Leishmania genus. We previously proposed a qPCR assay targeting kDNA, followed by high resolution melt (HRM) analysis (qPCR-ML) to distinguish L. (L.) infantum and L. (L.) amazonensis from L. Viannia species. Successively, this assay has been integrated with other qPCR assays, to differentiate L. (L.) infantum, L. (L.) amazonensis and L. (L.) mexicana. In this work, we tested the applicability of our qPCR-ML assay on L. (L.) donovani, L. (L.) major, L. (L.) tropica and L. (L.) aethiopica, showing that the qPCR-ML assay can also amplify Old World species, different from L. (L.) infantum, with good quantification limits (1 × 10-4-1 × 10-6 ng/pcr tube). Moreover, we evaluated 11 L. (L.) infantum strains/isolates, evidencing the variability of the kDNA minicircle target molecules among the strains/isolates of the same species, and pointing out the possibility of quantification using different strains as reference. Taken together, these data account for the consideration of qPCR-ML as a quantitative pan-Leishmania assay.


Ppp6c haploinsufficiency accelerates UV-induced BRAF(V600E)-initiated melanomagenesis.
According to TCGA database, mutations in PPP6C (encoding phosphatase PP6) are found in c. 10% of tumors from melanoma patients, in which they coexist with BRAF and NRAS mutations. To assess PP6 function in melanoma carcinogenesis, we generated mice in which we could specifically induce BRAF(V600E) expression and delete Ppp6c in melanocytes. In these mice, melanoma susceptibility following UVB irradiation exhibited the following pattern: Ppp6c semi-deficient (heterozygous) > Ppp6c wild-type > Ppp6c-deficient (homozygous) tumor types. Next-generation sequencing of Ppp6c heterozygous and wild-type melanoma tumors revealed that all harbored Trp53 mutations. However, Ppp6c heterozygous tumors showed a higher Signature 1 (mitotic/mitotic clock) mutation index compared with Ppp6c wild-type tumors, suggesting increased cell division. Analysis of cell lines derived from either Ppp6c heterozygous or wild-type melanoma tissues showed that both formed tumors in nude mice, but Ppp6c heterozygous tumors grew faster compared with those from the wild-type line. Ppp6c knockdown via siRNA in the Ppp6c heterozygous line promoted the accumulation of genomic damage and enhanced apoptosis relative to siRNA controls. We conclude that in the presence of BRAF(V600E) expression and UV-induced Trp53 mutation, Ppp6c haploinsufficiency promotes tumorigenesis.


Early morphological and functional changes in the GABAergic system of hippocampus in the rat lithium-pilocarpine model of epilepsy.
We studied early alterations in the GABAergic system of the rat hippocampus in the lithium-pilocarpine model of epilepsy. Twenty-four hours after the pilocarpine treatment, a decrease in the number of calretinin-positive interneurons was observed in the CA1 field of the hippocampus, whereas the number of parvalbumin-positive interneurons remained unchanged. The decreased levels of the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD67) and the membrane GABA transporter GAT1 were revealed using Western blot analysis. These data indicate an altered excitation/inhibition balance in the hippocampus with excitation dominance.


[Viability and angiogenic activity of mesenchymal stromal cells from adipose tissue and bone marrow in hypoxia and inflammation in vitro].
Adult progenitor stromal cells derived from adipose tissue (ADSC) and bone marrow (BMDSC) hold great promise for use in cell-based therapy of ischemic diseases. Both cell lines secrete a various number of angiogenic cytokines which are regulated by hypoxia and improve vascularization of ischemic tissues being injected in damaged muscle or intravenously. However, such factors as low oxygen level and inflammation may impair the viability and functional activity of these cells after delivery to the ischemic area. We directly compared the reactions of ADSCs and BMDSCs to hypoxic and inflammatory conditions in vitro. Cultured ADSCs and BMDSCs from Balb/c mice were cultivated for 48 h under 1% O2 (hypoxia), 20% O2 (normoxia) or in the presence of inflammatory cytokines. Cell viability analyzed by annexin V-PE binding and 7AAD storage (flow cytometry), and by quantitative TUNEL showed no decrease under hypoxic condition. But cell apoptotic rates significantly increased (up to 70 %) under inflammatory condition. Inflammatory cytokines did not stimulate gene expression of angiogenic growth factors. Otherwise, gene expression profiles of angiogenesis-related cytokines showed activation of pro-angiogenic and suppression of anti-angiogenic factors in the cells under hypoxic condition. In general this effect was higher for ADSCs than for BMDSCs. Using in vitro and in vivo models of angiogenesis we have demonstrated that incubation under hypoxic condition increases stromal cells ability to stimulate blood vessels growth.


9a-Hy-droxy-3,8a-dimethyl-5-methyl-ene-4,4a,5,6,9,9a-hexa-hydro-naphtho-[2,3-b]furan-2(8aH)-one.
The title compound, C(15)H(18)O(3), was isolated from Lacta-rius piperatus (Fr.) S. F. Gary collected from the Kunming area in Yunnan province, China. The central cyclo-hexyl ring adopts a chair conformation, while the furan-one ring is close to planar (r.m.s. deviation = 0.0174 Å). The remaining methyl-ene cyclo-hexene ring has a flattened chair conformation. In the crystal, mol-ecules are linked via inter-molecular O-H⋯O and C-H⋯O hydrogen bonds into zigzag chains along the a axis.


Fenugreek seed mucilage grafted poly methacrylate pH-responsive hydrogel: A promising tool to enhance the oral bioavailability of methotrexate.
This study aimed to develop the Fenugreek seed mucilage-based pH-responsive hydrogel system in order to improve the oral bioavailability of methotrexate (MTX). Fenugreek seed mucilage (FSM) was extracted from Trigonella foenum-graecum seeds. F1-F9 formulations of pH-responsive hydrogels were prepared using various FSM ratios, methacrylic acid (MAA), and methylene bis acrylamide (MBA) via free radical polymerization technique. Swelling behavior and in vitro drug release studies of prepared hydrogels were evaluated. Toxicity studies of prepared hydrogels were performed on normal cells and on Wistar rats (n = 6). Moreover, in vivo pharmacokinetics parameters were studied on albino rabbits. Hydrogels formation was confirmed by FTIR analysis, thermal analysis and SEM studies. The maximum swelling of hydrogel was found to be 384.7% at pH 7.4. MTX-loaded hydrogel showed the controlled release of MTX up to 24 h following Super Case II transport. Prepared hydrogels exhibited no toxicity in normal cells as well as in experimental subjects. MTX loaded hydrogels exhibited less inhibition compared to free MTX on Hela cells. In Vivo studies revealed 7.5-fold improved oral bioavailability of MTX with higher Cmax (928 ng/mL). These results indicate that the pH-responsive hydrogel system based on FSM is a promising tool for the controlled delivery of MTX.


Identification of defensin-encoding genes of Picea glauca: characterization of PgD5, a conserved spruce defensin with strong antifungal activity.
Plant defensins represent a major innate immune protein superfamily that displays strong inhibitory effects on filamentous fungi. The total number of plant defensins in a conifer species is unknown since there are no sequenced conifer genomes published, however the genomes of several angiosperm species provide an insight on the diversity of plant defensins. Here we report the identification of five new defensin-encoding genes from the Picea glauca genome and the characterization of two of their gene products, named PgD5 and endopiceasin. Screening of a P. glauca EST database with sequences of known plant defensins identified four genes with homology to the known P. glauca defensin PgD1, which were designated PgD2-5. Whereas in the mature PgD2-4 only 7-9 amino acids differed from PgD1, PgD5 had only 64% sequence identity. PgD5 was amplified from P. glauca genomic DNA by PCR. It codes for a precursor of 77-amino acid that is fully conserved within the Picea genus and has similarity to plant defensins. Recombinant PgD5, produced in Escherichia coli, had a molecular mass of 5.721 kDa, as determined by mass spectrometry. The PgD5 peptide exhibited strong antifungal activity against several phytopathogens without any effect on the morphology of the treated fungal hyphae, but strongly inhibited hyphal elongation. A SYTOX uptake assay suggested that the inhibitory activity of PgD5 could be associated with altering the permeability of the fungal membranes. Another completely unrelated defensin gene was identified in the EST library and named endopiceasin. Its gene codes for a 6-cysteine peptide that shares high similarity with the fungal defensin plectasin. Screening of a P. glauca EST database resulted in the identification of five new defensin-encoding genes. PgD5 codes for a plant defensin that displays non-morphogenic antifungal activity against the phytopathogens tested, probably by altering membrane permeability. PgD5 has potential for application in the plant biotechnology sector. Endopiceasin appears to derive from an endo- or epiphytic fungal strain rather than from the plant itself.


Determination of intracellular conductivity from electrical breakdown measurements.
The intracellular resistivity (conductivity) of cells can be easily calculated with high accuracy from electrical membrane breakdown measurements. The method is based on the determination of the size distribution of a cell suspension as a function of the electrical field strength in the orifice of a particle volume analyser (Coulter counter). The underestimation of the size distribution observed beyond the critical external field strength leading to membrane breakdown represents a direct access to the intracellular resistivity as shown by the theoretical analysis of the data. The potential and the accuracy of the method is demonstrated for red blood cells and for ghost cells prepared by electrical haemolysis. The average value of 180 omega X cm for the intracellular resistivity of intact red blood cells is consistent with the literature.


Arthroscopic Centralization Using Knotless Anchors for Extruded Medial Meniscus.
The load-distributing function is most critical in meniscal function, and meniscal extrusion suggests failure of this function, leading to the progression of osteoarthritis. The arthroscopic centralization technique has been developed to reduce meniscal extrusion; however, existing arthroscopic techniques sometimes fail to reduce the most extruded region, especially in cases with a medial meniscus (MM) posterior root tear, in which the most extruded region is on the posterior border of the medial collateral ligament, which is very difficult to approach. This Technical Note describes an arthroscopic technique for extrusion of the MM in which a centralization technique using knotless anchors efficiently reduces the MM extrusion at the posteromedial part and consequently restores the MM function. This technique efficiently reduces MM extrusion and restores its function, thus preventing the progression of osteoarthritis.


Immunogenicity and safety of combined intradermal recombinant Hepatitis B with BCG vaccines at birth.
This randomized, prospective, non-inferiority study aimed to quantify anti-HBs titers induced by recombinant Hepatitis B vaccine from healthy infants vaccinated with combined Hepatitis B and Bacillus Calmette-Guérin (BCG) vaccines (HbsAg 10 microg plus BCG suspension 0.1mg) and compare them to titers obtained with separated vaccines. Infants were immunized at birth either with combined intradermal (ID) BCG and Hepatitis B or ID BCG alone and intramuscular (IM) Hepatitis B. Both groups received IM Hepatitis B at 1 and 6 months of age. After the third dose anti-HBs titers > or =10 IU/mL were observed in 99% of vaccinees and > or =1000 IU/mL in 71%. There were no adverse events in both groups. Combination of HbsAg with BCG as first dose did not modify the profile of the humoral immune response for Hepatitis B indicating safety and immunogenicity of this vaccine in newborn.


[Response of thyrotropin (TSH) to the stimulation with hypothalamic releasing factor (TRF) in hypothyroid children].
The normal human serum thyrotropin (TSH) measured by radioimmunoassay in 36 children was 3.3 +/- 1.13 muUI/ml. No differences dependent on age or sex were found. Stimulation by intravenous injection of 100 mug of TRF has been carried out in 7 euthyroid children and in 14 with hypothyroidism. TSH was determined prior to, and during TRF stimulation. Among other thyroid tests included in this study were: serum T3 uptake, total thyroxine, serum protein-bound iodine, 131INa uptake and thyroid scanning. In the euthyroid children, TSH at zero time was 4.4 muUI/ml.; a significant increase was reached at 30-40 minutes. In 13 children with a light primary hypothyroidism, TSH was little increased in correlation with other parameters assessed. In two children with true primary hypothyroidism, TSH at zero time was X 465 muUI/ml., showing an increase at 30 minutes of X 550 muUL/ml. which was in agreement with the absence of thyroid tissue, showed by the scanning image and with the low concentration of thyroid hormones. The synthetic thyrotropin-releasing factor (TRF) should be a useful agent for clinical assessment of pituitary reserve. TSH, T3, T4 and T4I serum concentration measurement in pediatric age, makes possible to evaluate the physiopathology of the pituitary - thyroid axis.


[Electron microscopic demonstration of links between DNA filaments and fibrillar areas of nucleolar RNA in interphase nuclei of L 929 cells].
Electronic opacification by DAB, following ribonuclease digestion, allowed us to observe a network of DNA filaments which appeared in close relation with both intranucleolar clumps of DNA on one hand, and with the nucleolar fibrillar RNA structure on the other hand. It is inferred that the ribosomal genes may be located on these filaments of DNA.


Opportunities and limitations for studying neuropsychiatric disorders using patient-derived induced pluripotent stem cells.
Neuropsychiatric disorders affect a large proportion of the global population and there is an urgent need to understand the pathogenesis and to develop novel and improved treatments of these devastating disorders. However, the diverse symptomatology combined with complex polygenic etiology, and the limited access to disorder-relevant cell types in human brains represent a major obstacle for mechanistic disease research. Conventional animal models, such as rodents, are limited by inherent species differences in brain development, architecture, and function. Advances in human induced pluripotent stem cells (hiPSCs) technologies have provided platforms for new discoveries in neuropsychiatric disorders. First, hiPSC-based disease models enable unprecedented investigation of psychiatric disorders at the molecular, cellular, and structural levels. Second, hiPSCs derived from patients with known genetics, symptoms, and drug response profiles offer an opportunity to recapitulate pathogenesis in relevant cell types and provide novel approaches for understanding disease mechanisms and for developing effective treatments. Third, genome-editing technologies have extended the potential of hiPSCs for generating models to elucidate the genetic basis of rare monogenetic and complex polygenic psychiatric disorders and to establish the causality between genotype and phenotype. Here we review opportunities and limitations for studying psychiatric disorders using various hiPSC-derived model systems.


[Septic arthritis of the native joint and after ligamentoplasty : Diagnosis and treatment].
Septic arthritis is an acute emergency. It occurs more frequently in patients with pre-existing degenerative or chronic inflammatory joint diseases than in the general population. The causative microorganisms can be introduced in various ways. A rapid diagnosis is of great importance for the success of the therapy. In the clinical examination, the typical signs of inflammation are noticeable. The gold standard is the aspiration of synovial fluid and the subsequent laboratory and microbiological investigation. A prerequisite for successful therapy is the early initiation of an antimicrobial pathogen-specific treatment and the surgical alleviation of the joint.


Assessment of some factors of cellular and humoral immunity in radiology workers.
Radiation workers in medical diagnostic departments are occupationally exposed to long-term low-dose ionizing radiation, which may cause radiation-induced side effects. This study investigated subtypes of peripheral blood lymphocytes and immunoglobulin levels in workers who were occupationally exposed to X-ray radiation at the Department of Radiology. Seventeen radiology workers received low levels of ionizing radiation as the study group and 18 individuals who were not exposed to radiation were included as the control group. The percentage of lymphocyte subtypes (CD4+, CD8+ and CD4+/CD8+) and serum levels of immunoglobulins (IgA, IgG and IgM) were measured using peripheral blood samples. Considering all lymphocyte subtypes and serum levels of IgA, IgG and IgM, there was no significant difference between the study and control groups (P > 0.05). There were no significant differences in all mentioned parameters regarding gender (P > 0.05). For the length of employment period, there was a significant difference concerning CD4+/CD8+ (P < 0.05). The findings showed that exposure to low levels of ionizing radiation does not affect the immune system of workers in diagnostic radiology dose level. Because of relatively small samples of workers, it is suggested that these factors be investigated on larger samples of radiology workers.


The histopathology of acute renal failure.
The morphologic and immunohistologic features in acute renal failure (ARF) are discussed in this article. The focus is on tubular epithelial cell injury, the central pathologic finding in the majority of cases of ARF. Tubular damage may be due to sublethal or lethal cell injury. Cell necrosis, including classic coagulative necrosis and apoptosis, manifestations of sublethal injury, and tubular cell exfoliation are described in detail. Pathologic findings in native kidneys are described.


The recovery and identification of fluoroacetamide and fluoroacetic acid from tissues.
A rapid method is described for the extraction and identification of fluoroacetamide and fluoroacetic acid from tissues, by gas chromatography, using a mass spectrometer in the multiple ion monitoring mode as the detector. Recoveries from animal and human tissue macerates, after the addition of each compound, were approximately 30--35% for fluoroacetamide and 50--55% for fluoroacetic acid. Minimum detectable levels in tissues were found to be circa 0.7 mug/g for the amide and 0.1 mug/g for the acid. Visceral tissue levels were found to be approximately 3 mug/g for fluoroacetamide, and 2.5--10 mug/g fluoroacetic acid in guinea pigs and rabbits which had been fatally poisoned with these compounds.


Sympathetic yet painful: Autonomic innervation drives cluster firing of somatosensory neurons.
In this issue of Neuron, Zheng et al. (2021) report synchronized cluster firing of dorsal root ganglion (DRG) neurons that correlates with spontaneous pain in the setting of nerve injury. The authors' findings further suggest that sympathetic sprouting in the DRG plays a key role in this phenomenon.


Quantum chemical investigation of the thermal pyrolysis reactions of the carboxylic group in a brown coal model.
Different reaction pathways of the carboxylic group in a brown coal model were investigated by applying density function quantum chemical theory, examining the possible cross-linking and decomposition reactions between the hydrogen bonded carboxylic group-carboxylic group and the carboxylic group-hydroxyl group during the thermal pyrolysis process. The results show that bimolecular dehydration and decarboxylation of hydrogen bonded carboxylic groups have distinctly lower activation barriers and therefore, proceed preferentially at low temperature. The esterification reaction between the hydrogen bonded carboxylic group and hydroxyl group, together with unimolecular decarboxylation of isolated single carboxylic groups were also possible at moderate temperature. Aryl-aryl coupling is thought to occur via radical pyrolysis and recombination at relatively high temperature.


[Importance of structural characteristics of the esophagus and tracheo-esophageal fistula in surgical treatment of esophageal atresia in newborn infants].
The anatomical structure of the upper and lower segments of the esophagus and tracheoesophageal fistula in esophageal atresia in newborn in newborn infants conditions the clinical picture, character of surgical intervention and postoperative course. The gravity of pneumonic changes in the lung is dependent not only on terms of child's admission, but also on the width of esophagotracheal fistula, while bifurcation fistulas, sometimes, cause diagnostic erros.


Tuberculosis in Tanzania--a national survey of newly notified cases. Tanzanian/British Medical Research Council Collaborative Study.
A survey was carried out in 1978/80 in the same random sample of districts in Tanzania that were surveyed in 1969, to obtain information on: the proportions of patients with pulmonary and extra-pulmonary tuberculosis; the proportion of patients with bacteriologically-positive pulmonary tuberculosis; the proportion of patients with initial drug resistance; the radiographic extent of pulmonary disease; the changes that had occurred in these factors since 1969, and the results of therapy achieved by the routine treatment services. Of 2242 patients in this survey, 91.5% had pulmonary tuberculosis only, 0.6% had both extra-pulmonary and pulmonary tuberculosis and 7.9% had extra-pulmonary tuberculosis only. Of the 191 patients with extra-pulmonary tuberculosis, 39% had lymph node involvement, 26% bone or joint disease and 19% a pleural effusion. Of the 2064 patients with pulmonary (with or without extra-pulmonary) tuberculosis 96.2% had no history of previous chemotherapy. The proportion for the patients with extra-pulmonary tuberculosis only was similar. A sputum specimen was produced by 1006 (62%) of the patients aged 10 years or more with pulmonary tuberculosis; 77.2% of smear results were positive, as were 68.3% of the culture results. In every patient with identification test results the strain was Mycobacterium tuberculosis. Of 555 patients with no history of previous chemotherapy and a sensitivity test result available, 10.1% had a strain resistant to isoniazid and/or streptomycin, namely, 7.6% to isoniazid alone, 0.7% to streptomycin alone and 1.8% to both drugs. Of the 1161 patients with chest radiographs available and a diagnosis in Tanzania of pulmonary tuberculosis, a lung lesion or a pleural effusion was reported at independent assessment in London for 82% of patients aged 15 years or more and for 16% of patients under 15 years of age. For the patients with a pulmonary lesion, the total extent of disease was greater than 1 lung in 37% of adults and 39% of children and cavitation was present in 55% of adults and 21% of children. A comparison of the pretreatment data in 1978/80 with those from the 1969 survey showed that they were broadly similar in most respects. The results of therapy at one year or later were assessed for 413 patients who had had bacteriological confirmation of their disease in the reference laboratory on admission. At follow-up 40% had a negative culture and a further 9% were reported to be cured without bacteriological confirmation.(ABSTRACT TRUNCATED AT 400 WORDS)


Detrusor-external sphincter dyssynergia: a detailed electromyographic study.
We reviewed 54 cases of detrusor-external sphincter dyssynergia following complete urodynamic evaluations. All patients had well defined neurologic lesions of the suprasacral spinal cord. On the basis of urodynamic findings 3 types of dyssynergia were encountered: type 1 (30 per cent) was characterized by a crescendo increase in electromyographic activity that reached a maximum at the peak of the detrusor contraction, type 2 (15 per cent) consisted of clonic sphincter contractions interspersed throughout the detrusor contraction and type 3 (55 per cent) was characterized by a sustained sphincter contraction that coincided with the detrusor contraction. There was no correlation between the clinical neurologic level and the type of dyssynergia.


Local membrane source gathering by p62 body drives autophagosome formation.
Autophagosomes are double-membrane vesicles generated intracellularly to encapsulate substrates for lysosomal degradation during autophagy. Phase separated p62 body plays pivotal roles during autophagosome formation, however, the underlying mechanisms are still not fully understood. Here we describe a spatial membrane gathering mode by which p62 body functions in autophagosome formation. Mass spectrometry-based proteomics reveals significant enrichment of vesicle trafficking components within p62 body. Combining cellular experiments and biochemical reconstitution assays, we confirm the gathering of ATG9 and ATG16L1-positive vesicles around p62 body, especially in Atg2ab DKO cells with blocked lipid transfer and vesicle fusion. Interestingly, p62 body also regulates ATG9 and ATG16L vesicle trafficking flux intracellularly. We further determine the lipid contents associated with p62 body via lipidomic profiling. Moreover, with in vitro kinase assay, we uncover the functions of p62 body as a platform to assemble ULK1 complex and invigorate PI3KC3-C1 kinase cascade for PI3P generation. Collectively, our study raises a membrane-based working model for multifaceted p62 body in controlling autophagosome biogenesis, and highlights the interplay between membraneless condensates and membrane vesicles in regulating cellular functions.


A promising technique for easier single incision laparoscopic cholecystectomy: needle grasper traction of gallbladder.
Laparoscopic cholecystectomy (LC) is the primary treatment method for benign gallbladder diseases. Single incision laparoscopic cholecystectomy (SILC) was reported to be superior in terms of work return, cosmetic results, and post-operative pain, but limited maneuver capacity and overlapping of hand tools are technical difficulties associated with SILC that endanger patient safety. To perform SILC using a needle grasper for gallbladder traction, thus simplifying the dissection of Calot's triangle. The files of patients who underwent elective LC for gallbladder stone and polyps in general surgery clinics between December 2013 and December 2014 were analyzed retrospectively. The patients were divided into two groups: needle-grasper-assisted SILC (nSILC) and conventional laparoscopic cholecystectomy (CLC). Age, gender, height, weight, body mass index, visual analog scale (VAS) scores, ASA score, duration of operation, duration of post-operative hospital stay, complications, drain use, conversion to open and conventional technique, and oral feeding beginning time were analyzed. There were no per-operative or post-operative complications in either of the groups, and no significant differences were found between the groups in terms of complications. The mean duration of the operation was significantly longer in the nSILC group. There was no difference between the groups in terms of hospital stay. The mean visual analogue scale (VAS) scores in conventional nSILC were significantly lower for all hours. The patient satisfaction in terms of cosmetic results was better in the nSILC group. Needle-grasper-assisted SILC reduces the number of tools that need to be held by surgeons; it also provides safe dissection, better cosmetic results, and less post-operative pain in elective cases.


Stimuli-Responsive, Shape-Transforming Nanostructured Particles.
Development of particles that change shape in response to external stimuli has been a long-thought goal for producing bioinspired, smart materials. Herein, the temperature-driven transformation of the shape and morphology of polymer particles composed of polystyrene-b-poly(4-vinylpyridine) (PS-b-P4VP) block copolymers (BCPs) and temperature-responsive poly(N-isopropylacrylamide) (PNIPAM) surfactants is reported. PNIPAM acts as a temperature-responsive surfactant with two important roles. First, PNIPAM stabilizes oil-in-water droplets as a P4VP-selective surfactant, creating a nearly neutral interface between the PS and P4VP domains together with cetyltrimethylammonium bromide, a PS-selective surfactant, to form anisotropic PS-b-P4VP particles (i.e., convex lenses and ellipsoids). More importantly, the temperature-directed positioning of PNIPAM depending on its solubility determines the overall particle shape. Ellipsoidal particles are produced above the critical temperature, whereas convex lens-shaped particles are obtained below the critical temperature. Interestingly, given that the temperature at which particle shape change occurs depends solely on the lower critical solution temperature (LCST) of the polymer surfactants, facile tuning of the transition temperature is realized by employing other PNIPAM derivatives with different LCSTs. Furthermore, reversible transformations between different shapes of PS-b-P4VP particles are successfully demonstrated using a solvent-adsorption annealing with chloroform, suggesting great promise of these particles for sensing, smart coating, and drug delivery applications.


Glycosphingolipids of Echinococcus multilocularis metacestodes.
Neutral and acid glycosphingolipids of Echinococcus multilocularis metacestodes that were obtained after intraperitoneal infection of Meriones unguiculatus have been analyzed by thin layer chromatography. Neutral and acid glycosphingolipids accounted for 95% and 5% of total glycosphingolipids, respectively. 12 different fractions were observed in the neutral glycosphingolipids extracts of the parasite. The most important was a monohexosylceramide fraction accounting for 56.4% of neutral glycosphingolipids. 9 different fractions were detected in gangliosides (acid glycosphingolipids). The fact that these glycosphingolipids were specific to the parasite was established by the analysis of different cell populations of the host. Glycosphingolipids were purified from control and parasite-infected gerbil blood cells as well as from peritoneal exudate cells of healthy gerbils after a non-specific immunostimulation. The chromatograms obtained with these extracts were totally different from the parasite. In addition, parasitosis was found to have no effect on the host blood cell glycosphingolipids.


TANTIGEN: a comprehensive database of tumor T cell antigens.
Tumor T cell antigens are both diagnostically and therapeutically valuable molecules. A large number of new peptides are examined as potential tumor epitopes each year, yet there is no infrastructure for storing and accessing the results of these experiments. We have retroactively cataloged more than 1000 tumor peptides from 368 different proteins, and implemented a web-accessible infrastructure for storing and accessing these experimental results. All peptides in TANTIGEN are labeled as one of the four categories: (1) peptides measured in vitro to bind the HLA, but not reported to elicit either in vivo or in vitro T cell response, (2) peptides found to bind the HLA and to elicit an in vitro T cell response, (3) peptides shown to elicit in vivo tumor rejection, and (4) peptides processed and naturally presented as defined by physical detection. In addition to T cell response, we also annotate peptides that are naturally processed HLA binders, e.g., peptides eluted from HLA in mass spectrometry studies. TANTIGEN provides a rich data resource for tumor-associated epitope and neoepitope discovery studies and is freely available at http://cvc.dfci.harvard.edu/tantigen/ or http://projects.met-hilab.org/tadb (mirror).


Detection of Candida antigen in sera of patients with candidiasis by an enzyme-linked immunosorbent assay-inhibition technique.
A total of 37 serum samples from 27 cancer patients were tested by an enzyme-linked immunosorbent assay-inhibition technique for the detection of Candida antigen. In 20 randomly chosen sera from patients without clinical evidence of candidiasis and in 10 sera from patients proven by autopsy not to have candidiasis, the inhibition ranged up to 17%; in contrast, inhibition ranged from 22 to 56% in all seven patients proven by autopsy to have systemic candidiasis, indicating the presence of Candida antigen in the sera of these patients. This technique appears promising in diagnosing disseminated candidiasis in cancer patients.


Causal associations of serum matrix metalloproteinase-8 level with ischaemic stroke and ischaemic stroke subtypes: a Mendelian randomization study.
Elevated serum matrix metalloproteinase-8 (MMP-8) concentrations are associated with high risk of vascular disease, but the causality remains unclear. A two-sample Mendelian randomization (MR) study was performed to examine the causal effect of serum MMP-8 concentrations on the risk of ischaemic stroke, ischaemic stroke subtypes and coronary artery disease. Ten independent single-nucleotide polymorphisms related to serum MMP-8 concentrations were identified as instrumental variables from a genome-wide association study of 6049 European subjects. Genetic association estimates for ischaemic stroke were obtained from the Multiancestry Genome-wide Association Study of Stroke consortium with 446,696 European individuals. The inverse-variance weighted method was applied to assess the causal associations of serum MMP-8 with ischaemic stroke and its subtypes in the main analysis. No significant causal association was observed for MMP-8 levels with total ischaemic stroke, large artery stroke or cardioembolic stroke. Genetically determined 1 - unit higher log-transformed serum MMP-8 concentration was associated with an increased risk of small vessel stroke (odds ratio 1.25; 95% confidence interval 1.12-1.39; p < 0.001). In secondary analysis, a similar adverse impact was reported for MMP-8 on coronary artery disease (odds ratio 1.05; 95% confidence interval 1.01-1.10; p = 0.017). Sensitivity analyses further confirmed the relationship between serum MMP-8 level and small vessel stroke and coronary artery disease. Mendelian randomization Egger regression showed no evidence of pleiotropic bias. High serum MMP-8 concentrations were causally associated with increased risks of small vessel stroke and coronary artery disease. The mechanism underlying the effect of serum MMP-8 on the vascular system requires further investigation.


Interaction between poly(A)-binding protein PABPC4 and nuclear receptor corepressor NCoR1 modulates a metabolic stress response.
Mitochondria are organelles known primarily for generating ATP via the oxidative phosphorylation process. Environmental signals are sensed by whole organisms or cells and markedly affect this process, leading to alterations in gene transcription and, consequently, changes in mitochondrial function and biogenesis. The expression of mitochondrial genes is finely regulated by nuclear transcription factors, including nuclear receptors and their coregulators. Among the best-known coregulators is the nuclear receptor corepressor 1 (NCoR1). Muscle-specific knockout of NCoR1 in mice induces an oxidative phenotype, improving glucose and fatty acid metabolism. However, the mechanism by which NCoR1 is regulated remains elusive. In this work, we identified the poly(A)-binding protein 4 (PABPC4) as a new NCoR1 interactor. Unexpectedly, we found that silencing of PABPC4 induced an oxidative phenotype in both C2C12 and MEF cells, as indicated by increased oxygen consumption, mitochondria content, and reduced lactate production. Mechanistically, we demonstrated that PABPC4 silencing increased the ubiquitination and consequent degradation of NCoR1, leading to the derepression of PPAR-regulated genes. As a consequence, cells with PABPC4 silencing had a greater capacity to metabolize lipids, reduced intracellular lipid droplets, and reduced cell death. Interestingly, in conditions known to induce mitochondrial function and biogenesis, both mRNA expression and PABPC4 protein content were markedly reduced. Our study, therefore, suggests that the lowering of PABPC4 expression may represent an adaptive event required to induce mitochondrial activity in response to metabolic stress in skeletal muscle cells. As such, the NCoR1-PABPC4 interface might be a new road to the treatment of metabolic diseases.


First case of periprosthetic joint infection due to Clostridioides difficile in China.
Clostridioides difficile usually causes intestinal infections. However, a 75-year-old lady had a periprosthetic joint infection due to this microorganism. We report a C. difficile infection of a prosthetic hip joint. Such an infection is rarely reported around the world. The elder female patient presented with a 2-year history of right hip pain with movement restriction. Her right leg was shorter than another. The skin around the right hip joint was red and swollen without sinus. Her lab test result showed elevator ESR and CRP. Her X-ray film showed a massive bone defect. The patient had a total hip arthroplasty 16 years ago and had a revision 5 years ago. During this hospitalization, her cultures of the synovial fluid and tissue repeatedly grew C. difficile. She improved following two-stage revision surgery and antibiotic treatment. The patient has no recurrence of infection after a one-year follow-up. A rapid and accurate sample collection is significant for culture results, making an outstanding contribution to the successful treatment.


Ethyl pyruvate decreases proinflammatory gene expression in lipopolysaccharide-stimulated equine monocytes.
Monocytes are among the initial cells that interact with circulating LPS. Binding of LPS to monocyte surface receptors triggers an intracellular signaling cascade and results in the production of proinflammatory cytokines. Ethyl pyruvate, a stable derivative of pyruvate, has been effective in mitigating LPS induced alterations in isolated human monocytes. We hypothesized that ethyl pyruvate would suppress proinflammatory gene expression in LPS-stimulated equine monocytes without affecting cell viability. Equine monocytes were isolated from whole blood using a sediment-gradient centrifugation protocol and enriched to 76% purity by adhesion to tissue culture dishes. Isolated monocytes were incubated with 0, 1, 5, 10 and 50 mM ethyl pyruvate. Cell viability, production of caspase 3/7, and caspase-3 gene expression were determined. In a separate experiment, monocytes were stimulated with LPS (0.1 ng/ml for 1h) followed by incubation with 0, 1, 5, or 10 mM ethyl pyruvate for 1 h. Proinflammatory gene expression was determined by real-time PCR. Ethyl pyruvate at 50 mM adversely affected monocyte viability. Ethyl pyruvate at 10mM or less had no significant effect on monocyte viability, and did not increase activity of caspase 3/7 nor caspase-3 gene expression. Incubation with LPS alone induced a significant upregulation in proinflammatory gene expression. Subsequent treatment of monocytes with ethyl pyruvate significantly reduced IL-8 expression in LPS stimulated monocytes at 5 mM, and IL-8, TNF-α and COX-2 at 10 mM. No beneficial effect on expression of IL-1β or IL-6 was detected. Overall, 10 mM ethyl pyruvate did not adversely affect monocyte viability and suppressed LPS-induced proinflammatory gene expression. Ethyl pyruvate may be a beneficial anti-inflammatory therapy in equine endotoxemia.


Cell H2O2 steady-state concentration and mitochondrial nitric oxide.
For many years, mitochondrial respiration was thought to follow an "all or nothing" paradigm supporting the notion that in the normal O2 concentration range, respiration is mainly controlled by tissue demands. However, nitric oxide produced by cytosol or mitochondrial nitric oxide synthases adapts respiration to different physiologic conditions and increases the mitochondrial production of O2 active species that contributes to NO clearance. Because mitochondrial NO utilization is sensitive to environmental or hormonal modulation, and because diffusible active species, like H2O2, are able to regulate genes related to proliferation, quiescence, and death, we surmised that the two mechanisms converge to elicit the different responses in cell physiology.


On the human health benefits of microalgal phytohormones: An explorative in silico analysis.
Phytohormones represent a group of secondary metabolites with different chemical structures, in which belong auxins, cytokinins, gibberellins, or brassinosteroids. In higher plants, they cover active roles in growth or defense function, while their potential benefits for human health protection were noted for some phytohormones and little explored for many others. In this study, we developed a target fishing strategy on fifty-three selected naturally occurring phytohormones covering different families towards proteins involved in key cellular functions related to human metabolism and health protection/disease. This in silico analysis strategy aims to screen the potential human health-driven bioactivity of more than fifty phytohormones through the analysis of their interactions with specific targets. From this analysis, twenty-eight human targets were recovered. Some targets e.g., the proteins mitochondrial glutamate dehydrogenase (GLUD1) or nerve growth factor (NGF) bound many phytohormones, highlighting their involvement in amino acid metabolism and/or in the maintenance or survival of neurons. Conversely, some phytohormones specifically interacted with some proteins, e.g., SPRY domain-containing SOCS box protein 2 (SPSB2) or Inosine-5'-monophosphate dehydrogenase 1 (IMPDH1), both involved in human immune response. They were then investigated with a molecular docking analysis approach. Our bioprospecting study indicated that many phytohormones may endow human health benefits, with potential functional role in multiple cellular processes including immune response and cell cycle progression.


Associations between socioeconomic status and white matter microstructure in children: indirect effects via obesity and cognition.
Both neighborhood and household socioeconomic disadvantage relate to negative health outcomes and altered brain structure in children. It is unclear whether such findings extend to white matter development, and via what mechanisms socioeconomic status (SES) influences the brain. To test independent associations between neighborhood and household SES indicators and white matter microstructure in children, and examine whether body mass index and cognitive function (a proxy of environmental cognitive/sensory stimulation) may plausibly mediate these associations. This cross-sectional study used baseline data from the Adolescent Brain Cognitive Development (ABCD) Study, an ongoing 10-year cohort study tracking child health. School-based recruitment at 21 U.S. sites. Children aged 9 to 11 years and their parents/caregivers completed baseline assessments between October 1 st , 2016 and October 31 st , 2018. Data analysis was conducted from July to December 2022. Neighborhood disadvantage was derived from area deprivation indices at primary residence. Household SES indicators were total income and the highest parental education attainment. Thirty-one major white matter tracts were segmented from diffusion-weighted images. The Restriction Spectrum Imaging (RSI) model was implemented to measure restricted normalized directional (RND; reflecting oriented myelin organization) and isotropic (RNI; reflecting glial/neuronal cell bodies) diffusion in each tract. Obesity-related measures were body mass index (BMI), BMI z -scores, and waist circumference, and cognitive performance was assessed using the NIH Toolbox Cognition Battery. Linear mixed-effects models tested the associations between SES indicators and scanner-harmonized RSI metrics. Structural equation models examined indirect effects of obesity and cognitive performance in the significant associations between SES and white mater microstructure summary principal components. Analyses were adjusted for age, sex, pubertal development stage, intracranial volume, and head motion. The analytical sample included 8842 children (4299 [48.6%] girls; mean age [SD], 9.9 [0.7] years). Greater neighborhood disadvantage and lower parental education were independently associated with lower RSI-RND in forceps major and corticospinal/pyramidal tracts, and had overlapping associations in the superior longitudinal fasciculus. Lower cognition scores and greater obesity-related measures partially accounted for these SES associations with RSI-RND. Lower household income was related to higher RSI-RNI in almost every tract, and greater neighborhood disadvantage had similar effects in primarily frontolimbic tracts. Lower parental education was uniquely linked to higher RSI-RNI in forceps major. Greater obesity-related measures partially accounted for these SES associations with RSI-RNI. Findings were robust in sensitivity analyses and mostly corroborated using traditional diffusion tensor imaging (DTI). These cross-sectional results demonstrate that both neighborhood and household contexts are relevant to white matter development in children, and suggest cognitive performance and obesity as possible pathways of influence. Interventions targeting obesity reduction and improving cognition from multiple socioeconomic angles may ameliorate brain health in low-SES children. Question: Are neighborhood and household socioeconomic levels associated with children's brain white matter microstructure, and if so, do obesity and cognitive performance (reflecting environmental stimulation) mediate the associations?Findings: In a cohort of 8842 children, higher neighborhood disadvantage, lower household income, and lower parental education had independent and overlapping associations with lower restricted directional diffusion and greater restricted isotropic diffusion in white matter. Greater body mass index and poorer cognitive performance partially mediated these associations.Meaning: Both neighborhood and household poverty may contribute to altered white matter development in children. These effects may be partially explained by obesity incidence and poorer cognitive performance.


[Characteristics of the relationship between the circadian rhythm of the concentrations of corticosteroids and their precursors in the blood of baboon hamadryas and the initial functional state of the pituitary-adrenal cortex system].
The occurrence of a single-phase circadian rhythm in the content of cortisole, 11-deoxycortisole, 17-hydroxypregnenolone, 17-hydroxyprogesterone, pregnenolone and progesterone in the blood plasma of hamadryas baboons was established by radioimmunoassay with peak levels early in the morning and minimum ones in the evening. The time course of changes in aldosterone levels in the intact animals significantly differs from that in the adapted ones. The maintenance of animals under other ecological conditions results in the decreased levels of all steroids studied and in significant changes in circadian rhythms of the content of cortisol precursors.


[Influence of the natural history of disease on a previous diagnosis in patients with gout].
The clinical diagnosis of gout can be quite precise in clinically typical forms. However, in chronic or atypical forms, such precision tends to be diminished in clinical practice. A cohort of 248 patients with a diagnosis of urate crystal arthropathy was studied, sent with a definite clinical evaluation, and data such as severity of the disease, joint distribution and the presence of tophi were gathered. Precision data was analyzed with respect to the referral diagnosis according to the severity parameters and the type of physician sending the patient. The best diagnostic precision was seen in the monoarticular forms that were sent both by the emergency room as well as by family physicians, but not in those sent by other specialists. The presence of oligoarticular forms reduced significantly the diagnostic precision in all of the specialties referring patients. The presence of tophi did not improve diagnostic precision. Chronic and severe forms of gout are frequently wrongly evaluated from the clinical standpoint.


Immunoglobulin heavy G2 chain (IGHG2) gene restriction in the development of severe respiratory syncytial virus infection.
Respiratory syncytial virus (RSV) is a prominent cause of airway morbidity in children under 1 y of age. It is assumed that host factors influence the severity of disease presentation, and thus the need for hospitalization. The variation of IGHG genes from chromosome 14q32 are linked to serum IgG subclass levels but also to the variations in IgG responses to pneumococcal, meningococcal and Haemophilus influenzae antigens. The aim of this investigation was to clarify whether IGHG genes are involved in the development of severe RSV lower respiratory tract infection (LRTI). The alternative expressions of IGHG3(b) and (g), IGHG1(f) and (a), and IGHG2(n) and (-n) genes were studied in a cohort of 49 previously healthy children hospitalized for RSV LRTI. The gene frequencies were compared to a population of healthy individuals. The homozygous IGHG2(-n/-n) genotypes dominated in hospitalized children with severe RSV infection: 55.1%, compared with 34.2% in the healthy population (OR 2.3; p = 0.004). The IGHG2 genotypes containing (n/n) and (n/-n) were significantly decreased. The IGHG(bf-n) alleles were significantly increased (OR 1.7; p = 0.025) and the IGHG(bfn) alleles significantly decreased (OR 0.5; p = 0.005). The IGHG(bf-n) allele and homozygous IGHG2(-n/-n) genotypes are associated with the development of severe RSV LRTI.


[User Satisfaction in Social Media: How do Patients Rate their Hospital Stay Online?]
Patients are increasingly using social media and review sites to get information on hospitals, to rate them or to write a review about their own experiences during hospital stay. Hospitals have the opportunity to use these patient reviews for internal quality assurance as well as online reputation marketing. The objective is to analyze the satisfaction ratings of previous hospitals stays by social media users on platforms such as "Google", as well as reviews from the German site "Klinikbewertungen.de", and to compare them. User reviews (n=16,691) for all hospitals (n=180) in the federal state of Lower Saxony (complete survey) for 2013-2018 were extracted from the platforms Google and the German Klinikbewertungen.de. Subsequently, a descriptive and inductive analysis (Chi²-test) of the user reviews and a comparison specific for the platforms were completed. Most users are satisfied with their hospital stay, whereby users of the platform Google ratd their hospital stay worse (n=6,181; 57.6% satisfied) than users of the platform Klinikbewertungen.de (n=10,509; 65.4% satisfied). Features of the hospital (number of beds, specialist departments) and features of the users (author, type of insurance) were significantly associated with the rating for the hospital stay (star review). (Google: number of beds p<0.001; Klinikbewertungen.de: number of beds p<0.001, author p<0.001, specialist departments p<0.001, type of insurance p<0.001). The online review of a hospital stay is associated with the number of beds, the specialist departments of hospitals, whether the author is the patient or a relative, and the type of health insurance they have, namely private or statutory. Online reviews and ratings for hospitals may be used for internal quality assurance or improvements of company online reputation.


Pupal cases of three nearctic species of Machimus (Diptera: Asilidae).
The pupal cases of the Nearctic robber flies, Machimus latapex Martin, 1975, M. occidentalis (Hine, 1909), and M. prairiensis (Tucker, 1907) are described. They are compared with the previously described pupal cases of six other Nearctic species of Machimus. The pupal case of M. latapex and facial sheaths and posterior antennal processes of M. occidentalis are illustrated.


Distribution and clinical significance of Mycobacterium avium complex species isolated from respiratory specimens.
Mycobacterium avium complex (MAC) was originally composed of 2 species, M. avium and M. intracellulare. However, several new species closely related to M. intracellulare have recently been identified. In addition, M. avium has been further subdivided into 4 subspecies. The aim of this study was to determine the proportion of different MAC species recovered from respiratory specimens and to elucidate the clinical relevance of these species. Clinical isolates, from 219 patients, that had been initially identified as M. avium or M. intracellulare by non-sequencing methods were reidentified using multilocus sequence typing, and the clinical significance of the identified species was then investigated. Of 91 isolates originally identified as M. intracellulare, 75 (82%) were confirmed to be M. intracellulare, 8 (9%) isolates were identified as M. chimaera, and 4 (4%) isolates each were identified as "M. indicus pranii" and M. yongonense. The 128 isolates originally designated as M. avium were determined to be M. avium subsp. hominissuis. Of the 219 patients, 146 (67%) met the diagnostic criteria for MAC lung disease, and for each MAC species, the proportion of patients meeting these criteria was as follows: M. intracellulare (54/75, 72%), M. chimaera (3/8, 38%), "M. indicus pranii" (3/4, 75%), M. yongonense (2/4, 50%), and M. avium subsp. hominissuis (84/128, 66%). In summary, multilocus sequence typing of respiratory isolates initially identified as MAC revealed that, although most isolates were M. avium subsp. hominissuis or M. intracellulare, approximately 7% were newer MAC members, with clinical evidence supporting their potential pathogenicity for humans.


Three cases of the myelodysplastic syndrome with pericentric inversion of chromosome 16.
Inversion of chromosome 16, inv(16)(p13q22), is characteristic of acute myeloid leukaemia (AML) with eosinophilia and is rarely found in the myelodysplastic syndrome (MDS). We report three cases of MDS in which inv(16) was observed. They were classified to FAB subtypes RA, RARS and RAEBT; eosinophilia or abnormal eosinophils were not observed. The disease appeared to be stable in all three patients. MDS with inv(16) without eosinophilia may be a rare subgroup associated with a good prognosis.


Giant cyst formation in a fibrous capsule following breast augmentation. A case report and discussion on the pathology.
A report is presented of a patient who developed a giant cyst in the fibrous pseudosheath after removal of a silicone breast prosthesis. Emphasis is put on the histological features of the capsule which differentiate it from other cystic breast conditions and also preclude its spontaneous resolution. These features are capsular thickness, epidermal lining, calcification and chronic lymphocytic reaction.


The 2019 practice analysis of hand therapy and the use of orthoses by certified hand therapists.
In 2019, the Hand Therapy Certification Commission (HTCC), in consultation with Scantron Corporation, performed a practice analysis study of hand therapy, the sixth in a series of similar studies performed byHTCC over a 35-year period. The primary goal of this study was to update and validate the definition and delineation of hand therapy and to ensure that the test content outline for the Hand Therapy Certification Examination (HTCE) reflects the critical tasks, knowledge, and skills required in the practice of hand therapy. Additionally, HTCC explored specific trends in hand therapy practice, compared findings with previous studies, and gathered data about the frequency, criticality, and performance expectations for the use and fabrication of orthoses by hand therapists. Quantitative Descriptive. More than 40 subject matter experts from the United States and Canada, representing a broad range of experiences and perspectives, developed an updated delineation of the domains, tasks, knowledge, and techniques and tools used in hand therapy practice. A large-scale online survey of all certified hand therapists from the United States, Canada, and other countries was completed to test the profile within the practice of hand therapy. This large-scale online survey overwhelmingly validated the profile of hand therapy. The results affirmed the test specifications for the Hand Therapy Certification Examination; affirmed the definition of hand therapy; and refined the scope of hand therapy practice. New data was gathered regarding the use of orthotics in hand therapy. This study establishes content validity for the HTCE. It highlights that the specialty of hand therapy is a mature and stable specialty field of occupational therapy and physical therapy. Certified Hand Therapists frequently issue pre-fabricated and fabricate custom orthoses in the course of rehabilitation for clients with hand and arm injuries, and overall consider this a highly critical task in hand therapy practice. N/A.


The prevalence of multiple drug resistant urinary tract infections: A single-centered, observational retrospective study in King Abdulaziz Specialized Hospital, Taif, Saudi Arabia.
To determine the prevalence and patterns of antibiotic resistance, risk factors, and appropriate empiric therapy for multidrug-resistant Enterococcus (MDRE) urinary tract infections (UTIs) at King Abdulaziz Specialist Hospital (KAASH), Taif, Saudi Arabia. All patients attending KAASH with MDRE UTIs between January 2018 and December 2020 were enrolled in the study. After ethical approval, data were analyzed. The most common causative organisms were Gram-negative and anaerobes, and the most sensitive antibiotics were ciprofloxacin and ceftriaxone. Based on our analyses, regular culture and sensitivity should be made routine to gather information regarding susceptibility patterns, thereby reducing drug resistance in our setups.


[Incidental findings of liver, biliary system, pancreas and spleen in asymptomatic patients : Assessment and management recommendations].
The increased use of highly developed imaging procedures, such as multidetector-row computed tomography and magnetic resonance imaging has led to a substantial increase of asymptomatic and unexpected findings. Abdominal CT investigations are particularly affected with a large number of incidental findings. This valuable diagnostic procedure also entails the risk of complex and cost-intensive subsequent investigations with partly invasive procedures. For this reason radiologists are more often confronted with the difficult task of correctly assessing these lesions, to decide on the need for additional investigations and to inform the patient in detail about the clinical relevance. The aims of this article are to describe the most common abdominal incidentalomas, to assist with the interpretation and differential diagnosis and to give recommendations for further management.


Identification and characterization of enterococci from bryndza cheese.
To identify enterococci isolated from sheep milk cheese--bryndza, and to compare differences in the composition of enterococcal microflora affected by the season, and to evaluate the potential presence of vancomycin resistance and virulence determinants. Bacterial strains were isolated during analysis of bryndza cheese and identified on the genus and species level by phenotypic methods and with commercial biochemical sets. The identification of the species, Enterococcus faecium, Ent. durans and Ent. faecalis, was confirmed by PCR using species-specific primers for ddl genes. PCR was also used for assessment of presence of vanA and vanB genes and virulence determinants gelE, agg and cytolysin genes namely: cylL(L), cylL(S), cylM, cylB and cylA. Among 308 Enterococcus sp. strains, 177 isolates were proved to be Ent. faecium, 59 to be Ent. durans and 41 to be Ent. faecalis. Vancomycin resistance genes vanA and vanB were not detected. Agar plate testing confirmed their absence. Gene gelE, however, was found in 20 Ent. faecalis isolates, but only 13 of them showed gelatinase-positive phenotype. Seven isolates had five cytolysin genes, but none of the isolates exhibited a positive haemolytic phenotype. Four isolates possessed the agg gene. The prevalence of Ent. faecium species was highest in samples from the winter season harvest. Ent. faecium is the dominant enterococcal species in bryndza cheese and the most prevalent in the winter season product. None of the Enterococcus sp. strains was proved to have vanA or vanB genes and the vancomycin resistance. To our knowledge, this is the first report of enterococcal microflora in bryndza cheese and its evaluation for the presence of vanA and vanB genes as well as virulence determinants.


Regeneration of the Egyptian medicinal plant Artemisia judaica L.
An in vitro propagation system for Artemisia judaica L., a traditional Egyptian medicinal plant, has been developed. De novo shoot organogenesis was induced by culturing etiolated hypocotyls and intact seedlings on medium supplemented with thidiazuron [N-phenyl-N'-(1,2,3-thidiazol-yl) urea] via callusing at the cotyledonary notch region. Up to 16 shoots formed per seedling cultured on a medium containing 1 micro mol l(-1) thidiazuron for an optimal duration of exposure of 20 days. Regenerated shoots formed roots when subcultured onto a medium containing 1 micromol l(-1) indole-3-butyric acid. The regeneration protocol developed in this study provides a basis for germplasm conservation and for further investigation of medicinally active constituents of A. judaica.


Body mass index and absolute lymphocyte count predict disease-free survival in Korean breast cancer patients.
Our study evaluated the association between body mass index (BMI) and absolute lymphocyte count (ALC) in breast cancer patients and healthy females. Additionally, we determined the prognostic value of these factors in breast cancer. We retrospectively identified 1225 primary invasive breast cancer patients and 35,991 healthy females. Factors including BMI and complete blood count associated with disease-free survival (DFS) were assessed using a multi-variable Cox proportional hazard model. BMI and ALC were positively correlated in breast cancer patients and healthy females (both P < 0.001). In multi-variable analysis, overweight or obese participants had worse DFS (hazards ratio [HR], 1.98; 95% confidence interval [CI], 1.34-2.92; P = 0.001) than underweight or normal-weight individuals, but patients with high ALC had better DFS than those with low ALC (HR, 0.43; 95% CI, 0.29-0.65; P < 0.001). After risk stratification according to BMI/ALC, high-risk patients with high BMI/low ALC had worse DFS than others (HR, 2.48; 95% CI, 1.70-3.62; P < 0.001). BMI and ALC were positive correlated, but their effect on breast cancer prognosis was opposite. Patients with high BMI/low ALC had worse DFS than others. Underlying mechanisms for effect of BMI/ALC on breast cancer prognosis should be studied in the future.


Structural investigation of a self-assembled monolayer material 5-[(3-methylphenyl) (phenyl) amino] isophthalic acid for organic light-emitting devices.
The molecular structure and vibrations of 5-[(3-methylphenyl) (phenyl) amino] isophthalic acid (MePIFA) were investigated by infrared and Raman spectroscopies, UV-Vis, (1)H and (13)C NMR spectroscopic techniques and NBO analysis. FT-IR, FT-Raman and dispersive Raman spectra were recorded in the solid phase. (1)H and (13)C NMR spectra and UV-Vis spectrum were recorded in DMSO solution. HOMO-LUMO analysis and molecular electrostatic potential (MEP) analysis were performed. The theoretical calculations for the molecular structure and spectroscopies were performed with DFT (B3LYP) and 6-311G(d,p) basis set calculations using the Gaussian 09 program. After the geometry of the molecule was optimized, vibration wavenumbers and fundamental vibration wavenumbers were assigned on the basis of the potential energy distribution (PED) of the vibrational modes calculated with VEDA 4 program. The total (TDOS), partial (PDOS) density of state and overlap population density of state (OPDOS) diagrams analysis were made using GaussSum 2.2 program. The results of theoretical calculations for the spectra of the title compound were compared with the observed spectra.


Toward cognitivist ontologies : on the role of selective attention for upper ontologies.
Ontologies play a key role in modern information society although there are still many fundamental questions regarding their structure to be answered. In this paper, some of these are presented, and it is argued that they require a shift from realist to cognitivist ontologies, with ontology design crucially depending on taking both cognitive and linguistic aspects into consideration. A detailed discussion of central parts of a proposed cognitivist upper ontology based on qualitative representations of selective attention is presented.


Projections from the hypothalamus and its adjacent areas to the posterior pituitary in the rat.
Cholera toxin conjugated horseradish peroxidase was injected into the posterior pituitary and its afferents traced in 21 albino rats. The neuronal processes as well as the perikarya were elaborately displayed. The principal and retrochiasmatic supraoptic nuclei and the magnocellular paraventricular subnuclei were densely labelled. The accessory cell groups or nuclei labelled included: the medial preoptic and anterior hypothalamic areas, the anterior and posterior fornical nuclei, the lateral hypothalamic area, the nucleus circularis and nucleus of the forebrain bundle and hitherto unknown or not fully appreciated retrochiasmatic area, the dorsal accessory groups in an area between the stria medullaris and fornix, on the one hand, and the stria terminalis and internal capsule, on the other, and a well developed subependymalperiventricular zone. The medial preoptic nucleus, subfornical organ and organ vasculosum laminae terminalis were also weakly stained. Dendrites of the magnocellular paraventricular nucleus have been said by some to be largely confined to the subnuclei in which they lie. Immunohistochemical studies have proved that they extended beyond their nuclear confinement. The present study has found much wider extension of their dendritic fields. In fact, dendrites of the magnocellular neurosecretory cells in general were long and had a certain degree of directional bias. Several sites projecting to the posterior pituitary were closely related to the cerebrospinal fluid. Namely, the subependymal neuronal plexuses along the third ventricle and beneath the interventricular foramen, and the subpial dendritic plexuses of the supraoptic and retrochiasmatic supraoptic nuclei. Neurons were seen to squeeze in-between the ependymal cells, bringing themselves very close to the cerebrospinal fluid. No direct cerebrospinal fluid-contacting elements, either cell bodies or processes, however, could be ascertained. It is proposed that these plexuses may monitor changes in the cerebrospinal fluid. Besides the principal neurohypophysial tract the posterior pituitary was found in the present study to receive its afferents via two accessory fasciculi, one coursing in the medial forebrain bundle and the other running along the lateral wall of the infundibular recess subependymally.


The amino acid composition of proteins from anaerobic halophilic bacteria of the order Halanaerobiales.
We performed a comparative analysis of the genome sequences of three anaerobic halophilic fermentative bacteria belonging to the order Halanaerobiales: Halanaerobium praevalens, the alkaliphilic "Halanaerobium hydrogeniformans", and the thermophilic Halothermothrix orenii to assess the amino acid composition of their proteins. Members of the Halanaerobiales were earlier shown to accumulate KCl rather than organic compatible solutes for osmotic balance, and therefore the presence of a dominantly acidic proteome was predicted. Past reports indeed showed a large excess of acidic over basic amino acids in whole-cell hydrolysates of selected members of the order. However, the genomic analysis did not show unusually high contents of acidic amino acids or low contents of basic amino acids. The apparent excess of acidic amino acids in these anaerobic halophiles reported earlier is due to the high content in their proteins of glutamine and asparagine, which yield glutamate and aspartate upon acid hydrolysis. It is thus suggested that the proteins of the Halanaerobiales, which are active in the presence of high intracellular KCl concentrations, do not possess the typical acidic signature of the 'halophilic' proteins of the Archaea of the order Halobacteriales or of the extremely halophilic bacterium Salinibacter.


Apocynin stimulates osteoblast differentiation and inhibits bone-resorbing mediators in MC3T3-E1 cells.
Apocynin is a naturally occurring methoxy-substituted catechol, experimentally used as an inhibitor of NADPH-oxidase. In the present study, the effect of apocynin on the function of osteoblastic MC3T3-E1 cells was studied. Apocynin caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and mineralization in the cells (P<0.05). Antimycin A (AMA), which inhibits complex III of the electron transport system, has been used as a reactive oxygen species (ROS) generator in biological systems. We exposed cultured osteoblastic MC3T3-E1 cells to AMA with or without pretreatment with apocynin. Apocynin significantly (P<0.05) increased cell survival, calcium deposition, and osteoprotegerin release and decreased the production of ROS and osteoclast differentiation inducing factors such as TNF-α, IL-6, and receptor activator of nuclear factor-kB ligand (RANKL) in the presence of AMA. These results demonstrate that apocynin can protect osteoblasts from mitochondrial dysfunction-induced toxicity and may have positive effects on skeletal structure.


Evaluation of the impurity profile of alcuronium by means of capillary electrophoresis.
Alcuronium, a neuromuscular blocking drug, was recently introduced to the European Pharmacopoeia. A HPLC method is described to limit the impurities of alcuronium, namely the diallylcaracurine (DAC) and the allyl-Wieland-Gumlich-aldehyde (WCA), to less than 0.5%. Since alcuronium and all impurities are quaternary salts, capillary electrophoresis (CE) is highly suitable to evaluate the impurity profile. Using 12 mM heptakis-(2,6-di-O-methyl)-beta-cyclodextrin in a 50 mM phosphate buffer at pH 5.5 or 50 mM diethanolamine buffer (pH 9.2)-acetonitrile 19:1 containing heptakis-(2,3-O-diacetyl-6-sulfo)-beta-cyclodextrin the impurities could be baseline separated and quantified. The limit of detection for DAC and WCA was found to be in the same range as found with HPLC; thus, less than 0.1% of both DAC and WCA could be detected in the solution for injection in presence of alcuronium. In injection solutions of alcuronium which were stored at higher temperatures three additional, unidentified impurities were detected. In addition, the conversion of alcuronium to DAC, occurring under acidic condition, was monitored by means of the CE method developed.


Brevetoxin-B of Gymnodinium breve toxin-induced contractions of smooth muscles due to the transmitter release from nerves.
Brevetoxin-B (BTX-B), a Gymnodinium breve toxin, isolated from the dinoflagellate, produced contraction of guinea pig ileum and rabbit aorta. Atropine (10(-6) M) and phentolamine (10(-6) M) abolished the BTX-B (10(-7) or 10(-6)-induced contraction of the ileum or aorta, respectively. Tetrodotoxin and saxitoxin (both 5 x 10(-7) M) also abolished the responses to BTX-B in the both tissues. Results suggest that BTX-B produces smooth muscle contractions through the tetrodotoxin- and saxitoxin-sensitive release of transmitters from autonomic nerves.


ATR and ATM-dependent movement of BLM helicase during replication stress ensures optimal ATM activation and 53BP1 focus formation.
The BLM helicase, a deficiency that markedly increases cancer incidence in humans, is required for optimal repair during DNA replication. We show that BLM rapidly moves from PML nuclear bodies to damaged replication forks, returning to PML bodies several hours later, owing to activities of the DNA damage response kinases ATR and ATM, respectively. Immunofluorescence and cellular fractionation demonstrate that BLM partitions to different sub-cellular compartments after replication stress. Unexpectedly, fibroblasts lacking BLM were deficient in phospho-ATM (S-1981) and 53-binding protein-1 (53BP1), and these proteins failed to form foci following replication stress. Expression of a dominant p53 mutant or helicase-deficient BLM restored replication stress-induced 53BP1 foci, but only mutant p53 restored optimal ATM activation. Thus, optimal repair of damaged replication fork lesions likely requires both ATR and ATM. BLM recruits 53BP1 to these lesions independent of its helicase activity, and optimal activation of ATM requires both p53 and BLM helicase activities.


[3+3]-Annulation of Cyclic Nitronates with Vinyl Diazoacetates: Diastereoselective Synthesis of Partially Saturated [1,2]Oxazino[2,3-b][1,2]oxazines and Their Base-Promoted Ring Contraction to Pyrrolo[1,2-b][1,2]oxazine Derivatives.
A rhodium(II)-catalyzed reaction of cyclic nitronates (5,6-dihydro-4H-1,2-oxazine N-oxides) with vinyl diazoacetates proceeds as a [3+3]-annulation producing bicyclic unsaturated nitroso acetals (4a,5,6,7-tetrahydro-2H-[1,2]oxazino[2,3-b][1,2]oxazines). Optimization of reaction conditions revealed the use of Rh(II) octanoate as the preferred catalyst in THF at room temperature, which allows the preparation of target products in good yields and excellent diastereoselectivity. Under basic conditions, namely, the combined action of DBU and alcohol, these nitroso acetals undergo ring contraction of an unsaturated oxazine ring into the corresponding pyrrole. Both transformations can be performed in a one-pot fashion, thus constituting a quick approach to oxazine-annulated pyrroles from available starting materials, such as nitroalkenes, olefins, and diazo compounds.


Use of oral contraceptives and serum beta-carotene.
Antioxidants, in particular carotenoids, may influence the risk for cardiovascular disease. This study investigates the influence of oral contraceptives (OC) on the serum concentration of beta-carotene, which may in turn affect the risk of cardiovascular diseases due to its antioxidative impact. Cross-sectional epidemiologic study. Examinations included a detailed questionnaire on medical history and lifestyle factors, a 7 day food record, and blood samples. National health and nutrition survey among healthy people living in private homes in West Germany in 1987-1988. Nonpregnant and nonlactating women aged 18-44 (n = 610). Overall, the use of OC was negatively associated with serum beta-carotene concentration in bi- and multivariable analyses after adjustment for age, smoking, alcohol consumption, dietary intake of beta-carotene, use of vitamin supplements, body mass index, pregnancies, and serum concentrations of total triglyceride and cholesterol. A strong interaction between OC use and age on beta-carotene concentration was observed. While no relationship between OC use and serum beta-carotene was seen in the youngest age-group (18-24 y), there was a modest but significant negative association between OC use and beta-carotene levels among 25-34 y old women. The use of OC was associated with a strong decrease in beta-carotene levels among 35-44 y old women. The interaction between OC use and age could partly be explained by age dependent use of OC with higher estrogen content. OC use seems to be strongly related to serum beta-carotene levels, particularly among women above the age of 35. Further studies are needed to clarify the underlying mechanisms of this association and its implications for health risks of OC use.


Visualization of the subsarcolemmal cytoskeleton network of mouse skeletal muscle cells by en face views and application to immunoelectron localization of dystrophin.
The ultrastructural organization of the highly interconnected filamentous network underneath the sarcolemma as well as the role played by the muscle protein dystrophin within this cytoskeleton remain yet unclear. More accurate information has been obtained by using a method which provides three-dimensional en face views of large membrane areas applied to mouse cultured myotubes and isolated adult skeletal muscle fibres. Two levels have been distinguished in the cytoskeleton underlying the sarcolemma: the submembranous level, partly integrated into the membrane, and the cortical level, invading the proximal cytoplasmic space. Few differences have been found between the membrane cytoskeletons of myotubes issued from 14-day-old cultures and those of adult fibres. The comparison was done with cells where dystrophin is missing (mdx mouse muscle): surprisingly, the lack of dystrophin does not induce obvious or dramatic ultrastructural disorganization, either in the cortical cytoskeletal network or in the submembranous one. Immunogold labelling of either the central-rod or the C-terminal domain of dystrophin is not located among the cortical network. This study provides additional data on the spatial ordering of subsarcolemmal cytoskeletal elements: dystrophin does not appear as a filamentous structure entirely located among subsarcolemmal cytoskeleton but seems partly embedded in membranous material.


Prognostic value of total tumour volume, adding necrosis to metabolic tumour volume, in advanced or metastatic non-small cell lung cancer treated with first-line pembrolizumab.
Metabolic tumour volume (MTV) measured on fluorodeoxyglucose F18 (FDG) positron emission tomography coupled with computed tomography (PET/CT) is a prognostic factor of advanced non-small cell lung cancer (NSCLC) treated by first-line immunotherapy. However, these tumours are often necrotic and the necrosis, which is hypometabolic in PET FDG, is not included in the MTV. The aim of this study was to evaluate the prognostic value of total tumour volume (TTV), adding necrotic tumour volume (NTV) to metabolic tumour volume (MTV). We retrospectively included 65 patients with NSCLC treated with pembrolizumab as monotherapy. All patients had a pretreatment FDG PET/CT. PET/CT measured parameters were MTV, NTV and TTV. Clinical, biological and tumour parameters were also retrieved. Receiver operator characteristics (ROC) analysis was performed and overall survival at 1 year was studied using Kaplan-Meier and uni/multivariate Cox analysis. In the ROC analysis, MTV, NTV, TTV, age at diagnosis, polynuclear blood neutrophil, derived neutrophil/leukocyte ratio (dNLR), and haemoglobin had an area under the curve (AUC) significantly higher than 0.5. In Kaplan-Meier analysis, prognosis was worse for patients with high MTV (p = 0.02), high TTV (p = 0.003), high NTV (p = 0.014), low haemoglobin (p < 0.001), older people (p = 0.002), neutrophil polynucleosis (p < 0.001) and dNLR (p = 0.022). All these parameters, except age and neutrophil polynucleosis, were significant prognostic factors in univariate Cox analysis (p < 0.05). In a stepwise multivariate Cox analysis focused on PET parameters, the only significant parameter was TTV (HR = 3.66, p = 0.002) and in a stepwise multivariate Cox analysis exploring all the parameters, a model combining TTV, performance status and brain metastasis was found (p = 0.002). TTV and NTV measured on pretreatment FDG PET/CT are significant prognosis factor for stage III-IV NSCLC treated by pembrolizumab and TTV could have a higher prognostic value than MTV.


A pressure-affected headspace-gas chromatography method for determining calcium carbonate content in paper sample.
The present work reports on the development of a pressure-affected based headspace (HS) analytical technique for the determination of calcium carbonate content in paper samples. By the acidification, the carbonate in the sample was converted to CO2 and released into the headspace of a closed vial and then measured by gas chromatography (GC). When the amount of carbonate in the sample is significant, the pressure created by the CO2 affects the accuracy of the method. However, the pressure also causes a change in the O2 signal in the HS-GC measurement, which is a change that can be used as an indirect measure of the carbonate in the sample. The results show that the present method has a good precision (the relative standard deviation<2.32%), and good accuracy (the relative differences compared to a reference method was<5.76%). Coupled with the fact that the method is simple, rapid, and accurate, it is suitable for a variety of applications that call for the analysis of high carbonate content in paper samples.


Myositis and myopathy of sarcoidosis: A case report.
Muscular manifestations of sarcoidosis are commonly found on biopsy but rare on correlated imaging. We present a rare case of a 36-year-old male patient with sarcoid myositis and image findings of active myositis in the lower back and pelvic girdle musculature. This case suggests considering sarcoidosis as a differential diagnosis in the setting of chest findings and new lower back and lower extremity weakness.


Conversion of a homogeneously methicillin-resistant strain of Staphylococcus aureus to heterogeneous resistance by Tn551-mediated insertional inactivation.
Plasmid pRN3208, thermosensitive for replication, and carrying the erythromycin transposon Tn551, was used for insertional inactivation of methicillin resistance in a highly and homogeneously resistant strain of Staphylococcus aureus. Two kinds of insertionally inactivated cells were obtained. Cultures of the major class contained highly methicillin resistant cells with a frequency of about 10(-3) to 10(-4), produced DNA with methicillin resistance transforming activity, and also produced penicillin binding protein 2a, the 78 kd low affinity penicillin binding protein characteristic of methicillin resistant Staphylococcus aureus, in apparently normal quantities. The single member of class B had no detectable methicillin resistant cells (less than 10(-8)) with an MIC greater than 1 micrograms/ml, contained no DNA with methicillin resistant transforming activity and no penicillin binding protein 2a. The data suggest that in the class A cells insertional inactivation did not affect the structural gene(s) of methicillin resistance but a regulatory locus or loci needed for the homogeneous expression of resistance.


The influence of cardiac rehabilitation on heart rate variability indices in men with type 2 diabetes and coronary artery disease.
The study population included 141 male CAD patients prospectively and consecutively admitted to an outpatient comprehensive CR program. Twenty-seven patients with type-2 diabetes were compared with 114 males without diabetes. The participants performed a 45-min cycle ergometer interval training alternating 4-min workload and a 2-min active restitution three times a week for 8 weeks. The training intensity was adjusted so that the patient's heart rate achieved the training heart rate calculated according to the Karvonen formula. At the baseline and after 8 weeks, all the patients underwent the HRV assessment. HRV indices in the patients with diabetes were significantly lower as compared to the patients without diabetes in SDNN, TP, LF parameters, both at the baseline and after 8 weeks of CR. After 8 weeks of CR, a significant improvement of TP, SDNN, pNN50% and HF occurred in the patients without diabetes, whereas in the patients with diabetes only HF component improved significantly. As regards HRV indices, CR seems to be less effective in patients with CAD and type-2 diabetes.


Superparamagnetic iron oxide hepatic MR imaging: efficacy and safety using conventional and fast spin-echo pulse sequences.
The purpose of this study was to evaluate the technical efficacy and safety of iv ferumoxides (Feridex), a superparamagnetic iron oxide contrast agent for detection of hepatic lesions using conventional spin-echo and fast spin-echo MR images. Precontrast and postcontrast MR studies were performed on 25 patients with suspected focal hepatic lesions. Conventional T1- and T2-weighted MR images, as well as fast spin-echo and fat suppressed fast spin-echo MR images, were evaluated. Quantitative assessment of the contrast agent was performed obtaining region of interest measurements of the liver, spleen, and selected hepatic lesions. The pulse sequences were also evaluated subjectively for overall image quality and a subjective assessment of lesion detection. The use of a superparamagnetic iron oxide contrast agent led to a decrease in hepatic signal intensity on all pulse sequences. Lesion-to-liver contrast increased 41.1%, 36.5%, and 32.0% on the conventional T2, fast spin-echo, and fat suppressed fast spin echo pulse sequences, respectively. Lesion-to-liver contrast decreased on the T1-weighted postcontrast pulse sequence by 23.8%. Despite improvement in lesion-to-liver contrast, radiologists subjectively preferred the precontrast sequences because of overall better image quality. At a dose of 10 mumol/kg, ferumoxides favorably impacts lesion-to-liver contrast, and may be useful in hepatic imaging, more with conventional T2-weighted spin-echo pulse sequences than with T2-weighted fast spin-echo pulse sequence.


Characteristics of rhabdomyosarcoma cell lines derived from uterine carcinosarcomas.
Rhabdomyosarcoma (RMS) is occasionally found in the female genital tract, and mostly appears as one of the heterologous mesenchymal components in uterine carcinosarcoma designated as malignant mixed mullerian tumour (MMMT). We examined the biological properties of a pure rhabdomyosarcoma (RMS) cell line designated FU-MMT-3, which was newly established from a surgical specimen taken from a patient with uterine MMMT. We also evaluated c-myc and MYCN gene amplification in three RMS cell lines (including FU-MMT-3) derived from three MMMTs by Southern blot analysis. FU-MMT-3 cells were propagated continuously for 57 serial passages over a 2-year period in vitro. FU-MMT-3 was able to produce tumours demonstrating pure RMS in athymic nude mice. Cytogenetically, FU-MMT-3 showed a triploidy pattern, with complex karyotypic abnormalities including trisomy of chromosome 8. All three RMS cell lines, including FU-MMT-3, showed amplification of the c-myc gene (approximately fourfold to eightfold), while no cell lines demonstrated MYCN gene amplification. FU-MMT-3 is considered to provide a useful system for the study of the biological behaviour of RMS in MMMTs. Extra copies of chromosome 8 and c-myc gene amplification may be associated with the rhabdomyoblastic differentiation in MMMT.


Introducing potato starch-ecofriendly silver nanoparticles as a novel binary system for nanoencapsulation of riboflavin.
Riboflavin (Rf), an externally supplied nutrient, is highly photosensitive, and should be protected from sunlight once used in food and pharmaceutical manufacturing. The applications of encapsulated Rf have recently developed due to their therapeutic properties. In this study, the use of green silver nanoparticles (AgNPs) synthesized by Rosa damascena mill L. extract to control the encapsulation efficiency of Rf in potato starch was demonstrated for the first time. Starch/Rf, Starch/AgNPs/Rf and Starch/AgNPs nanocapsules were characterized by Fourier-transform infrared, field emission scanning electron microscopy, differential scanning calorimetry, and brunauer-Emmett-Teller techniques. The obtained results showed that the presence of AgNPs reduces Rf nanocapsules size (from 340 to 327 nm), increases the encapsulation efficiency (21.14 ± 0.62 to 92.52 ± 1.32 %) and improves the thermal stability, antibacterial and antioxidant activities. Moreover, UV-vis spectroscopy demonstrated the stronger association of AgNPs/Rf and AgNPs/Rf/Starch nanocapsules with BSA under physiological conditions.


Relations between periodontal status, oral health-related quality of life and perceived oral health and oral health consciousness levels in a Turkish population.
This study aimed to evaluate the interrelationship of periodontal status, socio-demographic characteristics, perceived oral health and oral health consciousness levels as well as the impact of these factors on quality of life using a questionnaire and the Oral Health Impact Profile-14(OHIP-14) scale. Seven hundred and fifty systemically healthy individuals aged ≥18 years referred to a Periodontology Department were included in the study. The OHIP-14 scale and survey were applied to identify socio-demographic characteristics, oral hygiene characteristics, perceived oral health and oral hygiene consciousness levels. Three groups were established based on periodontal status (periodontally healthy [H], gingivitis [G] and periodontitis [P]) determined using periodontal indexes, and the relationship between the above-mentioned factors and periodontal status with quality of life was assessed. Oral health-related quality of life differed significantly by gender, marital status, education level, oral hygiene habits and periodontal status. A statistically significant positive relationship was found between high OHIP-14 scores and unfavourable socio-demographic characteristics, increased severity of periodontal disease, and irregular dental care practices. The perceived oral health and oral health consciousness levels significantly differed due to periodontal status. Periodontal status, gender, marital status, education level, dental care practices, perceived oral health and oral hygiene consciousness levels are important determinants of oral health-related quality of life (Clinical Trial No. NCT03549247).


HsS versus a balanced salt solution as a corneal wetting agent during routine cataract extraction and lens implantation.
To evaluate HsS (elastoviscous hylan surgical shield, 0.45%) as an alternative to repeated corneal irrigation with a balanced salt solution during cataract surgery. York Finch General Hospital, Toronto, Ontario (Center A), and Centre Hospitalier de St. Laurent, St. Laurent, Quebec (Center B), Canada. This dual-center, randomized, prospective clinical trial comprised 60 patients (40 at Center A; 20 at Center B) who had routine small incision cataract surgery (Center A, endolenticular phacoemulsification; Center B, Khoury manual phacofragmentation) with in-the-bag implantation of a posterior chamber intraocular lens. The corneal irrigating fluid was randomly assigned to be HsS or a balanced salt solution. The frequency of required applications, duration of efficacy of each application, and assessment of corneal moisture, clarity, transparency, and reflection in the HsS and balanced salt solution groups were recorded and compared. Mean frequency of applications was 13.9 per procedure in the balanced salt solution group and 1.3 per procedure in the HsS group (P = .0001). Mean duration of effectiveness per application was 23.4 minutes for HsS and 2.0 minutes for balanced salt solution (P = .0001). No significant differences in safety or effect on corneal health were observed between the two solutions. The HsS was significantly more effective than a balanced salt solution in maintaining corneal moisture, clarity, and transparency. The use of HsS may be a safer, more effective option in ophthalmic surgery because it minimizes the obstructed visualization of the surgical field caused by frequent corneal irrigation and loss of surgeon concentration.


Does bulky disease at diagnosis influence outcome in childhood Hodgkin's disease and require higher radiation doses? Results from the German-Austrian Pediatric Multicenter Trial DAL-HD-90.
The identification of risk factors is required for risk-adapted treatment strategies in the treatment of Hodgkin's disease. To assess the influence of bulky disease at diagnosis as compared with other risk factors on event-free survival (EFS) in pediatric Hodgkin's disease such as stage, B-symptoms, number of involved lymph node regions, histology, and remission status after chemotherapy, we analyzed the outcome of 552 patients treated with a risk-adapted treatment strategy consisting of OPPA(OEPA)/COPP (vincristine, procarbazine, etoposide, prednisone, adriamycin, cyclophosphamide) and involved-field radiotherapy. Between 1990 and 1995, 578 patients with primary Hodgkin's disease (HD) were enrolled in the German/Austrian Pediatric Hodgkin's Disease Study Group (DAL) Multicenter Study (HD-90). Patients were stratified into three treatment groups (TGs) for early, intermediate, and advanced stage. All patients received induction chemotherapy (CT) with two cycles of OEPA for boys and two cycles of OPPA for girls. Patients in TG2 and TG3 received another two or four cycles, respectively, of COPP. Chemotherapy was followed by involved-field radiotherapy. The radiation field, which was prescribed by the study center, was treated with a dose of 25 Gy/25 Gy/20 Gy (TG1/TG2/TG3), and in case of insufficient remission with a local boost of 5 Gy to 10 Gy. The following prognostic factors were analyzed with regard to their impact on EFS: bulky disease, mediastinal tumor, number of involved lymph node regions, histology, treatment group, B-symptoms, sex, age, and remission status after chemotherapy. Significant univariate predictive factors for the EES were: nodular sclerosis type 2 (NS2) histology (relative risk [RR] 3.43; p = 0.0002), presence of B-symptoms (RR 2.70; p = 0.0014), number of involved regions (1.55; p = 0.019), and treatment groups (RR 1.33; p = 0.017). There was a higher risk (RR 1.92; p = 0.040) for patients with bulky compared with nonbulky disease (5-year EFS 89.6%/94.6%). In the multiple regression model, only NS2 and B-symptoms remained strong predictive factors. The remission status after chemotherapy did not correlate with EFS (p = 0.66). Treatment strategies in Hodgkin's disease have an impact on different risk factors. In the risk-adapted treatment strategy of the HD-90 study, tumor burden indicated as bulky disease or as number of involved lymph nodes loses its importance, whereas NS2 histology and B-symptoms have a major impact on treatment outcome. Bulky disease at diagnosis might require higher radiation doses only in case of insufficient remission.


Ligand modulation of KCNQ-encoded (KV7) potassium channels in the heart and nervous system.
KCNQ-encoded (KV7) potassium channels are diversely distributed in the human tissues, associated with many physiological processes and pathophysiological conditions. These channels are increasingly used as drug targets for treating diseases. More selective and potent molecules on various types of the KV7 channels are desirable for appropriate therapies. The recent knowledge of the structure and function of human KCNQ-encoded channels makes it more feasible to achieve these goals. This review discusses the role and mechanism of action of many molecules in modulating the function of the KCNQ-encoded potassium channels in the heart and nervous system. The effects of these compounds on KV7 channels help to understand their involvement in many diseases, and to search for more selective and potent ligands to be used in the treatment of many disorders such as various types of cardiac arrhythmias, epilepsy, and pain.


Targeted immunotherapy of cancer with CAR T cells: achievements and challenges.
The adoptive transfer of chimeric antigen receptor (CAR)-expressing T cells is a relatively new but promising approach in the field of cancer immunotherapy. This therapeutic strategy is based on the genetic reprogramming of T cells with an artificial immune receptor that redirects them against targets on malignant cells and enables their destruction by exerting T cell effector functions. There has been an explosion of interest in the use of CAR T cells as an immunotherapy for cancer. In the pre-clinical setting, there has been a considerable focus upon optimizing the structural and signaling potency of the CAR while advances in bio-processing technology now mean that the clinical testing of these gene-modified T cells has become a reality. This review will summarize the concept of CAR-based immunotherapy and recent clinical trial activity and will further discuss some of the likely future challenges facing CAR-modified T cell therapies.


Distal regulation, silencers, and a shared combinatorial syntax are hallmarks of animal embryogenesis.
The chromatin environment plays a central role in regulating developmental gene expression in metazoans. Yet, the ancestral regulatory landscape of metazoan embryogenesis is unknown. Here, we generate chromatin accessibility profiles for six embryonic, plus larval and adult stages in the sponge Amphimedon queenslandica These profiles are reproducible within stages, reflect histone modifications, and identify transcription factor (TF) binding sequence motifs predictive of cis-regulatory elements operating during embryogenesis in other metazoans, but not the unicellular relative Capsaspora Motif analysis of chromatin accessibility profiles across Amphimedon embryogenesis identifies three major developmental periods. As in bilaterian embryogenesis, early development in Amphimedon involves activating and repressive chromatin in regions both proximal and distal to transcription start sites. Transcriptionally repressive elements ("silencers") are prominent during late embryogenesis. They coincide with an increase in cis-regulatory regions harboring metazoan TF binding motifs, as well as an increase in the expression of metazoan-specific genes. Changes in chromatin state and gene expression in Amphimedon suggest the conservation of distal enhancers, dynamically silenced chromatin, and TF-DNA binding specificity in animal embryogenesis.


[Antibiotic and sulfanilamide resistance of the coli- and lactobacteria used for the production of bacterial preparations].
An antibiotic-resistant mutant recommended for the production of colibacterin preparation was isolated from the E. coli strain M-17 highly-sensitive to antibiotics. A possibility of complex use of bacterial preparations (coli- and lactobacterin) with antibiotics and sulphanylamides for prevention of dysbacteriosis was demonstrated.