p53 immunostaining is correlated with reduced survival and is not correlated with gene mutations in resected pulmonary large cell carcinomas

Malignancy of pulmonary large cell carcinomas (LCC) increases from classic LCC through LCC with neuroendocrine morphology (LCCNM) to large cell neuroendocrine carcinomas (LCNEC). However, the histological classification has sometimes proved to be difficult. Because the malignancy of LCC is highly dependent on proteins with functions in the cell cycle, DNA repair, and apoptosis, p53 has been targeted as a potentially useful biological marker. p53 mutations in lung cancers have been shown to result in expression and protein expression also occurs in the absence of mutations. To validate the importance of both p53 protein expression (by immunostaining) and p53 gene mutations in lung LCC (by PCR-single strand conformational polymorphism analysis of exons 5, 6, 7, and 8) and to study their relationships with clinical factors and sub-classification we investigated the correlation of p53 abnormalities in 15 patients with LCC (5 classic LCC, 5 LCNEC, and 5 LCCNM) who had undergone resection with curative intent. Of these patients, 5/15 expressed p53 and none had mutant p53 sequences. There was a negative survival correlation with positive p53 immunostaining (P = 0.05). After adjustment for stage, age, gender, chemotherapy, radiotherapy, and histological subtypes by multivariate analysis, p53 expression had an independent impact on survival. The present study indicates that p53 assessment may provide an objective marker for the prognosis of LCC irrespective of morphological variants and suggests that p53 expression is important for outcome prediction in patients with the early stages of LCC. The results reported here should be considered to be initial results because tumors from only 15 patients were studied: 5 each from LCC, LCNEC and LCCNM. This was due to the rarity of these specific diseases.

Synthesis and secretion of transferrin by a bovine trabecular meshwork cell line

The trabecular meshwork (TM) is the main outflow pathway in the mammalian eye. Oxidative damage to TM cells has been suggested to be an important cause of impairment of TM functions, leading to deficient drainage of aqueous humor, with deleterious consequences to the eye. Transferrin, a metalloprotein involved in iron transport, has been characterized as an intrinsic eye protein. Since transferrin is implicated in the control of oxidative stress, the objective of the present study was to determine if a bovine TM cell line (CTOB) synthesizes and secretes transferrin. The CTOB cell line was cultured in the presence of 35S-methionine and the incubation medium was submitted to immunoprecipitation. Total RNAs from CTOB and isolated bovine TM (freshly isolated, incubated or not) were subjected to the reverse transcription-polymerase chain reaction and the amplification products were sequenced. Also, both CTOB and histological TM preparations were processed for transferrin immunolocalization. A labeled peptide of about 80 kDa, the expected size for transferrin, was immunopurified from CTOB samples obtained from the incubation assays. The reverse transcription-polymerase chain reaction and sequencing experiments detected the presence of transferrin mRNA in CTOB and isolated bovine TM. Reactivity to antibodies against transferrin was observed both in CTOB and TM. The results obtained in all of these experiments indicated that the TM is capable of synthesizing and secreting transferrin. The possible implications for the physiology of the eye are discussed.

Acute exercise performed close to the anaerobic threshold improves cognitive performance in elderly females

The objective of the present study was to compare the effect of acute exercise performed at different intensities in relation to the anaerobic threshold (AT) on abilities requiring control of executive functions or alertness in physically active elderly females. Forty-eight physically active elderly females (63.8 ± 4.6 years old) were assigned to one of four groups by drawing lots: control group without exercise or trial groups with exercise performed at 60, 90, or 110% of AT (watts) and submitted to 5 cognitive tests before and after exercise. Following cognitive pretesting, an incremental cycle ergometer test was conducted to determine AT using a fixed blood lactate concentration of 3.5 mmol/L as cutoff. Acute exercise executed at 90% of AT resulted in significant (P < 0.05, ANOVA) improvement in the performance of executive functions when compared to control in 3 of 5 tests (verbal fluency, Tower of Hanoi test (number of movements), and Trail Making test B). Exercising at 60% of AT did not improve results of any tests for executive functions, whereas exercise executed at 110% of AT only improved the performance in one of these tests (verbal fluency) compared to control. Women from all trial groups exhibited a remarkable reduction in the Simple Response Time (alertness) test (P = 0.001). Thus, physical exercise performed close to AT is more effective to improve cognitive processing of older women even if conducted acutely, and using a customized exercise prescription based on the anaerobic threshold should optimize the beneficial effects.

Cognitive performance of young and elderly subjects on the free word recall memory test: effect of presentation order on recall order

The influence of aging on memory has been extensively studied, but the importance of short-term memory and recall sequence has not. The objective of the current study was to examine the recall order of words presented on lists and to determine if age affects recall sequence. Physically and psychologically healthy male subjects were divided into two groups according to age, i.e., 23 young subjects (20 to 30 years) and 50 elderly subjects (60 to 70 years) submitted to the Wechsler Adult Intelligence Scale-Revised and the free word recall test. The order of word presentation significantly affected the 3rd and 4th words recalled (P < 0.01; F = 14.6). In addition, there was interaction between the presentation order and the type of list presented (P < 0.05; F = 9.7). Also, both groups recalled the last words presented from each list (words 13-15) significantly more times 3rd and 4th than words presented in all remaining positions (P < 0.01). The order of word presentation also significantly affected the 5th and 6th words recalled (P = 0.05; F = 7.5) and there was a significant interaction between the order of presentation and the type of list presented (P < 0.01; F = 20.8). The more developed the cognitive functions, resulting mainly from formal education, the greater the cognitive reserve, helping to minimize the effects of aging on the long-term memory (episodic declarative).

Effect of blocking Rac1 expression in cholangiocarcinoma QBC939 cells

Cholangiocarcinomas (CCs) are malignant tumors that originate from epithelial cells lining the biliary tree and gallbladder. Ras correlative C3 creotoxin substrate 1 (Rac1), a small guanosine triphosphatase, is a critical mediator of various aspects of endothelial cell functions. The objective of the present investigation was to study the effect of blocking Rac1 expression in CCs. Seventy-four extrahepatic CC (ECC) specimens and matched adjacent normal mucosa were obtained from the Department of Pathology, Inner Mongolia Medicine Hospital, between 2007 and 2009. Our results showed that the expression of Rac1 was significantly higher (53.12%) in tumor tissues than in normal tissues. Western blotting data indicated a significant reduction in Rac1-miRNA cell protein levels. Rac1-miRNA cell growth rate was significantly different at 24, 48, and 72 h after transfection. Flow cytometry analysis showed that Rac1-miRNA cells undergo apoptosis more effectively than control QBC939 cells. Blocking Rac1 expression by RNAi effectively inhibits the growth of CCs. miRNA silencing of the Rac1 gene suppresses proliferation and induces apoptosis of QBC939 cells. These results suggest that Rac1 may be a new gene therapy target for CC. Blocking Rac1 expression in CC cells induces apoptosis of these tumor cells and may thus represent a new therapeutic approach.

Nitric oxide synthesis and biological functions of nitric oxide released from ruthenium compounds

During three decades, an enormous number of studies have demonstrated the critical role of nitric oxide (NO) as a second messenger engaged in the activation of many systems including vascular smooth muscle relaxation. The underlying cellular mechanisms involved in vasodilatation are essentially due to soluble guanylyl-cyclase (sGC) modulation in the cytoplasm of vascular smooth cells. sGC activation culminates in cyclic GMP (cGMP) production, which in turn leads to protein kinase G (PKG) activation. NO binds to the sGC heme moiety, thereby activating this enzyme. Activation of the NO-sGC-cGMP-PKG pathway entails Ca2+ signaling reduction and vasodilatation. Endothelium dysfunction leads to decreased production or bioavailability of endogenous NO that could contribute to vascular diseases. Nitrosyl ruthenium complexes have been studied as a new class of NO donors with potential therapeutic use in order to supply the NO deficiency. In this context, this article shall provide a brief review of the effects exerted by the NO that is enzymatically produced via endothelial NO-synthase (eNOS) activation and by the NO released from NO donor compounds in the vascular smooth muscle cells on both conduit and resistance arteries, as well as veins. In addition, the involvement of the nitrite molecule as an endogenous NO reservoir engaged in vasodilatation will be described.

Outcomes and organ dysfunctions of critically ill patients with systemic lupus erythematosus and other systemic rheumatic diseases

Our objective was to compare the pattern of organ dysfunctions and outcomes of critically ill patients with systemic lupus erythematosus (SLE) with patients with other systemic rheumatic diseases (SRD). We studied 116 critically ill SRD patients, 59 SLE and 57 other-SRD patients. The SLE group was younger and included more women. Respiratory failure (61%) and shock (39%) were the most common causes of ICU admission for other-SRD and SLE groups, respectively. ICU length-of-stay was similar for the two groups. The 60-day survival adjusted for the groups’ baseline imbalances was not different (P = 0.792). Total SOFA scores were equal for the two groups at admission and during ICU stay, although respiratory function was worse in the other-SRD group at admission and renal and hematological functions were worse in the SLE group at admission. The incidence of severe respiratory dysfunction (respiratory SOFA >2) at admission was higher in the other-SRD group, whereas severe hematological dysfunction (hematological SOFA >2) during ICU stay was higher in the SLE group. SLE patients were younger and displayed a decreased incidence of respiratory failure compared to patients with other-SRDs. However, the incidences of renal and hematological failure and the presence of shock at admission were higher in the SLE group. The 60-day survival rates were similar.

BC047440 antisense eukaryotic expression vectors inhibited HepG2 cell proliferation and suppressed xenograft tumorigenicity

The biological functions of the BC047440 gene highly expressed by hepatocellular carcinoma (HCC) are unknown. The objective of this study was to reconstruct antisense eukaryotic expression vectors of the gene for inhibiting HepG2 cell proliferation and suppressing their xenograft tumorigenicity. The full-length BC047440 cDNA was cloned from human primary HCC by RT-PCR. BC047440 gene fragments were ligated with pMD18-T simple vectors and subsequent pcDNA3.1(+) plasmids to construct the recombinant antisense eukaryotic vector pcDNA3.1(+)BC047440AS. The endogenous BC047440 mRNA abundance in target gene-transfected, vector-transfected and naive HepG2 cells was semiquantitatively analyzed by RT-PCR and cell proliferation was measured by the MTT assay. Cell cycle distribution and apoptosis were profiled by flow cytometry. The in vivo xenograft experiment was performed on nude mice to examine the effects of antisense vector on tumorigenicity. BC047440 cDNA fragments were reversely inserted into pcDNA3.1(+) plasmids. The antisense vector significantly reduced the endogenous BC047440 mRNA abundance by 41% in HepG2 cells and inhibited their proliferation in vitro (P < 0.01). More cells were arrested by the antisense vector at the G1 phase in an apoptosis-independent manner (P = 0.014). Additionally, transfection with pcDNA3.1(+)BC047440AS significantly reduced the xenograft tumorigenicity in nude mice. As a novel cell cycle regulator associated with HCC, the BC047440 gene was involved in cell proliferation in vitro and xenograft tumorigenicity in vivo through apoptosis-independent mechanisms.

Acute effect of alcohol intake on sine-wave Cartesian and polar contrast sensitivity functions

The aim of this study was to assess contrast sensitivity for angular frequency stimuli as well as for sine-wave gratings in adults under the effect of acute ingestion of alcohol. We measured the contrast sensitivity function (CSF) for gratings of 0.25, 1.25, 2.5, 4, 10, and 20 cycles per degree of visual angle (cpd) as well as for angular frequency stimuli of 1, 2, 4, 24, 48, and 96 cycles/360°. Twenty adults free of ocular diseases, with normal or corrected-to-normal visual acuity, and no history of alcoholism were enrolled in two experimental groups: 1) no alcohol intake (control group) and 2) alcohol ingestion (experimental group). The average concentration of alcohol in the experimental group was set to about 0.08%. We used a paradigm involving a forced-choice method. Maximum sensitivity to contrast for sine-wave gratings in the two groups occurred at 4 cpd sine-wave gratings and at 24 and 48 cycles/360° for angular frequency stimuli. Significant changes in contrast sensitivity were observed after alcohol intake compared with the control condition at spatial frequency of 4 cpd and 1, 24, and 48 cycles/360° for angular frequency stimuli. Alcohol intake seems to affect the processing of sine-wave gratings at maximum sensitivity and at the low and high frequency ends for angular frequency stimuli, both under photopic luminance conditions.

Serotonin regulates osteoblast proliferation and function in vitro

The monoamine serotonin (5-hydroxytryptamine, 5-HT), a well-known neurotransmitter, also has important functions outside the central nervous system. The objective of this study was to investigate the role of 5-HT in the proliferation, differentiation, and function of osteoblasts in vitro. We treated rat primary calvarial osteoblasts with various concentrations of 5-HT (1 nM to 10 µM) and assessed the rate of osteoblast proliferation, expression levels of osteoblast-specific proteins and genes, and the ability to form mineralized nodules. Next, we detected which 5-HT receptor subtypes were expressed in rat osteoblasts at different stages of osteoblast differentiation. We found that 5-HT could inhibit osteoblast proliferation, differentiation, and mineralization at low concentrations, but this inhibitory effect was mitigated at relatively high concentrations. Six of the 5-HT receptor subtypes (5-HT1A, 5-HT1B, 5-HT1D, 5-HT2A, 5-HT2B, and 5-HT2C) were found to exist in rat osteoblasts. Of these, 5-HT2A and 5-HT1Breceptors had the highest expression levels, at both early and late stages of differentiation. Our results indicated that 5-HT can regulate osteoblast proliferation and function in vitro.

Selenium plasma levels in hemodialysis patients: Comparison between North and Southeast of Brazil

Introduction: Patients with chronic kidney disease present selenium (Se) plasma deficiency which is an essential trace element with important biological functions and, the best known biological role is attributed to its presence in the antioxidant enzyme, glutathione peroxidase (GPx). The Se content of foods depends on soil and some authors have suggested that Amazon soil (North Brazilian region) has high Se concentrations when compared to other regions of Brazil. Objective: The objective of this work was to compare the Se status in hemodialysis (HD) patients from North and Southeast of Brazil. Methods: Thirty-eight patients from Southeast region (22 men and 16 women, 15% diabetic, 53.5 ± 26.4 yrs) were compared to 40 patients from North region (28 men and 12 women, 22.5% diabetic, 63.5 ± 11.9 yrs). Se in plasma was determined through atomic absorption spectrophotometry with hydride generation. Results: The plasma Se levels in patients from Southeast region were significantly lower (17.5 ± 11.9 μg/L) when compared to patients from the North (37.1 ± 15.8 μg/L) (p < 0.001). However, both patient groups presented low Se plasma levels when compared to recommended values (60- 120 μg/L). There was no correlation between plasma Se levels and analyzed parameters. Conclusion: We concluded that patients from North (Amazon) region present higher plasma Se levels when compared to the patients from Southeast of Brazil. However, independently of the region, HD patients presented Se deficiency.

Urana työllistyminen

Tutkimuksen tarkoituksena on ollut selvittää pitkään työttömänä olleiden ammatinvaihtajien ammatinvaihdon ja työllistymisen etenemistä. Tämän lisäksi tutkimuksessa on tarkasteltu kohderyhmän mielikuvaa ohjaavasta koulutuksesta. Pitkittäistutkimus on toteutettu seurannalla, jonka välietappina on ollut kohderyhmän osallistuminen ohjaavaan koulutukseen. Työvoimapoliittinen ohjaava koulutus edusti erityisesti vaikeasti työllistettäville kohdistettavaa interventiota, jolla pyrittiin puuttumaan työttömyyskierteeseen ja ehkäisemään syrjäytymiskehityksen jatkuminen. Tutkimuksen kohderyhmäksi valitut ihmiset (51) ovat suoranaisessa keskiössä tutkittaessa työelämästä syrjäytymistä. Heikentyneet työelämävalmiudet ja työmarkkinakelpoisuus ovat tehneet heistä vaikeasti työllistettäviä. Heidän valintansa kohderyhmäksi, työhallinnon tekemän kurssiosoituksen kriteereinä olevien pitkäaikaistyöttömyyden sekä epäonnistuneen ammatinvaihdon perusteella, osoittaa heidän olevan erityisen ohjauksen ja tuen tarpeessa. On kuitenkin huomattava, että heidät on valittu kohderyhmäksi paitsi kurssiosoituksen saaneina pitkäaikaistyöttöminä myös ammatinvaihtajina, jotka itse haluavat vaihtaa ammattiaan. Tutkimusaineistona ovat olleet työhallinnon asiakasrekisteritiedot ja seurantatutkimuksen perusteella muodostetuista ryhmistä valittujen neljän tapaushenkilön teemahaastatteluaineistot. Tutkimusaineistoa on käsitelty laadullisesti, koska tutkimus kuuluu laadullisen tutkimuksen piiriin sekä tutkimuksen kohteena olevan ilmiön, tutkimuskysymysten ja -aineistojen että tarkoituksensa perusteella. Tutkimuksessa on kuvattu pitkäaikaistyöttömien henkilöiden ammatinvaihtoa ja työllistymistä. Kuvatuissa tapauksissa ammatinvaihtotarve perustuu suurimmalla osalla terveydellisiin rajoitteisiin, joiden vuoksi aikaisemmassa työssä toimiminen on mahdotonta. Useimmiten vajaakuntoisuus on merkittävästi pitkittänyt heidän työttömyyttään. Tämän vuoksi seurantatutkimuksessa on keskitytty tarkastelemaan ammatinvaihtoprosessin etenemistä. Muun muassa työttömyyttä laajasti tutkineet Aho ja Vähätalo ovat todenneet vajaakuntoisuuden olevan merkittävä työttömyyden pitkittymiseen vaikuttava tekijä. Ahon mukaan vajaakuntoisiksi diagnosoituja on jopa viidennes kroonisesti pitkäaikaistyöttömistä työhallinnon asiakasrekisteriin kirjatuista henkilöistä. Monen tekijän summana pitkään työttömänä olleet henkilöt ovat vaarassa ajautua tilanteeseen, jossa he eivät enää kykene omin avuin löytämään ulospääsyä työttömyydestä Tämän seurantatutkimuksen perusteella vain hyvin harvalla kohderyhmään kuuluvista on ammatinvaihdossa ja työllistymisessä tapahtunut merkittävää edistymistä huolimatta useiden vuosien yrittämisestä. Ammatinvaihdon ja työllistymisen edistymistä on tarkasteltu sen perusteella, miten työelämään ja/tai koulutukseen sijoittumiset tai muut toiminnot ovat edesauttaneet tavoitteen toteutumista. Seurantatutkimuksessa on muodostettu neljä tyyppiryhmää ammatinvaihdon ja työllistymisen etenemisen perusteella. Ryhmät ovat: Ammatinvaihdossa onnistuneet (5), ammatinvaihdossa edistyneet (19), pitkäaikaistyöttömäksi jääneet (12) ja epätarkoituksenmukaisesti sijoittuneet (15). Ammatinvaihdossa onnistuneet ovat toteuttaneet ammatinvaihtonsa siirtymällä suoraan työelämään uudelle alalle, työllistymällä koulutusta vastaavaan työhön uudelleenkoulutuksen jälkeen tai kouluttautumalla uuteen ammattiin työllistymättä. Kuitenkin vain kolme heistä on sijoittunut seurannan päättyessä pysyväisluonteiseen terveydelleen soveltuvaan työhön muiden ollessa edelleen työttömänä. Ammatinvaihdossa edistyneet ovat suurimmaksi osaksi edistyneet ammatinvaihdossaan soveltuvien tukitöiden avulla. Työttömyys on kuitenkin jatkunut valtaosalla ja vain yksi heistä on sijoittuneena työelämään seurannan päättyessä. Tämä sijoittuminen ei ole kuitenkaan edistänyt ammatinvaihtoa. Pitkäaikaistyöttömäksi jääneet ovat olleet yhtäjaksoisesti työttömänä kokoseurantajakson ajan. Epätarkoituksenmukaisesti sijoittuneet ovat enimmäkseen sijoittuneet terveytensä kannalta haitallisiin ja/tai ammatinvaihtosuunnitelmilleen soveltumattomiin sijoitustöihin ja määräaikaisuuksiin. Seurannan päättyessä heovat joko työttömänä tai toimivat ammatinvaihtotavoitteensa kannalta haitallisissa työtehtävissä. Tämä on vahvistanut mielikuvaa, joka usealle heille on muodostunut ohjaavasta koulutuksesta työttömille suunnattuna sanktiona, joka ei edistä työllistymistä. Ohjaavassa koulutuksessa suurin osa heistä on kokenut hyötyneensä työharjoittelujaksosta, joka on antanut käyttökelpoista tietoa ammatinvalintaa varten. Lukuun ottamatta kolmea henkilöä, jotka ovat seurannan päättyessä pysyväisluonteisessa työssä terveydelleen soveltuvassa ammatissa, ovat kaikki muut kohderyhmään kuuluvat edelleen erityisen ohjauksen ja tuen tarpeessa johtuen ammatinvaihdon keskeneräisyydestä ja/tai työttömyyden jatkumisesta. Yhdistävänä tekijänä ammatinvaihdossa onnistuneilla vaikuttaa olleen tulevaisuususkon säilyminen pitkäaikaistyöttömyydessä. Sekä ammatinvaihdossa onnistuneet että edistyneet ovat kokeneet työttömyyteen katkoksia tuoneet työt merkityksellisinä riippumatta siitä, ovatko työtehtävät edistäneet ammatinvaihtoa.