Oxidative stress induces loss of pericyte coverage and vascular instability in PGC-1α-deficient mice.
Peroxisome proliferator-activated receptor γ co-activator 1α (PGC-1α) is a regulator of mitochondrial oxidative metabolism and reactive oxygen species (ROS) homeostasis that is known to be inactivated in diabetic subjects. This study aimed to investigate the contribution of PGC-1α inactivation to the development of oxygen-induced retinopathy. We analyzed retinal vascular development in PGC-1α(-/-) mice. Retinal vasculature of PGC-1α(-/-) mice showed reduced pericyte coverage, a de-structured vascular plexus, and low perfusion. Exposure of PGC-1α(-/-) mice to hyperoxia during retinal vascular development exacerbated these vascular abnormalities, with extensive retinal hemorrhaging and highly unstructured areas as compared with wild-type mice. Structural analysis demonstrated a reduction in membrane-bound VE-cadherin, which was suggestive of defective intercellular junctions. Interestingly, PGC-1α(-/-) retinas showed a constitutive activation of the VEGF-A signaling pathway. This phenotype could be partially reversed by antioxidant administration, indicating that elevated production of ROS in the absence of PGC-1α could be a relevant factor in the alteration of the VEGF-A signaling pathway. Collectively, our findings suggest that PGC-1α control of ROS homeostasis plays an important role in the regulation of de novo angiogenesis and is required for vascular stability.


Testosterone metabolism in patients with advanced carcinoma of the prostate: a comparative in vivo study of the effects of oestrogen and antiprolactin.
In the light of the high incidence of cardiovascular side effects with oestrogen therapy in patients with prostatic cancer, other medications altering androgen metabolism are under investigation. The influence of the anti-prolactin bromocriptin (CB157) on plasma kinetics of testosterone and on endogenous hormones was studied and compared with the effect of ethinyl oestradiol in 25 patients with prostatic carcinoma. Bromocriptine significantly suppressed both prolactin and testosterone, inhibited the transfer of androgen from the inner pool into the deep compartment and favoured its degradation. Ethinyl oestradiol decreased testosterone, LH and FSH, and prolonged the biological half-life of testosterone. The effects of bromocriptine on androgen metabolism might be of therapeutic value in patients with prostatic carcinoma.


Low-frequency earthquakes in Shikoku, Japan, and their relationship to episodic tremor and slip.
Non-volcanic seismic tremor was discovered in the Nankai trough subduction zone in southwest Japan and subsequently identified in the Cascadia subduction zone. In both locations, tremor is observed to coincide temporally with large, slow slip events on the plate interface downdip of the seismogenic zone. The relationship between tremor and aseismic slip remains uncertain, however, largely owing to difficulty in constraining the source depth of tremor. In southwest Japan, a high quality borehole seismic network allows identification of coherent S-wave (and sometimes P-wave) arrivals within the tremor, whose sources are classified as low-frequency earthquakes. As low-frequency earthquakes comprise at least a portion of tremor, understanding their mechanism is critical to understanding tremor as a whole. Here, we provide strong evidence that these earthquakes occur on the plate interface, coincident with the inferred zone of slow slip. The locations and characteristics of these events suggest that they are generated by shear slip during otherwise aseismic transients, rather than by fluid flow. High pore-fluid pressure in the immediate vicinity, as implied by our estimates of seismic P- and S-wave speeds, may act to promote this transient mode of failure. Low-frequency earthquakes could potentially contribute to seismic hazard forecasting by providing a new means to monitor slow slip at depth.


Perangiographic rupture of intracranial aneurysms.
The reports of perangiographic rupture of intracranial aneurysm are very few, but the actual incidence is doubtless higher than reported. We could find only 49 cases in the literature, all of which were recognized by extravasation of contrast media. Three of our own cases are documented; in all of them cerebral angiography was carried out soon after subarachnoid hemorrhages. The danger of cerebral angiography in patients with a recently hemorrhaged intracranial aneurysm is stressed.


Stable expression of human cytochrome P450IA2 cDNA in a human lymphoblastoid cell line: role of the enzyme in the metabolic activation of aflatoxin B1.
A human lymphoblastoid cell line stably expressing a human cytochrome P450IA2 cDNA was developed. This recombinant cell line displayed P450IA2 protein and estradiol 2-hydroxylase activity, neither of which was detected in the parental cell line. The recombinant cell line was also approximately 1000-fold more sensitive to the cytotoxicity and mutagenicity of the carcinogenic mycotoxin aflatoxin B1 than was the parent cell line. The increase in mutagenicity was supported by a corresponding increase in the level of aflatoxin B1 binding to DNA in cells expressing P450IA2 relative to control cells.


Virulence aspects of Listeria monocytogenes LO28 high pressure-resistant variants.
High pressure treatment is a novel food processing approach for reducing pathogens in foods and food ingredients. However, relatively little is known about the pathogenic potential of organisms that survive the treatment. Twelve previously isolated and characterized variants of Listeria monocytogenes LO28 obtained after a high pressure treatment were assessed for their virulence potential and antibiotic susceptibility. Ten variants showed attenuated virulence while two variants retained full virulence in a mouse model of infection. Seven of the attenuated variants demonstrated a reduction in virulence factor activity. Compared to the wild type, all variants exhibited similar or increased susceptibility to multiple antibiotics commonly used in listeriosis treatment.


A study of the hearing ability of horses.
The ability of 10 horses to hear frequencies between 14 and 25 Kc/s was tested. The horses appeared to perceive ultrasounds by showing either fright reactions or Pryer reflexes to all of the 12 frequencies. The highest frequencies were heard less by older animals, and elicited more reactions in geldings than in mares.


Medial preoptic and hypothalamic neuronal activity during sexual behavior of the male monkey.
Neuronal activity changes in the medial preoptic area of the male monkey were related to the commencement of sexual behavior, penile erection and the refractory period following ejaculation. Increased neuronal activity in the dorsomedial hypothalamic nucleus was found to be synchronized to each mating act. The involvement of medial preoptic neurons in sexual arousal, initial penile erection and that of dorsomedial hypothalamic neurons in the copulatory act are suggested by the present findings.


Sorption of uranium from aqueous solutions using palm-shell-based adsorbents: a kinetic and equilibrium study.
In this study adsorbents based on palm shell powder as well as modified and activated palm shell powder were studied to analyze their behavior in sorbing U(6+) by both batch and fixed column modes. Seven different two-parameter isotherm models were applied to the experimental data to predict the sorption isotherms. The ΔG(0) values from Langmuir and thermodynamic calculations indicate physisorption as the major mechanism for adsorption of uranium. Usefulness of various kinetic models like pseudo first order, pseudo second order, intraparticle diffusion, Bangham, Elovich and Liquid film diffusion were tested. The adsorption capacities were found to be greater than 200 mg/g for all the adsorbents under study. The column data were fitted by Thomas, Yoon and Nelson as well as Wolborska models. The Thomas and Yoon and Nelson models were best to fit the breakthrough curves under the experimental conditions studied.


Inter-rater reliability of four validated diabetic foot ulcer classification systems.
The need to classify, score and describe diabetic foot ulcerations is needed for accurate clinical documentation. An agreed wound classification can improve communication between health care professionals, and improve ease of discussion regarding potential outcomes. This study aimed to determine the inter-rater reliability between four validated wound classification systems. A prospective non-experimental comparative study was conducted. Forty patients with diabetes presenting with a new or long-standing ulceration were recruited. Each ulcer was graded/scored by three clinicians using four widely used classification systems; namely the University of Texas, SINBAD, Meggitt-Wagner and the PEDIS scoring systems. All classifications had a satisfactory inter-rater agreement when evaluated by three raters of various clinical experience, however, the strength of the agreement varied between classifications. The Meggitt-Wagner system had an almost perfect agreement, the SINBAD and UT systems had a strong inter-rater agreement whilst the PEDIS had a moderate inter-rater agreement. Results of study serve as assurance and provides the evidence that these systems may be reliably used by different clinicians, even those with varied experience in wound care.


Structural basis for HLA-A2 supertypes.
The human leukocyte antigen (HLA) alleles are extremely polymorphic among ethnic population, and the peptide-binding specificity varies for different alleles in a combinatorial manner. However, it has been suggested that majority of alleles can be covered within few HLA supertypes, where different members of a supertype bind similar peptides, yet exhibiting distinct repertoires. Nonetheless, the structural basis for HLA supertype-like function is not clearly known. Here, we use structural data to explain the molecular basis for HLA-A2 supertypes.


Conformational analysis in solution of protein kinase C betaII V5-1 peptide.
One of protein kinase C (PKC) isozymes, PKC beta binds to receptor for activated C kinase 1 (RACK1), and their complex is suggested to be translocated to melanosomes. The binding site of PKC beta for RACK1 is considered one of its catalytic domains, V5 domain which consists of three motifs such as V5-1, V5-2, and V5-3. Among these, V5-1 region, extreme C-terminal residues of PKC beta showed the highest RACK1-binding affinity. PKC beta can be classified into PKC betaI and PKC betaII based on their different V5 domains. RACK1-binding affinity of PKC betaII is five times greater than that of PKC betaI. The structures of PKC betaI, PKC betaII, and RACK1 are not known. However, the conformational study on PKC betaII V5-1 region showing high RACK1-binding selectivity may help us in understanding the interaction between RACK1 and PKC betaII.


Increasing trend of spina bifida and decreasing birth weight in relation to declining body-mass index of young women in Japan.
We analyzed the existing data from various surveys on the prevalence of spina bifida, birth weight, and body-mass index (BMI), dietary intake, and smoking habits of Japanese young women. We found that the increasing trend of the prevalence of spina bifida paralleled with the decrease in birth weight, and the decrease in BMI and dietary energy intake and the increase in smoking rate among women of childbearing age. The decreased energy intake is likely due to their distorted self-body image. Based on these findings, we hypothesize that lowering BMI in young women led to increasing prevalence of spina bifida and smaller babies in Japan. This low dietary intake possibly leads to poor folate nutritional status, a risk factor for the development of spina bifida, and increased smoking potentially accelerates these undesirable outcomes. Our hypothesis can be tested using two simultaneous approaches. The first step is an extensive educational campaign for young women to stop smoking, have a realistic body image and eat a sufficient balanced diet. Government officials and health-care workers are encouraged to establish an agenda to educate young women with emphasis on the importance of adequate nutrition during the critical period of reproduction. Together with this campaign, the distribution of folic acid tablets for the prevention of spina bifida can be tried in women of childbearing age through local health centers, which have been well established in Japan.


Laparoscopic versus open appendicectomy in pregnancy: experience from a single institution and meta-analysis.
Acute appendicitis remains an uncommon cause of non-obstetric abdominal pain during pregnancy, with surgery being the preferred management option. We examined our experience with the surgical management of appendicitis during pregnancy, particularly the risk of foetal loss during the 1st and 2nd trimester and performed a meta-analysis of the available literature. We performed a retrospective analysis of all patients who had an appendicectomy during pregnancy (January 2010 to December 2019) and a meta-analysis comparing foetal death in open appendicectomy versus laparoscopic appendicectomy during the 1st and 2nd trimester. Seventy pregnant patients were included in our study (57 laparoscopic, 13 open). There were 4 foetal deaths during the study period (7%), all of which occurred after the laparoscopic approach (P-value = 0.578). Open appendicectomies were associated with an increased risk of pre-term delivery (P-value = 0.038). The meta-analysis of 9 studies, which included 311 patients, showed that there was no significant difference between OA and LA in foetal deaths during the 1st and 2nd trimesters (1st trimester foetal deaths: 9/143 laparoscopic versus 4/57 open, M-H risk difference-0.02, 95% CI, -0.14 to 0.10): 2nd trimester foetal deaths: 7/159 laparoscopic versus 2/154 Open, M-H risk difference 0.03, 95% CI, -0.02 to 0.09). Our findings suggest there is no increased risk of foetal loss in pregnant patients undergoing a laparoscopic appendicectomy.


Biochemical and ultrasonic abnormalities of the pancreas in anorexia nervosa.
Seven of 10 patients with anorexia nervosa had ultrasonic and/or biochemical abnormalities of the pancreas. Seven patients had elevated amylase creatinine clearance ratios (greater than 4%), three patients had elevated serum amylase values (greater than 90 units/liter), and three patients had reduced echogenicity of the pancreas. There was no consistent association between presenting abdominal symptoms and abnormal ultrasonic and biochemical studies of the pancreas. After nutritional repletion, all studies reverted to normal. An eleventh patient, who was initially diagnosed as having anorexia nervosa but later found to have an astrocytoma of the medulla, had reduced echogenicity of the pancreas, suggesting malnutrition as the cause of these abnormal pancreatic studies. Pancreatic abnormalities due to protein-calorie malnutrition may be common in anorexia nervosa and must be differentiated from primary pancreatitis.


Something in the Blood? A history of the autoimmune hypothesis regarding myasthenia gravis.
From the first descriptions of myasthenia gravis (MG) in the late nineteenth century, speculation about the cause of MG has centered on the possibility of some curare-like factor circulating in the blood. The transfer of transient myasthenic symptoms from a myasthenic mother to her newborn reinforced this speculation. However, it was not until 1960, when William Nastuk and coworkers noted that serum complement correlated with the clinical course in MG, and Arthur Strauss and colleagues described antiskeletal muscle antibodies in the sera of some MG patients, that a paradigm shift occurred from prior exclusive focus on the neuromuscular junction to a broader consideration of the relevance of immunological mechanisms in myasthenia. These findings coincided with an even greater scientific revolution pioneered by Macfarlane Burnet towards cell-mediated and away from chemical immunology. The dominant immunological question of the decade 1955-1965, however, was whether human autoimmune diseases actually existed. During the next decade, 1965-1975, various diseases were accepted as being autoimmune in character, and although comparatively rare, MG became prominent among them because of a known antigen, the acetylcholine receptor, and an excellent experimental model.


Metastases-related genes in the classification of liver and peritoneal metastasis in human gastric cancer.
With the aim of identifying metastases-related genes in gastric cancer, we performed a broad analysis of differential gene expression between low-metastatic parental cell lines and established highly metastatic sublines. We established novel cell lines, AZ-H5c, NUGC-3H5, and TMK-1H7, with a high potential of liver metastasis, and AZ-P7a, NUGC-3P4T, and TMK-1P4a, with a high potential of peritoneal metastasis. These cell lines were derived from low-metastatic parental AZ-521, NUGC-3, and TMK-1 cell lines, respectively. Furthermore, to investigate different levels of gene expression implicated in metastatic potentials in gastric cancer, we investigated approximately 2000 expressed genes in each cell line using a DNA microarray. Varieties of genes were up-regulated or down-regulated in highly metastatic liver and peritoneal cell lines. Fifty-eight genes, including the transferrin receptor, ras-related rho, and osteopontin, and 22 genes, including apolipoprotein E and inhibin A-submit, were up-regulated and down-regulated in two or three liver metastatic sublines. On the other hand, 19 genes, the transferrin receptor, c-fos, and RANTES, and 26 genes, including MAC25, PISSLRE, and RNA polymerase, were up-regulated and down-regulated in two or three peritoneal metastatic sublines. How gene expression is implicated in gastric cancer metastasis has never been thoroughly explained, and further studies are necessary to understand the involvement of genes in cancer metastasis more thoroughly. We hope that our highly metastatic liver and peritoneal experimental models are helpful for further study and gene therapy of human gastric cancer.


Polygenic control of quantitative antibody responsiveness: restrictions of the multispecific effect related to the selection antigen.
Among the differences observed between the various high (H) and low (L) antibody responder lines of mice resulting from distinct bidirectional selective breedings, one of the most puzzling is the variation in the "multispecific effect," i.e., in the modification of antibody responses to antigens unrelated to those used during the selection. The best examples are the H and L lines of selection IV, selected on the basis of responses to somatic antigen of Salmonella which do not differ in their antibody responses to sheep erythrocytes (SE). However, a wide range of variability is observed in the responses of (HIV X LIV)F2 hybrids to this antigen, and it was therefore hypothesized that distinct groups of genes might regulate antibody responses to SE and the somatic antigen. Indeed, a new selection (IV-A) for anti-SE responsiveness started from these (HIV X LIV)F2 successfully produced a high and a low anti-SE responder line. The results of selection IV-A and the variance analysis of (HIV-A X LIV-A)F2 hybrids are reported. They are roughly similar to those in selection I, also carried out for anti-SE responsiveness. In vivo attempts to identify the major regulatory mechanism which contributes to the interline difference indicate that the efficiency of macrophage accessory function has been modified in selection IV-A, as was observed in selection I, whereas this function did not differ in HIV and LIV lines. Probably in relation to the involvement of macrophage function there is a notable increase of the multispecific effect in selection IV-A when compared with selection IV. The results of selection IV-A demonstrate that responsiveness to heterologous erythrocytes and to somatic antigen of Salmonella are under separate polygenic control operating through distinct regulatory mechanisms. The choice of the selection antigen and immunization procedure is of major importance for defining the gene interaction operating in each selective breeding experiment and the extent of its multispecific effect.


Zizyphus oenophlia: a potent substrate for lactic acid production.
The objective of the present investigation deals with optimization of L-lactic acid production from Zizyphus oenophlia, a potential low cost substrate. Central composite design (CCD) based on response surface methodology (RSM) was employed as a statistical tool to investigate the effect of substrate: media ratio, temperature, pH and inoculum volume on lactic acid production. Under optimum condition, the experimental yield was 95.09%, which matched well with the predictive yield of 95.45%. HPLC and circular dichroism (CD) analysis of the fermented product was carried out to confirm the presence of lactic acid in its L (+) seriospecificity respectively. For application of lactic acid in biotechnological sector polymerization studies were carried out. The PLA upon recovery through direct polycondensation reaction resulted in the yield of 85%.


Fulminant Anti-GAD antibody encephalitis presenting with status epilepticus requiring aggressive immunosuppression.
Antibodies against glutamic acid decarboxylase (GAD) are reported in association with numerous neurological conditions including temporal lobe epilepsy and limbic encephalitis. We report a case of Anti-GAD-Antibody associated encephalitis presenting with epilepsia partialis continua (EPC) progressing to a fulminant encephalopathy preferentially affecting the frontal lobes associated with coma and refractory status epilepticus. The abnormalities identified on MRI included marked bilateral frontal lobe involvement which has not been reported in other auto-immune encephalitides and may be specific for Anti-GAD-Antibody associated encephalitis. Similar to the majority of cases of Anti-GAD associated neurological disturbance no underlying malignancy was identified. Treatment with high dose corticosteriods, IVIG and plasmapheresis had minimal response, but escalation of treatment with rituximab and cyclophosphamide was associated with clinical improvement, reducing antibody titers and resolution of MRI changes.


Safety of Contrast Material Use in Children.
Iodinated and gadolinium-based contrast materials have excellent safety profiles for use in pediatric computed tomography and MR imaging. The rate of acute adverse reactions and contrast-induced nephropathy is lower than in adults. The relationship between nephrogenic systemic fibrosis and gadolinium administration is well established and only a small number of children suffer from this debilitating disorder. The recent finding of gadolinium deposition in the brain of patients with normal renal function has renewed concerns regarding its long-term toxicity. The necessity of gadolinium-based contrast agent use in children should be considered carefully for each examination, and more stable macrocyclic agents should be the primary choice.


Ability of Swept-source OCT and OCT-angiography to detect neuroretinal and vasculature changes in patients with Parkinson disease and essential tremor.
To evaluate the ability of swept-source optical coherence tomography (SS-OCT) implemented with angiography analysis (SS-OCTA) to detect neuro-retinal and vasculature changes in patients with Parkinson's disease (PD) and essential tremor (ET), and to distinguish between both pathologies. A total 42 PD and 26 ET patients and 146 controls underwent retinal evaluation using SS-OCT plus OCT-Angio™. The macular (m) and peripapillary (p) retinal nerve fiber layer (RNFL) and ganglion cell layer (GCL), and macular vasculature were assessed. A Linear discriminant function (LDF) was calculated to evaluate the diagnostic ability of SS-OCTA in both PD and ET. PD patients presented a reduction in mRNFL (p < 0.005), mGCL (all sectors, p < 0.05) and pRNFL (p < 0.005) vs healthy controls, and in mRNFL and pRNFL vs ET patients (p < 0.001). ET patients showed a significant reduction in mGCL vs controls (p < 0.001). No differences were observed in the macular vasculature between groups. Predictive diagnostic variables were significant only for PD and a LDF was obtained with an area under the ROC curve of 0.796. Neuro-retinal thinning is present in both diseases, being greater in PD. While SS-OCT could be useful in diagnosing ET and PD, the diagnostic potential for SS-OCTA based on an LDF applies only to PD, not ET.


MoMI-G: modular multi-scale integrated genome graph browser.
Genome graph is an emerging approach for representing structural variants on genomes with branches. For example, representing structural variants of cancer genomes as a genome graph is more natural than representing such genomes as differences from the linear reference genome. While more and more structural variants are being identified by long-read sequencing, many of them are difficult to visualize using existing structural variants visualization tools. To this end, visualization method for large genome graphs such as human cancer genome graphs is demanded. We developed MOdular Multi-scale Integrated Genome graph browser, MoMI-G, a web-based genome graph browser that can visualize genome graphs with structural variants and supporting evidences such as read alignments, read depth, and annotations. This browser allows more intuitive recognition of large, nested, and potentially more complex structural variations. MoMI-G has view modules for different scales, which allow users to view the whole genome down to nucleotide-level alignments of long reads. Alignments spanning reference alleles and those spanning alternative alleles are shown in the same view. Users can customize the view, if they are not satisfied with the preset views. In addition, MoMI-G has Interval Card Deck, a feature for rapid manual inspection of hundreds of structural variants. Herein, we describe the utility of MoMI-G by using representative examples of large and nested structural variations found in two cell lines, LC-2/ad and CHM1. Users can inspect complex and large structural variations found by long-read analysis in large genomes such as human genomes more smoothly and more intuitively. In addition, users can easily filter out false positives by manually inspecting hundreds of identified structural variants with supporting long-read alignments and annotations in a short time. MoMI-G is freely available at https://github.com/MoMI-G/MoMI-G under the MIT license.


Analysıs of gene copy number changes ın head and neck cancer.
Chromosomal alterations and copy number changes are frequent events in tumors, leading to amplification of focal regions containing several oncogenes. Gains and losses of several regions have been reported in head and neck cancer (HNC) but the copy number changes of the individual genes located in these regions have not been analyzed so far. In this study we aimed to analyze the copy number variations in patients with HNC. Prospective study SETTING: University hospital PARTICIPANTS: 50 patients with squamous cell carcinoma of the head and neck METHODS: Copy number changes and amplifications of 22 genes in tumors and matched tissue were analyzed by MLPA which allows simultaneous analysis of gene copy numbers in multiple genetic regions. Amplifications were observed in 52% and losses were detected in 20% of the samples. Chromosome 8 was found to harbor the most frequent copy number alterations. The most frequently amplified genes were CCND1 and the MED1 genes followed by the MTDH and MYC genes on the long arm and ZNF703 on the short arm of chromosome 8. Amplification of the ZNF703, PRDM14 and MYC genes were highly correlated suggesting that the genes displaying high copy number changes on chromosome 8 collaborate during carcinogenesis. The alterations found in our study supports the contribution of gene amplifications and indicate cooperation between certain oncogenes in the pathogenesis of HNSCC. Correlations between amplification of less familiar genes and known oncogenes warrant further investigation. This article is protected by copyright. All rights reserved.


Clonal Transitions and Phenotypic Evolution in Barrett's Esophagus.
Barrett's esophagus (BE) is a risk factor for esophageal adenocarcinoma but our understanding of how it evolves is poorly understood. We investigated BE gland phenotype distribution, the clonal nature of phenotypic change, and how phenotypic diversity plays a role in progression. Using immunohistochemistry and histology, we analyzed the distribution and the diversity of gland phenotype between and within biopsy specimens from patients with nondysplastic BE and those who had progressed to dysplasia or had developed postesophagectomy BE. Clonal relationships were determined by the presence of shared mutations between distinct gland types using laser capture microdissection sequencing of the mitochondrial genome. We identified 5 different gland phenotypes in a cohort of 51 nondysplastic patients where biopsy specimens were taken at the same anatomic site (1.0-2.0 cm superior to the gastroesophageal junction. Here, we observed the same number of glands with 1 and 2 phenotypes, but 3 phenotypes were rare. We showed a common ancestor between parietal cell-containing, mature gastric (oxyntocardiac) and goblet cell-containing, intestinal (specialized) gland phenotypes. Similarly, we have shown a clonal relationship between cardiac-type glands and specialized and mature intestinal glands. Using the Shannon diversity index as a marker of gland diversity, we observed significantly increased phenotypic diversity in patients with BE adjacent to dysplasia and predysplasia compared to nondysplastic BE and postesophagectomy BE, suggesting that diversity develops over time. We showed that the range of BE phenotypes represents an evolutionary process and that changes in gland diversity may play a role in progression. Furthermore, we showed a common ancestry between gastric and intestinal-type glands in BE.


Comparison of estrogen and progesterone receptor status to lymphocyte immunity against tumor antigens in breast cancer patients.
Estrogen (ER) and progesterone receptor (PgR) content of tumors were determined by both the dextran-coated charcoal (DCC) cytosol and immunocytochemical assays (ICA), and these hormone receptor results were compared to lymphocyte immunity against tumor antigen(s) for 52 breast carcinoma patients. Hormone receptor analysis by both methods demonstrated that 60% of the patients' tumors had ERs, while 44% were positive for PgRs. The ICA procedure was more sensitive than the cytosol technique for determining PgR content of the tumors. This increased sensitivity was not observed for ER by ICA. Patient age, tumor size, and nodal status were not related to the ER and PgR receptor status. A total of 21/52 (40%) of the patients had positive lymphocyte immunity against tumor antigen. This immunity was independent of patient age, tumor size, and nodal status. There was no significant relationship between lymphocytic immunity against tumor antigen and ER or PgR content of tumors, suggesting that patient lymphocyte immunity against tumor is independent of hormone receptor status. This is further evidence that lymphocyte immunity against tumor antigen status is an independent prognostic indicator that may be useful in the selection of a subset of node negative patients for adjuvant chemotherapy.


Neurotensin, schizophrenia, and antipsychotic drug action.
The search for the underlying pathophysiology of schizophrenia has been an active avenue of investigation since the disease was first recognized more than 100 years ago. Although a great deal of the research has been driven by the known pharmacology of effective antipsychotic drugs, i.e., overactivity of the dopamine system, recent advances in neurobiology provide evidence that reduced synaptic connectivity/neurotransmission may play a substantial role in this disorder. One neuropeptide long posited to play a role in the biology of schizophrenia is neurotensin (NT). Central nervous system administration of NT has been shown to produce a wide variety of effects. Because of its close association with the dopamine (DA) system, the role of the NT system in clinical disorders hypothesized to involve DA circuits such as schizophrenia, Parkinson's disease, and drug abuse has been closely scrutinized. In addition, NT neurotransmission has been implicated in regulation of the stress response, stress-induced gastric ulcers, temperature regulation, food consumption, and analgesia. NT also acts as a growth factor in a variety of human cancer cell lines derived from lung, colon, prostate, and pancreas. This review first provides a background of the NT system. Second, data indicating that NT may mediate the effects of antipsychotic drugs are summarized. Third, data implicating NT in the pathophysiology of schizophrenia are described. Finally, evidence suggesting the use of NTergic compounds as novel antipsychotic drugs are presented.


Class II MHC antigen induction on rat insulinoma (RINm5F) and colon carcinoma (TS) cells by co-culture with diabetic and normal xenogenic lymphocytes.
Two MHC Class II-negative rat epithelial cell lines (RINm5F beta-cells and TS colic cells) were co-cultured with xenogenic lymphocytes from Type I diabetic patients or from low-dose streptozotocin (SZ) diabetic mice. MHC Class II antigens (Ag) were easily induced on both cell lines in such co-culture conditions, representing an experimental approach to insulitis. Our data indicate that: (1) lymphocytes from diabetic patients or from SZ mice were more efficient than lymphocytes from healthy controls in inducing Class II Ag on RIN cells. Lymphocytes from patients with autoimmune thyroid diseases were also more efficient than control lymphocytes, indicating that the ability to induce Class II may be related to the activation of lymphocytes rather than being diabetes-specific. (2) Rat colon carcinoma cells (TS) were also induced to express high levels of Class II Ag upon co-culture with SZ or control mouse lymphocytes. (3) Class II+ RIN cells were observed after 24 h of co-culture; their number increased after 48 and 72 h. The number of class II+ RIN increased proportionally to the number of lymphocytes in the culture. (4) Induction of Class II Ag was obtained by cell-free supernatants of mouse lymphocytes/RIN co-cultures and was inhibited by cyclosporine A, suggesting that Class II induction in this model is mediated by lymphokines. (5) Depletion experiments indicate that both monocytes and lymphocytes play a role in this Class II induction.


[Determination of tin in canned foods with oscillographic polarography].
To develop an oscillographic polarography method for determining the concentration of tin in canned foods. Because the sensitive polarography wave of tin can be obtained in the system of H2SO4-KI-VitC- NH4VO3, the tin content of several canned foods was analyzed. The detection limit was 0.12 microgram/L. The calibration graph of Sn was linear from 0 to 2.0 micrograms/ml, and the relative standard deviation ranged from 4.22% to 8.33%. The recoveries were 94.7%-112.0%. The samples were analyzed by the proposed method and standard method. The results of the two methods agreed well. This method is simple, quick and almost free from interference, and it is available for use in analysis of the canned sample.


Multiple metastases from a meningeal hemangiopericytoma associated with severe hypoglycemia.
Hemangiopericytomas are rare vascular tumors that may be associated with profound hypoglycemia. We present correlative imaging findings for a 57-year-old woman who had multiple abdominal and muscular metastases with severe hypoglycemia 11 years after resection of meningeal hemangiopericytoma. The metastases were well-defined masses containing solid and cystic components, and they were extremely hypervascular in the solid component. Those characteristics on imaging associated with hypoglycemia and the past history of meningeal hemangiopericytoma may suggest the diagnosis and be helpful in differentiating the mass from other vascular tumors. Color and power Doppler ultrasound may play a valuable role in characterizing and diagnosing metastasis from hemangiopericytoma. A delay of several years between diagnosis of the primary hemangiopericytoma and the manifestation of metastasis appears to be a special feature of hemangiopericytoma.


Mortality in Patients With Ebstein Anomaly.
Low birth prevalence and referral bias constitute significant obstacles to elucidating the natural history of Ebstein anomaly (EA). An extensive 2-country register-based collaboration was performed to investigate the mortality in patients with EA. Patients born from 1970 to 2017 and diagnosed with EA were identified in Danish and Swedish nationwide medical registries. Each patient was matched by birth year and sex with 10 control subjects from the general population. Cumulative mortality and HR of mortality were computed using Kaplan-Meier failure function and Cox proportional regression model. The study included 530 patients with EA and 5,300 matched control subjects with a median follow-up of 11 years. In the EA cohort, 43% (228) underwent cardiac surgery. Cumulative mortality was lower for patients diagnosed in the modern era (the year 2000 and later) than for those diagnosed in the prior era (P < 0.001). Patients with isolated lesion displayed lower cumulative mortality than patients with complex lesions did (P < 0.001). Patients with a presumed mild EA anatomy displayed a 35-year cumulative mortality of 11% (vs 4% for the matched control subjects; P < 0.001), yielding an HR for mortality of 6.0 (95% CI: 2.7-13.6), whereas patients with presumed severe EA demonstrated an HR of 36.2 (95% CI: 15.5-84.4) compared with control subjects and a cumulative mortality of 18% 35 years following diagnosis. Mortality in patients with EA is high irrespective of presence of concomitant congenital cardiac malformations and time of diagnosis compared with the general population, but overall mortality has improved in the contemporary era.


Epigenetic regulation of neural N-glycomics.
Glycan expression is tightly regulated in a cell-type-specific manner, which is essential for the diverse functions of glycans. In particular, neural cells such as neurons and astrocytes are known to express unique functional glycans not found in other cells, and these glycans play critical roles in high-order brain functions and various neurological disorders. However, little is known about how the expression of these neural glycans is established and maintained. Here, we investigated which glycans are expressed in each primary neural cell and how epigenetics contributes to the expression of neural glycans. We first isolated primary neurons, astrocytes, and fibroblasts from mouse embryos and carried out N-glycomic and glycosyltransferase (GlycoT)-transcriptomic analyses to identify N-glycans specific to a particular neural cell type and to clarify the underlying transcriptional basis. We next treated the cells with epigenetic drugs (5-aza-2'-deoxycytidine (5-aza) and trichostatin A (TSA)) and characterized the changes in GlycoT-transcriptomes and N-glycomes. We found that the N-glycomes in neurons were highly stable and resistant to epigenetic stimulation. In contrast, astrocytes showed dynamic N-glycan changes after treatment, such as a shift in the linkages of sialic acid. These results provide novel insights into how the expression of neural glycans is maintained and epigenetically regulated.


The effects of DNIC-type inhibition on temporal summation compared to single pulse processing: does sex matter?
A few experimental observations have suggested that diffuse noxious inhibitory control (DNIC)-type inhibition acts preferentially on the pain system if this is in a sensitised state, e.g. after slow temporal summation (wind-up). However, firm evidence is still missing. Furthermore, sex-related factors, which seem to affect temporal summation as well as DNIC effects, might thus also modulate the interaction of these two processes. To answer these questions, we investigated 40 young and pain-free subjects (20 female and 20 male). The conditioning stimulus in our DNIC paradigm was realized by immersion of the hand into a water tub containing either 42 degrees C (non-painful heat) or 46 degrees C (painful heat) hot water. The test stimuli were either single pulses or series of five pulses (0.5 Hz repetition frequency) produced by a pressure algometer. The VAS ratings for the last stimulus in the series were significantly higher than for the single pulse (temporal summation). The ratings were significantly reduced by the 42 degrees C conditioning stimulus and even more by the 46 degrees C conditioning stimulus, suggesting DNIC-like inhibition. This was equally true both for the single pulse and for the series of pulses. Sex differences were not observed for temporal summation, DNIC inhibition or for the interaction of the two processes, although women exhibited significantly lower pressure pain thresholds and higher ratings for the tonic heat stimuli. In conclusion, DNIC-type inhibition apparently does not preferentially act on a sensitised pain system after slow temporal summation. Considering the sex of the subjects does not change this insight.


Extraction, purification, structural character and biological properties of propolis flavonoids: A review.
Propolis is an aromatic substance which is collected by bees and mixed with bee saliva. The plant sources of propolis are mainly consisted with plant exudates from bark, buds and etc. Flavonoids are secondary metabolites widely found in natural plants, which have a variety of health care functions and are the main active ingredients of propolis. This article summarized the types, active ingredients, pharmacological effects, extraction methods and applications of propolis flavonoids, the aim was to provide the theoretical basis for further research and development of propolis flavonoids.


Draft genome sequences of Pantoea agglomerans and Pantoea vagans isolates associated with termites.
The genus Pantoea incorporates many economically and clinically important species. The plant-associated species, Pantoea agglomerans and Pantoea vagans, are closely related and are often isolated from similar environments. Plasmids conferring certain metabolic capabilities are also shared amongst these two species. The genomes of two isolates obtained from fungus-growing termites in South Africa were sequenced, assembled and annotated. A high number of orthologous genes are conserved within and between these species. The difference in genome size between P. agglomerans MP2 (4,733,829 bp) and P. vagans MP7 (4,598,703 bp) can largely be attributed to the differences in plasmid content. The genome sequences of these isolates may shed light on the common traits that enable P. agglomerans and P. vagans to co-occur in plant- and insect-associated niches.


Morphometric analysis of sphenopalatine foramen on computed tomography images with clinical significance.
This study aimed to investigate the effect of anatomical variations in the sphenopalatine foramen and the lateral nasal wall on sphenopalatine foramen-related morphometric measurements. Paranasal sinus multidetector computed tomography records of 153 patients were evaluated. Morphometric measurements were made between the fixed bony landmarks and the sphenopalatine foramen. Number, shape, localization variations of the sphenopalatine foramen, concha bullosa, and septum deviation were noted and the results were compared with respect to sex, age, and laterality. No significant difference was detected with respect to laterality, whereas most of the measured distances were higher in males than females. There was a significant difference between the obtained morphometric data according to age groups. In our study, 91.2% single, 7.8% double, and 1% triple sphenopalatine foramen were detected and the most common irregularly shaped (37.3%). The location of sphenopalatine foramen was reported as the most common type II. Septum deviation types have no potential influence on the location of the foramen, but most of the measured parameters were found to be significantly smaller in the presence of concha bullosa, whereas the angle was found to be higher. This study revealed a significant relationship between the morphometric measurements of the sphenopalatine foramen and concha bullosa, while septum deviation types did not affect these results. For a safer and more effective surgery with prevention of iatrogenic complications, a surgeon should be aware of this correlation, especially in endoscopic transnasal approaches.


[Male genital examination: the usefulness of penis endoscopy and the acetic acid test for the detection of papillomavirus lesions].
This study was based on 2,400 genital examinations performed by means of the acetic acid test looking for human papillomavirus (HPV) lesions in the male partners of women with HPV genital lesions. These peniscopies demonstrated HPV lesions in 56% of the men examined. In 109 cases, histological and virological examination revealed that 30% of them had areas of intraepithelial neoplasia of the penis, associated with potentially oncogenic papillomavirus infection. The value of this examination is to detect HPV lesions and, in particular, dysplastic lesions. The treatment of these lesions appears to allow a reduction in the incidence of recurrence of HPV lesions in women after treatment.


Effect of Intralipid on clearance and organ uptake of bacteria in intra-abdominal sepsis in rats.
Although infusion of Intralipid has been reported to block reticuloendothelial system (RES) function, it is unclear if this is true at rates used clinically. This study investigates the effect of Intralipid, infused at a rate providing about half of the non-protein caloric needs, on survival and bacterial clearance in septic rats. Continuous infusion of Intralipid (0.9 g/100 g bw/d) or saline (controls) was started immediately after induction of hyperdynamic intra-abdominal sepsis (bacterial agents: E. coli, B. thetaiotamicron). RES function was studied by means of intravenous injection of Selenium-labelled, viable E. coli after 24 or 48 h. Compared to the saline-treated controls, Intralipid did not cause any change in clearance from blood or localisation to liver, spleen or lungs. The 24 and 48 h survival rates were about 80 and 65%, respectively, and similar in the two groups. It is concluded that infusion of Intralipid at a rate close to that used clinically did not impair survival or bacterial clearance in rats with gram-negative sepsis.


Barriers to conducting clinical research in reproductive medicine: Egypt.
There are many barriers to scientific research in Egypt, including lack of funding, lack of a scientific atmosphere, and "brain drain" of scientists who produce high quality research.


Histological typing and nuclear evaluation of endometrial adenocarcinoma.
On the basis of the nuclear pattern "homologous adenocarcinoma" of the endometrium, arising in women under 40, may be divided into three cytologically distinguishable entities: 1) atypical adenomatous hyperplasia Grade 1; 2) atypical adenomatous hyperplasia Grade 2; 3) adenocarcinoma; probably different also at biological level.


The relationship between glycosylated haemoglobin levels and various degrees of glucose intolerance.
To assess the use of glycosylated haemoglobin to discriminate between various degrees of glucose intolerance, glycosylated haemoglobin levels were determined in 107 subjects (48 males and 59 females, age range 18-80 years). Following a 75 g oral glucose tolerance test and according to World Health Organization criteria, subjects were classified as normal (n = 32), diabetic (n = 46) or as having impaired glucose tolerance (n = 29). Mean glycosylated haemoglobin levels were 5.8 +/- 1.3% (range 4%-9%) in normal subjects, 7.1 +/- 1.7% in subjects with impaired glucose tolerance (range 4.1%-10.1%) and 10.1 +/- 2.6% (range 4.7%-18.8%) in diabetic patients. The difference between the groups was highly significant (p less than 0.01). Twelve per cent of normal subjects exceeded and 52% of subjects with impaired glucose tolerance fell below 7.4% (mean +/- 2SD, considered as the upper limit of normal values). A significant correlation was observed between glycosylated haemoglobin values and fasting blood glucose (r = 0.68, p less than 0.01). These results provide evidence that glycosylated haemoglobin levels are influenced by slightly reduced carbohydrate tolerance. Glycosylated haemoglobin may be a useful test to improve the specificity of the oral glucose load to select and to follow-up subjects with impaired glucose tolerance.


Avenues for measuring and characterising violence in perinatal care to improve its prevention: A position paper with a proposal by the National College of French Midwives.
France is somewhat behind other countries in its consideration of the issue of violence in perinatal care. Its consequences on maternal, but also neonatal and infant health are recognised internationally. Nonetheless, research and data measuring its frequency and its determinants are inadequate, and the relevant definitions are not always consensual. In this context, we, as midwives and researchers in public health and as members of the National College of French Midwives, seek to propose a scientific and clinical contribution to this debate. We propose avenues for measuring and characterising violence in perinatal care. Our objective is to quantify and characterise the situations of violence in perinatal care in population-based studies and based on the perceptions of each woman questioned. This proposal for questions, simplified compared with those currently in used in the international scientific literature, has the advantage of focusing reflection around three categories: inappropriate medical care, inappropriate human behaviours in care, and sexual abuse. It should also allow the identification of the contexts of care during which violence may be experienced, as well as the categories of health-care workers concerned. It seems important to us to distinguish these situations, causal and context, for they require different responses if we hope to reduce the frequency and the effects of violence in perinatal care in the future. We propose questions that could also be used in clinical situations by midwives and other clinicians.


[Expression of Livin in laryngeal squamous cell carcinoma and relationship with bFGF].
To detect the roles of Livin and its relationship with bFGF in laryngeal squamous cell carcinoma (LSCC) through observing the expression of Livin and bFGF in laryngeal squamous cell carcinoma. Expression of Livin and bFGF in 41 cases of LSCC (11 cases with lymph node metastasis) and 20 cases of normal soft palate mucosa were detected by immunohistochemistry. Livin were positively detected in 29 (70.73%) cases of LSCC and negatively detected in all normal soft palate tissue. The positive rate of Livin was higher in LSCC than that in normal soft palate tissue and the expressions were significantly associated with lymph node metastasis status (P < 0.05) but not with histological grade, clinical stage and age (P > 0.05). The expression of Livin was positively correlated with the expression of bFGF. The elevated expression of Livin in LSCC might play an important role in the development of laryngeal squamous cell carcinoma and bFGF might be involved in the process.


Disease characteristics of diffuse large B-cell lymphoma predicting relapse and survival after autologous stem cell transplantation: A single institution experience.
While various tools such as the International Prognostic Index (IPI) and its derivatives exist for risk-stratification of diffuse large B-cell lymphoma (DLBCL) at diagnosis, patient and disease characteristics capable of predicting outcome after high-dose chemotherapy followed by autologous stem cell transplantation (HDC/ASCT) are not clearly defined. We retrospectively analyzed medical records of 111 DLBCL patients (78 relapsed and 33 refractory) who underwent HDC/ASCT at our institution from 2010-2015. After a median follow-up time of 4.6 years (interquartile range [IQR] 2.2-8.1), the likelihood of 5-year progression-free survival (PFS) was 62.2% (95% CI, 53.4%-72.4%) and the likelihood of 5-year overall survival (OS) was 68.9% (95% CI, 60.7%-78.2%). More than three chemotherapy regimens prior to ASCT was the only variable associated with lower likelihood of PFS (P = .004) and OS (P = 0.026). Male gender and high IPI score at time of ASCT were also associated with lower likelihood of PFS (P = .043; P = .013). NCCN IPI and age-adjusted IPI at time of ASCT were not predictive of outcome following ASCT. Patients with refractory and relapsed disease had similar outcomes post-ASCT (P = .207 for PFS, P = .073 for OS).


Modeling symptom drivers of oral intake in long-term head and neck cancer survivors.
This study examined the relationship between self-reported symptom severity and oral intake in long-term head and neck cancer (HNC) survivors. An observational survey study with retrospective chart abstraction was conducted. HNC patients who had completed an MD Anderson Symptom Inventory-Head and Neck (MDASI-HN) questionnaire and also had clinician graded oral intake ratings (Functional Oral Intake Scale [FOIS]) were included. Correlation coefficients were computed. FOIS scores were regressed on MDASI-HN symptom items using stepwise backwards elimination for multivariate models. One hundred and fifty-two survey pairings were included in the analysis (median 44 months follow-up, range 7-198). Per FOIS, 28% of survivors maintained a total oral diet with no restrictions, 67% reported a restricted oral diet (without tube), 3% were partially tube-dependent with some oral intake, and 2% were NPO. Of the 22 symptom items, the most severe items in decreasing order were dry mouth, difficulty swallowing\chewing, problems with mucus, tasting food, and choking/coughing. Significant bivariate correlations, after Bonferroni correction for multiple comparisons, were present for 8 of 22 symptoms with FOIS. On multivariate analysis, symptom severity for difficulty swallowing and problems with teeth/gums remained significantly associated with FOIS. Oral intake in HNC survivorship is a multidimensional issue and functional outcome that is impacted not only by dysphagia but also by dental status. Symptom drivers of oral intake likely differ in acute survivorship. Nonetheless, these findings highlight the lack of specificity in this end point and also the need for multidisciplinary supportive care to optimize oral intake in survivors.


Integrins regulate stemness in solid tumor: an emerging therapeutic target.
Integrins are the adhesion molecules and transmembrane receptors that consist of α and β subunits. After binding to extracellular matrix components, integrins trigger intracellular signaling and regulate a wide spectrum of cellular functions, including cell survival, proliferation, differentiation and migration. Since the pattern of integrins expression is a key determinant of cell behavior in response to microenvironmental cues, deregulation of integrins caused by various mechanisms has been causally linked to cancer development and progression in several solid tumor types. In this review, we discuss the integrin signalosome with a highlight of a few key pro-oncogenic pathways elicited by integrins, and uncover the mutational and transcriptomic landscape of integrin-encoding genes across human cancers. In addition, we focus on the integrin-mediated control of cancer stem cell and tumor stemness in general, such as tumor initiation, epithelial plasticity, organotropic metastasis and drug resistance. With insights into how integrins contribute to the stem-like functions, we now gain better understanding of the integrin signalosome, which will greatly assist novel therapeutic development and more precise clinical decisions.


Double deletion of PINK1 and Parkin impairs hepatic mitophagy and exacerbates acetaminophen-induced liver injury in mice.
Mitochondria damage plays a critical role in acetaminophen (APAP)-induced necrosis and liver injury. Cells can adapt and protect themselves by removing damaged mitochondria via mitophagy. PINK1-Parkin pathway is one of the major pathways that regulate mitophagy but its role in APAP-induced liver injury is still elusive. We investigated the role of PINK1-Parkin pathway in hepatocyte mitophagy in APAP-induced liver injury in mice. Wild-type (WT), PINK1 knockout (KO), Parkin KO, and PINK1 and Parkin double KO (DKO) mice were treated with APAP for different time points. Liver injury was determined by measuring serum alanine aminotransferase (ALT) activity, H&E staining as well as TUNEL staining of liver tissues. Tandem fluorescent-tagged inner mitochondrial membrane protein Cox8 (Cox8-GFP-mCherry) can be used to monitor mitophagy based on different pH stability of GFP and mCherry fluorescent proteins. We overexpressed Cox8-GFP-mCherry in mouse livers via tail vein injection of an adenovirus Cox8-GFP-mCherry. Mitophagy was assessed by confocal microscopy for Cox8-GFP-mCherry puncta, electron microscopy (EM) analysis for mitophagosomes and western blot analysis for mitochondrial proteins. Parkin KO and PINK1 KO mice improved the survival after treatment with APAP although the serum levels of ALT were not significantly different among PINK1 KO, Parkin KO and WT mice. We only found mild defects of mitophagy in PINK1 KO or Parkin KO mice after APAP, and improved survival in PINK1 KO and Parkin KO mice could be due to other functions of PINK1 and Parkin independent of mitophagy. In contrast, APAP-induced mitophagy was significantly impaired in PINK1-Parkin DKO mice. PINK1-Parkin DKO mice had further elevated serum levels of ALT and increased mortality after APAP administration. In conclusion, our results demonstrated that PINK1-Parkin signaling pathway plays a critical role in APAP-induced mitophagy and liver injury.


Recycling of sugarcane bagasse ash waste in the production of clay bricks.
This work investigates the recycling of sugarcane bagasse ash waste as a method to provide raw material for clay brick bodies, through replacement of natural clay by up 20 wt.%. Initially, the waste sample was characterized by its chemical composition, X-ray diffraction, differential thermal analysis, particle size, morphology and pollution potential. Clay bricks pieces were prepared, and then tested, so as to determine their technological properties (e.g., linear shrinkage, water absorption, apparent density, and tensile strength). The sintered microstructure was evaluated by scanning electron microscopy (SEM). It was found that the sugarcane bagasse ash waste is mainly composed by crystalline silica particles. The test results indicate that the sugarcane bagasse ash waste could be used as a filler in clay bricks, thus enhancing the possibility of its reuse in a safe and sustainable way.


Expression and characterization of recombinant IL-1Ra in Aspergillus oryzae as a system.
The interleukin-1 receptor antagonist (IL-1Ra) is a crucial molecule that counteracts the effects of interleukin-1 (IL-1) by binding to its receptor. A high concentration of IL-1Ra is required for complete inhibition of IL-1 activity. However, the currently available Escherichia coli-expressed IL-1Ra (E. coli IL-1Ra, Anakinra) has a limited half-life. This study aims to produce a cost-effective, functional IL-1Ra on an industrial scale by expressing it in the pyrG auxotroph Aspergillus oryzae. We purified A. oryzae-expressed IL-1Ra (Asp. IL-1Ra) using ion exchange and size exclusion chromatography (53 mg/L). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed that Asp. IL-1Ra is N-glycosylated and approximately 17 kDa in size. We conducted a comparative study of the bioactivity, binding kinetics, and half-life between Asp. IL-1Ra and E. coli IL-1Ra. Asp. IL-1Ra showed good bioactivity even at a low concentration of 0.5 nM. The in vitro half-life of Asp. IL-1Ra was determined for different time points (0, 24, 48, 72, and 96 h) and showed higher stability than E. coli IL-1Ra, despite exhibiting a 100-fold lower binding affinity (2 nM). This study reports the production of a functional Asp. IL-1Ra with advantageous stability, without extensive downstream processing. To our knowledge, this is the first report of a recombinant functional and stable IL-1Ra expressed in A. oryzae. Our results suggest that Asp. IL-1Ra has potential for industrial-scale production as a cost-effective alternative to E. coli IL-1Ra.


Cataract and metabolic disease.
In addition to the already recognized metabolic diseases which have been associated with cataract formation, e.g. galactosaemia, galactokinase deficiency, Lowe's syndrome and diabetes, several other disorders can also lead to the development of cataracts. They are sorbitol dehydrogenase deficiency, uridine diphosphate galactose-4-epimerase deficiency, marginal maternal transferase and galactokinase deficiency, galactitol and sorbitol accumulation of unknown origin, heterozygosity for galactosaemia and galactokinase deficiency as well as the carrier state for Lowe's syndrome. In this review these metabolic disorders have been divided into five groups according to the age at the first appearance of lens clouding and the possible means of treatment have been discussed.


The local physical structure of amorphous hydrogenated boron carbide: insights from magic angle spinning solid-state NMR spectroscopy.
Magic angle spinning solid-state nuclear magnetic resonance spectroscopy techniques are applied to the elucidation of the local physical structure of an intermediate product in the plasma-enhanced chemical vapour deposition of thin-film amorphous hydrogenated boron carbide (B(x)C:H(y)) from an orthocarborane precursor. Experimental chemical shifts are compared with theoretical shift predictions from ab initio calculations of model molecular compounds to assign atomic chemical environments, while Lee-Goldburg cross-polarization and heteronuclear recoupling experiments are used to confirm atomic connectivities. A model for the B(x)C:H(y) intermediate is proposed wherein the solid is dominated by predominantly hydrogenated carborane icosahedra that are lightly cross-linked via nonhydrogenated intraicosahedral B atoms, either directly through B-B bonds or through extraicosahedral hydrocarbon chains. While there is no clear evidence for extraicosahedral B aside from boron oxides, ∼40% of the C is found to exist as extraicosahedral hydrocarbon species that are intimately bound within the icosahedral network rather than in segregated phases.


3D visualization of XFEL beam focusing properties using LiF crystal X-ray detector.
Here, we report, that by means of direct irradiation of lithium fluoride a (LiF) crystal, in situ 3D visualization of the SACLA XFEL focused beam profile along the propagation direction is realized, including propagation inside photoluminescence solid matter. High sensitivity and large dynamic range of the LiF crystal detector allowed measurements of the intensity distribution of the beam at distances far from the best focus as well as near the best focus and evaluation of XFEL source size and beam quality factor M(2). Our measurements also support the theoretical prediction that for X-ray photons with energies ~10 keV the radius of the generated photoelectron cloud within the LiF crystal reaches about 600 nm before thermalization. The proposed method has a spatial resolution ~0.4-2.0 μm for photons with energies 6-14 keV and potentially could be used in a single shot mode for optimization of different focusing systems developed at XFEL and synchrotron facilities.


A first case of congenital TTP on the African continent due to a new homozygous mutation in the catalytic domain of ADAMTS13.
Hereditary thrombotic thrombocytopenic purpura (TTP) is a rare disorder characterized by occlusive microvascular thrombosis, consumptive thrombocytopenia, and microangiopathic hemolytic anemia. Homozygous or compound heterozygous mutations in the ADAMTS13 gene result in a congenital severe ADAMTS13 deficiency and subsequent accumulation of ultra-large von Willebrand factor multimers, which tend to form platelet thrombi in the microcirculation. We report a first case of congenital TTP on the African continent with a new, homozygous mutation in the metalloprotease domain of ADAMTS13. An initially oligo-symptomatic presentation was followed by acute exacerbation with ischemic stroke and acute renal failure highlighting the severity of this syndrome.


Ultrasonography of the temporomandibular joint: comparison of findings in patients with rheumatic diseases and temporomandibular disorders. A preliminary report.
The objective of the study was to compare findings from ultrasonography (US) of the temporomandibular joint (TMJ) in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), and temporomandibular disorders (TMD). US assessment of the temporomandibular joints was bilaterally performed in 68 patients (22 with RA, 11 with PsA, and 35 with TMD). All the TMJs were assessed for the presence of disc displacement, effusion, and changes of the condylar profile, and the prevalence of such abnormalities was compared across the 3 groups of patients. To confirm generalizability of results, US findings were also compared with those of magnetic resonance (MR), taken as the standard of reference. Prevalence of disc displacement and changes in condylar profile were similar between patients with rheumatic diseases and temporomandibular disorders, while effusion was significantly more present in TMJs of TMD patients. Sensitivity of US to detect TMJ abnormalities was acceptable, while specificity was low for condylar alterations. Temporomandibular joint involvement in patients with rheumatic diseases seems to be similar to that described in subjects with temporomandibular disorders. Ultrasonography confirmed to be an accurate technique to detect disc displacement and effusion within the temporomandibular joint, but not to detect condylar abnormalities.


Automated evoked potential monitoring.
We have developed a computer system for use in the operating room and intensive care unit for the automated acquisition and classification of flash evoked potentials. The project goals were to: (1) acquire evoked potentials without user intervention; (2) recognize, label, and trend important parameters; and (3) alarm if parameters deviate too far from normal. The system consists of a Nicolet Pathfinder, programmed for periodic flash visual evoked potential acquisition and a personal computer for analysis. Analysis software utilizes the C+ +language. The system has been tested on sixteen cases, determinations of specific waveforms were validated ex post facto by two experts. Three parameters in each of three channels were correctly recognized in more than 87% of all waveforms acquired. No signals at all were obtained in only 0.8% of all samples.


Changes of cardiac function in diabetic dogs.
This study aimed to evaluate cardiac function and compare the concentration of cardiac biomarkers including cardiac troponin I (cTnI), galectin-3 (Gal-3), and N-terminal pro B-type natriuretic peptides (NT-proBNP) in diabetic and control dogs. Thirty-nine dogs were included. The diabetic and control groups consisted of 19 and 20 dogs, respectively. Plasma cTnI, Gal-3, and NT-proBNP concentrations were measured in the diabetic and control groups. Echocardiography was performed in all dogs to evaluate cardiac structure and function. Echocardiographic values and cardiac biomarker concentrations between the two groups were compared with the Mann-Whitney U test. The p-value < 0.05 was considered statistical significance. No evidence of cardiac structural changes was detected in diabetic dogs on two-dimensional echocardiography. The echocardiographic values of diabetic and control dogs were within reference intervals. Echocardiographic changes indicating diastolic dysfunction assessed by spectral flow Doppler echocardiography and tissue Doppler imaging were found in diabetic dogs (42.10%) compared with control dogs (10.00%; p = 0.022). Diabetic dogs with durations of diabetes mellitus > 1 year had an increased left ventricular wall thickness and echocardiographic changes suggesting diastolic dysfunction compared with those with duration of diabetes mellitus < 1 year. No evidence of systolic dysfunction was detected in diabetic dogs. No significant difference in plasma cTnI, Gal-3, and NT-proBNP concentrations was found between the two groups. Echocardiographic changes suggested that left ventricular diastolic dysfunction was detected in diabetic dogs without changes in the concentration of cardiac biomarkers including cTnI, Gal-3, and NT-proBNP compared with the age- and breed-matched control dogs.


Hexanedione effects on protein phosphorylation in rat peripheral nerve.
Rats were treated with either 2,5-hexanedione (2,5-HD), 1,6-hexanediol (1,6-HDIOL), or saline for 7, 15 or 24 days. Protein phosphorylation was measured in proximal and distal sciatic nerve segments following incubation with [32P]orthophosphate. In proximal segments, 2,5-HD administration caused selective time-dependent increases in isotope incorporation in a 55 kDa protein, tentatively identified as tubulin, and a 180 kDa protein. Enhanced phosphorylation was highest at 24 days when motor function was most impaired. Administration of 1,6-HDIOL produced no consistent phosphorylation changes. Animals intoxicated with 3,4-dimethyl-2,5-hexanedione for 12 days showed proximal region increases in phosphorylation of the 55 and 180 kDa proteins and the major myelin proteins, Po and Pr.


Acute promyelocytic leukemia during pregnancy: report of 3 cases.
Acute promyelocytic leukemia (APL) is characterized by onset at a young age and a life-threatening hemorrhagic diathesis, which is attributed to a disseminated intravascular coagulation (DIC)-like coagulopathy. The discovery of all-trans-retinoic acid has changed the course of APL treatment by reducing the onset of DIC and inducing a complete and durable remission in more than 90% of patients. The occurrence of APL during pregnancy is not a frequent event, but the management of these patients raises many therapeutic and ethical dilemmas and requires a careful clinical case evaluation of fetal and maternal risk, coagulation status, the parents' wishes, and therapeutic options. Here we describe 3 patients with APL diagnosed during pregnancy. Clinical data and the therapeutic approaches are presented. In the discussion, we analyze clinical decisions and therapeutic options and compare our cases with those found in the literature.


Non-invasive Imaging of Antisense Oligonucleotides in the Brain via In Vivo Click Chemistry.
The treatment of complex neurological diseases often requires the administration of large therapeutic drugs, such as antisense oligonucleotide (ASO), by lumbar puncture into the intrathecal space in order to bypass the blood-brain barrier. Despite the growing number of ASOs in clinical development, there are still uncertainties regarding their dosing, primarily around their distribution and kinetics in the brain following intrathecal injection. The challenge of taking measurements within the delicate structures of the central nervous system (CNS) necessitates the use of non-invasive nuclear imaging, such as positron emission tomography (PET). Herein, an emergent strategy known as "pretargeted imaging" is applied to image the distribution of an ASO in the brain by developing a novel PET tracer, [18F]F-537-Tz. This tracer is able to undergo an in vivo "click" reaction, covalently binding to a trans-cyclooctene conjugated ASO. A novel small molecule tracer for pretargeted PET imaging of ASOs in the CNS is developed and tested in a series of in vitro and in vivo experiments, including biodistribution in rats and non-human primates. In vitro data and extensive in vivo rat data demonstrated delivery of the tracer to the CNS, and its successful ligation to its ASO target in the brain. In an NHP study, the slow tracer kinetics did not allow for specific binding to be determined by PET. A CNS-penetrant radioligand for pretargeted imaging was successfully demonstrated in a proof-of-concept study in rats, laying the groundwork for further optimization.


Ascending projections of long descending propriospinal tract (LDPT) neurons.
Ascending projections of long descending propriospinal tract (LDPT) cells were investigated using the technique of double retrograde labeling. In rat, injection of one fluorescent dye was made into either the reticular formation or the cerebellum, and a second dye was injected into the lumbosacral enlargement (LSE). In cat, injections were made into the reticular formation and into the lumbosacral enlargement. Using fluorescence microscopy, observation of neurons in the cervical enlargement (CE) revealed single- and double-labeled cells which were either spinoreticular or spinocerebellar tract cells and/or LDPT cells. In both cat and rat, the location of double-labeled LDPT-spinoreticular cells were in the ventromedial spinal gray matter of the CE and were coextensive with single-labeled LDPT and spinoreticular cells. The locations of double-labeled LDPT-spinocerebellar cells in rat were in the ventromedial gray and were coextensive with single-labeled LDPT cells, but not with single-labeled spinocerebellar cells. The latter group was located in central lamina VII and medial laminae V and VI. Overall, the mean number of double-labeled cells was 40% of rat and 7% of cat LDPT cells, indicating projections to either the brainstem reticular formation or cerebellum as well as to the lumbosacral enlargement. Thus, a subpopulation of LDPT cells apparently also serves as a spinoreticular (SR) and spinocerebellar (SC) projection system.


High-performance liquid chromatographic determination of diadenosine 5',5'"-p1,p4-tetraphosphate with precolumn fluorescence derivatization and its application to metabolism study in whole blood.
Diadenosine 5',5'"-p1,p4-tetraphosphate (Ap4A) was converted with chloroacetaldehyde to the fluorescent di-1,N6-ethenoadenosine derivative within 60 min at 80 degrees C. It was separated by reversed-phase HPLC and detected fluorimetrically (excitation and emission wavelengths of 275 and 410 nm, respectively). The detection limit of Ap4A was ca. 0.2 microgram/ml in plasma when 10 microliters of the sample was applied to the column. The rate of degradation of Ap4A added to whole blood (5 micrograms/ml) was examined using this method. Half-lives (means +/- S.E., n = 3) were 0.88 +/- 0.30 min (in rat blood), 13.7 +/- 3.6 min (in dog blood) and 17.2 +/- 1.4 min (in human blood). A marked species difference in the degradation rate of Ap4A in blood was observed.


Streptomyces formicae sp. nov., a novel actinomycete isolated from the head of Camponotus japonicus Mayr.
During a screening for novel and biotechnologically useful actinobacteria in insects, a novel actinomycete with antifungal activity, designated strain 1H-GS9(T), was isolated from the head of a Camponotus japonicus Mayr ant, which were collected from Northeast Agricultural University (Harbin, Heilongjiang, China). Strain 1H-GS9(T) was characterised using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain 1H-GS9(T) belongs to the genus Streptomyces with high sequence similarities to Streptomyces scopuliridis DSM 41917(T) (98.8 %) and Streptomyces mauvecolor JCM 5002(T) (98.6 %). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it forms a monophyletic clade with Streptomyces kurssanovii JCM 4388(T) (98.6 %), Streptomyces xantholiticus JCM 4282(T) (98.6 %) and Streptomyces peucetius JCM 9920(T) (98.5 %). Thus, a combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 1H-GS9(T) and the above-mentioned five strains, which further clarified their relatedness and demonstrated that strain 1H-GS9(T) could be distinguished from these strains. Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces formicae sp. nov. is proposed. The type strain is 1H-GS9(T) (=CGMCC 4.7277(T) = DSM 100524(T)).


Demographics, Management Practices and In-Hospital Outcomes: Results from the HRH Princess Maha Chakri Sirindhorn Medical Center Acute Coronary Syndrome Registry.
To determine the demographics and in-hospital outcomes of patients with acute coronary syndrome. It provides a real-life data to assess treatment strategies for acute coronary syndrome (ACS) patients. The prospective study included patients who were hospitalized with the diagnosis of ACS during January to December, 2013. The data were analyzed in terms of characteristics, clinical presentation, treatment, and in-hospital outcomes. A total of 105 patients were enrolled. Mean age of all patients was 73.8 years and half of the patients were older than 70 years old. Twenty patients were classified as ST-segment elevation myocardial infarction (STEMI) and 85 as unstable angina or non-ST-segment elevation myocardial infarction (UA/NSTEMI). Overall prevalence of diabetes was 52.4%. The STEMI group was predominantly male, with a fewer number of diabetes than in UA/NSTEMI group. Eighty percent of the STEMI patients received reperfusion therapy. Primary percutaneous coronary intervention (PCI) was performed in 70% of STEMI patients. The median door-to-needle and door-to-balloon time were 60 and 74 minutes, respectively. Six of fourteen primary PCI patients received it after 90 minutes. Nearly half of UA/NSTEMI patients went to coronary angiography and about one-third of them received revascularization with either PCI or coronary artery bypass grafting. The total mortality rate was 15% in both the STEMI patients and the UA/NSTEMI patients. This registry provided a detail of demographics, management practices, and in hospital outcomes of ACS patients. Door-to-needle time and door-to-balloon time were considered as suboptimal. In-hospital mortality was higher than the latest national average. Patient delay should be improved by giving more education to the general public and concerted effort to improve in-hospital time delay is warranted. These data have an impact on our patient care system and alert our team to improve patient care.


High glucose affected respiratory burst activity of peripheral leukocyte via G6PD and NOX inhibition in Megalobrama amblycephala.
High glucose levels are known to impair growth and immune function in fish. Here we investigated the role of glucose-6-phosphate dehydrogenase (G6PD) and NADPH oxidase (NOX) in high glucose-associated impairment of leukocyte respiratory burst activity in Megalobrama amblycephala. We cultured peripheral leukocytes isolated from M. amblycephala with media containing no glucose (non-glucose group), 11.1 mmol/L d-glucose (physiologic glucose group), 22.2 mmol/L d-glucose (high-glucose group), or 11.1 mmol/L d-glucose + 100 μmol/L dehydroepiandrosterone (DHEA) (DHEA-treated group). After 24 h, we assayed production of reactive oxygen species (ROS) as a measure of respiratory burst function as well as activity of G6PD and NOX. The high-glucose group and DHEA-treated group showed significantly reduced respiratory burst function, reduced production of ROS, and reduced G6PD and NOX activity at 24 h, compared to the non-glucose and physiologic glucose groups (P < 0.05). The degree of impairment was similar between high-glucose and DHEA-treated groups (P > 0.05). These findings suggest that reduced NADPH availability likely underlies the suppression of respiratory burst function in M. amblycephala leukocytes exposed to high glucose levels.


Difference in coronal curvature of the medial and lateral femoral condyle morphology by gender in implant design for total knee arthroplasty.
The purpose of this study was to investigate the morphometric data obtained from the three-dimensional magnetic resonance images of ethnic Korean knee osteoarthritis, and to evaluate the morphological differences between the coronal curvature of the female and male femoral condyles. The differences in coronal curvature of the femoral condyle morphology of 1990 patients (1689 females and 301 males) were evaluated in three dimensions. A close-fit diameter was, respectively, generated on the medial and lateral femoral condyle articular surfaces, and these diameters reflect the coronal diameter of the femoral condyle curvature. These measurements were compared with those of the femoral design of five different commonly used total knee arthroplasty (TKA) implant designs. The average diameter of the curvature of the medial condyles was significantly larger than that of the lateral condyles (P < 0.05). This trend was found in the male and female groups. In addition, the average diameter of the curvature of the femoral condyles was found to significantly differ between males and females (P < 0.05). For four TKA implant designs, the average diameter of the coronal curvature of femoral condyle was smaller than that obtained via our measurements, whereas one TKA implant design yielded a smaller average diameter. Furthermore, the medial and lateral coronal curvatures of the femoral condyle were symmetric in all TKA implant designs. The study provided a reliable and consistent evaluation of the coronal curvature of femoral condyles in the Korean population. These results showed that a gender-specific or asymmetric femoral component design is required to regenerate the coronal curvature of the femoral condyles for ethnically Korean males and females.


Exploring the Stability Effect of the Co-Substituted P2-Na0.67[Mn0.67Ni0.33]O2 Cathode for Liquid- and Solid-State Sodium-Ion Batteries.
Pursuing high-performance cathode materials for sodium-ion batteries (SIBs) has great significance in the modern green energy world. The P2-type sodium-based layered oxide Na0.67[Mn0.67Ni0.33]O2 with high operating potential upon 4.3 V and high theoretical capacity has emerged as the most promising cathode. However, the material suffers from severe capacity decay during the electrochemical reaction process. Herein, the P2-Na0.67[Mn0.67Ni0.21Li0.06Zn0.06]O2 cathode is gained by moderately substituting lithium/zinc for the nickel sites. The inactive Li/Zn co-substitution is endowed with the ability to stabilize the crystal structure, resulting in enhanced electrochemical kinetics and remarkable long cyclic performance in liquid- and solid-state electrolytes. Thus, the Li/Zn co-substituted cathode presents a specific capacity of 154 mAh g-1 at the first discharge process, excellent rate capability with 77 mAh g-1 at a high current density of 5 C, and long cyclic stability in liquid-state batteries. Excitingly, it is also endowed with a high capacity retention of 85% after 500 cycles in solid-state batteries. Furthermore, ex situ XRD, TEM, and ex situ XPS are applied to reveal the structural evolution and charge compensation mechanism of P2-Na0.67[Mn0.67Ni0.21Li0.06Zn0.06]O2, allowing a deep insight into the great significance of structural stability.


Analytical and bioluminescence-based non-invasive quality assessment of differentially grown strawberry (Fragaria x ananassa Duch. 'Asia') during household refrigeration storage.
Strawberry is a typical spring fruit, for which consumer demand is particularly high; however, information is scarce on the comparison of quality traits of strawberries of local and imported origin during household refrigeration storage. That is why we sought to answer the question of to what extent the quality of strawberries changes during 5 days of household refrigeration storage. The choice of methods was focused on fast and at the same time informative analytical methods: pH, total antioxidant capacity, and lipid oxidation. Furthermore, a non-invasive imaging technique: ultra-weak bioluminescence measurement was carried out along with the determination of fruit morphological (weight, height, width, and color) and analytical (antioxidant capacity and lipid oxidation) traits. The data indicate that the longer strawberries are cultivated in the same plantation, the smaller their fruit weight and the lower their fruit quality become. In addition, the results cast light on a controversial fact: despite the fact that the sample from the store had the finest appearance, as determined, its antioxidant capacity was the lowest for the duration of home refrigeration storage, indicating the lowest nutritional value.This was validated by the lipid oxidation levels, which were defined by the amount of malondialdehyde as well as by the rate of ultra-weak bioluminescence. The research underlines the exceptional value of local fruits over importand provides valuable information to customers, encouraging them to purchase strawberries from local farms in order to not only support the local economy but also to adopt a more health-conscious attitude. In addition, ultra-weak bioluminescence testing offers a non-invasive method for assessing fruit quality.


Temporary placement of covered self-expandable metallic stents in the management of benign biliary strictures.
Currently, endoscopic intervention is widely attempted as the first-line treatment of benign biliary strictures because of its convenience and low morbidity. Plastic tube stents (PS) are usually used for such treatment; however, covered self-expandable metallic stents (C-SEMS) are becoming more commonly used at some institutions. The temporary placement of C-SEMS may lead to better outcomes because of their larger diameter and, therefore, better dilation of the stricture, especially in refractory cases. The aim of the present study was to evaluate the efficacy of the temporary placement of C-SEMS in the management of benign biliary strictures. We retrospectively reviewed our endoscopic retrograde cholangiopancreatography (ERCP) database (May 1996 to December 2010), and extracted the data of patients who underwent endoscopic treatment for benign biliary strictures. Then, the follow-up data from patient charts were reviewed to determine the long-term outcomes of those procedures. All patients (n = 56) initially had a PS placed, with or without balloon dilation. However, C-SEMS placement was later attempted in 12 patients because the stricture was refractory to placement of the PS. During their follow-up periods, two patients died of unrelated diseases after 15 and 17 months, and another two still had the C-SEMS in place after 9 and 50 months. In the remaining eight patients, the C-SEMS was removed after a median placement period of 6 months (range, 2-15). Seven patients in this group have not experienced a recurrence at a median follow-up time of 48 months. However, in one patient, stenosis did recur 8 months after the C-SEMS was removed. Temporary placement of C-SEMS can be a treatment option for benign biliary strictures, especially in refractory cases.


Digital video image-shearing device for continuous microvessel diameter measurement.
A modular video device for the measurement of vessel diameter by image shearing is presented. The shearing line may be positioned anywhere and at any angle within the video field. The device produces a standard video signal so that the resulting sheared image can be viewed on any CCTV monitor.


Determination of paracetamol and its main impurity 4-aminophenol in analgesic preparations by micellar electrokinetic chromatography.
A micellar electrokinetic chromatographic (MEKC) method was developed for the quality control of paracetamol containing pharmaceutical preparations. The influence of several factors (surfactant concentration, buffer concentration, pH and applied voltage) was studied during development and optimisation of the method. Phosphate buffer (pH 9.0) containing sodium dodecyl sulphate (75 mM) was found as the ideal running buffer for the separation; the applied voltage was 25 kV and the analysis time was 10 min. The limit of quantitation for 4-aminophenol was 6 microgml(-1); the linearity of the method was studied in the concentration ranges 20-260 microgml(-1) for paracetamol and 20-150 microgml(-1) for 4-aminophenol. The method was successfully applied for the quality control of paracetamol containing products.


Amyloid β induces microglia to phagocytose neurons via activation of protein kinase Cs and NADPH oxidase.
Alzheimer's disease is characterized by brain plaques of amyloid beta and by neuronal loss, but it is unclear how amyloid beta causes neuronal loss and how to prevent this loss. We have previously shown that amyloid beta causes neuronal loss by inducing microglia to phagocytose neurons, and here we investigated whether protein kinase Cs and NADPH oxidase were involved in this. The loss of neurons induced by amyloid beta in co-cultures of primary glia and neurons was completely prevented by inhibiting protein kinase Cs with Gö6976 or Gö6983. Directly activating protein kinase Cs with phorbol myristate acetate stimulated microglial phagocytosis, and induced neuronal loss mediated by MFG-E8/vitronectin receptor pathway of microglial phagocytosis. Blocking phagocytosis by MFG-E8 knockout or receptor inhibition left live neurons, indicating microglial phagocytosis was the cause of neuronal death. Phorbol myristate acetate stimulated the microglial NADPH oxidase, and inhibiting the oxidase prevented neuronal loss. A physiological activator of NADPH oxidase, fMLP, also induced neuronal loss dependent on microglia. Amyloid beta-induced neuronal loss was blocked by NADPH oxidase inhibitors, superoxide dismutase or Toll-like receptor function-blocking antibodies. The results indicate that amyloid beta induces microglial phagocytosis of neurons via activating protein kinase Cs and NADPH oxidase, and that activating the kinases or oxidase is sufficient to induce neuronal loss by microglial phagocytosis. Thus inhibiting protein kinase Cs or NADPH oxidase might be beneficial in Alzheimer's disease or other brain pathologies involving inflammatory neuronal loss mediated by microglia.


Mammalian target of rapamycin signaling in diabetic cardiovascular disease.
Diabetes mellitus currently affects more than 170 million individuals worldwide and is expected to afflict another 200 million individuals in the next 30 years. Complications of diabetes as a result of oxidant stress affect multiple systems throughout the body, but involvement of the cardiovascular system may be one of the most severe in light of the impact upon cardiac and vascular function that can result in rapid morbidity and mortality for individuals. Given these concerns, the signaling pathways of the mammalian target of rapamycin (mTOR) offer exciting prospects for the development of novel therapies for the cardiovascular complications of diabetes. In the cardiovascular and metabolic systems, mTOR and its multi-protein complexes of TORC1 and TORC2 regulate insulin release and signaling, endothelial cell survival and growth, cardiomyocyte proliferation, resistance to β-cell injury, and cell longevity. Yet, mTOR can, at times, alter insulin signaling and lead to insulin resistance in the cardiovascular system during diabetes mellitus. It is therefore vital to understand the complex relationship mTOR and its downstream pathways hold during metabolic disease in order to develop novel strategies for the complications of diabetes mellitus in the cardiovascular system.


SW480, a p53 double-mutant cell line retains proficiency for some p53 functions.
During certain types of cellular stress, the p53 tumor suppressor protein binds to DNA and transactivates a variety of genes that regulate critical responses including apoptosis, cell cycle checkpoints, differentiation, and angiogenesis. In addition, functional p53 is known to be required for efficient nucleotide excision repair (NER) of bulky DNA adducts generated through exposure to environmental mutagens such as UV light. Nonetheless, we previously showed that the model p53-mutated human adenocarcinoma strain SW480 is proficient in the removal of UV-induced cyclobutane pyrimidine dimers (CPD) via NER. We undertook the present study to begin probing the molecular basis for this unexpected repair phenotype. Cytogenetic analysis indicated that SW480 is stable at the chromosomal level, i.e. manifests a karyotypic profile very similar to that revealed for this line as far back as 14 years ago. After fluorescence in situ hybridization (FISH), using a probe complementary to the p53 gene, we found that 98% of the SW480 interphase nuclei contains three copies of the gene, later revealed to be localized on intact short arms of three chromosomes 17. DNA sequence analysis further showed that all three p53 copies in SW480 carry two point mutations (R273H and P309S), and levels of the corresponding mutated p53 protein are about 20-fold higher than in the closely related p53 wild-type strain LoVo. Using an electrophoretic mobility shift assay (EMSA), we demonstrated that R273H/P309S p53 is able to bind with wild-type affinity to its consensus DNA sequence in vitro. Analysis of p21(Cip1/WAF1) expression and in vivo footprinting by ligation-mediated PCR (LMPCR) showed that, in wild-type LoVo cells, an exposure to cellular stress (e.g. UV or ionizing radiation) is necessary for p53 activation of the p21(Cip1/WAF1) promoter. In contrast, the R273H/P309S-mutated p53 protein in SW480 constitutively activates p21(Cip1/WAF1) in the absence of stress through an unknown mechanism. A similar phenomenon whereby mutated p53 in SW480 is able to induce NER-related proteins might explain the normal DNA repair phenotype previously observed in this strain. For now we conclude that, in general, results obtained using SW480 as a p53-deficient cell line should be interpreted very cautiously.


A clinical rating scale for Batten disease: reliable and relevant for clinical trials.
Batten disease (juvenile neuronal ceroid lipofuscinosis [JNCL]) is an autosomal recessive neurodegenerative disorder characterized by blindness, seizures, and relentless decline in cognitive, motor, and behavioral function. Onset is in the early school years, with progression to death typically by late adolescence. Development of a clinical instrument to quantify severity of illness is a prerequisite to eventual assessment of experimental therapeutic interventions. To develop a clinical rating instrument to assess motor, behavioral, and functional capability in JNCL. A clinical rating instrument, the Unified Batten Disease Rating Scale (UBDRS), was developed by the authors to assess motor, behavioral, and functional capability in JNCL. Children with verified JNCL were evaluated independently by three neurologists. Intraclass correlation coefficients (ICCs) were used to estimate the interrater reliability for total scores in each domain. Interrater reliability for scale items was assessed with weighted kappa statistics. Thirty-one children with confirmed JNCL (10 boys, 21 girls) were evaluated. The mean age at symptom onset was 6.1 +/- 1.6 years, and the mean duration of illness was 9.0 +/- 4.4 years. The ICCs for the domains were as follows: motor = 0.83, behavioral = 0.68, and functional capability = 0.85. The Unified Batten Disease Rating Scale (UBDRS) is a reliable instrument that effectively tests for neurologic function in blind and demented patients. In its current form, the UBDRS is useful for monitoring the diverse clinical findings seen in Batten disease.


Significance of coil orientation for motor evoked potentials from nasalis muscle elicited by transcranial magnetic stimulation.
In transcranial magnetic stimulation (TMS) of the motor cortex, the optimal orientation of the coil on the scalp is dependent on the muscle under investigation, but not yet known for facial muscles. Using a figure-of-eight coil, we compared TMS induced motor evoked potentials (MEPs) from eight different coil orientations when recording from ipsi- and contralateral nasalis muscle. The MEPs from nasalis muscle revealed three components: The major ipsi- and contra-lateral middle latency responses of approximately 10 ms onset latency proved entirely dependent on voluntary pre-innervation. They were most easily obtained from a coil orientation with posterior inducing current direction, and in this respect resembled the intrinsic hand rather than the masseter muscles. Early short duration responses of around 6 ms onset latency were best elicited with an antero-lateral current direction and not pre-innervation dependent, and therefore most probably due to stimulation of the nerve roots. Late responses (>18 ms) could inconsistently be elicited with posterior coil orientations in pre-innervated condition. By using the appropriate coil orientation and both conditions relaxed and pre-innervated, cortically evoked MEP responses from nasalis muscle can reliably be separated from peripheral and reflex components and also from cross talk of masseter muscle activation.