Clear cell ��sugar�� tumor of the lung �� case report

Background

The clear cell ??sugar?? tumor of the lung is an extremely rare benign mesenchymal tumor. Aim: To report a case of the sugar tumor and discuss diagnostic differentiation of the tumor.

Case report

A 53-year female presented with persisting cough. A CT scan revealed a round, 10 mm nodule located within the right lower lobe. The nodule was easily removed during thoracotomy. On the gross examination, the tumor was well circumscribed, and had a homogenous grayish-white appearance on the cut surface. The tumor consisted of round and oval cells with abundant clear cytoplasm, showing PAS pozitive abundant glycogen granules, which were removed by diastase pre-treatment before further staining with PAS. Immunohistochemical studies revealed the tumor cells were positive for HMB-45, vimentin, S-100 protein and very few cells for CD-117. The tumor cells were negative for ??SMA, CK-7, AE1/AE3, CD-10, chromogranin and TTF-1.

Conclusion

Based on the clinical, pathohistological and immunohistochemical data, the diagnosis of the primary clear cell sugar tumor of the lung was established.     

Low Doses of 17��-Estradiol and 17��-Estradiol Facilitate, Whereas Higher Doses of Estrone and 17��- and 17��-Estradiol Impair, Contextual Fear Conditioning in Adult Female Rats

Estrogens are known to exert significant structural and functional effects in the hippocampus of adult rodents. In particular, 17β-estradiol can improve, impair, or have no effect on hippocampus-dependent learning and memory depending on dose and time of administration. The effects of other forms of estrogen, such as estrone and 17α-estradiol, on hippocampus-dependent learning have not been as thoroughly investigated. Therefore, the purpose of this study was to investigate the effects of 17β-estradiol, estrone, and 17α-estradiol at three different doses on two different tasks: hippocampus-dependent contextual fear conditioning and hippocampus-independent cued fear conditioning. Adult ovariectomized female rats were injected with one of the estrogens at one of the three doses 30 mins before conditioning to assess the rapid effects of these estrogens on acquisition. Twenty-four hours later memory for the context was examined and 1 h later memory for the cue (tone) was assessed. Levels of synaptophysin were examined in the dorsal hippocampus of rats to identify a potential synaptic correlate of hormonal effects on contextual fear conditioning. Low 17β-estradiol and 17α-estradiol enhanced, whereas high 17β-estradiol and 17α-estradiol impaired, contextual fear conditioning. Only the middle dose of estrone severely impaired contextual fear conditioning. Estrogens did not alter performance in the hippocampus-independent cued task. Synaptophysin expression was increased by estrone (at a middle and high dose) and 17β-estradiol (at a middle dose) in the CA3 region of the hippocampus and was not correlated with cognition. The results of this study indicate that estradiol can positively or negatively influence hippocampus-dependent learning and memory, whereas estrone impairs hippocampus-dependent learning and memory in a dose-dependent manner. These results have important therapeutic implications, as estrone, a main component of a widely used hormone replacement therapy, was shown to have either a negative effect or no effect on learning and memory. It may be possible to use 17α-estradiol and lower doses of estrogens as potential alternatives in hormone replacement therapies.     

DNR declaration �� emergency medical system nurses�� opinions

Advance directive and other declarations of will made by patients in a case of mental illness still raise ethical and legal issues. In Poland there is no legal regulation, neither research about code of conduct in situation of do not attempt resuscitation. There are also not enough studies regarding Healthcare workers?? opinion about DNAR declaration (Do Not Attempt Resuscitation). The study is aimed at finding out emergency medical system nurses opinion on the subject of enforcing do not attempt resuscitation in situation of circulatory and respiratory arrest. Methods: The research was conducted by means of the diagnostic survey method applying a self ?? constructed questionnaire. The study was carried out among 82 (100%) nurses, from September to December 2011. Obtained information were analyzed statistically, Chi-square of independence with assumed p ?? 0.05. level of significance was used for statistical analysis. Results: The study of the respondents?? opinion shows that 67% Healthcare employees think that DNAR declaration should be obligatory in Poland. Contrary opinion has 7.3% of respondents. In their opinion the decision to refrain from resuscitating should be made by attending physician ? 46.3% and medical board ? 29.3%. Information enclosed within DNAR declaration, in most of respondents?? opinion ? 59.5%, should be only passed on in written form. Conclusions: Majority of respondents agree that patients have a right to refrain from resuscitating as a self ? determination act. Respondents concur the introduction of DNAR declaration in Polish Healthcare system. In respondents?? opinion that decision should be required in written form and an attending physician should decide about its implementation, what violates the existing rule. The execution of living will declaration raises ethical issues. Additionally, it also appears as public/social problem. The last stage of incurable disease is given as justifiable circumstances of DNAR.     

��4��2 Nicotinic acetylcholine receptors, willing if able

Li and Steinbach apply nonstationary noise analysis to the whole-cell current responses of low sensitivity α4β2 nAChR stably-expressed in HEK cells. These receptors represent one of the most important nAChR subtypes in brain, and also one of the most difficult to study in native tissues. They found the activating properties of the full agonists ACh and nicotine to be similar with regard to P_open and single channel conductance, whereas the weak α4β2 partial agonist cytisine caused channels to open with low probability but increased single channel conductance. When optimally stimulated by either of the full agonists, approximately 80% of the available receptors opened at the peak of the response. However, comparisons of whole-cell current to estimates of total cell surface receptors, indicated that only about 7% of the total receptor population can be activated. These observations provide important and intriguing new pieces of the brain nicotine receptor puzzle.     

Antibody-Mediated ��Universal�� Osteoclast Targeting Platform using Calcitonin as a Model Drug

Purpose

To generate and characterize a specific monoclonal antibody (mAb) against recombinant human RANK receptor and to develop an antiresorptive strategy using this mAb as an osteoclast-targeting platform that selectively targets osteoclast cells whilst delivering an attached (i.e. chemically conjugated) active drug cargo.

Methods

Using hybridoma technology, we generated a specific monoclonal antibody (mAb) against recombinant human RANK receptor and characterized by SDS PAGE, ELISA, Western Blot and immunocytochemistry, then synthesized osteoclast-targeting bioconjugates of salmon calcitonin (sCT) using this antibody by generating thiol groups on mAb using 2-Iminothiolane and subsequently reacting them with sCT-PEG-MAL synthesised from sCT and NHS-PEG-MAL. To test the efficacy of the conjugate in vitro, osteoclasts were generated from precursor RAW 264.7 cells by dosing with the cytokines macrophage-colony-stimulating factor (M-CSF), and RANK Ligand (RANKL) and TRAP activity assay, Resorption Pit Assay, TRAP staining were performed. Cytotoxicity of the mAb-sCT conjugate was also evaluated in RAW 264.7 cells; sCT bioactivity and CTR binding potential were evaluated by in vitro intracellular cAMP stimulation assay in human T47D breast cancer cells.

Results

Generation of antibody against human RANK receptor was confirmed by SDS PAGE, ELISA and Western Blot. Immunocytochemistry confirmed the osteoclast targeting potential of the antibody. Successful conjugation of the antibody with sCT was confirmed by SDS PAGE and ELISA.Multinucleated osteoclast formation was confirmed by staining for tartrate-resistant acid phosphatase (TRAP). Conjugate functionality was confirmed by TRAP activity and Resorption Pit assay, showing the inhibitory effect on osteoclast differentiation. cAMP assay confirmed the retention of calcitonin bioactivity after conjugation.

Conclusions

Our strategy offers the potential for a ??universal?? osteoclast-targeting platform??one that targets the RANK receptor on osteoclast cells by simply altering the conjugated cargo in order to affect the specific regulation of osteoclast cells.     

Chronic ��-9-tetrahydrocannabinol Administration Increases Lymphocyte CXCR4 Expression in Rhesus Macaques

Cannabinoids have been reported to produce various immunomodulatory effects, which could potentially impact the host response to bacterial or viral infection. We have recently demonstrated that chronic Δ-9-tetrahydrocannabinol (THC; 0.32 mg/kg i.m., BID) decreased early mortality in rhesus macaques infected with simian immunodeficiency virus (SIV). However, the possibility that prolonged THC administration affects lymphocyte counts, phenotype, and proliferation indices has not been addressed. We examined expression of proliferative and phenotypic markers in circulating lymphocytes of male young adult rhesus macaques chronically-treated with THC (i.m. twice daily 0.32 mg/kg) for 12 months. Chronic THC administration did not alter lymphocyte subtypes, na?ve and memory subsets, proliferation, or apoptosis of T lymphocytes when compared to time-matched vehicle-treated controls. However, chronic THC increased T lymphocyte CXCR4 expression on both CD4+ and CD8+ T lymphocytes compared to control. These results show that chronic THC administration produces changes in T cell phenotype, which can potentially contribute to host immunomodulation to infectious challenges.     

The ��apparent clearance�� of free phenytoin in elderly vs. younger adults

AIMS

To test the hypothesis that the ‘apparent clearance’ of free phenytoin is reduced in elderly patients.

METHODS

Two separate studies were conducted comparing free phenytoin ‘apparent clearance’ in elderly vs. younger adults. The first study was a retrospective analysis of free phenytoin concentrations measured at Christchurch Hospital from 1997 to 2006. In the second study free phenytoin concentrations were measured prospectively in ambulatory subjects who were taking phenytoin regularly.

RESULTS

In the retrospective study (n = 29), free phenytoin ‘apparent clearance’ was 0.27 ± 0.04 l kg⁻¹ day⁻¹ (95% CI 0.19, 0.34) in the elderly cohort vs. 0.37 ± 0.06 l kg⁻¹ day⁻¹ (95% CI 0.22, 0.52) in younger adults, but the difference was not statistically significant. In the prospective study, free phenytoin ‘apparent clearance’ showed a non-significant trend to being reduced in the elderly patients (0.12 ± 0.02 l kg⁻¹ day⁻¹, 95% CI 0.07, 0.17) compared with the younger cohort (0.18 ± 0.07 l kg⁻¹ day⁻¹, 95% CI 0.09, 0.26) in those not taking interacting drugs (n = 21).

CONCLUSIONS

This research does not prove the hypothesis that the ‘apparent clearance’ of free phenytoin is reduced in the elderly. However, the trends found in these two studies are supported by trends in the same direction in other published studies, suggesting an age effect.     

Activation of PPAR�� Attenuates IFN�� and IL-1��-induced Cell Proliferation in Astrocytes: Involvement of IL-6 Independent Pathway

The present study demonstrates the effect of fibrates, agonists of PPARα on cytokines-induced proliferation in primary cultured astrocytes. Alone or combination treatment with cytokines, such as IL-1β (10 ng/ml), IFNγ (10 ng/ml), and TNF-α (10 ng/ml) cause a significant increase of cell proliferation in a time-dependent manner. Treatment of astrocytes with bezafibrate and fenofibrate (0, 5, and 10 µM) reduced the IFNγ and IL-1β-induced cell proliferation in a dose-dependent manner. To address the involvement of IL-6 on the IFNγ and IL-1β-induced cell proliferation, released IL-6 level was measured. IFNγ and IL-1β cause an increase of released IL-6 protein level in a time-dependent manner. Furthermore, pretreatment with IL-6 antibody (0, 0.1, 1, 2.5, and 5 ng/ml) dose-dependently inhibited the IFNγ and IL-1β-induced cell proliferation. However, bezafibrate and fenofibrate did not affect increased mRNA and protein levels of IL-6 in IFNγ and IL-1β-stimulated astrocytes. Taken together, these results clearly suggest that activation of PPARα attenuates the IFNγ and IL-1β-induced cell proliferation through IL-6 independent pathway.     

Tuning in to the ��right�� calcium channel regulation in experimental models of diabetes

Elucidation of cellular and molecular mechanisms underlying vascular disease is of fundamental importance to the development of pharmacological agents to target these pathways. Pinho et al. in this issue of the BJP provide highly compelling evidence that the δ isoform of phosphatidyl inositol 3-kinase (PI3K δ) was upregulated and accounted for the increase in L-type, voltage-gated, Ca channel current in aortic vascular smooth muscle (VSM) cells of a mouse model of type 1 diabetes. There are several key issues of broad fundamental significance to this work. Firstly, what is the ‘right’ answer about calcium channel regulation in diabetes? Conflicting reports of increased and decreased Ca channel current may be due to specificity of the vascular bed and species. Then, the time course of diabetic vasculopathy may influence the expression of contractile versus proliferative phenotypes of VSM. Also the metabolic characterization of diabetes may enlighten or confound any study of diabetic vascular disease. These issues need attention to move forward work in this area.

LINKED ARTICLE

This article is a commentary on Pinho et al., pp. 1458–1471 of this issue. To view this paper visit http://dx.doi.org/10.1111/j.1476-5381.2010.00955.x     

Quercetin Inhibits ��3��4 Nicotinic Acetylcholine Receptor-Mediated Ion Currents Expressed in Xenopus Oocytes

Quercetin mainly exists in the skin of colored fruits and vegetables as one of flavonoids. Recent studies show that quercetin, like other flavonoids, has diverse pharmacological actions. However, relatively little is known about quercetin effects in the regulations of ligand-gated ion channels. In the previous reports, we have shown that quercetin regulates subsets of homomeric ligand-gated ion channels such as glycine, 5-HT_3A and α7 nicotinic acetylcholine receptors. In the present study, we examined quercetin effects on heteromeric neuronal α3β4 nicotinic acetylcholine receptor channel activity expressed in Xenopus oocytes after injection of cRNA encoding bovine neuronal α3 and β4 subunits. Treatment with acetylcholine elicited an inward peak current (I_ACh) in oocytes expressing α3β4 nicotinic acetylcholine receptor. Co-treatment with quercetin and acetylcholine inhibited I_ACh in oocytes expressing α3β4 nicotinic acetylcholine receptors. The inhibition of I_ACh by quercetin was reversible and concentration-dependent. The half-inhibitory concentration (IC₅₀) of quercetin was 14.9±0.8 µM in oocytes expressing α3β4 nicotinic acetylcholine receptor. The inhibition of I_ACh by quercetin was voltage-independent and non-competitive. These results indicate that quercetin might regulate α3β4 nicotinic acetylcholine receptor and this regulation might be one of the pharmacological actions of quercetin in nervous systems.     

Physicians�� perception of CPOE implementation

Objective To identify perceptions held by physicians of the benefits of computerized physician order entry (CPOE) and factors influencing its successful implementation in the context of the increased presence of a clinical pharmacist on ward. Setting A 2000-bed University Hospital. Method A cross-section opinion survey was conducted of all permanent physicians of the hospital to determine their perception on the benefits, or otherwise, of CPOE. Questionnaires, built upon the analysis of 10 preliminary semi-structured interviews with physicians, were sent to physicians by electronic and paper mail. It comprised three sections with a 4 level Likert scale: general perception of CPOE benefits (items 1.1?C1.8); opinion on the introduction of the CPOE system in the hospital (item 2); opinion on the presence of a pharmacist on ward (item 3). A fourth section recorded the respondent??s profile. Main outcome measures Level of agreement on the items describing the general perception of CPOE benefits; opinion on the introduction of a CPOE system in the hospital; and opinion on the pharmacist??s presence on ward. A Principal Component Analysis (PCA) was conducted on sections one and two. Analysis of this PCA representation in terms of the respondents?? profile was performed. Results One hundred and one physicians (18%) participated in the survey. Most (83%) physicians favoured the implementation of a CPOE (item 2). Among the advantages of CPOE, the greatest agreement concerned items related to safety and regulatory issues (from 80 to 76% agreement). Other items related to management issues were perceived as less tangible benefits (from 50 to 67% agreement). The increased presence of a pharmacist on the ward was supported by 94% of physicians. The PCA representation using profile items produced a 2-factor solution, accounting for 68% of the variance, with former experience of collaboration with a pharmacist (P = 0.002) and senior physician status (P = 0.013) positively influencing the perception of the CPOE. Conclusion Endorsement by senior physicians and the presence of a clinical pharmacist on ward promote a positive attitude towards CPOE and facilitate its implementation.     

The neuronal nicotinic ��4��2 receptor has a high maximal probability of being open

Background and purpose:

A fundamental property of transmitter-gated ion channels is the probability a channel will be open (P_open) when stimulated by a concentration of agonist that elicits a maximal response. This value is critical for interpreting steady-state concentration–response relationships in terms of channel activation, and for understanding the actions of drugs that potentiate responses. We used analysis of non-stationary noise to estimate the maximal probability the nicotinic α4β2 receptor is open.

Experimental approach:

HEK293 cells stably transfected to express human α4β2 nicotinic receptors were studied using whole-cell voltage clamp. Nicotinic agonists (acetylcholine, nicotine, cytisine and 5-iodo A-85380) were applied, and the relationship between variance of the elicited whole-cell current and mean current was analysed.

Key results:

The variance did not increase linearly with the mean current. For acetylcholine and nicotine the relationship between variance and mean indicates that the maximal P_open is greater than 0.8. The number of agonist-activatable channels was estimated to be about 1000 per cell. The mean single channel conductance at −60 mV was indistinguishable when currents were elicited by acetylcholine (18 pS), nicotine (17 pS) or 5-iodo A-85380 (17 pS), whereas the value for cytisine was larger (24 pS).

Conclusions and implications:

The neuronal nicotinic α4β2 receptor has a maximal probability of being open that is greater than 0.8. This conclusion applies to the receptor containing three α4 and two β2 subunits (the low-sensitivity stoichiometry), but may not apply to the receptor containing two α4 and three β2 subunits (the high-sensitivity stoichiometry).This article is commented on by Papke, pp. 1903–1905 of this issue. To view this commentary visit http://dx.doi.org/10.1111/j.1476-5381.2010.00868.x     

Cilostazol prevents amyloid �� peptide25-35-induced memory impairment and oxidative stress in mice

BACKGROUND AND PURPOSE

Cilostazol may be effective in dementia associated with a cerebral ischaemia. In this study, we examined whether it exerts beneficial effects on learning and/or memory impairment induced by Aβ_25-35 in mice, and compared its effects with those of aspirin.

EXPERIMENTAL APPROACH

Aβ_25-35 (9 nmol) was administered to mice i.c.v. Learning and memory behaviour were evaluated by measuring spontaneous alternation in a Y-maze and a step-down type passive avoidance test, on the 5th and 8th days after injection respectively. Levels of lipid peroxidation (malondialdehyde) and cytokines in the frontal cortex and hippocampus were measured 2, 3, 5 and 7 days after the Aβ_25-35 injection. The effects of repeated administration of cilostazol and aspirin (both at 30 and 100 mg·kg⁻¹, p.o.) on any changes induced by Aβ_25-35 were evaluated.

KEY RESULTS

Repeated administration of cilostazol significantly attenuated the impairment of spontaneous alternation and the shortened step-down latency induced by Aβ_25-35. Aspirin did not show any beneficial effect. A significant increase in the levels of malondialdehyde (MDA) and IL-1β (only measured in hippocampus) was observed 2, 3 and 5 days after the Aβ_25-35 injection in the frontal cortex and hippocampus. Repeated administration of cilostazol (100 mg·kg⁻¹) completely prevented the increase in MDA levels but failed to antagonize the increase in the expression of IL-1β induced by Aβ_25-35.

CONCLUSIONS AND IMPLICATIONS

These results suggest that the protective effect of cilostazol on Aβ_25-35-induced memory impairment may be related to oxidative stress in the frontal cortex and the hippocampus.     

Design, synthesis and in vitro antibacterial and antifungal activities of some novel spiro[azetidine-2,3��-indole]-2,4(1��H)-dione

The present study deals with the synthesis of novel spiro[azetidine-2,3′-indole]-2′,4(1′H)-dione derivative from the reactions of 3-(phenylimino)-1,3-dihydro-2H-indol-2-one derivatives with chloracetyl chloride in the presence of triethylamine (TEA). All the compounds were characterized using IR, ¹H-NMR, MS, and elemental analysis. They were screened for their antibacterial and antifungal activities. The bacterial strains used were Gram-positive Staphylococcus aureus (MTCC-96) and Gram-negative Escherichia coli (MTCC-521) and Pseudomonas aeruginosa (MTCC-647). The antifungal screening was done on Candida albicans (MTCC-183) and Asperigillus niger (MTCC-343) fungal strains. Results revealed that, compounds (7a), (7b), (7c), (7d), and (7e) showed very good activity with MIC value of 6.25–12.5 μg/ml against three evaluated bacterial strains and the remaining compounds showed good to moderate activity comparable to standard drugs as antibacterial agents. Compounds (7c) and (7h) displayed equipotent antifungal activity in comparison to standard drugs. Amoxicillin, gentamycin, and streptomycin were used as standard drugs for antibacterial activity while fluconazole and itraconazole were used as standard drugs for antifungal activity. Structure–activity relationship study of the compounds showed that the presence of electron withdrawing group substitution at 5′ and 7′ positions of indoline ring and on ortho or para position of phenyl ring increases both antibacterial and antifungal activity of the compound. Henceforth, our findings will have a good impact on chemists and biochemists for further investigations in search of spiro-fused antimicrobial agents.     

In vitro antimycobacterial activity of novel N��-(4-(substituted phenyl amino)-6-(pyridin-2-ylamino)-1,3,5-triazin-2-yl)isonicotinohydrazide

A variety of N′-(4-(substituted phenyl amino)-6-(pyridin-2-ylamino)-1,3,5-triazin-2-yl)isonicotinohydrazide, 7a–r were synthesized by using 2-aminopyridine, isonicotic acid hydrazide and cyanuric chloride, and the structures of these compounds were confirmed by IR and NMR (¹H and ¹³C) spectral analyses. Newly synthesized compounds were tested for their in vitro anti-tubercular activity against Mycobacterium tuberculosis H37Rv using the BACTEC 460 radiometric system.