Reduction of osteopontin in vivo inhibits tubular epithelial to mesenchymal transition in rats with chronic allograft nephropathy.
Chronic allograft nephropathy (CAN) is an important etiological factor causing graft loss. However, the mechanism of CAN is unclear. Osteopontin (OPN), a proinflammatory and profibrosis molecule, plays a key role in late stages of renal diseases. We investigated the potential role of OPN in the pathogenesis of CAN. Using a F344 to Lewis rat CAN model, we injected short hairpin RNA (shRNA) constructs targeting OPN or negative control plasmids through the renal vein following electroporation. At 12 weeks after the transplantation, we determined interstitial fibrosis (IF) and tubular atrophy (TA) of the tubular epithelial cells (TECs). OPN expression was examined using Western blots and immunohistochemistry (IHC). Molecules involved in epithelial to mesenchymal transition (EMT) of TECs were examined using IHC and Western blots. OPN expression in kidney grafts was decreased by the RNA interference (RNAi) group. Histology observations showed IF and TA to be mild with stable renal function in the RNAi-treated group. EMT of TECs was significantly lessened after reducing OPN. Reduction of OPN in vivo inhibited progression of CAN. OPN may be of therapeutic value in transplantation settings.


Effects of quercetin on production and release of endothelin and cGMP from cultured endothelial cells.
To evaluate the effects of quercetin (Que) on the formation and release of endothelin (ET) and cyclic guanosine monophosphate (cGMP) by endothelial cells. Radioimmunoassay (RIA) was used to assess the amounts of ET and cGMP produced by unstimulated human umbilical vein cells (HUVEC) or HUVEC stimulated with activated platelets and thrombin. Following the treatment of the cultured HUVEC with activated platelets, the amount of ET in the medium was increased, but no significant effect on cGMP within the cells was seen. Thrombin not only increased the amount of ET in the medium, but also raised the level of cGMP within the cells. Pretreatment of HUVEC with Que 0.5, 5, and 50 mumol.L-1, the release of ET by HUVEC stimulated with thrombin and thrombin-activated platelets was much inhibited. Que 0.5-50 mumol.L-1 markedly increased the amount of cGMP in HUVEC stimulated with thrombin and activated platelets. Que also inhibited the release of ET from unstimulated HUVEC, but increased the production of cGMP from unstimulated EC. Thrombin and activated platelets stimulated the release of ET from HUVEC and Que inhibited the release of ET and promoted the formation of cGMP from HUVEC.


Moore prosthesis in the treatment of fresh femoral neck fractures. A critical review with special attention to secondary acetabular degeneration.
A consecutive series of 106 patients with displaced femoral neck fractures was treated initially with Moore prostheses. The patients' mean age was 82.5 years (range, 67-91). At follow-up after two years, 71 patients attended. Thirty-seven per cent of these were in need of a conversion of their one-piece device to a total hip replacement on account of hip pain. When only active patients living in their own homes were considered, 55 per cent were found to need total hip replacements. The main reason for the development of hip pain was acetabular derangement. It was concluded that active patients, disregarding their age, should not be treated with one piece prostheses of their fractures could be satisfactorily reduced and pinned. If this cannot be accomplished, a primary total hip replacement may be considered.


Hybrid flotation--membrane filtration process for the removal of heavy metal ions from wastewater.
A promising process for the removal of heavy metal ions from aqueous solutions involves bonding the metals firstly to a special bonding agent and then separating the loaded bonding agents from the wastewater stream by separation processes. For the separation stage, a new hybrid process of flotation and membrane separation has been developed in this work by integrating specially designed submerged microfiltration modules directly into a flotation reactor. This made it possible to combine the advantages of both flotation and membrane separation while overcoming the limitations. The feasibility of this hybrid process was proven using powdered synthetic zeolites as bonding agents. Stable fluxes of up to 80l m(-2)h(-1) were achieved with the ceramic flat-sheet multi-channel membranes applied at low transmembrane pressure (<100 mbar). The process was applied in lab-scale to treat wastewater from the electronics industry. All toxic metals in question, namely copper, nickel and zinc, were reduced from initial concentrations of 474, 3.3 and 167mg x l(-1), respectively, to below 0.05 mg x l(-1), consistently meeting the discharge limits.


"Hit the primary": A paradigm shift in the treatment of metastatic prostate cancer?
Patients with metastatic prostate cancer (PC) represent a heterogeneous group with survival rates varying between 13 and 75 months. The current standard treatment in this setting is hormonal therapy, with or without docetaxel-based chemotherapy. In the era of individualized medicine, however, maximizing treatment options, especially in long-term surviving patients with limited disease burden, is of capital importance. Emerging data, mainly from retrospective surgical series, show survival benefits in men diagnosed with metastatic PC following definitive therapy for the prostate. Whether the irradiation of primary tumor in a metastatic disease might improve the therapeutic ratio in association with systemic treatments remains investigational. In this scenario, modern radiation therapy (RT) can play a significant role owing to its intrinsic capability to act as a more general immune response modifier, as well as to the potentially better toxicity profile compared to surgery. Preclinical data, clinical experience, and challenges in local treatment in de novo metastatic PC are reviewed and discussed.


Detection of glucose in synthetic tear fluid using dually functionalized gold nanoparticles.
A simple fluorescent sensing of glucose in aqueous fluids (e.g. tear fluid) using dually functionalized gold nanoparticles is presented. As a first step gold nanoparticles (AuNPs) were synthesized using oxidised dextran which acted both as reducing and stabilizing agent. Aminophenyl boronic acid was conjugated onto AuNPs by Schiff's base formation and the formed Schiff's base was stabilized by sodium borohydride reduction. Rhodamine B isothiocyanate (RBITC) was then assembled onto the modified AuNPs. The fluorescence of RBITC was nearly quenched and found to be revived when glucose was added. It is reasoned that the glucose binding induces restructuring of the surface assembly resulting in an overall increase in the size and thereby enhancing the distance between the gold core and fluorophore. TEM image and size measurements using dynamic light scattering (DLS) in fact, reflected this possibility. The increase in fluorescence was proportional with the concentration of glucose enabling quantitative detection. A good linearity was observed between the fluorescence intensity and glucose concentration in a range of 0.025-0.125 μM with detection limit of 0.005 ± 0.002 μM. The potential of the method was demonstrated by measuring glucose in real tear fluids collected from volunteers. The method is extremely sensitive and can be employed to measure low concentration of glucose in aqueous fluids such as tear.


Cytoplasmic, but not nuclear, expression of the neuronal nuclei (NeuN) antibody is an exclusive feature of Dogiel type II neurons in the guinea-pig gastrointestinal tract.
This study aimed to reveal if NeuN, a neuronal nuclei (NeuN) antibody, is a selective marker of intrinsic primary afferent neurons (IPANs) in the guinea-pig gastrointestinal tract as previously hypothesised. The NeuN immunoreactivity was found in the enteric nervous system with exception of the esophagus. Two groups of NeuN-expressing neurons were observed: neurons with immunostained nuclei and cytoplasm (NeuN(NC)) and neurons only expressing immunoreactivity in their nuclei (NeuN(N)). The NeuN(N)-immunoreactive neurons were found in the myenteric plexus of the stomach and the colon. In the stomach, none of the NeuN(N)-expressing neurons, of which 55+/-3% co-expressed calbindin, had a Dogiel type I or II morphology. The NeuN(N)-positive neurons of the colon, which did not express calbindin, did not resemble a Dogiel type II morphology either, but were small-sized neurons. The NeuN(NC)-immunoreactive neurons were observed in both the small and large intestine. These neurons were smooth-contoured and bigger-sized, resembling a Dogiel type II morphology. Some of these neurons co-expressed calbindin. The present data reveal the existence of two populations of Dogiel type II neurons, exhibiting NeuN(NC)+/calbindin+ or NeuN(NC)+/calbindin- immunoreactivity, in the intestine. Assuming that all IPANs exhibit a Dogiel type II morphology, we conclude that the cytoplasmic expression of NeuN is an exclusive feature of IPANs.


ST-YOLOA: a Swin-transformer-based YOLO model with an attention mechanism for SAR ship detection under complex background.
A synthetic aperture radar (SAR) image is crucial for ship detection in computer vision. Due to the background clutter, pose variations, and scale changes, it is a challenge to construct a SAR ship detection model with low false-alarm rates and high accuracy. Therefore, this paper proposes a novel SAR ship detection model called ST-YOLOA. First, the Swin Transformer network architecture and coordinate attention (CA) model are embedded in the STCNet backbone network to enhance the feature extraction performance and capture global information. Second, we used the PANet path aggregation network with a residual structure to construct the feature pyramid to increase global feature extraction capability. Next, to cope with the local interference and semantic information loss problems, a novel up/down-sampling method is proposed. Finally, the decoupled detection head is used to achieve the predicted output of the target position and the boundary box to improve convergence speed and detection accuracy. To demonstrate the efficiency of the proposed method, we have constructed three SAR ship detection datasets: a norm test set (NTS), a complex test set (CTS), and a merged test set (MTS). The experimental results show that our ST-YOLOA achieved an accuracy of 97.37%, 75.69%, and 88.50% on the three datasets, respectively, superior to the effects of other state-of-the-art methods. Our ST-YOLOA performs favorably in complex scenarios, and the accuracy is 4.83% higher than YOLOX on the CTS. Moreover, ST-YOLOA achieves real-time detection with a speed of 21.4 FPS.


Matricellular proteins at the crossroad of inflammation and cancer.
Some proteins of the extracellular matrix known as matricellular proteins have regulatory function in all aspects of physiological and pathological stroma rearrangement. Many aspects of their activity are related to inflammation and immune response suggesting their role in bridging inflammation and cancer.


Targeting oncogenic KRAS in non-small cell lung cancer with EGFR aptamer-conjugated multifunctional RNA nanoparticles.
KRAS mutations are one of the most common oncogenic driver mutations in human cancers, including non-small cell lung cancer (NSCLC), and have established roles in cancer pathogenesis and therapeutic resistance. The development of effective inhibitors of mutant KRAS represents a significant challenge. Three-way junction (3WJ)-based multi-functional RNA nanoparticles have the potential to serve as an effective in vivo siRNA delivery platform with the ability to enhance tumor targeting specificity and visualize biodistribution through an imaging moiety. Herein, we assembled novel EGFRapt-3WJ-siKRASG12C mutation targeted nanoparticles to target EGFR-expressing human NSCLC harboring a KRASG12C mutation to silence KRASG12C expression in a tumor cell-specific fashion. We found that EGFRapt-3WJ-siKRASG12C nanoparticles potently depleted cellular KRASG12C expression, resulting in attenuation of downstream MAPK pathway signaling, cell proliferation, migration/invasion ability, and sensitized NSCLC cells to chemoradiotherapy. In vivo, these nanoparticles induced tumor growth inhibition in KRASG12C NSCLC tumor xenografts. Together, this study suggests that the 3WJ pRNA-based platform has the potential to suppress mutant KRAS activity for the treatment of KRAS-driven human cancers, and warrants further development for clinical translation.


Melatonin for the masses. Antiaging entrepreneurs peddle therapies that promise to cheat the clock.
The come-ons for a new wave of antiaging clinics may be over the top, but they speak to boomer anxieties about flagging energy levels and flabby midsections. And they just may be the next hot trend in health care.


Alkylamides from Zanthoxylum armatum DC. and their neuroprotective activity.
Zanthoxylum armatum DC. is an important medicinal plant, and its pericarps are commonly used as a natural spice in Asian countries. In this study, fifteen alkylamides were isolated and elucidated from the pericarps of Z. armatum, including five undescribed alkylamides (1-5) and ten known compounds (6-15). The molecular structures of all compounds were elucidated by 1D and 2D NMR spectroscopic analysis and mass spectrometry, among which the absolute configuration of compound 15 was determined by the Mo2(OAc)4-induced circular dichroism method. Moreover, all compounds were screened for their neuroprotective activity against H2O2-induced oxidative stress in human neuroblastoma SH-SY5Y cells for the evaluation of their neuroprotective activity. Especially, compounds 2-4 expressed potential neuroprotective activity, and further research showed that the cell viability was significantly enhanced in a concentration dependent manner when the cells were treated for 6 h. Moreover, compounds 2-4 could decrease reactive oxygen species accumulation. This paper enriched structure types of alkylamides in Zanthoxylum armatum.


Immunomodulatory effect of picroliv on the efficacy of paromomycin and miltefosine in combination in experimental visceral leishmaniasis.
Combination therapy for the treatment of visceral leishmaniasis has increasingly been advocated as a way to increase treatment efficacy and tolerance, to reduce treatment duration and cost, and to limit the emergence of drug resistance. In the present work, we have adopted a rational approach, which can modulate the immune response to overcome the negative control systems and to boost the positive killing responses. This study was designed to investigate the immunomodulatory effect of picroliv (standardized fraction from the alcoholic extract of root and rhizome of Picrorhiza kurroa) on a combination of paromomycin and miltefosine using Leishmania donovani/hamster model. Picroliv has significantly enhanced antileishmanial efficacy and lymphocyte proliferation when given in combination with paromomycin and miltefosine. Increased toxic oxygen metabolite generation and phagocytosis were also witnessed. Present study thus establishes the possible use of picroliv as adjunct to antileishmanial chemotherapy.


Autophagy in mouse hepatocytes induced by lysine acetylsalicylate.
I.v. administration of lysine acetylsalicylate inces autophagy in mouse liver cells. Single and multiple membrane-bounded vacuoles were found. The latter seems to be an unusual morphological form of the sequestration process. These findings could express a transitory sublethal liver cell injury induced by the drug.


Regulation of thyrotropin biosynthesis. Discordant effect of thyroid hormone on alpha and beta subunit mRNA levels.
We have studied the regulation of the biosynthesis of thyrotropin (TSH) and its alpha and beta subunits by thyroid hormone in thyrotropic tumors carried in hypothyroid mice. Treatment with 3,5,3'-triiodo-L-thyronine (T3) (20 micrograms/100 g, body weight) daily for 4 or 10 days reduced serum TSH to 3 and 0.3% of control, respectively. Serum levels of free alpha subunit were reduced to 60 and 11% of control at 4 days and 10 days, respectively, and serum free TSH-beta was undetectable at both time points. There was no significant decrease in tumor TSH content after 4 days of treatment and, after 10 days, TSH content was reduced to 15% of control levels. There was no significant effect of T3 on tumor alpha subunit levels at either 4 or 10 days. In contrast, tumor TSH-beta content was markedly reduced after 4 days and 10 days of T3 treatment, to 29 and 10% of control levels, respectively. Translation of tumor poly(A) mRNA in a rabbit reticulocyte lysate system showed that thyroid hormone decreased translatable TSH-beta mRNA to undetectable levels at both 4 and 10 days, whereas translatable alpha mRNA was reduced strikingly only at 10 days in one of two tumors. RNA blot hybridization with 32P-labeled plasmid probes containing alpha or TSH-beta cDNAs showed that TSH-beta mRNA was reduced to less than 10% of control after both 4 and 10 days of T3 treatment, whereas, again, alpha mRNA was only reduced in one of two tumors at 10 days. Our data thus show that thyroid hormone affects alpha and TSH-beta mRNA and protein levels discordantly and suggest that regulation of TSH biosynthesis may occur predominantly at the level of TSH-beta mRNA.


Palliative Cryotherapy of Aggressive Endobronchial Parotid Carcinoma ex Pleomorphic Adenoma: A Case Report.
Carcinoma ex pleomorphic adenoma is a rare salivary gland malignancy with a poor prognosis that can metastasize to the lungs. Endobronchial metastasis and associated bronchoscopic characteristics have not been described in the literature. Cryotherapy is a safe and efficacious palliative treatment option for obstructive bronchial tumors that can be performed with flexible bronchoscopy. We report the successful use of cryotherapy for the palliation of an aggressive parotid malignancy with extraordinary endobronchial metastases and describe bronchoscopic characteristics of this unique entity.


Sensitivity to pre- and post-reinforcer delays in self-control choice.
Rats were exposed to a concurrent-chains schedule in which reinforcer amounts, pre-reinforcer and post-reinforcer delays were varied between alternatives. Two independent variable-interval 60-s schedules were in effect during the initial links. Pre- and post-reinforcer delays were defined by fixed-time schedules. In one condition, rats chose between a larger reinforcer delayed by 0, 40, 80, or 120-s and an immediate smaller reinforcer. In the other condition, rats chose between an immediate larger reinforcer terminated with 0, 40, 80, or 120-s post-reinforcer delays and an immediate smaller reinforcer terminated with no post-reinforcer delay. In both conditions, choice was predicted from overall reinforcement density (i.e. reinforcer amount divided by total trial time). The results indicate that the preference for the larger reinforcer systematically decreased as both pre- and post-reinforcer delays increased. Although the effect of pre-reinforcer delay was greater than that of post-reinforcer delay, post-reinforcer delay itself influenced rats' choices in the present self-control situation. In conclusion, the present study demonstrated the limitation of local reinforcement density model of the matching law and all time components that constitute a choice alternative need to be weighted and incorporated into the general model of choice behavior.


Evidence for long-range glycosyl transfer reactions in the gas phase.
A long-range glycosyl transfer reaction was observed in the collision-induced dissociation Fourier transform (CID FT) mass spectra of benzylamine-labeled and 9-aminofluorene-labeled lacto-N-fucopentaose I (LNFP I) and lacto-N-difucohexaose I (LNDFH I). The transfer reaction was observed for the protonated molecules but not for the sodiated molecules. The long-range glycosyl transfer reaction involved preferentially one of the two L-fucose units in labeled LNDFH I. CID experiments with labeled LNFP I and labeled LNFP II determined the fucose with the greatest propensity for migration. Further experiments were performed to determine the final destination of the migrating fucose. Molecular modeling supported the experiments and reaction mechanisms are proposed.


Subarachnoid neurolytic block (L5-S1) and unilateral percutaneous cervical cordotomy in the treatment of pain secondary to pelvic malignant disease.
The present study deals with the immediate and long-term results of subarachnoid neurolytic block (L5-S1) with 7.5%, 10% and 15% concentrations of phenol in glycerine and/or unilateral percutaneous cervical cordotomy in 73 patients (follow-up in 56 patients) suffering from perineal, perineopelvisacral or pelvisacral pain secondary to malignant diseases of the pelvic cavity. Subarachnoid neurolytic block (L5-S1) produced satisfactory, long-lasting relief of perineal pain when the higher concentrations of phenol (10 and 15%) were used. The only sequela reported was urinary retention. Percutaneous cervical cordotomy used for the treatment of pelvisacral or predominantly unilateral perineal pain gave complete pain relief until death in 76.7% of patients, either alone (36.7%) or in conjunction with pharmacological therapy (40%). Analysis of the data enabled us to establish the respective indications for the two procedures and to identify those cases in which their use may be complementary.


Inductive teaching method-an alternate method for small group learning.
Different forms of small group interactive teaching have found an increasing role in medical education in the recent past. This study aimed to evaluate the efficacy of topic-based small group tutorials and compare between two such tutorials, the directed tutorial where the tutor takes the initiative and the inductive tutorial which places the initiative on the student. Final-year medical students were exposed to alternate sessions of directed and inductive teaching during their rotation in paediatric surgery. For inductive teaching, the topic was announced and the students took turns in asking questions and answering them, the tutor stepping in only when required. The tutor both initiated and guided the discussion in the directed method. Students assessed the tutorials and compared the two learning methods evaluating eight different parameters using a Likert scale. Statistical significance was tested using the Wilcoxon signed ranks test. Feedback was available from 49 out of 52 students. The tutorials were ranked highly for all parameters studied. The inductive method was better for comfort, interest, memory, inspiration to read more, comprehension and for correlation with previous knowledge (P < 0.05). The directed method was superior for the content covered in a given time (P < 0.001). The topic-based tutorials were effective. The inductive method by adding a reflective component, appeared to facilitate deep learning. Further research will determine its suitability for formal introduction into the curriculum.


Conversion of sterols and triterpenes by mycobacteria. II. Transformation of 7-oxygenated sterols into androstane derivatives via a 7-deoxygenation.
The bioconversion of 7-oxygenated sterols by Mycobacterium aurum was studied in a preliminary investigation of the microbial conversion of wool wax. 7-Oxocholesterol was found to be transformed mainly into 3,17-dioxygenated androstane derivatives. 7 xi-Hydroxylated sterols were formed in an initial reduction step, and the C-7 hydroxyl group was then eliminated in a dehydration reaction. This was thought to take place during the isomerisation of cholest-4-en-3-one to cholest-5-en-3-one. Deuterium labelling experiments showed that this elimination proceeded faster for the C-7 alpha isomer, although it was not stereospecific. The C-7 alpha and C-7 beta-hydroxy isomers were weakly interconverted via the 7-oxo derivatives. Cholest-4-en-3-one, cholest-1,4-dien-3-one and cholest-4,6-dien-3-one all lost their side chains following a hydrogenation/dehydrogenation reaction. The resulting 3,17-dioxoandrostene or 3,17-androstadiene derivatives were mainly hydrogenated into 5 alpha-androstane-3,17-dione and 5 alpha-androstane-3 beta-ol-17-one. Elimination of the 3 beta-hydroxyl groups giving cholesta-3,5-dien-7-one, and subsequent microbial degradation of the side chain was not observed to any significant extent. The convergence of the bioconversion pathways of cholesterol and the 7-oxygenated cholesterols enabled crude, partially auto-oxidised cholesterol to be used as a substrate for the production of 3,17-dioxygenated androstane derivatives by M. aurum.


[The effects of nm23-H1 on metastases ability and chemo-sensitivity in Tca8113 cell lines].
The purpose of this study was to establish a stable, high-efficient and low-toxic way of transfecting nm23-H1 into Tca8113 line cells, and then to find out whether nm23-H1 could affect the invasion and metastases ability of Tca8113 line cells. nm23-H1 was transfected into Tca8113 line cells with Lipofect. The different expressions of nm23-H1 between transfected and non-transfected line cells were detected by the method of immunohistochemistry. The difference of the invasion and metastases ability between transfected and non-transfected line cells was detected by transwell-room and wash techniques. The change of chemo-sensitivity was evaluated by MTT. Using pCMV-NEO-BAM system to keep stable expression of nm23-H1, the significant difference of NDPKA expression between transfected and non-transfected Tca8113 line cells was discovered; The metastases ability of transfected Tca8113 line cells decreased significantly; The chemo-sensitivity of transfected Tca8113 line cells to CDDP increased significantly. nm23-H1 can inhibit the metastases of Tca8113 line cells and increase the chemo-sensitivity to CDDP significantly.


High-performance liquid chromatographic assay for acid and alkaline phosphatase in serum.
High-performance liquid chromatography was used to assay serum acid and alkaline phosphatase. Samples were incubated with adenosine-5'-monophosphoric acid (AMP) in a buffer of required pH, 5'-nucleotidase was inhibited with Ni2+ ions, and the phosphatase activity was determined by measuring the concentration of the reaction product, adenosine. The analysis time, after the incubation is terminated, is short (7 min), and the assay is quantitative and reproducible. Complete separation of the reaction product from the substrate and the naturally occurring serum constituents and the high sensitivity of the ultraviolet detection system eliminate some of the problems commonly encountered in spectrophotometric assays.


Role of interleukin 15 in colitis induced by dextran sulphate sodium in mice.
Interleukin (IL)-15 is a member of the IL-2 family, stimulating dendritic cells, natural killer (NK) cells, NK T cells and memory CD8+ T cells. IL-15 levels were elevated in the intestinal mucosa of inflammatory bowel diseases. Here we investigated the involvement of IL-15 in the pathogenesis of acute and chronic dextran sulphate sodium (DSS) induced colitis. IL-15 knockout (KO) mice and control C57BL/6 mice were used to induce colitis with DSS in their drinking water. Survival rate, clinical activity of diseases, extent of tissue damage, leucocyte population, and cytokine production of lamina propria (LP) cells of the large intestines were assessed. IL-15 KO mice exhibited resistance to DSS induced acute colitis, as reflected by lower lethality, weight loss, clinical scores, and histological scores compared with those in control mice (p<0.05). The proportions of CD44(high) CD8+ T cells and NK cells in LP cells and levels of interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, and IL-12p40 in culture supernatants of LP cells were reduced in IL-15 KO mice (p<0.05). In vivo depletion of CD8+ T cells and NK cells decreased levels of IFN-gamma, TNF-alpha, and IL-12p40 in culture supernatants of LP cells in C57BL/6 mice (p<0.01). In chronic colitis, weight loss and clinical scores were improved and levels of IFN-gamma, TNF-alpha, and IL-12p40 in culture supernatants of LP cells were also reduced in IL-15 KO mice (p<0.05). IL-15 plays an important role in the pathogenesis of both acute and chronic colitis induced by DSS in mice.


Analyses of combined effects of cytostatic drugs on micronucleus formation in the Tradescantia.
Recent experiments showed that 5-fluorouracil (5FU), cisplatin (CDDP), etoposide (ET), and imatinib mesylate (IM), which are currently among the most widely used anticancer drugs, cause damage of the genetic material in higher plants. The aim of the present study was to determine whether mixtures of these drugs cause synergistic or antagonistic effects which may have an impact on their environmental safety. Therefore, the effects of binary mixtures of these anticancer drugs on the induction of micronuclei (MN) which reflect structural and numerical chromosomal aberrations were assessed in Tradescantia tetrads. Synergistic/antagonistic effects were determined by comparison with single exposures that would be equally effective in a reference model of independent action. This comparison was performed at two distinct effect sizes. We found clear evidence for synergisms in combination experiments with IM and antagonism in a high-dose experiment with ET and 5FU. Our findings indicate that IM increases the genotoxic effects of other anticancer drugs. The maximal effects which we found were in the range between 19 and 38 % in the excess of effect sizes predicted under independent action. These effects may have an impact on the overall genotoxic activities of untreated hospital waste waters but not on the environment in general as the predicted environmental concentrations of the studied drugs are several orders of magnitude lower as the levels which are required to cause induction of MN in higher plants.


Insulin action on brain microvessels; effect on alkaline phosphatase.
The effects of insulin on brain alkaline phosphatase activity have been examined. Insulin inhibited the activity of alkaline phosphatase on brain microvessels in in vitro experiments. The inhibition observed was of the non-competitive type. These observations indicate that the hormone is able to induce neurochemical modifications revealed in this case as changes in the phosphate transfer enzymes in brain microvessels.


dBre1/dSet1-dependent pathway for histone H3K4 trimethylation has essential roles in controlling germline stem cell maintenance and germ cell differentiation in the Drosophila ovary.
The Drosophila ovarian germline stem cells (GSCs) constantly experience self-renewal and differentiation, ensuring the female fertility throughout life. The balance between GSC self-renewal and differentiation is exquisitely regulated by the stem cell niche, the stem cells themselves and systemic factors. Increasing evidence has shown that the GSC regulation also involves epigenetic mechanisms including chromatin remodeling and histone modification. Here, we find that dBre1, an E3 ubiquitin ligase, functions in controlling GSC self-renewal and germ cell differentiation via distinct mechanisms. Removal or knock down of dBre1 function in the germline or somatic niche cell lineage leads to a gradual GSC loss and disruption of H3K4 trimethylation in the Drosophila ovary. Further studies suggest that the defective GSC maintenance is attributable to compromised BMP signaling emitted from the stem cell niche and impaired adhesion of GSCs to their niche. On the other hand, dBre1-RNAi expression in escort cells causes a loss of H3K4 trimethylation and accumulation of spectrosome-containing single germ cells in the germarium. Reducing dpp or dally levels suppresses the germ cell differentiation defects, indicating that dBre1 limits BMP signaling activities for the differentiation control. Strikingly, all phenotypes observed in dBre1 mutant ovaries can be mimicked by RNAi-based reduced expression of dSet1, a Drosophila H3K4 trimethylase. Moreover, genetic studies favor that dBre1 interacts with dSet1 in controlling GSC maintenance and germ cell differentiation. Taken together, we identify a dBre1/dSet1-dependent pathway for the H3K4 methylation involved in the cell fate regulation in the Drosophila ovary.


The non-affine fiber network solver: A multiscale fiber network material model for finite-element analysis.
Multiscale mechanical models in biomaterials research have largely relied on simplifying the microstructure in order to make large-scale simulations tractable. The microscale simplifications often rely on approximations of the constituent distributions and assumptions on the deformation of the constituents. Of particular interest in biomechanics are fiber embedded materials, where simplified fiber distributions and assumed affinity in the fiber deformation greatly influence the mechanical behavior. The consequences of these assumptions are problematic when dealing with microscale mechanical phenomena such as cellular mechanotransduction in growth and remodeling, and fiber-level failure events during tissue failure. In this work, we propose a technique for coupling non-affine network models to finite element solvers, allowing for simulation of discrete microstructural phenomena within macroscopically complex geometries. The developed plugin is readily available as an open-source library for use with the bio-focused finite element software FEBio, and the description of the implementation allows for the adaptation to other finite element solvers.


Sickle cell anemia. Pathophysiology and treatment.
Current concepts of the pathophysiology of the sickling process are reviewed. New approaches to treatment - including induced hyponatremia, membrane-active antisickling agents, and gene manipulation - are described.


Ultrastructural study of a patient with progressive dementia showing special intraplasmic inclusions in diffuse cerebral cortex. Mechanism of inclusions production.
Numerous inclusions were diffusely observed in the neurons and glial cells in the cerebral cortex of a patient with progressive dementia. These inclusions were initially produced by rough ER in the cells and gradually enlarged destroying the rough ER membranes. The nature of these inclusions was neutral mucopolysaccaride including protein histochemically. In the nuclei of the nerve cells and glial cells, some types of intranuclear virus-like inclusions were found. The first type was papova virus-like structure with high electron density measured 30--35 nm in diameter. The second type was paramyxovirus-like structure with granulo-fibrilar shape. The third type was large hexogonal core with high electron density measured 100 nm in diameter. These intranuclear virus-like bodies were not specific in the present case; however, it was speculated that intranuclear virus-like inclusions might have a deep relation to change the function of the nuclei and produced large intraplasmic inclusion bodies.


Good neurologic recovery after cardiac arrest using hypothermia through continuous renal replacement therapy.
Therapeutic hypothermia (TH) is becoming a standard of care to mitigate neurologic injury in cardiac arrest survivors. Several cooling methods are available for use in TH. For maintaining a target temperature, intravascular cooling is superior to, more efficacious than, and safer than surface cooling methods. The use of an intravenous cooling catheter is independently associated with a higher odds ratio for survival. However, many techniques use commercially developed equipment that is expensive to purchase and use. The application and popularization of the intravascular cooling method have been difficult. In patients with pulmonary edema or cardiac insufficiency, liquid is restricted, so intravascular cooling systems cannot be used. Studies have shown abnormalities mimicking the immunologic and coagulation disorders observed in severe sepsis. Continuous renal replacement therapy has been widely used in the intensive care unit to improve clinical parameters and survival in patients with multiple-organ dysfunction of septic origin. Continuous renal replacement therapy can also be used as another type of core cooling method. We used continuous renal replacement therapy as a cooling method to induce TH in a patient who had a cardiac arrest, and the patient regained consciousness 52 hours later.


Applications of Dynamic Contrast-Enhanced Ultrasound in Differential Diagnosis of Hepatocellular Carcinoma and Intrahepatic Cholangiocarcinoma in Non-cirrhotic Liver.
The aim of the work described here was to investigate the value of dynamic contrast enhanced ultrasound (DCE-US) and quantitative analysis in pre-operative differential diagnosis of intrahepatic cholangiocarcinoma (ICC) and hepatocellular carcinoma (HCC) in non-cirrhotic liver. In this retrospective study, patients with histopathologically proven ICC and HCC lesions in non-cirrhotic liver were included. All patients underwent contrast-enhanced ultrasound (CEUS) examinations with an Acuson Sequoia unit (Siemens Healthineers, Mountain View, CA, USA) unit or LOGIQ E20 (GE Healthcare, Milwaukee, WI, USA) within 1 wk before surgery. SonoVue (Bracco, Milan, Italy) was used as the contrast agent. B-mode ultrasound (BMUS) features and CEUS enhancement patterns were analyzed. DCE-US analysis was performed by VueBox software (Bracco). Two regions of interest (ROIs) were set in the center of the focal liver lesions and their surrounding liver parenchyma. Time-intensity curves (TICs) were generated, and quantitative perfusion parameters were obtained and compared between the ICC and HCC groups using the Student t-test or Mann-Whitney U-test. From November 2020 to February 2022, patients with histopathologically confirmed ICC (n = 30) and HCC (n = 24) lesions in non-cirrhotic liver were included. During the arterial phase (AP) of CEUS, ICC lesions exhibited heterogeneous hyperenhancement (13/30, 43.3%), heterogeneous hypo-enhancement (2/30, 6.7 %) and rim-like hyperenhancement (15/30, 50.0%), whereas all HCC lesions exhibited heterogeneous hyperenhancement (24/24, 100.0%) (p < 0.05). Subsequently, most of the ICC lesions exhibited AP wash-out (83.3%, 25/30), whereas a few cases exhibited wash-out in the portal venous phase (PVP) (15.7%, 5/30). In contrast, HCC lesions exhibited AP wash-out (41.7%, 10/24), PVP wash-out (41.7%, 10/24) and a small part of late phase wash-out (16.7%, 4/24) (p < 0.05). Compared with those of HCC lesions, TICs of ICCs revealed earlier and lower enhancement during the AP, faster decline during the PVP and reduced area under the curve. The combined area under the receiver operating characteristic curve (AUROC) of all significant parameters was 0.946, with 86.7% sensitivity, 95.8% specificity and 90.7% accuracy in differential diagnosis between ICC and HCC lesions in non-cirrhotic liver, which improved the diagnostic efficacy of CEUS (58.3% sensitivity, 90.0% specificity and 75.9% accuracy). ICC and HCC lesions in non-cirrhotic liver might exhibit some overlap of CEUS features in diagnosis. DCE-US with quantitative analysis would be helpful in pre-operative differential diagnosis.


Striatal dopamine release in vitro: a beta-adrenoceptor-regulated response not mediated through cyclic AMP.
The effects of (-)isoproterenol (10(-6) M), dibutyryl cyclic AMP (10(-3) M), and the phosphodiesterase inhibitor 3-isobutyl-l-methylxanthine (IBMX) (10(-4) M) on in vitro [3H]dopamine ([3H]DA) efflux and synthesis were studied in rat striatal slices continuously superfused with [3H]tyrosine. The beta-adrenoceptor agonist (-)isoproterenol induced an immediate and significant facilitation of [3H]DA efflux but did not alter [3H]DA synthesis as measured by [3H]H2O formation. In contrast, both dibutyryl cyclic AMP and IBMX enhanced [3H]DA synthesis as well as efflux. The presence of IBMX in the superfusing medium did not potentiate the augmentation of [3H]DA efflux caused by (-)isoproterenol. Additionally, the blockade of [3H]DA synthesis by alpha-methyl-p-tyrosine (10(-4) M) completely prevented the action of dibutyryl cyclic AMP on [3H]DA efflux. However, under similar conditions, (-)isoproterenol was still able to increase [3H]DA efflux. The results suggest that (-)isoproterenol can modify striatal DA release through a mechanism not involving cyclic AMP.


The water extracts of Euonymus alatus (Thunb.) Siebold attenuate diabetic retinopathy by mediating angiogenesis.
Euonymus alatus (Thunb.) Siebold (family Celastraceae) is a deciduous woody shrub that is recorded in ShenNong BenCaoJing. It has been widely used for diabetes in traditional Chinese medicine. This study aimed to identify the most effective extract of Euonymus alatus (EA) against high glucose-induced endothelial cells in vitro, evaluate its pharmacological effect on retinopathy in diabetic mice and explore its underlying mechanism by RNA sequencing. Retinal vascular endothelial cells (RF/6A) were treated with normal glucose (5.5 mmol/L glucose), high glucose (25 mmol/L glucose) or high glucose plus methanol extracts of EA (MEA), ethyl acetate extracts of EA (EEA) or water extracts of EA (WEA). The cytotoxicity and cell viability were determined by Cell Counting Kit-8 (CCK-8) assay. Cell migration was examined using the Transwell assay, and tube formation ability was measured using the Matrigel assay. Then, the KK-Ay mice were administered WEA or water for 12 weeks. The velocities of ocular blood flow were determined by Doppler ultrasound. RNA sequencing and reverse transcription quantitative PCR (RT-qPCR) were performed on WEA-stimulated RF/6A cells to reveal the underlying mechanism. The cytotoxicity assay found that 30 μg/mL MEA, 20 μg/mL EEA and 30 μg/mL WEA had no toxic effect on RF/6A cells. The cell viability results showed that MEA, EEA and WEA all decreased cell viability. Compared with the high-glucose group, both MEA and WEA decreased the number of migrated cells, while the inhibition rate of WEA was higher. The Matrigel results showed that 30 μg/mL WEA effectively reduced the total tube length. Moreover, WEA improved the haemodynamics of the central retinal artery. RNA sequencing coupled with RT-qPCR verified that WEA regulated angiogenesis-related factors in high glucose-stimulated RF/6A cells. WEA inhibits the migration and tube formation of RF/6A cells and improves diabetic retinopathy (DR) by mediating angiogenesis.


Rash: annoying but prognostic.
Targeted therapies are commonly utilised in modern oncology practice. Skin toxicity is a frequently encountered problem associated with them. Some of the skin toxicites carry prognostic significance and it is important to identify them. Though annoying, treatment interruption is seldom required.


Dynamic movement of intracanal gas bubbles during cleaning and shaping procedures: the effect of maintaining apical patency on their presence in the middle and cervical thirds of human root canals-an in vivo study.
The vapor lock effect has been shown to limit the penetration of irrigating solutions into the apical third of root canals both in vivo and in vitro; however, little attention has been given to gas bubbles in the remainder of the root canal system. Seventy-one human root canals were randomly divided into 4 groups. Apical patency was maintained in 2 groups in both small (PS) (n = 21) and big canals (PB) (n = 15) during shaping and cleaning procedures with a 10 K-file 1 mm beyond working length but not in the other 2 groups: no patency, small canals (NPS) (n = 19) and no patency, big canals (NPB) (n = 16). Irrigation was performed with 1 mL of a solution prepared with a radiopaque contrast medium and sodium hypochlorite 5.25%. Digital images were taken, and a calibrated reader determined the presence or absence of gas bubbles in the middle and cervical thirds of the root canal system. When a patency file was not used, 40% of the root canals presented gas bubbles; when a patency file was used, 25% of the cases exhibited gas bubbles. The presence of gas bubbles in the middle and cervical thirds of the root canal system during cleaning and shaping procedures is a common finding regardless of whether a patency file was used, although maintaining apical patency significantly leads to minimizing the presence of gas bubbles in big canals (P < .05).


Estimation of the determinants of transcutaneous oxygen tension using a dynamic computer model.
A dynamic model for oxygen transport within the outer layers of the skin and a tcPO2 sensor has been developed. By comparing model simulations with clinical measurements from adults, it is possible to analyze some of the physiologic and physical determinants of the tcPO2 measurement. The results indicate that the permeability of the epidermis is a significant parameter when using a microcathode sensor. Using this model, the values obtained for normalized stratum papillare blood flow and metabolic oxygen consumption of the dermis are higher than those seen for other models.


How physicians use array comparative genomic hybridization results to guide patient management in children with developmental delay.
Array comparative genomic hybridization is an emerging test used clinically to identify the etiology of children with developmental delay, yet little data are available regarding how physicians use these results. This pilot study evaluated how positive test results were used to influence patient management. We surveyed 14 physicians of 48 patients who had copy number changes detected by microarray technology. Of 48 patients, 34 (70.8%) had 65 management changes after receiving the test result (with individual patients having 1-3 changes). Most commonly, physicians provided patients' families with a recurrence risk for affected subsequent pregnancies (35% of patients). Patients avoided other forms of testing (35%) and had improved access to services (25%). In 27% of patients, physicians altered medical management by referring patients to a specialist or recommending medical screening. Patients with known syndromes had multiple changes, but patients with novel copy number changes also had recommendations made based on the array result. Overall, physicians reported making changes in management among most patients with positive test results, in ways similar to abnormalities detected by conventional cytogenetics. Our study demonstrates that this testing, in our clinical setting, is affecting management of children with developmental delay.


The Association between Taking Dietary Supplements and Healthy Habits among Korean Adults: Results from the Fifth Korea National Health and Nutritional Examination Survey (2010-2012).
Recently, the number of people interested in health in South Korea has increased, and the rate of dietary supplement use is rising. Researchers have hypothesized that the rate of practicing healthy habits is higher among those who use dietary supplements than those who do not. Therefore, this study aimed to discover the association between taking dietary supplements and practicing various healthy habits in the Korean, adult population. The sample included 15,789 adults over 19 years old who participated in the fifth Korea National Health and Nutrition Examination Survey. The user group was defined as those taking dietary supplements for more than 2 weeks during the previous year or once during the past month. Measures for the seven healthy habits were based on those included in the Alameda study and were analyzed accounting for the complex sampling design. The rate of taking dietary supplements was significantly higher in women, middle aged participants, urban residents, those with a higher income, those with a higher education level, and nonsmokers as well as among women with a moderate subjective health status, women who limited their alcohol content, and women with dyslipidemia. In the adjusted analysis, the rate of performing three of the 'Alameda 7' habits-eating breakfast regularly, restricting snacking, and limiting drinking-was higher in the female dietary supplement user group than in the other groups. Women practiced more healthy habits and had a higher dietary supplement intake rate than men. We found that taking dietary supplements in Korean adults is highly associated with demographic and social factors. Taking dietary supplements had a relationship with dietary habits, and there was no significant association between dietary supplement and other healthy habits. Thus in the health clinic, we suggest that taking dietary supplements complements a patient's healthy habits, with the exception of dietary habits, for health promotion.


Thoracoscopic "cut-through" segmentectomy for small-sized lung cancer in a deep central location.
The use of segmentectomy and subsegmentectomy for the management of lung lesions is well established. However, the use of subsegmentectomy for deep seated lesions in the upper lobe is difficult because of sufficient surgical margins. Here, we present a patient whose lung lesion was in a deep central area and at the borders of three segments in the upper lobe of the right lung. We used combined subsegmentectomy (S1b + S3a) video-assisted thoracoscopic surgery for this small-sized lung cancer in a deep central location.


Diphenyl diselenide prevents hepatic alterations induced by paraquat in rats.
This study aimed to investigate the beneficial effect of diphenyl diselenide (PhSe)₂ on paraquat (PQ) induced alterations in rats liver. Adult male Wistar rats received (PhSe)₂ at 10 mg kg(-1), by oral administration (p.o.), during five consecutive days. Twenty-four hours after the last (PhSe)₂ dose, rats received PQ at 15 mg kg(-1), in a single intraperitoneally injection (i.p.). Seventy-two hours after PQ exposure, animals were sacrificed by decapitation for blood and liver samples obtainment. Histological alterations induced by PQ exposure, such as inflammatory cells infiltration and edema, were prevented by (PhSe)₂ administration. Moreover, (PhSe)₂ prevented hepatic lipid peroxidation (LPO) induced by PQ and was effective in reducing the myeloperoxidase (MPO) activity in liver, which was enhanced by PQ exposure. (PhSe)₂ also was effective in protecting against the reduction in ascorbic acid and non-protein thiols (NPSH) levels induced by PQ. The inhibition of glutathione S-transferase (GST) activity, in rats exposed to PQ, was normalized by (PhSe)₂ pre-treatment, whereas the inhibition of catalase (CAT) activity was not prevented by (PhSe)₂. The serum alkaline phosphatase (ALP) inhibition, induced by PQ administration, was also prevented by (PhSe)₂ pre-treatment. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were not modified by PQ and/or (PhSe)₂ administration. Therefore, (PhSe)₂ pre-treatment was effective in protecting against the hepatic alterations induced by PQ in rats. This protective effect can involve the antioxidant and anti-inflammatory properties of (PhSe)₂.


Orange carotenoid protein burrows into the phycobilisome to provide photoprotection.
In cyanobacteria, photoprotection from overexcitation of photochemical centers can be obtained by excitation energy dissipation at the level of the phycobilisome (PBS), the cyanobacterial antenna, induced by the orange carotenoid protein (OCP). A single photoactivated OCP bound to the core of the PBS affords almost total energy dissipation. The precise mechanism of OCP energy dissipation is yet to be fully determined, and one question is how the carotenoid can approach any core phycocyanobilin chromophore at a distance that can promote efficient energy quenching. We have performed intersubunit cross-linking using glutaraldehyde of the OCP and PBS followed by liquid chromatography coupled to tandem mass spectrometry (LC/MS-MS) to identify cross-linked residues. The only residues of the OCP that cross-link with the PBS are situated in the linker region, between the N- and C-terminal domains and a single C-terminal residue. These links have enabled us to construct a model of the site of OCP binding that differs from previous models. We suggest that the N-terminal domain of the OCP burrows tightly into the PBS while leaving the OCP C-terminal domain on the exterior of the complex. Further analysis shows that the position of the small core linker protein ApcC is shifted within the cylinder cavity, serving to stabilize the interaction between the OCP and the PBS. This is confirmed by a ΔApcC mutant. Penetration of the N-terminal domain can bring the OCP carotenoid to within 5-10 Å of core chromophores; however, alteration of the core structure may be the actual source of energy dissipation.


Bringing resistance modulation to methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) strains using a quaternary ammonium compound coupled with zinc oxide nanoparticles.
Nowadays, there are concerns about the inadequacy of new antimicrobials and the rise of antimicrobial resistance. Hence, novel antibacterial agents need to be discovered. In this respect, the use of nanoparticles (NPs) seems promising. Zinc oxide nanoparticles (ZnONPs) are functional and inexpensive NPs that possess antimicrobial characteristics, stability, microbial selectivity, and an easy manufacturing procedure. Imidazolium is one of the quaternary ammonium compounds (QACs) frequently employed as antimicrobial materials in industrial and clinical fields. The present study successfully employed imidazolium to couple with ZnONPs to improve their antimicrobial properties. The antimicrobial activities of ZnONPs doped with imidazolium (IM@ZnONPs) compared to ZnONPs and zinc (Zn) ions against some pathogen microorganism species including Streptococcus aureus (S. aureus), Enterococcus faecalis (E. faecalis), methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and Candida albicans (C. albicans) were evaluated by the microdilution method. The minimum inhibitory concentration (MIC) results revealed that the antimicrobial activities of Zn ions, ZnONPs, and IM@ZnONPs were concentration-dependent. Moreover, we found that the nanoparticulate forms of Zn had considerably stronger antibacterial activities, particularly against VRE and MRSA, compared to Zn ions which failed to restrain the microbial strains at the tested microdilutions of this experiment (MIC: ≥512 µg/mL). Interestingly, the incorporation of imidazolium into ZnONPs resulted in significant inhibition of microbial growth in antimicrobial-resistant pathogens at low concentrations (MIC: 32 µg/mL) and effectively improved the monodispersity of the final coated NPs in terms of size and morphology. To sum up, IM@ZnONPs can be a favorable substitute for conventional antimicrobial agents to combat antimicrobial resistance in many fields, including pharmaceuticals, dental materials, and cosmetic products.


Chocolate consumption modulates cytokine production in healthy individuals.
Epidemiological studies suggest that chocolate increases the incidence and severity of acne. Here we demonstrate that chocolate consumption primes human blood mononuclear cells from volunteers to release more interleukin-1β (IL-1β) and IL-10 upon stimulation with Propionibacterium acne or Staphylcoccus aureus, the two microorganisms involved in the pathogenesis of acne. In contrast, production of the Th17-derived cytokine IL-22 was inhibited by chocolate. Modulation of inflammation could represent an important mechanism through which chocolate consumption influences acne.


[Prognostic value of syncope in the presentation of pulmonary embolism].
Although the prognostic value of syncope has not been specifically addressed, it has generally been considered an indicator of poor prognosis in pulmonary embolism. The objective of this study was to carry out a prospective evaluation of the risk of recurrence and/or death in patients with pulmonary embolism that presents with syncope. A total of 168 patients had a confirmed diagnosis of pulmonary embolism. Twelve were lost to follow up and did not enter statistical analysis. The mean follow-up period was 5 months. The prevalence of syncope in the patients studied was 22%. Of the 34 patients who presented syncope, objectively confirmed recurrence occurred in 2 (5.9%). In the patients who did not present syncope, recurrence was confirmed in 8 (6.6%; P=.8). Death occurred in 2 patients (5.9%) from the group presenting syncope and 15 (12.3%) from the remaining patients in the series (P=.4). The relative risk of recurrence and/or death associated with presentation of syncope was 0.5 (95% confidence interval, 0.2-1.8). A similar risk was obtained following adjustment for the presence or absence of cancer or deep vein thrombosis. Patients with pulmonary embolism that presents with syncope do not have an increased risk of recurrence and/or death.


Possible integration of upstream signals at Cdc42 in filamentous differentiation of S. cerevisiae.
Various environmental stimuli (such as nitrogen starvation, short-chain alcohols and slowed DNA synthesis) induce filamentous differentiation in S. cerevisiae. Genetic mutations (such as deletion of the mitotic cyclin gene CLB2) cause constitutive filamentous differentiation. Although different stimulus-induced filamentous differentiation involves different signalling pathways, Cdc42 has been identified as a common regulator. We show here that Cdc42 is also required for hydroxyurea (HU)-induced and clb2Delta-caused filamentous growth. We show that the mitotic CDK Clb2/Cdc28 functions upstream of Cdc42 in regulating filamentous differentiation. This result points to possible existence of a Cdc42-MAPK-Clb2/Cdc28 positive feedback loop in the signalling of filamentous differentiation. We report isolation of a cdc42-Y40F allele that blocks HU-induced, but not nitrogen starvation-induced, short-chain alcohol-induced or clb2Delta-caused, filamentation. Based on these results, we propose a model in which Cdc42 functions as a possible integrator for the upstream signals of filamentous differentiation (from the filamentous growth MAPK pathway, the cAMP pathway and the Mec1/Rad53 checkpoint pathway). We also show evidence that the mitotic CDK inhibitor Swe1 may mediate the cross-talk between the cAMP and MAPK pathways.


Central nervous system physiology.
This is the second chapter of the series on the use of clinical neurophysiology for the study of movement disorders. It focusses on methods that can be used to probe neural circuits in brain and spinal cord. These include use of spinal and supraspinal reflexes to probe the integrity of transmission in specific pathways; transcranial methods of brain stimulation such as transcranial magnetic stimulation and transcranial direct current stimulation, which activate or modulate (respectively) the activity of populations of central neurones; EEG methods, both in conjunction with brain stimulation or with behavioural measures that record the activity of populations of central neurones; and pure behavioural measures that allow us to build conceptual models of motor control. The methods are discussed mainly in relation to work on healthy individuals. Later chapters will focus specifically on changes caused by pathology.


Phase behavior and structural characteristics of hydrated bovine brain gangliosides.
Hydrated bovine brain gangliosides have been studied by differential scanning calorimetry, X-ray diffraction, and polarized light microscopy. Over the hydration range 18-50 wt.% H2O, mixed brain gangliosides exhibit a hexagonal mesophase structure, in which the ganglioside molecules form hexagonally packed rod-like structures. The apolar lipid chains radiate from the center of the rods, with the sugar groups on the cylinder surface in contact with water. At higher water contents, an isotropic micellar solution is formed. Over the hydration range 20-30 wt.% H2O, two small thermal transitions with peak maxima at 30 degrees C and 46 degrees C are observed by differential scanning calorimetry. These transitions broaden and move apart in temperature as the hydration is increased to 50 wt.% H2O. X-ray diffraction data indicate that this double transition is associated with a hydrocarbon chain rearrangement from a disordered state to another, possibly more disordered, state. Thus, the gangliosides, although membrane lipid components, have physical characteristics which are very different from those of the membrane phospholipids.


Technique of continuous collection of parotid saliva in the rat.
A method is described, which is suitable to collect continuously and quantitatively saliva from both parotid glands in rats under in vivo conditions and which allows to investigate salivary electrolyte composition in relation to flow rate during several hours. This might be of expanding scientific interest in disturbances of transepithelial electrolyte transport in various pathophysiological situations (for instance different forms of experimental hypertension, endstage renal failure and so on).


Tubular Pouch Technique Using Aortic Tissues for Single Coronary Artery Transfer.
A single coronary artery with a single ostium from sinus 2 (2 R L Cx) is rare and difficult to transfer. A 6-day-old male neonate was diagnosed with dextro-transposition of the great arteries with an intact ventricular septum at birth. Computed tomography revealed the presence of a single coronary artery with a single ostium from sinus 2 (2 R L Cx). We performed a new coronary transfer technique involving a tubular pouch using the original aortic tissue for this single coronary artery. The postoperative course was uneventful, and postoperative computed tomography revealed a smooth coronary route with no stenosis.


α1AMP-activated protein kinase mediates vascular protective effects of exercise.
We investigated whether AMP-activated protein kinase (AMPK) may be involved in the signaling processes leading to exercise-mediated vascular protection. The effects of voluntary exercise on AMPK activity, endothelial NO synthase expression and phosphorylation, vascular reactive oxygen species formation, and cell senescence were tested in α1AMPK knockout and corresponding wild-type mice. Exercise significantly improved endothelial function, and increased plasma nitrite production in wild-type mice, associated with an activation of aortic AMPK assessed by its phosphorylation at threonine 172. In addition, regular physical activity resulted in an upregulation of endothelial NO synthase protein, serine 1177 endothelial NO synthase phosphorylation, and an increase of circulating Tie-2(+)Sca-1(+)Flk-1(+) myeloid progenitor cells. All these changes were absent after α1AMPK deletion. In addition, exercise increased the expression of important regulators of the antioxidative defense including heme oxygenase-1 and peroxisome proliferator-activated receptor γ coactivator 1α, decreased aortic reactive oxygen species levels, and prevented endothelial cell senescence in an α1AMPK-dependent manner. Intact α1AMPK signaling is required for the signaling events leading to the manifestation of vascular protective effects during exercise. Pharmacological AMPK activation might be a novel approach in the near future to simulate the beneficial vascular effects of physical activity.


Transcriptomic Analysis of mRNA-lncRNA-miRNA Interactions in Hepatocellular Carcinoma.
Fully elucidating the molecular mechanisms of non-coding RNAs (ncRNAs), including micro RNAs (miRNAs) and long non-coding RNAs (lncRNAs), underlying hepatocarcinogenesis is challenging. We characterized the expression profiles of ncRNAs and constructed a regulatory mRNA-lncRNA-miRNA (MLMI) network based on transcriptome sequencing (RNA-seq) of hepatocellular carcinoma (HCC, n = 9) patients. Of the identified miRNAs (n = 203) and lncRNAs (n = 1,090), we found 16 significantly differentially expressed (DE) miRNAs and three DE lncRNAs. The DE RNAs were highly enriched in 21 functional pathways implicated in HCC (p < 0.05), including p53, MAPK, and NAFLD signaling. Potential pairwise interactions between DE ncRNAs and mRNAs were fully characterized using in silico prediction and experimentally-validated evidence. We for the first time constructed a MLMI network of reciprocal interactions for 16 miRNAs, three lncRNAs, and 253 mRNAs in HCC. The predominant role of MEG3 in the MLMI network was validated by its overexpression in vitro that the expression levels of a proportion of MEG3-targeted miRNAs and mRNAs was changed significantly. Our results suggested that the comprehensive MLMI network synergistically modulated carcinogenesis, and the crosstalk of the network provides a new avenue to accurately describe the molecular mechanisms of hepatocarcinogenesis.


Vividness as the similarity between generated imagery and an internal model.
Vividness in visual mental imagery has been relatively under-explored compared to imagery's representational format and neural mechanisms. In this paper, we take a deeper look at vividness and suggest that in re-framing it, we can potentially reconcile disparate findings regarding visual cortex activation during imagery. Unlike traditional views of vividness that define the concept in terms of perception, we frame vividness in terms of imagery's relation to an internal model; the closer the generated imagery is to this model, the more vivid it is. This view is considered alongside existing neuroscientific, psychological, and philosophical research, as well as directions for future research.


Assessment of sequence homology and cross-reactivity.
Three aspects of allergenicity assessment and are discussed: IgE immunogenicity, IgE cross-reactivity and T cell cross-reactivity, all with emphasis on in-silico predictability: from amino acid sequence via 3D structure to allergenicity. (1) IgE immunogenicity depends to an overwhelming degree on factors other than the protein itself: the context and history of the protein by the time it reaches the immune system. Without specification of these two factors very few foreign proteins can be claimed to be absolutely non-allergenic. Any antigen may be allergenic, particularly if it avoids activation of TH2-suppressive mechanisms (CD8 cells, TH1 cells, other regulatory T cells and regulatory cytokines). (2) IgE cross-reactivity can be much more reliably assessed by a combination of in-silico homology searches and in vitro IgE antibody assays. The in-silico homology search is unlikely to miss potential cross-reactivity with sequenced allergens. So far, no biologically relevant cross-reactivity at the antibody level has been demonstrated between proteins without easily-demonstrable homology. (3) T cell cross-reactivity is much more difficult to predict compared to B cell cross-reactivity, and its effects are more diverse. Yet, pre-existing cross-reactive T cell activity is likely to influence the outcome not only of the immune response, but also of the effector phase of the allergic reaction.


Improving psychotherapeutic medication prescribing in Florida: implementation of the Florida Medicaid Drug Therapy Management Program (MDTMP).
This paper describes a program that was established by Florida Medicaid to improve the quality of prescribing of psychotherapeutic medications. It relates the process used for defining quality medication treatment including the definitions of unusual psychotherapeutic medication indicators (UPMI). It details the results of analysis of FY 2007-2008 pharmacy claims data using these indicators that enabled the Program to identify practices and prescribers that required targeted interventions. The most frequently triggered UPMI for adults involved the use of 2 or more antipsychotics for greater than 60 days; high doses of psychotherapeutic medications was the indicator most frequently triggered for children closely followed by the use of 2 or more antipsychotics for more than 45 days. Prescriptions that triggered UPMI were concentrated in a small number of prescribers. These results led to the Program focusing on these high frequency practices and on the prescribers most associated with them. They also led to the implementation of new quality improvement initiatives like the implementation of a psychiatric telephone consultation line for pediatricians who are treating children with serious emotional disturbances who do not have access to child psychiatrists.


Survey on the contamination of Toxoplasma gondii oocysts in the soil of public parks of Wuhan, China.
Toxoplasma gondii of warm-blooded animals and humans is an important pathogenic agent throughout the world. Soil is increasingly recognized as an important source in the transmission of Toxoplasma. To attain the contamination status of T. gondii in the soil of public parks, a total of 252 soil samples were collected from September 2009 to August 2010 at different sites located in 6 public parks of Wuhan, Hubei, China and detected by PCR and loop-mediated isothermal amplification (LAMP). The detection limit of PCR/B1, PCR/529 and LAMP was determined to be 50, 5, and 5 tachyzoites in soil, respectively. Forty-one samples were found positive for Toxoplasma DNA by PCR on both genes, whereas LAMP products were generated in 58 samples (χ(2)=3.6328, P=0.0567). All parks were found contaminated and no significant difference was found among the parks (PCR: χ(2)=0.0072, P=0.9325; LAMP: χ(2)=0.6101, P=0.4347). However, contamination was found with significantly different among the four seasons (PCR: χ(2)=11.6066, P=0.0007; LAMP: χ(2)=12.4636, P=0.0004), with a gradual decrease in the prevalence from spring to winter on both analyses. This is the first investigation on soil contamination of public parks in China by T. gondii oocysts. The results indicate that the soil of public parks contaminated with T. gondii oocysts may play a role in the epidemiology of toxoplasmosis and effective preventive measures should be considered. Moreover, the conventional PCR and LAMP used in the present study are applicable to detect T. gondii oocysts in soil samples.


Contrast-enhanced voiding urosonography (CEVUS) as a safe alternate means of assessing vesicoureteral reflux in pediatric kidney transplant patients.
Although voiding cystourethrogram (VCUG) is currently the gold standard in VUR evaluation, there is ionizing radiation exposure. Contrast-enhanced voiding urosonography (CEVUS) uses ultrasound contrast agents to visualize the urinary tract and has been reported to be safe and effective in VUR evaluation in children. CEVUS application has yet to be specifically described in VUR evaluation in the pediatric kidney transplant population. The purpose of this study was to report the use of CEVUS and VCUG in evaluating and managing VUR in pediatric renal transplant patients. Retrospective review was conducted for pediatric kidney transplant patients (18 years and younger) who underwent VCUG or CEVUS to assess for transplant VUR from July 2019 through June 2021. Demographic information, reason for VUR evaluation, fluoroscopy time, and postimaging complications were evaluated. Costs of imaging modalities were also considered. Eight patients were evaluated for transplant VUR during the study period. Of the 3 patients who underwent VCUG, all 3 had VUR (median grade 3). Median fluoroscopy time was 18 s and dose-area product was 18.7 uGy*m2 . Of the 5 patients who underwent CEVUS, 4 had VUR (median grade 4). There were no complications for either modality. Based on clinical and radiographic findings, patients were recommended no intervention, behavioral modification, or ureteral reimplantation. The total cost of CEVUS was $800 less than that of VCUG. CEVUS can provide an alternate means of safely evaluating VUR in kidney transplant patients with similar outcomes, potentially lower costs, and no exposure to ionizing radiation.


Folate deficiency in patients seeking treatment of alcohol use disorder.
Nutritional deficiency is frequent in patients with an alcohol use disorder (AUD). We aimed to analyze serum and erythrocyte folate deficiency in a case series of patients that initiated treatment of AUD. A cross-sectional study in patients admitted for detoxification between 2007 and 2015 was performed. Sociodemographic characteristics, history of alcohol consumption, type of alcohol, and medical co-morbidity were assessed at admission. Blood samples for biochemistry and hematological parameters were collected at admission. Logistic regression models were used to establish predictors of folate deficiency. 211 patients (79.1% men) were eligible; age at admission was 46 years [IQR:40-51], and the amount of alcohol consumption was of 160g/day [IQR:120-200]. Thirty four percent of patients had macrocytosis (MCV>100fL), 12.8% had anemia, 23% of cases presented with serum folate deficiency and 7% presented with erythrocyte folate deficiency. Most (69%) of the patients with serum folate deficiency had normal erythrocyte folate levels. In univariate analysis, macrocytosis (OR=3.4, 95%CI:1.7-6.6), alcohol-related liver disease (ARLD) (OR=2.5, 95%CI:1.0-6.1) and drinking alcoholic beverages other than beer (OR=3.3, 95%CI:1.5-7.3) were associated with folate deficiency. However, only macrocytosis was significantly associated with serum folate deficiency in multivariate analysis (OR=3.1, 95%CI:1.1-8.9). Macrocytosis (P<0.001), ARLD (P=0.01) and the type of alcohol consumption (P<0.001) were factors associated with erythrocyte folate deficiency in univariate analysis. In multivariate analysis only macrocytosis remained significantly associated to erythrocyte folate deficiency (P=0.037). Folate deficiency is a relatively frequent finding in contemporary, middle-aged patients with AUD, and macrocytosis is significantly associated with the deficiency.


The Zurich Study--a prospective epidemiological study of depressive, neurotic and psychosomatic syndromes. IV. Recurrent and nonrecurrent brief depression.
How common and how significant are brief depressive episodes (BDE) lasting less than 2 weeks? The authors propose splitting the BDE into two groups: one occurring monthly over 1 year of observation, termed 'recurrent brief depression' (RBD), and those occurring less frequently, labeled 'nonrecurrent brief depression' (NRBD). From a medical point of view, the RBD are a relevant group. Different thresholds of definition are tested, the narrowest of which (including occupational impairment and predetermined minimum number of symptoms) is accepted for 'case'-definition. The such defined RBD (SYM) group differs from major depression only by length and frequency of episodes. In a young cohort, its 1-year prevalence rate was found to be 4.4% (males 3.9%, females 4.9%). One-third of these cases needed treatment, a fourth suffered from pronounced subjective and social impairment as well as from persistent suicidal ideation. The self-reporting of subjective impairment, assessed with the SCL-90 symptom inventory and an analog-rating, yields high scores which are in no way inferior to major depression diagnosed with RDC, DSM-III or EDE (SYM) criteria. The RBD (SYM) demonstrate less hypomania than the major depressive disorders. On the other hand, a family history of depression is equally frequent across all groups. The validity of the RBD (SYM) group has yet to be confirmed by a follow-up study, and further research is needed to delineate it from secondary depression. The findings largely support the hypothesis of a continuum from mild and short to more severe, longer lasting depressive syndromes, but they do not exclude heterogeneity of RBD (Angst and Dobler-Mikola 1984b).(ABSTRACT TRUNCATED AT 250 WORDS)


Mast cell infiltration around gastric cancer cells correlates with tumor angiogenesis and metastasis.
BACKGROUND: Increased numbers of mast cells are found in various solid tumors. To investigate the role of mast cells in the vicinity of gastric cancer cells, we used special staining and an immunohistochemical technique.METHODS: Specimens were surgically obtained from 102 patients with gastric cancer. Mast cells around the tumor edge of gastric cancer nests were counted by staining with 0.05% toluidine blue solution. Blood vessels in these areas were also counted, by immunohistochemical staining of endothelial cells for factor VIII.RESULTS: The average number of mast cells and blood vessels in gastric cancer specimens was significantly higher than that in normal gastric tissue. Specimens from patients with advanced disease with metastases to lymph nodes had more mast cells than specimens from patients with early-stage disease. Mast cells in specimens from patients with metastatic lymph nodes were significantly increased in comparison with numbers in specimens from those without nodal metastases. Mast cell numbers in the specimens of patients with lymphatic or blood vessel invasion were significantly higher than numbers in specimens from patients without such invasion. Mast cells were localized near the new vessels around gastric cancer cells. Mast cell numbers increased as the number of blood vessels increased (correlation coefficient, 0.783). Postoperative survival curves revealed that patients with increased numbers of mast cells had a poor prognosis.CONCLUSIONS: All these results suggest that mast cell accumulation at the tumor site may lead to increased rates of tumor vascularization and, consequently, increased rates of tumor growth and metastasis.


Metabolism of nitroxide spin labels in subcellular fraction of rat liver. I. Reduction by microsomes.
As part of an ongoing study of the role of subcellular fractions on the metabolism of nitroxides, we studied the metabolism of a set of seven nitroxides in microsomes obtained from rat liver. The nitroxides were chosen to provide information on the effects of the type of charge, lipophilicity and the ring on which the nitroxide group is located. Important variables that were studied included adding NADH, adding NADPH, induction of enzymes by intake of phenobarbital and the effects of oxygen. Reduction to nonparamagnetic derivatives and oxidation back to paramagnetic derivatives were measured by electron-spin resonance spectroscopy. In general, the relative rates of reduction of nitroxides were similar to those observed with intact cells, but the effects of the various variables that were studied often differed from those observed in intact cells. The rates of reduction were very slow in the absence of added NADH or NADPH. The relative effect of these two nucleotides changed when animals were fed phenobarbital, and paralleled the levels of NADPH cytochrome c reductase, cytochrome P-450, cytochrome b5 and NADH cytochrome c reductase; results with purified NADPH-cytochrome c reductase were consistent with these results. In microsomes from uninduced animals the rate of reduction was about 10-fold higher in the absence of oxygen. The products of reduction of nitroxides by microsomes were the corresponding hydroxylamines. We conclude that there are significant NADH- and NADPH-dependent paths for reduction of nitroxides by hepatic microsomes, probably involving cytochrome c reductases and not directly involving cytochrome P-450. From this, and from parallel studies now in progress in our laboratory, it seems likely that metabolism by microsomes is an important site of reduction of nitroxides. However, mitochondrial metabolism seems to play an even more important role in intact cells.


Surface Plasmon Resonance Analysis for Quantifying Protein-Carbohydrate Interactions.
During the past two decades, surface plasmon resonance (SPR) analysis has emerged as an important tool for studying protein-carbohydrate interactions, with several commercial instruments available. Binding affinities in the nM to mM range can be determined; however, there are pitfalls that require careful experimental design to avoid. Here we give an overview of each step in the SPR analysis from immobilization to data analysis, providing key points of consideration that will allow practitioners to achieve reliable and reproducible results.


A simple noise clipping circuit for neurophysiological audio monitors.
A very simple circuit is described which removes the low amplitude components of audio frequency signals, leaving the positive and negative peaks superimposed on a flat baseline. The clipping is symmetrical and adjustable in gain. Thus the baseline noise in audio monitors used in extracellular single-unit recording can be attenuated. This enhances the aural discrimination of single units and reduces aural fatigue.


Regioselective C3-H Alkylation of Imidazopyridines with Donor-Acceptor Cyclopropanes.
Alkylated imidazopyridines are crucial structures for medicinal chemistry. Here, an efficient method for the C3-H alkylation of imidazopyridines was devised. Under Lewis acid-catalyzed conditions, reactions of imidazopyridines with different donor-acceptor cyclopropanes were carried out. Finally, 1,3-bisfunctionalizaton of donor-acceptor cyclopropanes was performed. In addition, synthesized C3-alkylated imidazopyridines are amenable for diverse synthetic applications.


PROTAC targeting cyclophilin A controls virus-induced cytokine storm.
Cytokine storms caused by viruses are associated with elevated cytokine levels and uncontrolled inflammatory responses that can lead to acute respiratory distress syndrome. Current antiviral therapies are not sufficient to prevent or treat these complications. Cyclophilin A (CypA) is a key factor that regulates the production of multiple cytokines and could be a potential therapeutic target for cytokine storms. Here, three proteolysis targeting chimeras (PROTACs) targeting CypA were designed. These PROTACs bind to CypA, enhance its ubiquitination, and promote its degradation in both cell lines and mouse organs. During influenza B virus (IBV) infection, PROTAC-mediated CypA depletion reduces P65 phosphorylation and NF-κB-mediated proinflammatory cytokine production in A549 cells. Moreover, Comp-K targeting CypA suppresses excessive secretion of proinflammatory cytokines in bronchoalveolar lavage fluid, reduces lung injury, and enhances survival rates of IBV-infected mice. Collectively, we provide PROTACs targeting CypA, which are potential candidates for the control of cytokine storms.


Successful endovascular embolization of the common hepatic artery for pseudoaneurysm associated with pancreatic fistula after liver transplantation: a case report.
After orthotopic liver transplantation (OLT), complications such as hepatic artery stenosis, thrombosis, and bleeding are possible. Hepatic artery pseudoaneurysms (HAP) are prone to rupture, rupture hemorrhage, and increased mortality risk. Endovascular treatment of HAP may result in recurrence, even after successful embolization with thrombin. Formation of a HAP in the common hepatic artery (CHA) is challenging because the CHA is the only artery in the liver graft after OLT. Therefore, CHA embolization in HAP is not an initial option. We report a case of HAP at the CHA after OLT that was treated with endovascular therapy, resulting in the occlusion of the CHA with coil embolization, achieving a radical cure. A 59-year-old man with decompensated hepatitis C virus cirrhosis underwent deceased donor whole-liver transplantation after graft failure of a living donor liver transplantation. After the second transplantation, the patient developed infectious narrow-necked HAP at the CHA associated with postoperative pancreatic fistula. Repeated transcatheter arterial embolization with thrombin and n-butyl-2-cyanoacrylate was unsuccessful, as confirmed by postprocedure angiography, which revealed recanalization and regrowth of the HAP. Eight months after the first transcatheter arterial embolization, the patient presented with a chief complaint of abdominal pain due to an enlarged HAP. Angiography of the superior mesenteric artery (SMA) revealed a collateral bypass around the bile duct from the SMA to the liver graft. Coil embolization of the HAP in the CHA completely occluded the HAP without complications. More than 2 years after coil embolization, the liver graft function test results remained within normal limits without HAP recurrence. HAP at the CHA after liver transplantation can be fatal if ruptured. Because the liver is a highly angiogenic organ, even if initial treatment is not successful, radical treatment to occlude the CHA with HAP is possible if sufficient collateral vessels are developed.


Enhancement of heavy metal phytoremediation by Alnus firma with endophytic Bacillus thuringiensis GDB-1.
Phytoremediation shows potential for remediating mine tailing sites contaminated with heavy metals. Our aim was to isolate, characterize, and assess the potential of endophytic bacteria to enhance growth and metal accumulation by the hyperaccumulator Alnus firma. A bacterial strain isolated from roots of Pinus sylvestris had the capacity to remove heavy metals from mine tailing and was identified as Bacillus thuringiensis GDB-1 based on 16S ribosomal DNA sequencing. GDB-1 exhibited plant growth-promoting traits, including 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, indole acetic acid (IAA) and siderophore production, and P solubilization. The efficiency of GDB-1 to remove heavy metals was influenced by pH and initial metal concentration. Removal capacity (mg/l) was 77% for Pb (100), 64% for Zn (50), 34% for As (50), 9% for Cd (10), 8% for Cu (10), and 8% for Ni (10) during the active growth cycle in heavy metal-amended, mine tailing extract medium. Inoculating soil with GDB-1 significantly increased biomass, chlorophyll content, nodule number, and heavy metal (As, Cu, Pb, Ni, and Zn) accumulation in A. firma seedlings. Results indicate that inoculating the native plant A. firma with B. thuringiensis GDB-1 improves its efficiency for phytoremediation of soil containing mine tailings contaminated with heavy metals.


A novel mutation af Cln3 associated with delayed-classic juvenile ceroid lipofuscinois and autophagic vacuolar myopathy.
Juvenile neuronal-ceroid-lipofuscinosis (JNCL) is a lysosomal storage disease caused by mutations in CLN3. The most frequent mutation is a 1.02-kb deletion that, when homozygous, causes the classical clinical presentation. Patients harboring mutations different than the major deletion show a marked clinical heterogeneity, including protracted disease course with possible involvement of extraneuronal tissues. Cardiac involvement is relatively rare in JNCL and it is usually due to myocardial storage of ceroid-lipofuscinin. Only recently, histopathological findings of autophagic vacuolar myopathy (AVM) were detected in JNCL patients with severe cardiomyopathy. We describe a 35-year-old male showing a delayed-classic JNCL with visual loss in childhood and neurological manifestations only appearing in adult life. He had an unusual CLN3 genotype with an unreported deletion (p.Ala349_Leu350del) and the known p.His315Glnfs*67 mutation. Autophagic vacuolar myopathy was shown by muscle biopsy. At clinical follow-up, moderately increased CPK levels were detected whereas periodic cardiac assessments have been normal to date. Adult neurologists should be aware of protracted JNCL as cause of progressive neurological decline in adults. The occurrence of autophagic vacuolar myopathy necessitates periodic cardiac surveillance, which is not usually an issue in classic JNCL due to early neurological death.


Treatment of heavy menstrual bleeding of endometrial origin: randomized controlled trial of medroxyprogesterone acetate and tranexamic acid.
This study aimed at comparing the efficacy of medroxyprogesterone acetate (MPA) and tranexamic acid (TA) for treating heavy menstrual bleeding of endometrial origin (HMB). A randomized controlled trial was carried out in three gynecology clinics in Tehran, Iran. Ninety women with the HMB of endometrial origin were randomized into the study: 44 patients took MPA for 21 days from day 5 and 46 patients took tranexamic acid for 5 days from day 1 of menses for three consecutive menstrual cycles. Blood loss was measured using the pictorial blood loss assessment chart (PBAC); hematological assessments were made before intervention and after treatment. SF-36 and HMB Questionnaire (MQ) were given to assess quality of life. Statistical analysis was performed using t test, Paired t test, χ(2), Mann-Whitney, Wilcoxon signed-rank test, and repeated measure analysis. PBLC mean score, duration of bleeding and Hb values as well as quality of life were significantly improved in both groups (P < 0.05). But there was no significant deference between groups. More drug complication and less satisfaction were reported by MPA group (P = 0.003 and P = 0.002, respectively). Long-term use of MPA is as effective as Tranexamic acid in treating HMB and increasing quality of life. However, bleeding irregularity side effects of MPA might limit its use.


Spectrophotometric determination of cimetidine in pharmaceuticals and urine using batch and flow-injection methods.
Two sensitive and fast spectrophotometric methods using batch and flow-injection procedures for the determination of cimetidine (CMT) are proposed. The methods are based on the formation of a green complex between this drug and Cu(II) in acetic/acetate medium of pH 5.9. The calibration graphs resulting from measuring the absorbance at 330 nm are linear over the ranges 2.5 x 10(-6)-1.0 x 10(-3) and 5 x 10(-6)-2.0 x 10(-3) M with detection limits of 9.5 x 10(-7) and 2.1 x 10(-6) for batch and flow-injection methods, respectively. The methods are applied to the routine analysis of CMT in pharmaceuticals and human urine.


T8 antigen density on peripheral blood lymphocytes remains unchanged during exacerbations of multiple sclerosis.
Fluorescence-activated cell sorter (FACS) analysis was used to quantitate the antigen density of the lymphocyte markers T3, T4, T8, and Leu 7 on the surface of peripheral blood mononuclear cells (MNC) from exacerbating/remitting multiple sclerosis (MS) patients. Serial studies showed that the mean fluoresce intensity for all the markers studied did not change significantly during exacerbations. There was a tendency for a generalized decline in T-cells, evidenced by a drop in both T8+- and T4+-cells; however, these changes were not statistically significant. It appears, therefore, that the lymphocyte markers studied here are not useful as markers of disease activity in exacerbating/remitting MS.


[Testing pararosanilines and fuchsins histophotometrically for their usefulness in the quantitative PAS-reaction (author's transl].
Seven pararosanilines and nine fuchsins of different sources have been compared histophotometrically to "Pararosanilin acridinfrei" (Bayer). With respect to the quantitative evaluation of the PAS-reaction three problems are investigated: 1) Wavelength of extinction maxima of the reaction products; 2) Influence of the disulfite concentration; 3) Possibility to differentiate between long time and short time oxidized liver sections. The reaction products of the tested reagents are not comparable with the control substance. Partly there are many reaction products with different maxima. In all reaction products, except Pararosaniline Chloride (Gurr), the extinction maxima are shifting with variations in the disulfite concentration. The difference of the oxidation time is reproduced by five substances half as good as by the control substance, some substances reproduce the difference worse, and some substances do not differentiate in any way between different oxidation times. It is impossible to compare the results of PAS-reactions quantitatively using different colouring substances.


Neuropilin 2 Is a Novel Regulator of Distal Colon Contractility.
Appropriate coordination of smooth muscle contraction and relaxation is essential for normal colonic motility. The impact of perturbed motility ranges from moderate, in conditions such as colitis, to potentially fatal in the case of pseudo-obstruction. The mechanisms underlying aberrant motility and the extent to which they can be targeted pharmacologically are incompletely understood. This study identified colonic smooth muscle as a major site of expression of neuropilin 2 (Nrp2) in mice and humans. Mice with inducible smooth muscle-specific knockout of Nrp2 had an increase in evoked contraction of colonic rings in response to carbachol at 1 and 4 weeks following initiation of deletion. KCl-induced contractions were also increased at 4 weeks. Colonic motility was similarly enhanced, as evidenced by faster bead expulsion in Nrp2-deleted mice versus Nrp2-intact controls. In length-tension analysis of the distal colon, passive tension was similar in Nrp2-deficient and Nrp2-intact mice, but at low strains, active stiffness was greater in Nrp2-deficient animals. Consistent with the findings in conditional Nrp2 mice, Nrp2-null mice showed increased contractility in response to carbachol and KCl. Evaluation of selected proteins implicated in smooth muscle contraction revealed no significant differences in the level of α-smooth muscle actin, myosin light chain, calponin, or RhoA. Together, these findings identify Nrp2 as a novel regulator of colonic contractility that may be targetable in conditions characterized by dysmotility.


Substrate selectivity APPLies to Akt.
The protein kinase Akt occupies a central position in multiple signaling pathways. Although numerous Akt substrates have been identified, less is known about the factors that regulate specific cellular responses to Akt signaling. In this issue, Schenck et al. (2008) demonstrate that the endosomal protein Appl1 modulates Akt's substrate selectivity to promote cell survival during zebrafish development.


Autologous Single Semitendinosus Anatomical Posterolateral Corner Reconstruction With Adjustable-Loop Cortical Suspension Devices.
We present our surgical technique for the reconstruction of the posterolateral corner of the knee. It is a tibia- and fibular-based reconstruction technique. Most of these procedures require the use of 2 tendons (autograft or allograft). In our technique, a single semitendinosus tendon is required, making the procedure more suitable if the surgeon prefers the use of autograft or when there is no access to a tissue bank. This is even more important in the setting of multiligament knee injuries. The most defining feature of this modification is the possibility of achieving the desired graft tension in a progressive and independent way, due to the use of 3 adjustable-loop cortical suspension devices.


Effect of repeated application of fenthion as a mosquito larvicide on Nile tilapia (Oreochromis niloticus) inhabiting selected water canals in Sri Lanka.
Health status of feral Nile tilapia following repeated applications of fenthion as a mosquito larvicide to selected water canals in Sri Lanka was assessed. With three spray applications of fenthion to the study sites at weekly intervals at the concentration recommended for mosquito control, condition factor and brain acetylcholinesterase activity of the fish were depressed in a time dependent manner. Prominent histopathological alterations displayed were gill hyperplasia and telangiectasis and vacoulation of hepatocytes. Observed ill health effects of fenthion on the fish demonstrate probable ecological risk to the fish populations inhabiting the water canals which receive repeated inputs of fenthion.


Gene regulation by O2 deprivation: an anoxia-regulated novel gene in Drosophila melanogaster.
Organisms, across the animal kingdom, vary in their tolerance or susceptibility to cell injury from O2 deprivation. In this study we have taken advantage of the genetically well studied fruit fly to dissect basic mechanisms underlying their ability to tolerate lack of O2. Using differential display and molecular techniques, we cloned and characterized a novel gene, named fau, which is up-regulated considerably following anoxia in Drosophila melanogaster. Northern blot analysis revealed that the transcript of this gene is approximately 0.9 kb in length with an open reading frame encoding a small hydrophilic protein ( approximately 14.4 kDa). This protein has no homology to previously described gene products but has many potential phosphorylation sites. In situ hybridization showed that this gene is located in region 7C-D on the Drosophila X-chromosome and its transcript concentrated in the lamina and cortical neurons of the Drosophila central nervous system (CNS). Transgenic flies showed that over-expression of fau significantly reduced the recovery time of the flies from anoxia. We conclude that (1) this study provided a framework on which the mechanisms underlying anoxia tolerance can be dissected in the fruit fly and (2) fau gene plays an important role in the regulation of tissue responsiveness to O2 deprivation.


Attitudes of general practitioners and midwives towards ethnicity-based haemoglobinopathy-carrier screening.
Haemoglobinopathies (HbP) are severe autosomal recessive disorders with high prevalence among certain ethnic groups. World Health Organisation (WHO) advises implementing screening programmes for risk groups. Research in the Netherlands has shown that general practitioners and midwives do not perceive ethnicity as a risk factor for HbP. Moreover, registration of ethnicity is a controversial societal issue, which may complicate the introduction of a national preconception or antenatal carrier screening programme. This study investigates attitudes, intention and behaviour of general practitioners and midwives towards ethnicity-based HbP-carrier screening in general. A structured questionnaire based on the Theory of Planned Behaviour was sent by mail to a random selection of 2100 general practitioners and 1800 primary care midwives. Response was 35% (midwives 44.2%; GPs 27.6%). Although 45% of respondents thought that offering a carrier test on the basis of ethnicity alone should become national policy, it is currently not carried out. The main factor explaining lack of intention towards ethnicity-based HbP-carrier screening was subjective norm, the perception that their peers do not think they should offer screening (52.2% variance explained). If ethnicity-based HbP-carrier screening would become national policy, most professionals report that they would carry this out. Most respondents favoured ethnicity registration for health purposes. As most practitioners look for role models among peers, debate among general practitioners and midwives should be encouraged when new policy is to be developed, articulating the voices of colleagues who already actively offer HbP-carrier screening. Moreover, primary care professionals and professional organisations need support of policy at national level.