Habilidade competitiva de plantas de arroz com biótipos de capim-arroz resistente ou suscetível ao quinclorac

Objetivou-se com este trabalho avaliar a habilidade competitiva de plantas de arroz cultivar BRS Pelota com biótipos de capim-arroz resistente ou suscetível ao herbicida quinclorac. Para isso, foi instalado experimento em casa de vegetação, em delineamento completamente casualizado com quatro repetições, sendo os tratamentos dispostos em esquema fatorial 2 x 6. As unidades experimentais constaram de vasos plásticos contendo 10 dm³ de solo, cujo pH e nível de nutrientes foram previamente corrigidos. Os tratamentos consistiram na competição entre uma planta de arroz, cultivar BRS Pelota, com populações (0, 1, 2, 3, 4 ou 5 plantas por vaso) dos biótipos de capim-arroz resistente (ECH-13) ou suscetível (ECH-12) ao herbicida quinclorac. As avaliações foram realizadas aos 40 dias após emergência, sendo avaliados; massa seca da parte aérea (MSPA), taxa de crescimento (TC), área foliar específica (AFE), razão de massa foliar (RMF), razão de área foliar (RAF) e índice de área foliar (IAF). A interferência no desenvolvimento do cultivar de arroz BRS Pelota foi proporcionalmente maior com o aumento da população de ambos os biótipos de capim-arroz. Os biótipos apresentaram, em geral, habilidade competitiva similar.

Crescimento inicial de Euterpe edulis Mart. em diferentes regimes de luz

Plantas sob forte sombreamento (2% ou 6% da luz solar direta) apresentaram, em relação às plantas sob maior nível de luz, menor biomassa, menores taxas de crescimento, menor razão raiz/parte aérea, menor massa foliar específica (MFE), menor razão clorofila a/b e maior razão de área foliar (RAF). Com o aumento da irradiância as plantas apresentaram três tipos de comportamento, dependendo da quantidade de luz dada: 1) até cerca de 20% da luz solar direta as plantas apresentaram, com aumento da luz, aumento de biomassa, das taxas de crescimento relativo (TCR) e de assimilação líquida (TAL), maior alocação de biomassa para a raiz, maior número de folhas, maior MFE, maior razão clorofila a/b e menor razão de peso foliar (RPF) e RAF; 2) entre 20% e 70% de luz as plantas não mostraram alterações morfológicas ou fisiológicas com aumento na quantidade de luz, à exceção de um aumento na razão clorofila a/b; e 3) plantas crescendo em luz solar plena apresentaram uma redução do crescimento em massa seca. As plantas transferidas de 4% para 20 ou 30% de luz mostraram respostas similares àquelas das plantas crescidas sempre em mais luz. A densidade de estômatos mostrou uma leve tendência ao aumento em plantas transferidas para maior quantidade de luz. O menor crescimento em níveis mais fortes de sombreamento e o maior crescimento com aumento de irradiância até 20-30% da luz solar total sugere que a espécie possa se beneficiar do aparecimento de clareiras para sua regeneração. O menor desempenho das plantas em condições de luz plena ou forte sombreamento sugere menor capacidade competitiva da espécie em grandes clareiras ou sob dossel fechado.

Crescimento inicial de três espécies arbóreas da Floresta Atlântica em resposta à variação na quantidade de luz

Verificou-se a resposta de crescimento à variação na intensidade de luz de plântulas de três espécies arbóreas da Floresta Tropical Atlântica, Cecropia glazioui Sneth., Cedrela fissilis Vell. e Bathysa australis (A. St.-Hil.) Hook. ex Sch., respectivamente de estádios inicial, intermediário e final de sucessão. As três espécies mostraram, dentro de um determinado gradiente de luz, plasticidade para aumentar a captação de luz quando em baixa irradiância (através de aumento da razão de área foliar -RAF e diminuição da razão entre raiz e parte aérea - R/PA) e plasticidade para aumentar o ganho de carbono e diminuir a transpiração quando em alta irradiância (através dos aumentos da razão R/PA e densidade estomática, e da diminuição da RAF). A plasticidade das espécies em variar determinado parâmetro em função da intensidade de luz foi dependente do gradiente de intensidade de luz aplicado. A plasticidade foi maior nas intensidades mais baixas de luz tanto para C. glazioui quanto para C. fissilis. Para a maior parte dos parâmetros analisados, C. glazioui mostrou maior parte plasticidade para aclimatar-se à maior irradiância, que C. fissilis. As variações apresentadas pelas espécies na morfologia e fisiologia em relação à variação na intensidade de luz são consistentes com o local de ocorrência de cada espécie.

Spitfirestä Bloodhoundiin : Kuninkaallisten Ilmavoimien hävittäjäilmavoimien kehitys 1945–1958

Tässä tutkielmassa käsitellään Kuninkaallisten Ilmavoimien hävittäjäilmavoimien kehitystä vuosina 1945 - 1958. Ison-Britannian hävittäjäilmavoimat kehittyi suurin harppauksin toisen maailmansodan jälkeisinä vuosina. Kehitys käsitti teknisiä, strategisia ja organisaatiota kos-kevia muutoksia. Suurvaltojen ilmavoimien pelikenttä laajentui yhä suuremmaksi nopeuksien kasvaessa ja asejärjestelmien kehittyessä. Kuninkaallisten Ilmavoimien oli tärkeä pysyä mu-kana tässä kehityksessä niin strategisella, kuin teknisellä tasolla. Kylmän sodan edetessä RAF tasapainoili myös sen faktan kanssa, että puolustusmäärärahoja leikattiin toisen maailmanso-dan jälkeen. Demobilisaatio käynnistyi heti sodan loputtua ja tärkeimmät investoinnit kohdis-tettiin sodan runteleman Ison-Britannian talouden elvyttämiseen. Politiikassa ja sotilasjoh-dossa oltiin yleisesti sitä mieltä, että laajamittaiset sotatoimet eivät olleet todennäköisiä aina-kaan seuraavaan kymmenen vuoden aikana. Lähdeaineistona tutkimuksessa on käytetty pääosin englanninkielisiä kirjallisuus- ja internet-lähteitä. Vertailussa on otettu huomioon teoksia mahdollisimman monelta eri aikakaudelta. Eri aikakausien lähteiden vertailu ja niiden avulla hävittäjäilmavoimien kehityksen selvittä-minen on tutkimuksen ydin. Tärkeimpänä aikalaislähteenä käytetään Ison-Britannian asevoi-mien vuosittaista julkaisusarjaa Brassey’s Annual: The Armed Forces Year-Book. Muut kir-jallisuusläheet jakautuvat tasaisesti eri vuosikymmenille vanhimpien ollessa 1950-luvulta ja uusimmat 2010-luvulta Tutkimuksessa on käytetty myös saatavilla olevia suomenkielisiä tut-kimuksia kuten Petteri Joukon väitöskirjaa Strike Hard, Strike Sure – Operation Musketeer: British Military Planning during the Suez Crisis, 1956. Tutkimuksen johtopäätöksissä todetaan hävittäjäilmavoimien roolin kasvaneen osana sodan-käyntiä toisen maailmansodan jälkeen. Hävittäjätoiminnan kehityksen suuntaan on vaikutta-nut etenkin toisessa maailmansodassa ensimmäisen kerran käytetty ydinase. Hävittäjäilma-voimien tehtävät muuttuivat vuosien 1945–1958 aikana sen moton mukaisesta hyökkäyksestä ja puolustuksesta (Offence, Defence) lähemmäs valvontaa ja turvaamista. Taustalla tärkeim-pänä säilyi kuitenkin aina Ison-Britannian ilmatilan puolustaminen. Hävittäjäilmavoimat ei osallistunut laajamittaisesti operaatioihin edellä mainittuina vuosina. Tämä ei kuitenkaan tar-koittanut, että se olisi ollut toimettomana. Ison-Britannian ilmatilaa oli valvottava idästä nousseen uhan takia vuoden jokaisena päivänä 24 tuntia vuorokaudessa sääolosuhteista riip-pumatta.

The interplay of JNK and SCG10 during cortical development and in excitotoxic responses in brain

Initially identified as stress activated protein kinases (SAPKs), the c-Jun Nterminal kinases (JNKs) are currently accepted as potent regulators of various physiologically important cellular events. Named after their competence to phosphorylate transcription factor c-Jun in response to UVtreatment, JNKs play a key role in cell proliferation, cell death or cell migration. Interestingly, these functions are crucial for proper brain formation. The family consists of three JNK isoforms, JNK1, JNK2 and JNK3. Unlike brain specific JNK3 isoform, JNK1 and JNK2 are ubiquitously expressed. It is estimated that ten splice variants exist. However, the detailed cellular functions of these remain undetermined. In addition, physiological conditions keep the activities of JNK2 and JNK3 low in comparison with JNK1, whereas cellular stress raises the activity of these isoforms dramatically. Importantly, JNK1 activity is constitutively high in neurons, yet it does not stimulate cell death. This suggests a valuable role for JNK1 in brain development, but also as an important mediator of cell wellbeing. The aim of this thesis was to characterize the functional relationship between JNK1 and SCG10. We found that SCG10 is a bona fide target for JNK. By employing differential centrifugation we showed that SCG10 co-localized with active JNK, MKK7 and JIP1 in a fraction containing endosomes and Golgi vesicles. Investigation of JNK knockout tissues using phosphospecific antibodies recognizing JNK-specific phosphorylation sites on SCG10 (Ser 62/Ser 73) showed that phosphorylation of endogenous SCG10 was dramatically decreased in Jnk1-/- brains. Moreover, we found that JNK and SCG10 co-express during early embryonic days in brain regions that undergo extensive neuronal migration. Our study revealed that selective inhibition of JNK in the cytoplasm significantly increased both the frequency of exit from the multipolar stage and radial migration rate. However, as a consequence, it led to ill-defined cellular organization. Furthermore, we found that multipolar exit and radial migration in Jnk1 deficient mice can be connected to changes in phosphorylation state of SCG10. Also, the expression of a pseudo-phosphorylated mutant form of SCG10, mimicking the JNK1- phopshorylated form, brings migration rate back to normal in Jnk1 knockout mouse embryos. Furthermore, we investigated the role of SCG10 and JNK in regulation of Golgi apparatus (GA) biogenesis and whether pathological JNK action could be discernible by its deregulation. We found that SCG10 maintains GA integrity as with the absence of SCG10 neurons present more compact fragmented GA structure, as shown by the knockdown approach. Interestingly, neurons isolated from Jnk1-/- mice show similar characteristics. Block of ER to GA is believed to be involved in development of Parkinson's disease. Hence, by using a pharmacological approach (Brefeldin A treatment), we showed that GA recovery is delayed upon removal of the drug in Jnk1-/- neurons to an extent similar to the shRNA SCG10-treated cells. Finally, we investigated the role of the JNK1-SCG10 duo in the maintenance of GA biogenesis following excitotoxic insult. Although the GA underwent fragmentation in response to NMDA treatment, we observed a substantial delay in GA disintegration in neurons lacking either JNK1 or SCG10.

Regulation of gap junctions by protein phosphorylation

Gap junctions are constituted by intercellular channels and provide a pathway for transfer of ions and small molecules between adjacent cells of most tissues. The degree of intercellular coupling mediated by gap junctions depends on the number of gap junction channels and their activity may be a function of the state of phosphorylation of connexins, the structural subunit of gap junction channels. Protein phosphorylation has been proposed to control intercellular gap junctional communication at several steps from gene expression to protein degradation, including translational and post-translational modification of connexins (i.e., phosphorylation of the assembled channel acting as a gating mechanism) and assembly into and removal from the plasma membrane. Several connexins contain sites for phosphorylation for more than one protein kinase. These consensus sites vary between connexins and have been preferentially identified in the C-terminus. Changes in intercellular communication mediated by protein phosphorylation are believed to control various physiological tissue and cell functions as well as to be altered under pathological conditions.

Using green fluorescent protein to understand the mechanisms of G-protein-coupled receptor regulation

G protein-coupled receptor (GPCR) activation is followed rapidly by adaptive changes that serve to diminish the responsiveness of a cell to further stimulation. This process, termed desensitization, is the consequence of receptor phosphorylation, arrestin binding, sequestration and down-regulation. GPCR phosphorylation is initiated within seconds to minutes of receptor activation and is mediated by both second messenger-dependent protein kinases and receptor-specific G protein-coupled receptor kinases (GRKs). Desensitization in response to GRK-mediated phosphorylation involves the binding of arrestin proteins that serve to sterically uncouple the receptor from its G protein. GPCR sequestration, the endocytosis of receptors to endosomes, not only contributes to the temporal desensitization of GPCRs, but plays a critical role in GPCR resensitization. GPCR down-regulation, a loss of the total cellular complement of receptors, is the consequence of both increased lysosomal degradation and decreased mRNA synthesis of GPCRs. While each of these agonist-mediated desensitization processes are initiated within a temporally dissociable time frame, recent data suggest that they are intimately related to one another. The use of green fluorescent protein from the jellyfish Aqueora victoria as an epitope tag with intrinsic fluorescence has facilitated our understanding of the relative relationship between GRK phosphorylation, arrestin binding, receptor sequestration and down-regulation.

Perturbation of EGF-induced MAP kinase activation by TGF-ß1

TGF-ß1 regulates both cellular growth and phenotypic plasticity important for maintaining a growth advantage and increased invasiveness in progressively malignant cells. Recent studies indicate that TGF-ß-1 stimulates the conversion of epitheliod to fibroblastoid phenotype which presumably leads to the inactivation of growth-inhibitory effects by TGF-ß1 (Portella et al. (1998) Cell Growth and Differentiation, 9: 393-404). Therefore, the investigation of TGF-ß1 signaling that leads to altered growth and migration may provide novel targets for the prevention of increased cell growth and invasion. Although much attention has been paid to TGF-ß1 responses in epithelial cells, the above studies suggest that examination of signal transduction pathways in fibroblasts are important as well. Data from our laboratory are consistent with the concept that TGF-ß1 can act as a regulatory switch in density-dependent C3H 10T1/2 fibroblasts capable of either promoting or delaying G1 traverse. The regulation of this switch is proposed to occur prior to pRb phosphorylation, namely prior to activation of cyclin-dependent kinases. The current study is concerned with the evaluation of a key cyclin (cyclin D1) which activates cdk4 and p27KIP1 which in turn inhibit cdk2 in the proliferative responses of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) and their modulation by TGF-ß1. Although the molecular events that lead to elevation of cyclin D1 are not completely understood, it appears likely that activation of p42/p44MAPK kinases is involved in its transcriptional regulation. TGF-ß1 delayed EGF- or PDGF-induced cyclin D1 expression and blocked the induction of active p42/p44MAPK. The mechanism by which TGF-ß1 induces a block in p42/p44MAPK activation is being examined and the possibility that TGF-ß1 regulates phosphatase activity is being tested.

Angiotensin II-mediated vascular smooth muscle cell growth signaling

The mechanism by which Ang II stimulates the growth of vascular smooth muscle cells was investigated by measuring the phosphorylation of mitogen-activated protein kinases ERK 1 and ERK 2. Ca2+ ionophore was found to have effects practically analogous to Ang II. We found that the signaling pathway involves the activation of epidermal growth factor receptor (EGFR) kinase, activation of the adaptor proteins Shc and Grb2, and the small G-protein Ras. Although the mechanism of AT1- (or Ca2+)-induced activation of EGFR is not yet clear, we have found that calcium-dependent protein kinase CAKß/PYK2 and c-Src are involved in this process. These studies indicate a transactivation mechanism that utilizes EGFR as a bridge between a Gq-coupled receptor and activation of phosphotyrosine generation.

Proliferative signaling initiated in ACTH receptors

This article reviews recent results of studies aiming to elucidate modes of integrating signals initiated in ACTH receptors and FGF2 receptors, within the network system of signal transduction found in Y1 adrenocortical cells. These modes of signal integration should be central to the mechanisms underlying the regulation of the G0->G1->S transition in the adrenal cell cycle. FGF2 elicits a strong mitogenic response in G0/G1-arrested Y1 adrenocortical cells, that includes a) rapid and transient activation of extracellular signal-regulated kinases-mitogen-activated protein kinases (ERK-MAPK) (2 to 10 min), b) transcription activation of c-fos, c-jun and c-myc genes (10 to 30 min), c) induction of c-Fos and c-Myc proteins by 1 h and cyclin D1 protein by 5 h, and d) onset of DNA synthesis stimulation within 8 h. ACTH, itself a weak mitogen, interacts with FGF2 in a complex manner, blocking the FGF2 mitogenic response during the early and middle G1 phase, keeping ERK-MAPK activation and c-Fos and cyclin D1 induction at maximal levels, but post-transcriptionally inhibiting c-Myc expression. c-Fos and c-Jun proteins are mediators in both the strong and the weak mitogenic responses respectively triggered by FGF2 and ACTH. Induction of c-Fos and stimulation of DNA synthesis by ACTH are independent of PKA and are inhibited by the PKC inhibitor GF109203X. In addition, ACTH is a poor activator of ERK-MAPK, but c-Fos induction and DNA synthesis stimulation by ACTH are strongly inhibited by the inhibitor of MEK1 PD98059.

ERBB4 mutations in cancer and amyotrophic lateral sclerosis

ErbB receptor tyrosine kinases, epidermal growth factor receptor (EGFR, also known as ErbB1), ErbB2 (HER2 or NEU), ErbB3 (HER3), and ErbB4 (HER4), transduce signals borne by extracellular ligands into central cellular responses such as proliferation, survival, differentiation, and apoptosis. Mutations in ERBB genes are frequently detected in human malignant diseases of epithelial and neural origin, making ErbB receptors important drug targets. Targeting EGFR and ErbB2 has been successful in eg. lung and breast cancer, respectively, and mutations in these genes can be used to select patients that are responsive to the targeted treatment. Although somatic ERBB4 mutations have been found in many high-incidence cancers such as melanoma, lung cancer, and colorectal cancer and germ-line ERBB4 mutations have been linked to neuronal disorders and cancer, ErbB4 has generally been neglected as a potential drug target. Thus, the consequences of ERBB4 mutations on ErbB4 biology are largely unknown. This thesis aimed to elucidate the functional consequences and assess the clinical significance of somatic and germ-line ERBB4 mutations in the context of cancer and amyotrophic lateral sclerosis. The results of this study indicated that cancer-associated ERBB4 mutations can promote aberrant ErbB4 function by activating the receptor or inducing qualitative changes in ErbB4 signaling. ERBB4 mutations increased survival or decreased differentiation in vitro, suggesting that ERBB4 mutations can be oncogenic. Importantly, the potentially oncogenic mutations were located in various subdomains in ErbB4, possibly providing explanation for the characteristic scattered pattern of mutations in ERBB4. This study also demonstrated that hereditary variation in ERBB4 gene can have a significant effect on the prognosis of breast cancer. In addition, it was shown that hereditary or de novo germ-line ERBB4 mutations that predispose to amyotrophic lateral sclerosis inhibit ErbB4 activity. Together, these results suggest that ErbB4 should be considered as a novel drug target in cancer and amyotrophic lateral sclerosis.

Effects of lipopolysaccharide on low- and high-density cultured rabbit vascular smooth muscle cells: differential modulation of nitric oxide release, ERK1/ERK2 MAP kinase activity, protein tyrosine phosphatase activity, and DNA synthesis

Previous studies have shown that exogenously generated nitric oxide (NO) inhibits smooth muscle cell proliferation. In the present study, we stimulated rabbit vascular smooth muscle cells (RVSMC) with E. coli lipopolysaccharide (LPS), a known inducer of NO synthase transcription, and established a connection between endogenous NO, phosphorylation/dephosphorylation-mediated signaling pathways, and DNA synthesis. Non-confluent RVSMC were cultured with 0, 5, 10, or 100 ng/ml of the endotoxin. NO release was increased by 86.6% (maximum effect) in low-density cell cultures stimulated with 10 ng/ml LPS as compared to non-stimulated controls. Conversely, LPS (5 to 100 ng/ml) did not lead to enhanced NO production in multilayered (high density) RVSMC. DNA synthesis measured by thymidine incorporation showed that LPS was mitogenic only to non-confluent RVSMC; furthermore, the effect was prevented statistically by aminoguanidine (AG), a potent inhibitor of the inducible NO synthase, and oxyhemoglobin, an NO scavenger. Finally, there was a cell density-dependent LPS effect on protein tyrosine phosphatase (PTP) and ERK1/ERK2 mitogen-activated protein (MAP) kinase activities. Short-term transient stimulation of ERK1/ERK2 MAP kinases was maximal at 12 min in non-confluent RVSMC and was prevented by preincubation with AG, whereas PTP activities were inhibited in these cells after 24-h LPS stimulation. Conversely, no significant LPS-mediated changes in kinase or phosphatase activities were observed in high-density cells. LPS-induced NO generation by RVSMC may switch on a cell density-dependent proliferative signaling cascade, which involves the participation of PTP and the ERK1/ERK2 MAP kinases.

Recent advances in angiotensin II signaling

Angiotensin II (Ang II)* is a multifunctional hormone that influences the function of cardiovascular cells through a complex series of intracellular signaling events initiated by the interaction of Ang II with AT1 and AT2 receptors. AT1 receptor activation leads to cell growth, vascular contraction, inflammatory responses and salt and water retention, whereas AT2 receptors induce apoptosis, vasodilation and natriuresis. These effects are mediated via complex, interacting signaling pathways involving stimulation of PLC and Ca2+ mobilization; activation of PLD, PLA2, PKC, MAP kinases and NAD(P)H oxidase, and stimulation of gene transcription. In addition, Ang II activates many intracellular tyrosine kinases that play a role in growth signaling and inflammation, such as Src, Pyk2, p130Cas, FAK and JAK/STAT. These events may be direct or indirect via transactivation of tyrosine kinase receptors, including PDGFR, EGFR and IGFR. Ang II induces a multitude of actions in various tissues, and the signaling events following occupancy and activation of Ang receptors are tightly controlled and extremely complex. Alterations of these highly regulated signaling pathways may be pivotal in structural and functional abnormalities that underlie pathological processes in cardiovascular diseases such as cardiac hypertrophy, hypertension and atherosclerosis.

Interruption of the blood-stage cycle of the malaria parasite, Plasmodium chabaudi, by protein tyrosine kinase inhibitors

Malaria is a devastating disease caused by a unicellular protozoan, Plasmodium, which affects 3.7 million people every year. Resistance of the parasite to classical treatments such as chloroquine requires the development of new drugs. To gain insight into the mechanisms that control Plasmodium cell cycle, we have examined the effects of kinase inhibitors on the blood-stage cycle of the rodent malaria parasite, Plasmodium chabaudi. In vitro incubation of red blood cells for 17 h at 37ºC with the inhibitors led to a decrease in the percent of infected cells, compared to control treatment, as follows: genistein (200 µM - 75%), staurosporine (1 µM - 58%), R03 (1 µM - 75%), and tyrphostins B44 (100 µM - 66%) and B46 (100 µM - 68%). All these treatments were shown to retard or prevent maturation of the intraerythrocytic parasites. The diverse concentration ranges at which these inhibitors exert their effects give a clue as to the types of signals that initiate the transitions between the different developmental stages of the parasite. The present data support our hypothesis that the maturation of the intraerythrocytic cycle of malaria parasites requires phosphorylation. In this respect, we have recently reported a high Ca2+ microenvironment surrounding the parasite within red blood cells. Several kinase activities are modulated by Ca2+. The molecular identification of the targets of these kinases could provide new strategies against malaria.

The role of calcium, calcium-activated K+ channels, and tyrosine/kinase in psoralen-evoked responses in human melanoma cells

8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 µM). The intracellular Ca2+ chelator BAPTA/AM (1 µM) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 µM protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92% increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.