Overexpression of annexin 1 in the development and differentiation of urothelial carcinoma.
This study investigates the expression of annexin 1 in urothelial carcinoma (UC) and its relation with clinicopathologic factors, and evaluates its potential clinical significance. Annexin 1 expression was analyzed by immunohistochemical staining with manual tissue microarrays and Western blot in UC. Immunohistochemical analysis of UC in tissue microarrays showed that annexin 1 protein was 76.5% (150/196) positive, which was markedly increased compared with that in the normal urothelium 20.8% (5/24) (p < 0.01). In addition, the positive expression rate of annexin 1 was higher in the high-grade UC (81.7%; 143/175) than in the low-grade UC (33.3%; 7/21). Western blot revealed that the expression of annexin 1 was low in low-grade UC, and markedly increased in high-grade UC. In conclusion, annexin 1 overexpression is observed in UC, which suggests it may be associated with tumorigenesis and its expression correlates with the differentiation of UC.


Subtypes of social phobia and comorbidity with depression and other anxiety disorders.
Epidemiological studies have identified two subtypes of social phobia: speaking-only social phobia which is characterized by the fear of public speaking situations and complex social phobia which is characterized by the fear of multiple social situations. Speaking-only social phobia most closely corresponds to the DSM-IV's 'nongeneralized social phobia' while complex social phobia resembles 'generalized social phobia'. In contrast to the speaking-only social phobia, the complex form is usually more disabling, familial and longer-lasting. In addition, the complex form has a lower chance of spontaneous recovery and carries a higher risk of comorbidity and impairment. Overall, both types of social phobia tend to be underdiagnosed and under-treated. Effective treatments which can manage not only complex social phobia, but also its spectrum of comorbid conditions, are required.


Posterior fossa reconstruction using titanium plate for the treatment of cerebellar ptosis after decompression for Chiari malformation.
We describe our use of a perforated titanium plate to perform a partial posterior fossa cranioplasty in the treatment of cerebellar ptosis and dural ectasia after posterior fossa decompression (PFD). Twelve patients who had undergone PFD underwent posterior fossa reconstruction using a titanium plate. Symptoms were related to either descent of the cerebellum into the decompression or to dural ectasia into the craniectomy defect. Twelve patients who had undergone large suboccipital craniectomies and who presented with persistent headaches and some with neurological symptoms related to syringomyelia, underwent reoperation with placement of a small titanium plate. Ten of 12 patients showed symptomatic improvement after reoperation. Placement of a titanium plate appears to be an effective method of treatment of cerebellar ptosis and dural ectasia after PFD for Chiari malformation.


Undifferentiated thyroid tumors of diffuse small cell type. Histological and immunohistochemical evidence for their lymphomatous nature.
A histological review of 72 undifferentiated thyroid tumors was performed in order to discover small cell anaplastic carcinomas and Non-Hodgkin lymphomas. Cases suspected to be lymphoma were examined for the presence of Ig and keratin and lectins with a PAP-procedure. Among the 72 cases, 68 (94,5%) were anaplastic carcinomas of various types. Four cases (5,5%) were diffuse small cell tumors, which had previously been regarded as anaplastic carcinomas. All four could be identified as Non-Hodgkin lymphomas by histology, immunohistochemistry, repeat biopsy or autopsy. The findings suggest that the majority of small cell anaplastic thyroid tumors are lymphomas and that true anaplastic small cell carcinoma of the thyroid must be extremely rare. Its diagnosis requires electronmicroscopy and/or immunohistochemistry to demonstrate the epithelial nature of tumor cells.


Identification of lncRNA-associated competing endogenous RNA networks for occurrence and prognosis of gastric carcinoma.
Gastric cancer (GC) is one of the common digestive malignancies worldwide and causes a severe public health issue. So far, the underlying mechanisms of GC are largely unclear. Thus, we aim to identify the long non-coding RNA (lncRNA)-associated competing endogenous RNA (ceRNA) for GC. TCGA database was downloaded and used for the identification of differentially expressed (DE) lncRNAs, miRNAs, and mRNAs, respectively. Then, the ceRNA network was constructed via multiple online datasets and approaches. In addition, various in vitro assays were carried out to validate the effect of certain hub lncRNAs. We constructed a ceRNA network, including 76 lncRNAs, 18 miRNAs, and 159 mRNAs, which involved multiple critical pathways. Next, univariate and multivariate analysis demonstrated 11 lncRNAs, including LINC02731, MIR99AHG, INHBA-AS1, CCDC144NL-AS1, VLDLR-AS1, LIFR-AS1, A2M-AS1, LINC01537, and LINC00702, and were associated with OS, and nine of those lncRNAs were considered as hub lncRNAs involved in the sub-ceRNA network. The in vitro assay indicated two lncRNAs, INHBA-AS1 and CCDC144NL-AS1, which were positively related to the GC aggressive features, including proliferation, invasion, and migration. We identified nine hub lncRNAs and the associated ceRNA network related to the prognosis of GC, and then validated two out of them as promising oncogenes in GC.


Strain-rate sensitive ductility in a low-alloy carbon steel after quenching and partitioning treatment.
We investigate an extraordinarily high ductility in a low alloy carbon steel at an elevated temperature after a quenching and partitioning (Q&P) treatment. The conventional (quenched and tempered) reference material does not show similar behavior. Interestingly, the Q&P treated material's ductility is considerably reduced at increasing strain rates while strength remains almost constant. These results indicate the presence of a diffusion-controlled deformation mechanism at elevated temperatures. Our research shows that interlath retained austenite is more stable during deformation at higher temperatures, resulting in a delayed transformation to martensite and therefore to a more pronounced contribution to plastic deformation at (and in the vicinity of) the many interfaces inherently present in this multi-phase steel.


Molecular characterization of the 3-phosphoglycerate kinase gene (PGK1) from the methylotrophic yeast Pichia pastoris.
We report the cloning of the 3-phosphoglycerate kinase gene (PGK1) from the methylotrophic yeast Pichia pastoris by a PCR approach. The coding sequence of the PGK1 gene comprises 1251 bp with the potential to encode a polypeptide of 416 amino acid residues, which shows high identity to homologous proteins from other yeasts. The promoter region of this gene (P(PGK1)) contains regulatory cis-elements found in other PGK1 genes, such as TATA box, CT-rich block and a heat shock element. In the 3' downstream region we identified a tripartite element 5'-TAG-TAGT-TTT-3', which is supposed to be important for transcription termination. As in other yeasts, the PGK1 gene from P. pastoris is present as a single-copy gene. Northern blot analysis revealed that the gene is transcribed as a 1.5 kb mRNA; when cells are grown on glucose the levels of this mRNA are increased two-fold in comparison to cells grown on glycerol. The transcriptional regulation of this gene by the carbon source was further confirmed when the alpha-amylase gene from Bacillus subtilis was placed under the control of P(PGK1): higher levels of expression were obtained when cells were grown on glucose as compared to glycerol and methanol. Preliminary results related to the strength of P(PGK1) show that it represents a potential alternative to constitutive heterologous expression in P. pastoris.


Low Risk of Birth Defects for Infants Whose Mothers Are Treated With Anti-Tumor Necrosis Factor Agents During Pregnancy.
Safety data on anti-tumor necrosis factor (anti-TNF) treatment during pregnancy are limited. We studied the risk of birth defects after anti-TNF treatment in early pregnancy. We collected data on 1,272,424 live-born infants identified from the Danish (2004-2012) and Swedish (2006-2012) population-based health registers. We determined the prevalence of birth defects among infants born to women with chronic inflammatory disease (inflammatory bowel disease, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, or psoriasis), with (n = 683) and without (n = 21,549) anti-TNF treatment during early pregnancy, and in the general population. We compared the risk of any major birth defect and birth defect by organ system for infants born to women with chronic inflammatory disease, with and without anti-TNF treatment. Risks were presented as odds ratios (ORs) with 95% confidence intervals (CIs). We adjusted for maternal age, parity, smoking, body mass index, multiple gestation, country, and chronic inflammatory diagnosis. Birth defects were more prevalent among infants born to women with chronic inflammatory disease, regardless of anti-TNF treatment status, than in the general population (4.8% vs 4.2%). Birth defects occurred in 43 of the infants born to the 683 women who received anti-TNF treatment (6.3%), and 1019 of the infants born to women with chronic inflammatory disease (4.7%). The OR for any defect in women receiving anti-TNF therapy was 1.32 (95% CI, 0.93-1.82); the OR for a cardiovascular defect was 1.60 (95% CI, 0.93-2.58), and the OR for a urinary defect was 2.22 (95% CI, 0.86-4.71). Based on an analysis of data from the health registries in Denmark and Sweden, women who received anti-TNF agents during pregnancy had a slightly (but not significantly) higher risk of having children with birth defects. Although larger studies are needed, the heterogeneity of the observed birth defects did not indicate a common etiology.


SMARCE1-related Coffin-Siris Syndrome: Case report and otolaryngologic manifestations of the syndrome.
Coffin-Siris Syndrome (CSS) is a genetic syndrome associated with multiple congenital anomalies due to mutations in the BAF-complex or SOX gene. Although well characterized overall, the subunits of the BAF-complex or SOX gene affected demonstrate phenotypic differences which are continuing to be defined. Among the variants is the SMARCE1 mutation, the least common identified genotype. This case report presents a pediatric patient with SMARCE1-related CSS, the seventh case reported in the literature. The congenital anomalies are discussed and compared to the reported cases of SMARCE1-related CSS and CSS overall with an emphasis on otolaryngologic manifestations.


Subunit identity of the dimeric 17 beta-hydroxysteroid dehydrogenase from human placenta.
Human placental 17 beta-hydroxysteroid dehydrogenase has been purified with a new rapid procedure based on fast protein liquid chromatography, yielding quantitatively a homogeneous preparation with high specific activity catalyzing the oxidation of 7.2 mumol of estradiol/min/mg of enzyme protein at 23 degrees C, pH 9.2. This preparation was shown to have a subunit mass of 34.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis while having a molecular mass of 68 kDa by both Superose-12 gel-filtration and native pore gradient gel electrophoresis. When 17 beta-hydroxysteroid dehydrogenase was expressed in HeLa cells or overproduced in insect cells using the baculovirus expression system, both from its cDNA encoding a protein of 34 kDa, the enzyme had the same migration in native and sodium dodecyl sulfate-gel electrophoresis as the purified one from human placenta and eluted from the Superose-12 column at the same elution volume. Moreover, all the above forms of this enzyme have similar specific activity. These results clearly demonstrate the identity of the three enzyme forms. The enzyme produced from the cDNA is expressed as a dimer, and its two subunits are identical. 17 beta-Hydroxysteroid dehydrogenase subunit identity is thus proved. The NH2-terminal analysis revealed a unique sequence of Ala-Arg-Thr-Val-Val-Leu-Ile for the purified enzyme from placenta, further confirming the above conclusion.


Exploring the need for a new UK occupational therapy intervention for people with dementia and family carers: Community Occupational Therapy in Dementia (COTiD). A focus group study.
In the Netherlands, Graff et al. found Community Occupational Therapy in Dementia (COTiD) demonstrated benefits to people with dementia and family carers. In this study, focus groups took place with people with dementia and family carers to explore how to make COTiD relevant to the UK context. Six focus groups (three with people living with dementia (n = 18) and three with family carers (n = 21)) took place. Participants were asked for their impressions of the intervention, the extent to which it could meet their needs, and what modifications were needed. Audio-recordings of the groups were transcribed and analysed. Three key themes emerged covering 'loss and living with dementia', 'what helped us', and 'consistency and continuity'. People with dementia and family carers spoke about the impact of their diagnosis on them and their family and what strategies helped. Issues such as timing, follow-up, and the importance of an early intervention in preventing crises were highlighted. There was some concern over the length of the intervention and the disruption it might cause to current schedules. Overall, participants were optimistic about COTiD being used in the United Kingdom if it was to be introduced in a flexible and timely manner, incorporating the needs and existing strategies of the person with dementia. These outcomes have led to changes, such as incorporating more flexibility into COTiD, being made to the intervention prior to its implementation in the United Kingdom.


Centromeric DNA hypomethylation as an epigenetic signature discriminates between germ and somatic cell lineages.
Germ cells have unique features strikingly distinguishable from somatic cells. The functional divergence between these two cell lineages has been postulated to result from epigenetic mechanisms. Here we show that the chromosomal centric and pericentric (C/P) regions in male and female germline cells are specifically DNA-hypomethylated, despite the hypermethylation status in somatic cells. In multipotent germline stem cells, the C/P region was initially hypomethylated and then shifted to the hypermethylation status during differentiation into somatic lineage in vitro. Moreover, the somatic-type hypermethylation pattern was maintained in the somatic cell-derived nuclear transfer embryos throughout preimplantation development. These results imply that the identity of germ cell lineage may be warranted by the hypomethylation status of the C/P region as an epigenetic signature.


Medium term moderate to low-level air pollution exposure is associated with higher C-reactive protein among healthy Danish blood donors.
Air pollution is a significant contributor to the global burden of disease with a plethora of associated health effects such as pulmonary and systemic inflammation. C-reactive protein (CRP) is associated with a wide range of diseases and is associated with several exposures. Studies on the effect of air pollution exposure on CRP levels in low to moderate pollution settings have shown inconsistent results. In this cross-sectional study high sensitivity CRP measurements on 18,463 Danish blood donors were linked to modelled air pollution data for NOx, NO2, O3, CO, SO2, NH3, mineral dust, black carbon, organic carbon, sea salt, secondary inorganic aerosols and its components, primary PM2.5, secondary organic aerosols, total PM2.5, and total PM10 at their residential address over the previous month. Associations were analysed using ordered logistic regression with CRP quartile as individuals outcome and air pollution exposure as scaled deciles. Analyses were adjusted for health related and socioeconomic covariates using health questionnaires and Danish register data. Exposure to different air pollution components was generally associated with higher CRP (odds ratio estimates ranging from 1.11 to 1.67), while exposure to a few air pollution components was associated with lower CRP. For example, exposure to NO2 increased the odds of high CRP 1.32-fold (95%CI 1.16-1.49), while exposure to NH3 decreased the odds of high CRP 0.81-fold (95%CI 0.73-0.89). This large study among healthy individuals found air pollution exposure to be associated with increased levels of CRP even in a setting with low to moderate air pollution levels.


The spectrum concept of schizophrenia: evidence for a genetic-environmental continuum.
Family data from 84 chronic schizophrenic, 11 schizotypal and 90 normal control probands were analyzed by multivariate-multifactorial genetic models using morbid risk statistics. The results were consistent with multifactorial inheritance whereby chronic schizophrenia and schizotypal personality disorder represent different phenotypic manifestations of the same underlying process; that is, the two disorders were found to have different thresholds on a single continuum of genetic-environmental liability. When subclassified according to Taylor and Abrams' criteria, chronic schizophrenic subjects who met these criteria (narrow schizophrenia) had a higher threshold of liability than those who did not (broad schizophrenia). The hypothesis of separate liabilities for the different disease states was rejected. Overall, the results suggest a gradation in multifactorial liability from schizotypal personality disorder (mild) to broad schizophrenia (moderate) to narrow schizophrenia (severe).


Gastric secretory effect of amino acids given enterally and parenterally in dogs.
To investigate the possible role of absorbed amino acids in the "intestinal phase" of gastric secretion, experiments were carried out in 5 mongrel dogs with Heidenhain pouches, gastrostomies, and Roux en-Y jejunostomises. A 3% solution of mixed amino acids was infused at a constant rate of 120 ml per hr throughout a 2-hr period either intravenously or intrajejunally, and the secretory response of the pouches was compared to that obtained during similar infusions of 0.9% NaCl. Intravenous administration of saline did not stimulate gastric secretion. The mean secretory responses (muEq per 2 hr) were: intrajejunal saline 37, intravenous amino acids 330, and intrajejunal amino acids 301. These results suggest that amino acids, absorbed from the gut, may play a role in the stimulation of gastric secretion which occurs when food passes through the intestine.


Colony-forming cells in chronic granulocytic leukemia--I. Proliferative responses to growth factors.
Peripheral blood cells from 2 patients with chronic granulocytic leukemia were separated by density centrifugation. Mononuclear cells of low density (d less than 1.062g cm-3) with blast-cell morphology were cryopreserved before culture in vitro. Upon culture in conventional colony assays, up to 20% of the cells formed hemopoietic colonies. Although the spectrum of colony types resembled that of normal bone-marrow cells, there were large differences between the patients with respect to the number, type and size of colonies that were observed. Colony formation required the addition of hemopoietic growth factors, such as colony-stimulating activity and erythropoietin to the culture medium. The cells were used to assay hemopoietic regulatory molecules. Both erythropoietin and colony-stimulating activity induced a strong proliferative response as measured by thymidine incorporation. Maximal stimulation was observed when erythropoietin and the supernatant of mixed lymphocyte cultures were added simultaneously. The difference between the cells from the 2 patients in clonal assays was reflected by the different response to individual hemopoietic regulators. The time course of maximal stimulation followed distinct patterns dependent on the source of stimulator. The stimulation was linearly dependent on the input cell number. Taken together, cryopreserved blast cells from patients with chronic granulocytic leukemia appear to be very useful for the characterization of factors regulating hemopoiesis, as well as for studies of hemopoiesis in general.


Trends in youth antidepressant dispensing and refill limits, 2000 through 2009.
Antidepressant (AD) dispensing for depression in youth declined in the years following the 2003-04 Food and Drug Administration actions regarding increased risk of suicidal behavior. To extend observation of youth AD dispensing and associated characteristics through 2009 to determine if AD dispensing continues to decline, has stabilized, or has rebounded. Retrospective time series design. Youth (n=57,782) ages 10 to 17 inclusive. Both new (incident) and refill AD dispensing continued to decline through 2009, with no sign of leveling off. However, among youth who started AD treatment the cumulative supply of AD medication remained consistent across the pre- and postperiods, suggesting that cumulative treatment episode duration has not been degraded--possibly as a function of greater days supply with each new refill in the postperiod. Prescribers dramatically curtailed preauthorized refills in the postwarning period. Declines in AD dispensing to depressed youth may not reflect less intensive treatment for those youth who persist beyond the initial dispense. Lower rates of preauthorized refills may have been an attempt by prescribers to encourage return visits to evaluate response and adverse consequences.


Intravenous immunoglobulin treatment in painful sensory neuropathy without sensory ataxia associated with Sjögren's syndrome.
Patients having neuropathy associated with Sjögren's syndrome may present with pain and superficial sensory involvement in the absence of sensory ataxia. Treatment for this form of associated neuropathy has not been established. The case of a patient with painful sensory neuropathy associated with Sjögren's syndrome, whose symptoms, particularly pain, responded well to intravenous immunoglobulin both at onset and in a relapse, is reported. Other patients with painful sensory neuropathy associated with Sjögren's syndrome may also be candidates for intravenous Ig treatment.


The rising phenomenon of circular cities in Japan. Case studies of Kamikatsu, Osaki and Kitakyushu.
Japan has a culture of appreciating nature, but Japan is the 7th biggest waste producer of OECD (Organisation for Economic Co-operation and Development) countries, as it only recycles 19,6 %, deposits 1 % in landfills, and incinerates the rest. Japan also has one of the lowest rates (20 %) of renewable energy of OECD countries, and recently has decided to reactivate nuclear power plants. In contrast, cases of local circular economy initiatives are increasing in Japan. This suggests an opportunity for progress towards more ambitious circular economy goals. Circular Cities (CC) embrace the principles of the circular economy (CE) by managing resources to minimize waste, maximize efficiency, and promote sustainability. The idea of circular cities is taking hold in the country. This work assesses in a top-down approach CC from a general perspective considering the current state of the art, and then, its application to the Japanese scheme to be contrasted within three particular case studies. CC in Japan represents a holistic and sustainable approach that enhances well-being and the economy by using CE principles. It incorporates Japanese cultural attributes and the necessary stakeholders' involvement to maximize closed loops, reduce resource consumption, and establish a regenerative society. The paper recognizes current progress towards circularity in Japan's cities, towns, and villages, even though they may not call themselves "circular cities" per se. The paper proposes a definition for circular cities specifically for Japan. It analyses three case studies: Kamikatsu, Osaki, and Kitakyushu, urban areas with very different circumstances as examples of circular cities, with the final goal to provide recommendations to policymakers to help increase the circular cities phenomenon.


Effect of human TGF-β on the gene expression profile of Schistosoma mansoni adult worms.
Schistosoma mansoni is responsible for schistosomiasis, a parasitic disease that affects 200 million people worldwide. Molecular mechanisms of host-parasite interaction are complex and involve a crosstalk between host signals and parasite receptors. TGF-β signaling pathway has been shown to play an important role in S. mansoni development and embryogenesis. In particular human (h) TGF-β has been shown to bind to a S. mansoni receptor, transduce a signal that regulates the expression of a schistosome target gene. Here we describe 381 parasite genes whose expression levels are affected by in vitro treatment with hTGF-β. Among these differentially expressed genes we highlight genes related to morphology, development and cell cycle that could be players of cytokine effects on the parasite. We confirm by qPCR the expression changes detected with microarrays for 5 out of 7 selected genes. We also highlight a set of non-coding RNAs transcribed from the same loci of protein-coding genes that are differentially expressed upon hTGF-β treatment. These datasets offer potential targets to be explored in order to understand the molecular mechanisms behind the possible role of hTGF-β effects on parasite biology.


How the COVID-19 pandemic shaped adolescents' future orientations: Insights from a global scoping review.
Throughout adolescence, youth imagine what the future holds and determine plans to achieve their educational, professional, and personal goals. In this article, we review research that explores how adolescents' future orientations were shaped by the societal unpredictability, physical and mental health risks, and educational disruptions brought on by the COVID-19 pandemic. Findings show that the pandemic, which exacerbated existing societal inequities, also heightened adolescents' social awareness, provoked feelings of uncertainty, and altered adolescents' short- and long-term plans for educational and career prospects. Throughout school building closures and program cancellations, families played a greater role in fostering adolescents' plans. With prospects for more societal uncertainty on the horizon, future directions point toward supporting adolescents in developing adaptable and flexible future orientations.


Alleviation of drug withdrawal symptoms by treatment with a potent mixture of natural lipids.
Acute treatment with morphine, as well as with other addictives, causes an initial decrease in brain membrane lipid microviscosity. Upon chronic exposure to the addictive drug, the original fluidity of brain membranes is restored, mostly due to an increase in the cholesterol content. Withdrawal of the drug leaves the membranes hyperviscous and the central nervous system supersensitive, as manifested in the abstinence syndrome. Based on this scheme of events, we have designed treatments of addicted mice with a special lipid mixture extracted from hen egg-yolk (Active-lipid, AL), which is a highly potent membrane fluidizer. In vivo treatment of morphine-addicted mice with AL, either by injection or as a diet supplement, reversed the brain membrane hyperviscosity and, concomitantly, markedly reduced, or even completely abolished the withdrawal symptoms precipitated by naloxone. Treatments with the fully saturated, rigidifying phospholipid dipalmitoyl-lecithin, exacerbated the withdrawal symptoms. These results could be the basis for a novel innocuous treatment which may facilitate the rehabilitation of drug and alcohol addicts.


Evaluation of the performance of four rapid tests for detection of hepatitis B surface antigen in Antananarivo, Madagascar.
Four rapid immunochromatographic assays--Determine HBsAg, Virucheck HBsAg, Cypress HBsAg Dipstick and Hexagon HBsAg--for human hepatitis B surface antigen (HBsAg) detection in human serum were evaluated. A collection of reference serum samples (91 HBsAg positive and 109 HBsAg negative) stored at -80 degrees C was used. Sensitivity and positive predictive value (PPV) exceeded 95%, and specificity and negative predictive value (NPV) exceeded 96% for all tests. The Determine HBsAg test performed best in this study, with a sensitivity of 97.8%, a specificity and PPV of 100%, a NPV of 98.2% and an accuracy rate of 99.0%. However, the differences between the tests were not significant. Other factors should therefore also be taken into account by the Ministry of Health in its decision to recommend a particular test: price, availability, delivery time and feasibility of whole-blood testing. The Determine test appears to be the most suitable for Madagascar, based on all these criteria. The use of this test, despite its lower sensitivity, could prevent blood-borne transmission of hepatitis B virus (HBV) in areas with limited resources.


Phenolic modification as an approach to improve the pharmacology of the 3-acyloxy-2-benzylpropyl homovanillic amides and thioureas, a promising class of vanilloid receptor agonists and analgesics.
In order to improve the analgesic activity and pharmacokinetics of thioureas 2 and 3, which we previously developed as potent vanilloid receptor (VR) agonists, we prepared and characterized phenolic modifications of them and of their amide surrogates (7, 8). The aminoethyl analogue of the amide template 13 was a potent analgesic with an EC50=0.96 microg/kg in the AA-induced writhing test and with better in vivo stability than the parent phenol.


Anterior pituitary gland T1 signal intensity is influenced by time delay after injection of gadodiamide.
To test the hypothesis of washout from the anterior pituitary (AP) gland after serial injections of gadodiamide. We included 59 patients with history of at least 5 injections of gadodiamide. Values of mean signal intensity of the AP and of the central pons were measured on unenhanced sagittal T1-weighted images. AP-to-pons signal intensity ratios were calculated dividing the values of the AP by those of the pons. The measurements were performed using MR images acquired at four different time points including baseline (prior to any gadodiamide injection), minimum post-injection time delay, maximum post-injection time delay, and last available MR scans. Normalized ratios (i.e. ratios divided total volume of injected gadodiamide) were also calculated. To assess the difference between ratios, non-parametric Wilcoxon signed-rank test was applied. The correlations were tested with non-parametric Spearman correlation coefficient. A p-value < 0.05 was considered as statistically significant. A statistically significant increase of AP signal intensity was found by comparing the baseline scans with both the minimum time delay (p = 0.003) and maximum time delay scans (p = 0.005). We found significant higher normalized ratios for minimum post-injection time delay with respect to maximum post-injection time delay (p < 0.001). The normalized ratios demonstrated a statistically significant negative correlation with the post-injection time delay (r = - 0.31; p = 0.006). The findings of this study suggest that washout phenomena of retained/deposited gadolinium from the AP are influenced by the total injected volume and post-injection time delay.


Group II mGlu receptor antagonist LY341495 enhances the antidepressant-like effects of ketamine in the forced swim test in rats.
Numerous preclinical and clinical studies have reported the rapid and sustained antidepressant effects of the NMDA receptor antagonist ketamine. Because ketamine induces several undesirable and dangerous effects, a variety of strategies have been suggested to avoid such effects. Here, we propose to enhance the sub-effective doses of ketamine by co-administration with the group II metabotropic glutamate (mGlu) receptor antagonist LY341495. This compound potentially acts as an antidepressant via a mechanism similar to that of ketamine. To investigate the rapid and sustained antidepressant-like effects of these drugs, we administered ketamine and LY341495 individually or in combination, 40 min and 24 h before the forced swim test (FST). We found that sub-effective doses of ketamine and LY341495, given jointly, induce significant antidepressant-like effects, at both 40 min and 24 h after administration. The results obtained using Western blot technique indicate that mammalian target of rapamycin (mTOR) pathway activation may be involved in the mechanism of this action. The effects of drugs, used at identical ranges of times and doses, on spontaneous locomotor activity in rats were excluded. Furthermore, the results obtained from the rota-rod test and the ketamine-induced hyperlocomotion test suggest a lack of potentially adverse effects from the combined administration of ketamine and LY341495 at doses previously used in the FST. Altogether, these data suggest that the joint administration of ketamine and LY341495 might be a noteworthy alternative to the use of solely ketamine in the therapy of depression.


To test or not to test? Moderators of the relationship between risk perceptions and interest in predictive genetic testing.
The moderating effects of motivational factors (illness prevention vs. emotional reassurance), regulatory focus (health vs. illness orientations), and cancer anxiety on the relationship between risk perceptions and women's interest in predictive genetic testing for breast cancer were studied among 102 women with no history of breast cancer. Risk perceptions per se were unrelated to testing interests. Perceptions of higher personal risk for developing breast cancer were positively related to women's interest in testing only among women whose dominant motivation was not emotional reassurance, who were not oriented towards ruling-out disease, and who were not highly anxious about breast cancer. These findings pointed to conditions under which risk perceptions may enhance screening behaviors, and other conditions under which they may not.


Spatio-temporal analysis of malaria incidence and its risk factors in North Namibia.
Millions of dollars have been spent in fighting malaria in Namibia. However, malaria remains a major public health concern in Namibia, mostly in Kavango West and East, Ohangwena and Zambezi region. The primary goal of this study was to fit a spatio-temporal model that profiles spatial variation in malaria risk areas and investigate possible associations between disease risk and environmental factors at the constituency level in highly risk northern regions of Namibia. Malaria data, climatic data, and population data were merged and Global spatial autocorrelation statistics (Moran's I) was used to detect the spatial autocorrelation of malaria cases while malaria occurrence clusters were identified using local Moran statistics. A hierarchical Bayesian CAR model (Besag, York and Mollie's model "BYM") known to be the best model for modelling the spatial and temporal effects was then fitted to examine climatic factors that might explain spatial/temporal variation of malaria infection in Namibia. Average rainfall received on an annual basis and maximum temperature were found to have a significant spatial and temporal variation on malaria infection. Every mm increase in annual rainfall in a specific constituency in each year increases annual mean malaria cases by 0.6%, same to average maximum temperature. The posterior means of the time main effect (year t) showed a visible slightly increase in global trend from 2018 to 2020. The study discovered that the spatial temporal model with both random and fixed effects best fit the model, which demonstrated a strong spatial and temporal heterogeneity distribution of malaria cases (spatial pattern) with high risk in most of the Kavango West and East outskirt constituencies, posterior relative risk (RR: 1.57 to 1.78).


Prospective direct comparison of non-invasive liver tests in outpatients with type 2 diabetes using intention-to-diagnose analysis.
No prospective diagnostic studies have directly compared widespread non-invasive liver tests in patients with type 2 diabetes (T2D) using the intention-to-diagnose method for each of the three main histological features of metabolic dysfunction associated steatotic liver disease - namely fibrosis, metabolic dysfunction-associated steatohepatitis (MASH), and steatosis. To compare the performance of nine tests using the intention-to-diagnose rather than the standard method, which would exclude non-evaluable participants METHODS: Biopsy was used as the reference with predetermined cut-offs, advanced fibrosis being the main endpoint. The Nash-FibroTest panel including FibroTest-T2D, SteatoTest-T2D and MashTest-T2D was optimised for type 2 diabetes. FibroTest-T2D was compared to vibration-controlled transient elastography stiffness (VCTE), two-dimensional shear-wave elastography stiffness (TD-SWE), and Fibrosis-4 blood test. NashTest-T2D was compared to aspartate aminotransferase. SteatoTest-T2D was compared to the controlled attenuation parameter and the hepatorenal gradient. Among 402 cases, non-evaluable tests were 6.7% for VCTE, 4.0% for hepatorenal gradient, 3.2% for controlled attenuation parameter, 1.5% for TD-SWE, 1.2% for NashTest-T2D, and 0.02% for Fibrosis-4, aspartate aminotransferase and SteatoTest-T2D. The VCTE AUROC for advanced fibrosis was over-estimated by 6% (0.83 [95% CI: 0.78-0.87]) by standard analysis compared to intention-to-diagnose (0.77 [0.72-0.81] p = 0.008). The AUROCs for advanced fibrosis did not differ significantly in intention-to-diagnose between FibroTest-T2D (0.77; 95% CI: 0.73-0.82), VCTE (0.77; 95% CI: 0.72-0.81) and TD-SWE(0.78; 0.74-0.83) but were all higher than the Fibrosis-4 score (0.70; 95% CI all differences ≥7%; p ≤ 0.03). For MASH, MashTest-T2D had a higher AUROC (0.76; 95% CI: 0.70-0.80) than aspartate aminotransferase (0.72; 95% CI: 0.66-0.77; p = 0.035). For steatosis, AUROCs did not differ significantly between SteatoTest-T2D, controlled attenuation parameter and hepatorenal gradient. In intention-to-diagnose analysis, FibroTest-T2D, TD-SWE and VCTE performed similarly for staging fibrosis, and out-performed Fibrosis-4 in outpatients with type 2 diabetes. The standard analysis over-estimated VCTE performance. gov: NCT03634098.


Treatment response to Janus kinase inhibitor in a child affected by Aicardi-Goutières syndrome.
Baricitinib, a Janus kinase inhibitor (JAK-inhibitor), seems to contribute to an improvement of a child affected by Aicardi-Goutières syndrome (AGS), reducing the interferon score and determining a recovery of cognitive, communicative, and relational dysfunctions, while the gross motor deficit persisted. We report the treatment response to baricitinib, a JAK-inhibitor, in a 4-year-old girl affected by Aicardi-Goutières syndrome (AGS2, RNASEH2B mutation). Using quantitative measures, we detected a significant amelioration characterized by a complete recovery of cognitive, communicative, and relational skills after 8 and 16 months from the beginning of therapy.


Calcified encapsulating peritoneal sclerosis associated with peritoneal dialysis: A case report.
Encapsulating peritoneal sclerosis (EPS) is a rare, but sometimes fatal, complication of peritoneal dialysis characterized by diffuse thickening and encapsulation of the bowel and peritoneum. In more advanced cases, the peritoneum will gradually calcify. EPS usually presents as partial small bowel obstruction and diagnosed on imaging studies. We present a case of a 19-year-old female on long-term peritoneal dialysis with EPS and diffuse peritoneal calcifications.


Heart rate, arterial pressure and propofol-sparing effects of guaifenesin in dogs.
To evaluate the heart rate (HR) and systemic arterial pressure (sAP) effects, and propofol induction dose requirements in healthy dogs administered propofol with or without guaifenesin for the induction of anesthesia. Prospective blinded crossover experimental study. A total of 10 healthy adult female Beagle dogs. Dogs were premedicated with intravenous (IV) butorphanol (0.4 mg kg-1) and administered guaifenesin 5% at 50 mg kg-1 (treatment G50), 100 mg kg-1 (treatment G100) or saline (treatment saline) IV prior to anesthetic induction with propofol. HR, invasive sAP and respiratory rate (fR) were recorded after butorphanol administration, after guaifenesin administration and after propofol and endotracheal intubation. Propofol doses for intubation were recorded. Repeated measures analysis of variance (anova) was used to determine differences in propofol dose requirements among treatments, and differences in cardiopulmonary values over time and among treatments with p < 0.05 considered statistically significant. Propofol doses (mean ± standard deviation) for treatments saline, G50 and G100 were 3.3 ± 1.0, 2.7 ± 0.7 and 2.1 ± 0.8 mg kg-1, respectively. Propofol administered was significantly lower in treatment G100 than in treatment saline (p = 0.04). In treatments G50 and G100, HR increased following induction of anesthesia and intubation compared with baseline measurements. HR was higher in treatment G100 than in treatments G50 and saline following induction of anesthesia. In all treatments, sAP decreased following intubation compared with baseline values. There were no significant differences in sAP among treatments. fR was lower following intubation than baseline and post co-induction values and did not differ significantly among treatments. When administered as a co-induction agent in dogs, guaifenesin reduced propofol requirements for tracheal intubation. HR increased and sAP and fR decreased, but mean values remained clinically acceptable.


Therapeutic anti-psoriatic effects of myeloid-derived suppressor cells in combination with systemic tacrolimus (FK-506) in an imiquimod-induced mouse model of psoriasis.
Although tacrolimus (FK-506) has been shown to be an effective monotherapy for psoriasis, it does not always work well. Currently, combination therapy is frequently used to manage psoriasis because clinical trials have shown it may provide additive or synergistic benefits and reduce risks of adverse effects. Myeloid-derived suppressor cells (MDSCs) have potent immunomodulatory and anti-inflammatory properties in autoimmune diseases. We previously reported that MDSCs had protective effects in a murine model of imiquimod (IMQ)-induced psoriasis. The present study was undertaken to investigate the systemic immunomodulatory and therapeutic efficacy effects of MDSC plus FK-506 in an IMQ-induced mouse model of psoriasis and to investigate the immunomodulatory mechanisms involved. Systemic MDSC plus FK-506 therapy was found to have a significant anti-psoriatic effect in the murine model, to reduce levels of pro-inflammatory cytokines Th1 cytokines (TNF-α and IFN-γ) and Th17 cytokines (IL-17A and IL-23) in serum and skin. However, treatment with MDSCs or FK-506 alone had little impact. Furthermore, the anti-psoriatic effects of MDSC plus FK-506 were associated with histopathological reductions in inflammatory infiltration, epidermal hyperplasia, and hyperkeratosis. In addition, this combined treatment also attenuated IMQ-induced splenomegaly, and increased the proportion of CD4+CD25+FoxP3+ regulatory T (Treg) cells and decreased the proportions of CD4+IFN-γ+ Th1 cells and CD4+IL-17+ Th17 cells in spleen. Taken together, our results show systemic combination therapy with MDSCs and FK-506 had a better therapeutic effect in our IMQ-induced psoriasis model than either agent alone, and suggest that this combinatorial therapy might be useful for the management of autoimmune skin diseases like psoriasis.


The effect of two different local anaesthetic solutions on pain experience following apicectomy.
Post-operative pain experience following apicectomy of a single maxillary anterior tooth was compared in two groups of patients having this treatment under local anaesthesia. Twenty patients received 3.6 ml of 2% lignocaine with 1:80,000 adrenaline and 23 patients the same volume of 1.5% etidocaine with 1:200,000 adrenaline, as the local anaesthetic agent. Although soft tissue anaesthesia lasted significantly longer when etidocaine with adrenaline was used, pain experience and analgesic intake did not differ between regimens. Lignocaine with adrenaline produced better operating conditions as haemorrhage control was more effective and the quality of operative anaesthesia was more satisfactory than with etidocaine and adrenaline. The use of etidocaine with adrenaline offered no advantages over lignocaine with adrenaline when administered as infiltration anaesthesia for apical surgery.


Development of a HILIC-MS/MS method for the quantification of histamine and its main metabolites in human urine samples.
An LC-MS/MS method was developed enabling the separation and quantification of histamine and its main metabolites (imidazole acetaldehyde, imidazole acetic acid, methyl imidazole acetic acid, methyl histamine, acetyl histamine) in urine samples. A fast separation was achieved in 10 min on two HILIC columns connected in series by adopting a linear gradient followed by an isocratic hold. The sample preparation consisted of a simple dilution step wherein 10 μL of urine was diluted with acetonitrile (ACN) to a final volume comprising 95% ACN. For methyl imidazole acetic acid, an additional dilution step was incorporated due to its high natural levels. Hereafter, the samples were stored at -20 °C and centrifuged prior to injection. Matrix matched calibrators were unavailable due to the endogenous occurrence of the compounds of interest. The occurrence of matrix effects and the lack of labeled internal standards prompted the use of the standard addition method as a viable alternative to solvent calibration. The validation of the method entailed matrix effects, accuracy and precision and was performed in compliance with the recent guidelines on endogenous compounds issued by the International Conference of Harmonization (ICH). The method was then adopted for the quantification of histamine and its metabolites in human urine samples collected from healthy volunteers and patients suffering from gastrointestinal discomfort.


Influence of growth structures and fixed appliances on automated cephalometric landmark recognition with a customized convolutional neural network.
One of the main uses of artificial intelligence in the field of orthodontics is automated cephalometric analysis. Aim of the present study was to evaluate whether developmental stages of a dentition, fixed orthodontic appliances or other dental appliances may affect detection of cephalometric landmarks. For the purposes of this study a Convolutional Neural Network (CNN) for automated detection of cephalometric landmarks was developed. The model was trained on 430 cephalometric radiographs and its performance was then tested on 460 new radiographs. The accuracy of landmark detection in patients with permanent dentition was compared with that in patients with mixed dentition. Furthermore, the influence of fixed orthodontic appliances and orthodontic brackets and/or bands was investigated only in patients with permanent dentition. A t-test was performed to evaluate the mean radial errors (MREs) against the corresponding SDs for each landmark in the two categories, of which the significance was set at p < 0.05. The study showed significant differences in the recognition accuracy of the Ap-Inferior point and the Is-Superior point between patients with permanent dentition and mixed dentition, and no significant differences in the recognition process between patients without fixed orthodontic appliances and patients with orthodontic brackets and/or bands and other fixed orthodontic appliances. The results indicated that growth structures and developmental stages of a dentition had an impact on the performance of the customized CNN model by dental cephalometric landmarks. Fixed orthodontic appliances such as brackets, bands, and other fixed orthodontic appliances, had no significant effect on the performance of the CNN model.


Microbial fuel cell (MFC) for bioelectricity generation from organic wastes.
Microbial fuel cells (MFCs) have gained a lot of attention recently as a mode of converting organic matter into electricity. In this study, a compost-based microbial fuel cell that generates bioelectricity by biodegradation of organic matter is developed. Grass cuttings, along with leaf mold, rice bran, oil cake (from mustard plants) and chicken droppings (waste from chickens) were used as organic waste. The electric properties of the MFC under anaerobic fermentation condition were investigated along with the influence of different types of membranes, the mixing of fly ash, and different types of electrode materials. It is observed that the maximum voltage was increased by mixing fly ash. Cellophane showed the highest value of voltage (around 350mV). Bamboo charcoal is good for anode material; however carbon fiber is better for the cathode material in terms of optimization of power generated. This developed MFC is a simple cell to generate electricity from organic waste.


The comparative costs of laparoscopic and open liver resection: a report for the 2nd International Consensus Conference on Laparoscopic Liver Resection.
The techniques of laparoscopic liver resection (LLR) have developed rapidly in selected centers, while global adoption of this approach has been cautious. The costs of LLR compared to open resection (OLR) are considered an important metric in evaluating this approach and may be a barrier to adoption in some centers. To formulate a consensus statement using the Zurich-Danish consensus model to the question of "What are the comparative outcomes of cost for LLR and OLR, minor and major?" a systematic search of the literature was conducted. Results were presented to the jury in September 2014 and updated in August 30, 2015. Adjustments for currency conversions and inflation were not performed due to limitations in available data. Thirty-four studies were reviewed, and 11 relevant papers were selected for inclusion. No randomized control studies were found. Five studies were case-matched comparisons, while the remaining studies were retrospective reviews. The number of patients in each study ranged from 28 to 74, and the cumulative number of patients was 643 comparing 350 OLR to 293 LLR. Overall median hospital stay was lower for LLR at 4.6 versus 7.4 days. This remained valid when only the case-matched studies were analyzed, 4.6 (n = 178) versus 6.6 days (n = 266). The median overall total costs were 16.3 % lower (range 0 to -22 %) for LLR compared to OLR. This remained valid in the subgroup analysis of the case-matched studies, with a median 17.4 % lower costs for the LLR. Median OR costs were 3 % higher for LLR (range -9 to 40 %) but 32.9 % lower for hospital ward costs (range 0 to -60 %) when compared to OLR. Currently, the published literature indicates that overall hospital costs are less for LLR when compared to OLR (Level of evidence 3a and 3b). This evidence is strongest for minor hepatic resections. The decreased cost is based on savings in hospital ward costs and likely related to a significantly shorter hospital stay for LLR.


Stereoscopic computed three-dimensional surface displays.
Stereoscopic displays of certain anatomic areas are said consistently to provide clinical information that is not recognized in monoscopic viewing. Computed 3-D displays entail no extra patient irradiation.


Salvage Endoscopic Nasopharyngectomy for Locally Recurrent T1 and T2 Nasopharyngeal Carcinoma.
Assessment of salvage endoscopic nasopharyngectomy for recurrent nasopharyngeal carcinoma (rNPC). This is a retrospective study of ten rNPC who underwent salvage endoscopic nasopharyngectomy. Recurrent status for each recurrence was determined before surgery and only recurrent T1 (rT1) and recurrent T2 (rT2) were taken up for surgery after review with radiation oncology colleagues. There were seven rT2 and three rT1 patients. Two patients have undergone simultaneous radical neck dissection (RND) together with endoscopic nasopharyngectomy for associated neck nodes. Outcome of the study was done in turn of disease free, disease residual and disease recurrence. Locally disease free and overall survival rates were 40% (4/10) and 50% (5/10) respectively. Locally disease free till the last follow up was achieved in 4 patients while one patient is on palliative chemotherapy post-surgery for locally positive disease for the last three years. Of the four patients that are locally disease free, two patients received chemoradiation (CTRT) post-surgery while two patients only underwent endoscopic nasopharyngectomy for rT1. There were no any major operative complications except nasal crusting. Recurrent T1 and T2 can be manage with endoscopic nasopharyngectomy and post-surgery ctrt should be given in feasible patients. To detect early recurrent and improve the survival, regular endoscopic follow up is needed.


Experience with International Neuroblastoma Staging System and Pathology Classification.
The International Neuroblastoma Staging System and Pathology Classification were proposed in 1988 and in 1999, respectively, but their clinical value has not yet been fully studied in new patients. Six hundred and forty-four patients with neuroblastoma treated between January 1995 and December 1999 were analysed by these classifications. The 4-year overall survival rate of patients <12 months of age with INSS stages 1, 2A, 2B, 3 and 4S disease was 98.5%, which was significantly higher than the 73.1% rate in stage 4 patients <12 months (P<0.0001). When patients were > or = 12 months, the 4-year overall survival rate of patients with neuroblastoma at 1, 2A, 2B and 3 stages was 100% and that of patients at stage 4 was 48.5% (P<0.0001). As to the International Neuroblastoma Pathology Classification histology, the 4-year overall survival rate was 98.8% in patients with favourable histology and 60.7% in those with unfavourable histology in the <12 months group (P<0.0001). In the > or = 12 months group, the 4-year oral survival of patients with favourable histology was 95.3% and that of patients with unfavourable histology was 50.6% (P<0.0001). Among biological factors, MYCN amplification, DNA diploidy and 1p deletions were significantly associated with poor prognosis in patients <12 months, as were MYCN amplification and DNA diploidy in patients > or = 12 months of age. Multivariate analysis showed that the INSS stage (stage 4 vs other stages) and International Neuroblastoma Pathology Classification histology (unfavourable vs favourable) were significantly and independently associated with the survival of patients undergoing treatment, stratified by age, stage and MYCN amplification (P=0.0002 and P=0.0051, respectively).


Development of a Modular and Equitable Surgical Simulator.
There is a tremendous need for affordable and accessible surgical simulators in the United States and abroad. Our group developed a portable, modular, inexpensive surgical simulator designed for all levels of surgical trainees, from medical students to cardiothoracic surgery fellows, and adaptable to a variety of surgical specialties. Our goal is to provide a platform for innovative surgery simulation that applies to any learner or resource setting. We describe the development, assembly, and future directions for this simulator.


A longitudinal seizure outcome following the COVID-19 pandemic in 2020 and 2021: Transient exacerbation or sustainable mitigation.
To study the longitudinal seizure outcomes of people with epilepsy (PWE) following the acute and chronic phases of the coronavirus disease 2019 (COVID-19) pandemic. Consecutive PWE who were treated at the epilepsy center of Hiroshima University Hospital between 2018 and 2021 were enrolled. We evaluated the incidence of seizure frequency increase or decrease following the pandemic during observational periods in 2020 and 2021. Data between 2018 and 2019 were used as a control set. The sustainability of the altered seizure frequency condition was evaluated throughout the study period. We analyzed the clinical, psychological, and social factors associated with PWE with seizure exacerbation or amelioration. Among the 223 PWE who were evaluated (mean age 37.8 ± 16.3 years), seizure frequency increased for 40 (16.8%) and decreased for 34 (15.2%) after the pandemic began. While seizure exacerbation tended to be a transient episode during 2020, seizure amelioration was likely to maintain excellent status over the observation periods; the sustainability of the altered seizure frequency condition was more prominent for amelioration than exacerbation (p < 0.001). Seizure exacerbation was significantly associated with "no housemate" (odds ratio [OR] 3.37; p = 0.045) and "comorbidity of insomnia" (OR 5.80; p = 0.004). Conversely, "structural abnormality of MRI" (OR 2.57; p = 0.039) and "two-generation householding" (OR 3.70; p = 0.004) were independently associated with seizure amelioration. This longitudinal observation confirmed that seizure exacerbation and amelioration emerged during the COVID-19 pandemic. The COVID-19 pandemic has shed light on the stark difference that social support systems can make on outcomes for PWE.


Effects of antihypertensive therapy on cardiovascular response to exercise.
The effects of alpha methyldopa and hydrochlorothiazide therapy on exercise induced changes in arterial pressure, heart rate, rate-pressure product and total duration of exercise were evaluated with graded treadmill exercise. Although both agents were equally effective in reducing resting arterial pressure, alpha methyldopa appeared to provide slower build-up, a lower ceiling and more rapid recovery of arterial pressure in response to exercise stress than did hydrochlorothizide. Alpha methyldopa significantly reduced the degree of rise of the rate-pressure product (an index of myocardial oxygen consumption) during the exercise and recovery phases, whereas hydrochlorothiazide failed to do so. The total duration of exercise (an index of work capacity) was unchanged with either hydrochlorothiazide or alpha methyldopa. Therapy with alpha methyldopa presumably reduced myocardial oxygen demand without reducing work capacity.


Mechanism of intra-oral transport in a herbivore, the hyrax (Procavia syriacus).
Movements in the jaw, tongue and hyoid during feeding behaviour were recorded with cine-X-ray. Food was moved through the mouth by anterior/posterior motion of the tongue surface relative to the hard palate. This was true for both stage I transport, from the front of the mouth to the molar tooth row and stage II transport, from the molar tooth row to the vallecular area of the oropharynx. During a series of chew cycles, processed food collected in the oropharynx prior to a swallow. Swallows occurred as discrete events punctuating chew sequences and were characterized by coordinated movements of tongue and soft palate. Similar mechanisms of transport have been observed in the opossum and the cat, indicating a common mammalian behaviour. Differences between this herbivore and anthropoid primates can be attributed to differences in anatomy of the oral apparatus.


Neurochemical correlates of behavioural responses to frustrative nonreward in the rat: implications for the role of central noradrenergic neurones in behavioural adaptation to stress.
We tested whether the stress of nonreward has neurochemical effects on noradrenergic neurones which resemble those reported for other forms of stress. Rats trained to run in a straight runway for food reward were subjected to either 1 or 10 extinction trials. Half the rats in each group were injected before the start of acquisition with IP 6-hydroxydopamine to deplete peripheral noradrenaline stores. All animals were killed immediately after their final test in the runway, together with untrained controls. Noradenaline depletion had no behavioural or neurochemical effects. The rate of extinction in the 10-trial group, which was indexed by the slope of the linear regression of running time on trial, correlated negatively with both alpha 2- and beta-adrenoceptor number (Bmax). There were no differences between groups in cerebral cortical noradrenaline content, or alpha 2- or beta-adrenoceptor binding. These results substantially conflict with those predicted from Stone's hypothesis relating beta-adrenoceptor sensitivity to the behavioural response to stress. A further finding was that alpha 2-, but not beta-adrenoceptor number, negatively correlated with levels of noradrenaline in the tissue, suggesting that noradrenaline is less involved in the regulation of beta- than in that of alpha 2-adrenoceptors.


Acute diuretic effect of continuous intravenous infusion of an aqueous extract of Coriandrum sativum L. in anesthetized rats.
The present investigation was carried out to evaluate the acute diuretic activity of continuous intravenous infusion of an aqueous extract of the seed of Coriandrum sativum L. Apiaceae (coriander) in rats. The aqueous extract of coriander seed was administered by continuous intravenous infusion (120 min) at two doses (40 and 100mg/kg) to anesthetized Wistar rats. Furosemide (10mg/kg), a standard diuretic was used as the reference drug. Excretion of water and electrolytes (sodium, potassium and chloride) in urine was measured, and glomerular filtration rate (equal to creatinine clearance) was determined. The crude aqueous extract of coriander seeds increased diuresis, excretion of electrolytes, and glomerular filtration rate in a dose-dependent way; furosemide was more potent as a diuretic and saluretic. The mechanism of action of the plant extract appears to be similar to that of furosemide. The aqueous extract of coriander seed possesses diuretic and saluretic activity, thus, validating the use of coriander as a diuretic plant in Moroccan pharmacopoeia.


Coordination of eye and head components of movements evoked by stimulation of the paramedian pontine reticular formation.
Constant frequency microstimulation of the paramedian pontine reticular formation (PPRF) in head-restrained monkeys evokes a constant velocity eye movement. Since the PPRF receives significant projections from structures that control coordinated eye-head movements, we asked whether stimulation of the pontine reticular formation in the head-unrestrained animal generates a combined eye-head movement or only an eye movement. Microstimulation of most sites yielded a constant-velocity gaze shift executed as a coordinated eye-head movement, although eye-only movements were evoked from some sites. The eye and head contributions to the stimulation-evoked movements varied across stimulation sites and were drastically different from the lawful relationship observed for visually-guided gaze shifts. These results indicate that the microstimulation activated elements that issued movement commands to the extraocular and, for most sites, neck motoneurons. In addition, the stimulation-evoked changes in gaze were similar in the head-restrained and head-unrestrained conditions despite the assortment of eye and head contributions, suggesting that the vestibulo-ocular reflex (VOR) gain must be near unity during the coordinated eye-head movements evoked by stimulation of the PPRF. These findings contrast the attenuation of VOR gain associated with visually-guided gaze shifts and suggest that the vestibulo-ocular pathway processes volitional and PPRF stimulation-evoked gaze shifts differently.


The influence of ATP on sugar uptake mediated by the constitutive glucose carrier of Saccharomyces cerevisiae.
The glucose carrier of Saccharomyces cerevisiae transports the phosphorylatable sugars glucose, mannose, fructose and 2-deoxy-D-glucose (2-dGlc) and the non-phosphorylatable sugar 6-deoxy-D-glucose (6-dGlc). Reduction of the ATP concentration by, for example, incubating cells with antimycin A, results in a decrease in uptake of 2-dGlc and fructose. These uptake velocities can be increased again by raising the ATP level. These results establish a role of ATP in sugar transport. Transport of glucose and mannose is less affected by changes in the ATP concentration than 2-dGlc and fructose uptake, while the 6-dGlc transport is independent of the amount of ATP in the cells. Also, reduction of the kinase activity by incubation with xylose diminished transport of 2-dGlc and fructose, while the uptake of glucose and mannose remained unchanged. It is discussed that these results are due to transport-associated phosphorylation with ATP as substrate and the hexokinases and the glucokinase as phosphorylating enzymes.


Inactivating mutations of the caspase-10 gene in gastric cancer.
We have analysed the genetic alteration of the entire coding region and all splice sites of caspase-8 and -10 genes in 99 gastric cancers by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and sequencing. We found LOH of the caspase-8 and -10 in nine (28%) of 32 and in four (15%) of 26 informative cases, respectively. Overall, three of 99 gastric cancers (3%) were found to have the caspase-10 mutations, which were identified in the coding regions of the death effector domain (codon 147) and the p17 large protease domain (codons 257 and 410), whereas no mutation was detected in caspase-8. In vitro expression studies, the M147T and Q257stop mutants severely impaired caspase-10-mediated apoptosis, whereas the V410I which was the same mutation detected in ALPS patient had a significant, albeit less severe, effect on apoptosis. The data presented here suggest that somatic alterations of the caspase-10 gene might contribute to the pathogenesis in a subset of gastric cancers through the loss of their apoptotic function.


How the immunoassay transformed C-peptide from a duckling into a swan.
This edition of 'Then and now' re-examines Lise Heding's very highly cited paper 'Radioimmunological determination of human C-peptide in serum', which was published in Diabetologia in 1975. We show how this article and other related articles by Heding contributed to heightened respect for C-peptide (and transformation of Heding's research programme). Initially thought of as an inert discard, C-peptide in blood is now recognised as an excellent surrogate measure of insulin secretion under a wide range of conditions. The assay is especially valuable for acute ascertainment of the insulin secretory capabilities of patients with type 1 diabetes or of transplanted beta cells. The assay is also being used to monitor endogenous beta cell loss or in vivo expansion of beta cell mass over the long term. We conclude with two promising future applications: (1) measurements of C-peptide in blood (along with insulin, glucose, and HbA(1c)) at annual intervals as a potential approach to earlier diagnosis of diabetes; and (2) among many recent advances in recognising properties of C-peptide (including status as a candidate hormone), most promising is C-peptide as a possible therapy for diabetic neuropathy and nephropathy.


A novel likely pathogenic CLCN5 variant in Dent's disease.
The majority of cases of Dent's disease are caused by pathogenic variants in the CLCN5 gene, which encodes a voltage-gated chloride ion channel (ClC-5), resulting in proximal tubular dysfunction. We present three members of the same family and one unrelated paediatric patient with the same insertion-deletion CLCN5 variant. The identification of these patients and positive familial segregation led to the re-classification of this variant from one of unknown significance to one of likely pathogenicity. A 41 year old male presented with end stage kidney failure, proteinuria and haematuria. Whole genome sequencing identified an insertion-deletion variant in CLCN5, resulting in a missense change (c.1744_1745delinsAA p.(Ala582Lys)). His brother and nephew, who both exhibited renal impairment, haematuria, proteinuria, glycosuria and nephrocalcinosis, were found to have the same variant. In addition, genetic testing of an unrelated paediatric patient who presented with proteinuria and hypercalciuria, demonstrated the same variant. The identification of this novel variant in four individuals with features of Dent's disease, has led to the re-classification of the variant to one of likely pathogenicity. As a result, our patients and any future patients with the same variant can be offered a likely diagnosis, without the need for kidney biopsy, and their family members can be offered genetic screening.


Anti-ischemic effects of first and second dose of 20 mg isosorbide dinitrate administered 5 hours apart: attenuation of effects despite rising plasma concentration.
Based on evidence that there may be early tolerance development even within the first daily cycle of treatment, this study was undertaken to evaluate the duration and extent of the antiischemic effects of two 20 mg doses of isosorbide dinitrate as used in a well-established regimen documented to maintain effectiveness during long-term treatment. Ischemia parameters were analyzed at 2 and 4 1/2 hours after the first dose as well as at 2 and 7 hours after the second dose given 5 hours later. The studies were performed in 10 male patients with documented coronary artery disease using bicycle ergometry and a double-blind, randomized, placebo-controlled, crossover protocol. ST-segment depression was reduced by 59% (p < 0.0005) at 2 hours and by 42% (p < 0.01) at 4 1/2 hours after the first tablet and by 38% (p < 0.005) at 2 hours and by 15% (p < 0.05) at 7 hours after the second tablet. Increments in ischemia-free workload capacity amounted to 112% (p < 0.005) and to 41% (p < 0.05) after the first tablet and 68% (p < 0.05) and 38% (p < 0.05) at 2 and 7 hours after the second tablet. At 2 and 4 1/2 hours after the first tablet, plasma concentrations of isosorbide dinitrate were 8.4 and 5.9 ng/ml, and those of isosorbide-5-mononitrate were 166.6 and 130.3 ng/ml. At 2 and 7 hours after the second tablet, the concentrations of isosorbide dinitrate were 9.1 and 5.9 ng/ml, and those of isosorbide-5-mononitrate were 224.5 and 148.1 ng/ml.(ABSTRACT TRUNCATED AT 250 WORDS)


The CIOMS view on the use of placebo in clinical trials.
Based on worldwide consultations with experts in science and ethics the revised CIOMS 2002 International Ethical Guidelines for Biomedical Research Involving Human Subjects provide guidance on when the use of placebo as a comparator in clinical research is ethically acceptable. The article reviews the main points of the CIOMS Guidelines and commentaries including the use of placebo in situations where the best current method is available and the relation of placebo to established effective intervention. It discusses the use of placebo in externally sponsored research in low-resource countries and requirements for informed consent related to placebo studies.


[Resection of the deep dorsal penile vein in the treatment of vascular-induced impotence].
In 10 patients aged from 26 to 58 years suffering from impotence caused by abnormal venous outflow a resection of the deep dorsal vein of the penis was performed. In all patients an improvement of the sexual function was observed.


Histopathological effects of delayed reperfusion after middle cerebral artery occlusion in the anesthetized baboon.
In patients with middle cerebral artery (MCA) territory stroke, attempts to recanalize the brain are currently being extended beyond the classic 3-h time window. Mechanical thrombectomy is particularly attractive as it may carry lesser risks of severe hemorrhagic transformation than thrombolysis. However, whether late reperfusion per se promotes hemorrhagic transformation and increases infarct volume as compared to permanent occlusion is unclear. There is no study of the histopathologic sequelae of late reperfusion following MCA occlusion (MCAo) in the non-human primate. Five young adult baboons completed a specially designed protocol of 20-h MCAo (under etomidate anaesthesia), followed by 4-week survival and finally perfusion-fixation. Infarct volume was measured histologically using validated stereological methods. The results were compared to our previously published series of 6 h and permanent MCAo performed with identical experimental and post mortem procedures. An infarct was present in each baboon, consistently involving the caudate head, internal capsule and putamen; the adjacent inferior frontal cortex was involved in one subject. Infarct volume was significantly larger than with 6 h MCAo, as expected, but did not differ from permanent MCAo. There was no evidence of hemorrhage around the infarcted area in any animal. We found that following a 20 h ischemic episode, the infarct volume was similar to that found with permanent occlusion, with no evidence of hemorrhagic transformation. Cautiously extrapolating to the human situation, our findings suggest that even late mechanical recanalization may not promote brain damage and could be considered in selected cases.


Enhancing soil aggregation and acetamiprid adsorption by ecofriendly polysaccharides hydrogel based on Ca2+- amphiphilic sodium alginate.
Soil aggregation plays an important role in agricultural production activities. However, the structure of soil aggregation is destroyed by the natural environment and unreasonable farming management, resulting in the loss of water, fertilizers and pesticides in soil. At present, hydrogels have been widely reported to promote the formation of soil aggregation. In this paper, amphiphilic calcium alginate (ASA/Ca2+) was applied to promote the formation of soil aggregation and enhance pesticide retention. Initially, an ASA was obtained through the one-pot Ugi condensation (a four-component green chemical reaction). Then, ASA/Ca2+ hydrogel is prepared by Ca2+ cross-linking. The formation of soil aggregation was determined through the Turbiscan Lab Expert stability analyzer, Confocal Laser Scanning Microscope (CLSM), and Transmission Electron Microscope (TEM). And the effect of soil aggregation on acetamiprid environmental behavior was investigated by adsorption kinetics, adsorption isotherms, and leaching. The results shown that the three-dimensional network structure of ASA/Ca2+ hydrogel can promote the formation of soil aggregation. Aggregate durability index (ADI) was 0.55 in the presence of ASA/Ca2+ hydrogel, indicating that amphiphilic hydrogel can enhance the stability of soil aggregation. The adsorbing capacity of acetamiprid was 1.58 times higher than pure soil, and the release of acetamiprid only about 20% in the presence of ASA/Ca2+ hydrogel. These results would be helpful for the formation of soil aggregation and pesticides adsorption on soil aggregation. Thus, ASA/Ca2+ hydrogel is likely to improve soil quality, simultaneously it can minimize the mobility of pesticides in the agricultural system.


Regulation of Rho proteins by phosphorylation in the cardiovascular system.
The small G protein Rho signaling pathways are recognized as major regulators of cardiovascular functions, and activation of Rho proteins appears to be a common component for the pathogenesis of hypertension and vascular proliferative disorders. Rho proteins are tightly regulated, and recent evidence suggests that modulation of Rho protein signaling by phosphorylation of Rho proteins provides an additional simple mechanism for coordinating Rho protein functions. This regulation by phosphorylation is particularly important in the arterial wall, where RhoA protein expressed in vascular smooth muscle cells is controlled by the endothelium through the nitric oxide/cGMP-dependent kinase pathway.


Study of human radius construction systematics: evaluation by DXA in dry bone.
This study has been undertaken in order to describe the bone mass distribution of the dry human radius via dual x-ray absorptiometry (DXA) with a Norland XR-800 densitometer machine. A sample of 39 dry radius bones was used. Two projections were made: antero-posterior and lateral, and five regions of interest were selected. The bone densities and the bone mineral contents of the various regions of the radius in the two projections were compared using Student's t tests for paired samples, with statistically significant differences being found in all of the values, except in the proximal extremity (P Ext). The area of greatest bone mineral content (BMC) was the medial diaphysis (M Diaph), followed by the distal extremity (D Ext), with the lowest value being found in the proximal extremity (P Ext). As for bone mineral density (BMD), a great symmetry is observed if we take the mean point of the longitudinal axis as a reference, with it being distributed from highest to lowest from the central part to the extremities. A correlation study of the BMD and BMC values between the segments themselves and with the total, in both positions, provides us with a high correlation (p ≤ 0.01), with the highest correlation value being found for the proximal diaphysis (P Diaph) region, indicating the heterogeneous nature of the distribution of the radius bone mass. Bone densitometry via DXA is useful in order to establish an overview of the structural construction of the human radius.


Low molecular weight protamine as an efficient and nontoxic gene carrier: in vitro study.
The structural similarity between low molecular weight protamine (LMWP), prepared by enzymatic digestion of protamine, and HIV-TAT protein transduction peptide suggested the feasibility of LMWP as an efficient carrier for delivering therapeutic genes while alleviating the cytotoxicity of currently employed gene carriers. LMWP was prepared by enzymatic digestion of protamine with thermolysine. The prepared LMWP peptide and TAT peptide, as well as their complexes with plasmid DNA (pDNA), were examined for cellular uptake behaviors by using confocal microscopy and flow cytometry. The complexation of pDNA and LMWP was monitored by gel retardation test as well as size and zeta potential measurements, and was then further assessed by DNase I protection assay. The transfection efficiency of pDNA/LMWP was examined by varying the pDNA content and charge ratio in the complex, and then compared with that of pDNA/PEI. Cytotoxicity induced by pDNA/LMWP and pDNA/PEI was also examined. Prepared LMWP showed similar transcellular localization behavior and kinetics to those of TAT, and efficiently transferred the pDNA into nucleus and cytoplasm in a short time period. The size and zeta potential of the pDNA/LMWP complex were 120 nm and 30 mV, respectively, which were adequately suitable for cellular uptake. After forming the complex, LMWP appeared to effectively protect pDNA against DNase I attack. The pDNA/LMWP complex showed significantly enhanced gene transfer than both naked pDNA and the pDNA/PEI complex, while exhibiting a markedly reduced cytotoxicity than that of the pDNA/PEI complex. The present study suggested that LMWP could be a useful and safe tool for enhancing delivery of bioactive molecules and therapeutic DNA products into cells when applied in gene therapy.


Silencing in yeast rDNA chromatin: reciprocal relationship in gene expression between RNA polymerase I and II.
About half of approximately 150 rRNA genes are transcriptionally active in Saccharomyces cerevisiae. Chromatin structures in the inactive, and not the active, copies were previously thought to silence both rRNA genes and reporter Pol II genes. Contrary to this belief, we found that silencing of reporters is much stronger in a mutant with approximately 25 rDNA copies, all of which are transcriptionally active. By integrating reporter gene mURA3 with an inactive rDNA copy missing the Pol I promoter, we found that mURA3 is not silenced in chromosomal rDNA repeats. Together with the demonstration of a requirement for active Pol I in silencing, these results show a reciprocal relationship in gene expression between Pol I and Pol II in rDNA.


Intravenous immunoglobulin replacement treatment does not alter polymorphonuclear leukocytes function and surface receptors expression in patients with common variable immunodeficiency.
The study of the expression of CD16, CD11b and Siglec 9 receptors and the oxidative burst provides insights on polymorphonuclear leukocytes (PMN) functionality in common variable immunodeficiency (CVID) and on the possible effects of intravenous immunoglobulin (IVIg) infusion. We evaluated in vivo before and soon after IVIg administration the CD16, CD11b and Siglec 9 expression on unstimulated and Escherichia coli-stimulated PMN and the oxidative burst induced by Escherichia coli and PMA. The E. coli stimulation up-regulated CD16 and Siglec 9 expression and it induced a strong CD11b up-regulation at baseline and soon after IVIg. The oxidative burst overlapped that observed in healthy donors when induced by Escherichia coli while it increased when induced by PMA. Soon after IVIg infusion, the oxidative burst decreased only when induced by PMA. Our results showed that the IVIg infusion in vivo had a minimal effect on CVID's PMN.


Positive and Negative Affect in Clinic-Referred Youth With ADHD.
To examine self-reported positive affect (PA) and negative affect (NA) among youth with ADHD (only and comorbid) and other non-ADHD-referred youth in an ethnically diverse clinical sample. Semi-structured interviews identified 80 pure ADHD, 284 ADHD plus one or more comorbidities, and 730 non-ADHD youth (e.g., other diagnoses or no diagnosis). The Positive and Negative Affect Scale-Children (PANAS-C) was used to assess affective states. Even after controlling for the influence of potential confounds, youth with only ADHD reported higher PA and lower NA than other clinic-referred youth. The ADHD-comorbid group reported higher PA than the "non-ADHD" group, but these groups did not differ on level of NA. ADHD subtype did not influence results. Among clinic-referred youth, ADHD is associated with higher levels of PA and when there are no comorbid disorders, lower levels of NA.


Impact of Hydroalcoholic Extract of Humulus Lupulus L. on Sperm Quality, Reproductive Organs and Hormones in Male Rats.
To investigate the impact of Humulus Lupulus L. hydroalcoholic extract on the body weights, reproductive organs, sperm quality and hormone levels in male rats. By simple random sampling method, seventy male Sprague-Dawley rats were randomly assigned to 7 groups including control group [distilled water, 1 mL/(kg•d)], Tween 80 group [25% Tween 80 solution, 1 mL/(kg•d)], olive oil group [olive oil, 1 mL/(kg•d)], diethyl stilbestrol (DES) group [DES, 100 μg/(kg•body weight)], H50, H150 and H450 [50, 150 and 400 mg/(kg•d) of Humulus Lupulus L extract, respectively]. The administration was performed via gavage once daily for 7 weeks. Body and reproductive organs weights including testes, seminal vesicles, epididymis and prostate were weighted and epididymal sperm quality were determined by digital balance. Blood samples were collected and serum free testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estrogen (E2) levels were measured by rat specific enzyme-linked immunosorbent assay. The percentage increase in mean body weights of rats in the DES and H50, H150 and H450 groups decreased significantly compared to olive oil and Tween 80 groups (all P<0.05). The weights of seminal vesicle, epididymis and testes in rats receiving H50 were significantly higher than the control group (P<0.05 or P<0.01). The sperm count in the rats receiving H50 was significantly lower than the control group (P<0.05). The sperm motile characteristics of the rats receiving hydroalcoholic extract at and DES were significantly lower than those of the control or rats receiving vehicles (all P<0.05). In H50, H150, H450 and DES groups, T and LH levels were decreased, and E2 was significantly increased compared to the control (P<0.05 or P<0.01). The FSH level did not change in all groups (P>0.05). Humulus Lupulus L. extract significantly increased the seminal vesicle and testes weights and reduced the sperm motility.


A Retrospective Study of Rapid Symptom Response in Bleeding Gynecologic Malignancies With Short Course Palliative Radiation Therapy: Less is More.
Advanced gynecologic malignancies can cause significant vaginal bleeding. Radiotherapy (RT) is often used to palliate symptoms, but limited data exist concerning the optimal dose and expected time to bleeding hemostasis in this population. 1) To investigate the overall hemostasis response and kinetics of hemostasis in women with gynecologic malignancies receiving palliative RT. 2) To compare the efficacy of short-course RT (SCRT, less than or equal to five fractions, >3.5 Gy per fraction) vs. conventionally fractionated long-course regimens (greater than five fractions). We identified women receiving palliative RT for bleeding gynecologic malignancies. Initial and maximal hemostasis responses (IHR and MHR) were recorded and categorized as progressive bleeding (PD), stable disease (SD), partial response (PR), or complete response (CR). Clinical variables were correlated with response or toxicity using binary logistic regression statistical methods. Thirty-three women (median age 63) were identified between 2010 and 2019. Median follow-up and survival after RT were 131 days. About 54.5% (18 of 33) received SCRT. Median time to IHR was five days (two-and-a-half days with SCRT) and 78.8% (26 of 33) responded during treatment. Median time to MHR was 13 days. About 100% achieved PR or CR at MHR. Rates of CR were similar between SCRT (83%) and conventionally fractionated schedules (87%). Average durability of hemostatic control was 5.4 months. Overall rate of rebleeding and Grade 3+ toxicity was 9.1% (3 of 33 each). Women receiving SCRT for bleeding gynecologic malignancies achieved rapid symptom control (often during treatment) with minimal rebleeding. In a population whose median survival is four months, SCRT effectively addresses symptomatic disease while minimizing patient burden and toxicity.


Multiple activities of RNA-binding proteins S1 and Hfq.
In all organisms, RNA-binding proteins participate in modulating all the steps in the life cycle of RNA, including transcription, folding, translation and turnover. In bacteria, RNA-binding proteins may be specific for a few RNA targets (e.g., several ribosomal proteins that recognize both rRNA during ribosome assembly and their own mRNAs when acting as highly specific autogenous repressors) or function as global regulators implicated in numerous regulatory networks. Some RNA-binding proteins combine all these features, and this particularly concerns the ribosomal protein S1 and the Sm-like protein Hfq. S1 is a key mRNA-binding protein in gram-negative bacteria; it recognizes mRNA leaders and provides binding of diverse mRNAs to the ribosome at the initiation step of translation. Moreover, S1 is a highly specific autogenous repressor that is able to distinguish its own mRNA from all the others. Hfq is recognized as a global regulator that facilitates small RNA-mRNA interactions in bacteria; it thereby controls the expression of many mRNAs either positively or negatively. In addition, these two proteins were reported to affect transcription, RNA degradation and other processes. Although they have no sequence specificity, Hfq and S1 preferentially bind A/U-rich single-stranded RNA regions; despite this, they nevertheless carry out very different tasks in the cell. This review is focused on the diversity of functions that can be performed by these abundant RNA-binding bacterial proteins.


Development of peptide-based biosensors for detecting cross-linking and deamidation activities of transglutaminases.
Transglutaminases (TGs) are a protein family that catalyzes isopeptide bond formation between glutamine and lysine residues of various proteins. There are eight TG isozymes in humans, and each is involved in diverse biological phenomena due to their characteristic distribution. Abnormal activity of TG1 and TG2, which are major TG isozymes, is believed to cause various diseases, such as ichthyosis and celiac disease. To elucidate TGs' mechanisms of action and develop new therapeutic strategies, it is essential to develop bioprobes that can specifically examine the activity of each TG isozyme, which has not been sufficiently studied. We previously have identified several substrate peptide sequences containing Gln residues for each isozyme and developed a method to detect isozyme-specific activities by incorporating a labeled substrate peptide into lysine residues of proteins. We prepared the fluorescein isothiocyanate (FITC)-labeled Gln substrate peptide (FITC-K5 and FITC-T26) and Rhodamine B-labeled Lys substrate peptide (RhoB-Kpep). Each TG reaction specifically cross-linked these probe pairs, and the proximity of FITC and Rhodamine B significantly decreased the fluorescence intensity of FITC depending on the concentration and reaction time of each TG. In this study, we developed a peptide-based biosensor that quickly and easily measures TG isozyme-specific activity. This probe is expected to be helpful in elucidating TG's physiological and pathological functions and in developing compounds that modulate TG activity.


Genome architecture of the human beta-globin locus affects developmental regulation of gene expression.
To test the role of gene order in globin gene expression, mutant human beta-globin locus yeast artificial chromosome constructs were used, each having one additional globin gene encoding a "marked" transcript (epsilon(m), gamma(m), or beta(m)) integrated at different locations within the locus. When a beta(m)-globin gene was placed between the locus control region (LCR) and the epsilon-globin gene, beta(m)-globin expression dominated primitive and definitive erythropoiesis; only beta(m)-globin mRNA was detected during the fetal and adult definitive stages of erythropoiesis. When an (A)gamma(m)-globin gene was placed at the same location, (A)gamma(m)-globin was expressed during embryonic erythropoiesis and the fetal liver stage of definitive erythropoiesis but was silenced during the adult stage. The downstream wild-type gamma-globin genes were not expressed. When an epsilon(m)-globin gene was placed between the delta- and beta-globin genes, it remained silent during embryonic erythropoiesis; only the LCR-proximal wild-type epsilon-globin gene was expressed. Placement of a beta(m)-globin gene upstream of the (G)gamma-globin gene resulted in expression of beta(m)-globin in embryonic cells and in a significant decrease in expression of the downstream wild-type beta-globin gene. These results indicate that distance from the LCR, an inherent property of spatial gene order, is a major determinant of temporal gene expression during development.


Isolation, characterization and chemical synthesis of a new insect defensin from Chironomus plumosus (Diptera).
Injection of low doses of bacteria into the aquatic larvae of the dipteran insect Chironomus plumosus induces the appearance in their hemolymph of a potent antibacterial activity. We have isolated two 36-residue peptides from this hemolymph which are active against Gram-positive bacteria. The peptides are novel members of the insect defensin family and their sequences present marked differences with those of insect defensins isolated from other dipteran species. We have developed a method for efficient renaturation of this cysteine-rich molecule and obtained a highly pure synthetic Chironomus defensin.


FTY720, a novel immunosuppressant, induces sequestration of circulating mature lymphocytes by acceleration of lymphocyte homing in rats, III. Increase in frequency of CD62L-positive T cells in Peyer's patches by FTY720-induced lymphocyte homing.
FTY720, a novel immunosuppressant, sequesters circulating mature lymphocytes, especially T cells, within lymph nodes and Peyer's patches by accelerating lymphocyte homing, and thereby causes lymphocyte depletion in the blood. The FTY720-induced acceleration of lymphocyte homing appears to be mediated by lymphocyte homing receptors including CD62L, CD49d/beta7, and CD11a/CD18. In this study, expressions of CD62L, CD49d and CD11a on T cells in the peripheral blood, lymph nodes and Peyer's patches were analysed by flow cytometry in rats given FTY720 (1 mg/kg) orally. FTY720 markedly decreased the number of peripheral blood T cells, while not affecting CD62L, CD49d and CD11a expressions at 1-3 hr after administration. In contrast, both the frequency of CD62L-positive T cells and intensity of CD62L expression on T cells were increased in Peyer's patches but not lymph nodes at 3 hr after administration of FTY720. CD49d and CD11a expressions on T cells were unaffected by FTY720 in both Peyer's patches and lymph nodes at the same point in time. On the other hand, analysis of lymphocyte homing with calcein-labelled lymphocytes and anti-CD62L monoclonal antibody (mAb) confirmed that FTY720 predominantly increased CD62L-dependent lymphocyte homing to Peyer's patches. These findings indicate that FTY720 increases the frequency of CD62L-positive T cells by accelerating CD62L-predominant homing in Peyer's patches.


Response rates in a case-control study: effect of disclosure of biologic sample collection in the initial contact letter.
Participants in epidemiology studies are often asked to complete an interview and to provide biospecimens. In a population-based case-control study of kidney cancer involving an interview and optional biospecimens, we examined whether mentioning the biospecimens in the initial contact letter adversely affects willingness to be interviewed. Eligible cases (n = 434) and controls (n = 775) in Detroit were alternately assigned to receive one of two versions of the contact letter. Both explained that the study involves an interview with 100 dollars compensation plus an optional component with additional compensation; only one disclosed that the optional component involved biospecimens. There were no meaningful differences between the groups in willingness to be interviewed. However, among 303 cases and 351 controls already interviewed, the proportion providing biospecimens was higher in the fully informed group: for blood, the differences were 10.8 (95% CI, 2.0, 19.5) for cases and 6.7% (95% CI, -1.7, 15.1) for controls. Findings were similar for saliva. In a study involving an interview and optional biospecimens, informing people about the samples in the contact letter seems preferable to a non-specific reference to a second study component. Both approaches yielded similar interview participation rates, but biospecimen participation rates were higher among those informed about the samples in the contact letter.


Allosteric oscillatory enzymes: influence of the number of protomers on metabolic periodicities.
The study of a concerted allosteric model for an enzyme activated by the reaction product shows that this system can generate sustained metabolic oscillations regardless of the number of protomers constituting the enzyme. The analysis extends the results previously obtained in a dimeric model for the phosphofructokinase reaction which produces glycolytic periodicities. When the substrate and product concentrations evolve on comparable time scales, the amplitude of oscillations significantly drops as the number of enzyme subunits evolves from 2 to 8. The width of the domain of substrate injection rates which produce oscillations and the periodic variation in enzyme activity also depend on the number of protomers and on the time scale structure of the system. Theoretical predictions are compared with the experiments on glycolytic oscillations in yeast and muscle, and with the structural characteristics of phosphofructokinase. The results are also discussed in relation with the mechanism of cyclic AMP oscillations in the slime mold Dictyostelium discoideum.


Epidemiological implications of the use of various methods for the diagnosis of canine leishmaniasis in dogs with different characteristics and in differing prevalence scenarios.
Our aim was to establish the influence on the prevalence of canine leishmaniasis (CanL) of the following: (1) the use of different diagnostic techniques; (2) different positivity thresholds; (3) selection of animals either at random from a population or focused on symptomatic individuals, (4) the function which the dog performs; and (5) scenarios with differing epidemiological characteristic. Three groups of dogs were analysed (416 sampled at random from an endemic area, 71 with symptomatology compatible with CanL and 15 from a non-endemic area) using three serological techniques (indirect fluorescence antibody test (IFAT), Kalazar Detect(TM) and Q Letitest ELISA) and a PCR-ELISA. The diagnostic technique had a considerable influence on the CanL prevalence value obtained. Uncertain antibody titres were more representative in dogs sampled at random and with the IFAT technique. Although employing different capture antigens, correlation of results was higher between the two commercial techniques in the group of dogs with symptomatology compatible with CanL. The sensitivity and specificity values of the different diagnostic techniques were affected by the epidemiological characteristic of the area under study, the presence of clinical signs and the function which the dog performs. This must be taken into account when comparing endemicity in different geographical areas, such as in studies carried out for the construction of risk maps. Using more than one technique, and adopting the criterion of considering an animal to be positive only when it has been diagnosed as such by more than one technique, considerably raises the prevalence values but maintains the differences between areas with different characteristics.


A new and improved synthesis of 19-iodocholesterol 3-acetate.
19-Iodocholesterol 3-acetate (VI) was synthesized in a single step by iodo group substitution for hydroxyl using either one of two different reagents: (1) carbodiimidonium methiodide (VIII) or (2) triphenyl-phosphine/N-iodosuccinimide (IX). The yields were as satisfactory as those obtained from the two step iodide replacement of a 19-hydroxy group via the 19-tosyloxy group. The principal intermediate, 19-hydroxy cholesterol 3-acetate (V), was derived in appreciable quantities, and relatively inexpensively, through the Pb (OCOCH3) 4 photolytic oxidation of the bromohydrin of cholesterol 3-acetate (III) to the epoxide (IV) thence Zn reduction to the 19-hydroxy compound. A specially designed 12 liter flask was of aid in accomplishing the photolysis reaction. Dry column chromatography with the supportive puncture sampling was integral to achieving the good yields and high purity of 19-iodocholesterol (VIII).