Recognition of pathogen-derived sphingolipids in Arabidopsis.

In plants, many invading microbial pathogens are recognized by cell-surface pattern recognition receptors, which induce defense responses. Here, we show that the ceramide Phytophthora infestans-ceramide D (Pi-Cer D) from the plant pathogenic oomycete P. infestans triggers defense responses in Arabidopsis. Pi-Cer D is cleaved by an Arabidopsis apoplastic ceramidase, NEUTRAL CERAMIDASE 2 (NCER2), and the resulting 9-methyl-branched sphingoid base is recognized by a plasma membrane lectin receptor-like kinase, RESISTANT TO DFPM-INHIBITION OF ABSCISIC ACID SIGNALING 2 (RDA2). 9-Methyl-branched sphingoid base is specific to microbes and induces plant immune responses by physically interacting with RDA2. Loss of RDA2 or NCER2 function compromised Arabidopsis resistance against an oomycete pathogen. Thus, we elucidated the recognition mechanisms of pathogen-derived lipid molecules in plants.

Influence of ABCB1 and ABCG2 polymorphisms on the antiemetic efficacy in patients with cancer receiving cisplatin-based chemotherapy: a TRIPLE pharmacogenomics study.

Resistance to antiemetic treatment with 5-hydroxytryptamine-3 receptor antagonist is an issue. This study evaluated the potential roles of ABCB1 and ABCG2 polymorphisms in antiemetic treatment resistance in patients with cancer previously enrolled in a randomized controlled trial. A total of 156 patients were evaluated for their responses to antiemetic therapy and then subdivided into granisetron or palonosetron groups. The genotypes were evaluated for their association with antiemetic efficacy in each treatment groups. Additional risk factors associated with complete response (CR) were examined using a multivariate regression analysis. No significant associations were identified for genetic polymorphisms in the palonosetron group. In the granisetron group, patients with ABCB1 2677TT and 3435TT genotypes had higher proportion of CR. In addition to ABCB1 polymorphisms, gender and cisplatin dose were associated with granisetron response by univariate analysis. Multivariate logistic regression analysis revealed that the ABCB1 3435C>T polymorphism and cisplatin dose were significant predictors of CR.

Expression of CD44 variant isoforms, CD44v3 and CD44v6, are associated with prognosis in nasopharyngeal carcinoma.

OBJECTIVE: The clinical and prognostic significance of CD44 variant isoform expression in nasopharyngeal carcinoma is not well known. This study aimed to clarify whether CD44 variant isoform expression serves as a prognostic factor in nasopharyngeal carcinoma. METHODS: Forty-two nasopharyngeal carcinoma patients, who underwent concurrent chemoradiotherapy as the initial treatment, were the subjects of investigation. Expression of CD44 variant isoforms, CD44v3, CD44v4, CD44v5, CD44v6 and CD44v7, in nasopharyngeal carcinoma was assessed in relation to concurrent chemoradiotherapy resistance and disease-specific survival of the patients. RESULTS AND CONCLUSION: The patients with CD44v6 high expression showed a clinically incomplete response to concurrent chemoradiotherapy at the primary site. The disease-specific survival rate was lower in patients with high expression of CD44v3 than in those with low expression. These results suggest that analysis of CD44v6 and CD44v3 expression is useful in estimating prognosis and determining effective treatment strategies in nasopharyngeal carcinoma.

Particle therapy using carbon ions or protons as a definitive therapy for patients with primary sacral chordoma.

OBJECTIVE: This study retrospectively evaluated the efficacy and toxicity of particle therapy using carbon ions or protons for primary sacral chordomas. METHODS: We evaluated 23 patients with primary sacral chordoma treated with carbon ion therapy (CIT) or proton therapy (PT) between July 2005 and June 2011 at the Hyogo Ion Beam Medical Center, Hyogo, Japan. The median patient age was 72 years. 14 patients were treated with 70.4 Gy equivalents (GyE) in 16 fractions and 9 were treated with 70.4 GyE in 32 fractions. CIT was used for 16 patients, and PT was used for 7 patients. RESULTS: The median follow-up period was 38 months. At 3 years, local control (LC), overall survival (OS) and progression-free survival (PFS) for all patients were 94%, 83% and 68%, respectively. The log-rank test revealed that male sex was significantly related to better PFS (p=0.029). No other factors, including dose fractionation and ion type, were significant for LC, OS or PFS. In nine patients, ≥ Grade 3 acute dermatitis was observed, and ≥ Grade 3 late toxicities were observed in nine patients. The 32-fraction protocol reduced severe toxicities in both the acute and late phases compared with the 16-fraction protocol. CONCLUSION: Particle therapy for patients with sacral chordoma showed favourable LC and OS. Severe toxicities were successfully reduced by modifying the dose fractionation and treatment planning in the later treatment era. Thus, this therapeutic modality should be considered useful and safe. ADVANCES IN KNOWLEDGE: This is the first study including both CIT and PT for sacral chordomas.

Particle therapy for mucosal melanoma of the head and neck. A single-institution retrospective comparison of proton and carbon ion therapy.

PURPOSE: To retrospectively analyze treatment outcomes after particle therapy using protons or carbon ions for mucosal melanoma of the head and neck (HNMM) at the Hyogo Ion Beam Medical Center, as well as to compare proton therapy (PT) and carbon ion therapy (CIT). PATIENTS AND METHODS: Data from 62 HNMM patients without metastasis, treated with PT or CIT between October 2003 and April 2011 were analyzed. Median patient age was 70.5 years (range 33-89 years). Of the total patients, 33 (53 %) had received PT and 29 (47 %) had undergone CIT. Protocols for 65 or 70.2 GyE in 26 fractions were used for both ion types. RESULTS: Median follow-up was 18.0 months (range 5.2-82.7 months). The 1-/2-year overall survival (OS) and local control (LC) rates were 93 %/61 % and 93 %/78 % for all patients, 91 %/44 % and 92 %/71 % for the PT patients and 96 %/62 % and 95 %/59 % for the CIT patients, respectively. No significant differences were observed between PT and CIT. Local recurrence was observed in 8 patients (PT: 5, CIT: 3) and 29 (PT: 18, CIT: 11) experienced distant metastases. Acute reactions were acceptable and all patients completed the planned radiotherapy. Regarding late toxicity, grade 3 or greater events were observed in 5 patients (PT: 3, CIT: 2), but no significant difference was observed between PT and CIT. CONCLUSION: Our single-institution retrospective analysis demonstrated that particle therapy for HNMM achieved good LC, but OS was unsatisfactory. There were no significant differences between PT and CIT in terms of either efficacy or toxicity.

Posterior maximization and averaging for Bayesian working model choice in the continual reassessment method.

The continual reassessment method (CRM) is a method for estimating the maximum tolerated dose in a dose-finding study. Traditionally, use is made of a single working model or 'skeleton' idealizing an underlying true dose-toxicity relationship. This working model is chosen either by discussion with investigators or published data, before the beginning of the trial or simply on the basis of operating characteristics. To overcome the arbitrariness of the choice of such a single working model, Yin and Yuan (biJ. Am. Statist. Assoc. 2009; 104:954-968) propose a model averaging over a set of working models. Here, instead of averaging, we investigate some alternative Bayesian model criteria that maximize the posterior distribution. We propose three adaptive model-selecting CRMs using the Bayesian model selection criteria, in which we specify in advance a collection of candidate working models for the dose-toxicity relationship, especially initial guesses of toxicity probabilities, and adaptively select the only one working model among the candidates updated by using the original CRM for each working model, based on the posterior model probability, the posterior predictive loss or the deviance information criteria, during the course of the trial. These approaches were compared via a simulation study with the model averaging approach.

Transgenic analysis of the BmBLOS2 gene that governs the translucency of the larval integument of the silkworm, Bombyx mori.

The larval integument of the silkworm, Bombyx mori, is opaque because urate granules accumulate in the epidermis. Although the biosynthetic pathway of uric acid is well studied, little is known about how uric acid accumulates as urate granules in epidermal cells. In the distinct oily (od) mutant silkworm, the larval integument is translucent because of the inability to construct urate granules. Recently, we have found that the od mutant has a genomic deletion in the B. mori homologue of the human biogenesis of lysosome-related organelles complex1, subunit 2 (BLOS2) gene (BmBLOS2). Here, we performed a molecular and functional characterization of BmBLOS2. Northern blot analysis showed that BmBLOS2 was ubiquitously expressed in various tissues. We analysed the structure of a newly isolated mutant (od(B) ) allelic to od and found a premature stop codon in the coding sequence of BmBLOS2 in this new mutation. Moreover, the translucent phenotype was rescued by the germ-line transformation of the wild-type BmBLOS2 allele into the od mutant. Our results suggest that BmBLOS2 is responsible for the od mutant phenotype and plays a crucial role in biogenesis of urate granules in the larval epidermis of the silkworm. The relationships amongst Hermansky-Pudlak syndrome (HPS) genes in mammals, granule group genes in Drosophila and translucent mutant genes in B. mori are discussed.

Genetic polymorphisms in folate pathway enzymes as a possible marker for predicting the outcome of methotrexate therapy in Japanese patients with rheumatoid arthritis.

BACKGROUND: Low-dose methotrexate (MTX) therapy is widely used in the treatment of rheumatoid arthritis (RA). Though the difference in response to MTX between patients with RA is large, the factors that contribute to this variability remain unclear. OBJECTIVE: We aimed to identify those factors with a particular emphasis on the pharmacogenetics of MTX. METHOD: We evaluated the association of possible factors, including genetic polymorphisms of folate metabolic pathway enzymes, with the cumulative value of C-reactive protein, an index of MTX anti-inflammatory efficacy, in 87 Japanese patients with RA. RESULTS: Polymorphisms of the reduced folate carrier gene (RFC) G80A and of the gamma-glutamylhydrolase gene (GGH) C-401T were more closely associated (beta = 2.1194, P = 0.0017) than other polymorphisms, with the anti-inflammatory response to MTX. CONCLUSION: Patients with RA having RFC 80A and GGH-401T alleles were less responsive to MTX than those with RFC 80A and without GGH-401T alleles. Thus, this data may be useful for guiding treatment of RA patients with MTX.

Reproducibility of temporal volume change in CT of lung cancer: comparison of computer software and manual assessment.

The purpose of this study was to investigate the reproducibility of volumetric software evaluation and manual evaluation of tumour growth. Three observers manually evaluated whether tumour volume was increasing, if it was unchanged, or if it had decreased in size in 2 serial CT examinations of 45 solid lung cancers. The tumour volumes were calculated 3 times using volumetric software and were evaluated using the same classifications as for manual evaluation. Both data sets were divided into three groups: growth or reduction with consistency among all three evaluations (group A), growth or reduction with consistency between only two evaluations (group B), and others (group C). The volume variation and relative volume variation were calculated from the median volumes measured by volumetric software. Although all 45 tumours were categorised in group A by volumetric software, only 21 tumours were categorised in group A by manual assessment. The relative volume variation of the manual assessment was 88.5 +/- 76.5%, 20.8 +/- 28.3% and 12.9 +/- 12.8% in group A, B and C, respectively. Significant differences were found between groups A and B (p<0.01) and between groups A and C (p<0.001). Inconsistency is often seen in manual assessment; in contrast, evaluation using volumetric software has good reproducibility, even when the relative change in tumour volume is small.

Identifying patients with peripheral-type early non-small cell lung cancer (T1N0M0) for whom irradiation of the primary focus alone could lead to successful treatment.

We investigated the indication of radiotherapy in operable patients with peripheral-type early non-small cell lung cancer (T1N0M0 (TNM staging in 1997)). The subjects comprised 396 patients with non-small cell lung cancer in whom the clinical stage was evaluated as IA. We examined age, gender, Brinkmann's index, histopathological type, the grade of histopathological differentiation, tumour diameter and the level of carcinoembryonic antigen as factors involved in lymph node metastasis. Lymph node metastasis was detected in 79 patients (20%). Factors such as the grade of histopathological differentiation and tumour diameter were involved in lymph node metastasis. In well-differentiated lesions, the probability of metastasis was <10% even when the tumour diameter exceeded 2 cm. However, the probability rapidly increased with tumour size in moderately and poorly differentiated lesions. Among the patients with peripheral-type early non-small cell lung cancer (T1N0M0), the risk of lymph node metastasis was low in those with well-differentiated carcinoma and those with moderately differentiated lesions measuring

Intra-arterial chemoradiotherapy for locally advanced oral cavity cancer: analysis of therapeutic results in 134 cases.

The objective of this study was to investigate the therapeutic results of arterial injection therapy via the superficial temporal artery for 134 cases of stages III and IV (M0) oral cavity cancer retrospectively, and to clarify the prognostic factors. We administered intra-arterial chemoradiotherapy by continuous infusion of carboplatin in 65 cases from January 1993 to July 2002. Systemic chemotherapy was performed on 26 cases at the same time. We administered intra-arterial chemoradiotherapy by cisplatin with sodium thiosulphate in 69 cases from October 2002 to December 2006. Systemic chemotherapy was performed on 48 cases at the same time. The 3-year local control rate was 68.6% (T2-3: 77.9%; T4: 51.3%), and the 3-year survival rate was 53.9% (stage III: 62.9%; stage IV: 45.3%). Regarding the results of multivariate analysis of survival rates, age (<65), selective intra-arterial infusion, and the use of cisplatin as an agent for intra-arterial infusion were significant factors. The therapeutic results of intra-arterial chemoradiotherapy via the superficial temporal artery were not inferior to the results of surgery. In particular, the results of arterial injection therapy by cisplatin with sodium thiosulphate were excellent, so we believe that it will be a new therapy for advanced oral cavity cancer.

Functional characterization of chitinase from Cydia pomonella granulovirus.

Baculovirus chitinases (V-CHIAs) play a crucial role in the terminal liquefaction of virus-infected larvae after death. Although v-chiAs from nucleopolyhedroviruses (NPVs) have been well characterized, little is known about v-chiAs from granuloviruses (GVs). We characterized the v-chiA of Cydia pomonella GV (CpGV) by constructing a recombinant Bombyx mori NPV (BmNPV) in which BmNPV v-chiA was replaced by CpGV v-chiA (103CpGV virus). CpGV v-chiA encoded an approximately 70-kDa chitinase with an exo-type substrate preference. CpGV V-CHIA lacked a C-terminal KDEL endoplasmic reticulum retention motif and was suggested to be a secretory protein. Terminal host liquefaction of B. mori larvae and proper folding of BmNPV-encoded cysteine protease (BmNPV V-CATH) were observed following infection with 103CpGV, indicating that CpGV v-chiA is able to compensate for the absence of its BmNPV counterpart. Our data suggest that the molecular interaction between V-CHIA and V-CATH may be conserved across a broad range of lepidopteran GVs and NPVs.

Relationship between the growth pattern of nasopharyngeal cancer and the cervical lymph nodes based on MRI findings: can the cervical radiation field be reduced in patients with nasopharyngeal cancer?

To identify patients with nasopharyngeal cancer in whom the cervical radiation field can be reduced, we classified the growth patterns of nasopharyngeal cancer based on MRI findings into 4 types and performed an evaluation. Based on MRI findings, we classified the growth patterns of primary cancer in 94 patients with nasopharyngeal cancer into Type 1 (superficial type), Type 2 (lateral invasive type), Type 3 (upward invasive type), and Type 4 (anterior extension type), and further classified Type 2, based upon nasopharyngoscopic findings, into Type 2a (unilateral invasive type) and Type 2b (bilateral invasive type). The cervical lymph node metastasis areas were evaluated according to these types. Type 2 showed a significantly higher incidence of cervical lymph node metastasis only on the ipsilateral side than the other types (p = 0.0024). In particular, all patients with Type 2a had cervical lymph node metastasis only on the ipsilateral side (p = 0.0212). This study suggests that the distribution of metastasised cervical lymph nodes depends on the pattern of tumour extent of the primary site.

The use of electrolyzed solutions for the cleaning and disinfecting of dialyzers.

Recently, the use of electrolyzed solutions has attracted considerable interest in Japan. This study investigates the efficiency of electrolyzed solutions as disinfecting agents (DA) in the reuse of dialyzers and compares their efficiency to that of other disinfectants currently in use. The following 3 methods were employed. First, the rinsing time and rebound release of reused dialyzers were measured and compared after electrolyzed solutions, electrolyzed strong acid aqueous solution (ESAAS) and electrolyzed strong basic aqueous solution (ESBAS), made from reverse osmosis (RO) water (ESAAS, ESBAS; Generating apparatuses: Super Oxseed alpha 1000, Amano Corporation, Yokohama, Japan), 2% Dialox-cj (Teijin Gambro Medical, Tokyo, Japan), and 3.8% formalin were used as DAs. This involved performing dialysis with 2 types of dialyzers: a cellulose acetate membrane (CAM) dialyzer and a polysulfone membrane (PSM) dialyzer. The dialyzers were cleaned and disinfected using the different DA and left for 48 h. Next, after performing dialysis the dialyzer membranes were cleaned with a saline solution (0.9% NaCl) and RO water and then cleaned with the various DA. These membranes were observed using a scanning electron microscope (SEM) to check for the presence of physical and biological contaminants. Finally, in vitro tests were performed to determine the level of dialyzer clearance when PSM dialyzers were reused after having been cleaned and disinfected with the electrolyzed solutions. The rinsing time results for both the CAM and PSM dialyzers showed the electrolyzed solutions (ESBAS and ESAAS) as being undetectable within 10 min. With regard to the rebound release, for both the CAM and PSM dialyzers, the electrolyzed solutions were undetectable at all checking times between 30 and 240 min. Observation by SEM showed that cleaning with both ESAAS and ESBAS left the fewest contaminants, and cleaning with 2% Dialox-cj left the highest level of contaminants in the CAM dialyzers. With regard to experiments concerning use in vitro, no major changes in the dialyzer clearance were noticed after 6 uses. In every experiment, the previous investigations showed the electrolyzed solutions to be superior to 3. 8% formalin and 2% Dialox-cj DA for the reuse of dialyzers.

Use of a Technovit 7200 VLC to facilitate integrated determination of aluminum by light and electron microscopy.

Technovit 7200 VLC is an excellent embedding medium for both inorganic histochemistry by light microscopy and X-ray microanalysis by scanning and transmission electron microscopy. Liver samples from rats after intraperitoneal treatment with aluminum chloride were fixed in glutaraldehyde and embedded in the resin. Thick sections were easily cut on an ultramicrotome and stained with aluminon for aluminum (Al). An intense positive reaction with aluminon was observed in the Kupffer cells by light microscopy. The surface structures of the same resin block cut for light microscopy were observed under a scanning electron microscope fitted with an energy dispersive X-ray spectrometer. The Kupffer cells appeared white in the backscattered mode. Localization of Al in the Kupffer cells was confirmed by an X-ray distribution map in the scanning electron microscope. Subcellular localization of Al in the Kupffer cells was performed on the same semithin sections using a transmission electron microscope equipped with an energy dispersive X-ray spectrometer. Most Al was found in lysosomes of the Kupffer cells. The resin was stable in the electron beam and chlorine-free.

The cleaning and disinfecting of hemodialysis equipment using electrolyzed strong acid aqueous solution.

In general, sodium hypochlorite, formalin, and Dialox (Teijin Gambro Medical, Ltd., Tokyo, Japan [main ingredients: H2O2, CH3CHOOOH, CH3COOH, H2O]) are used to clean and disinfect hemodialysis pipelines. In this study, the suitability of electrolyzed strong acid aqueous solution (ESAAS), which has attracted considerable interest in Japan because of its strong disinfecting properties, was examined. The crossover method was used to investigate the effectiveness of ESAAS in disinfecting the dialysis pipelines in comparison to that of sodium hypochlorite (200 ppm) used alternately with 1% acetic acid. The number of bacteria and the concentration of endotoxin (Et) were measured over an approximately 3 year period, starting in September 1994. Until then, 200 ppm sodium hypochlorite had been used alternately with 1% acetic acid, and the contamination of the pipeline had been marked. However, after switching to the ESAAS disinfection method, the dialysis pipelines very rapidly became cleaner. Therefore, the decision to develop an automated ESAAS cleaning system for long-term use was made. During the development period, the original disinfectants (200 ppm sodium hypochlorite used alternately with 1% acetic acid) were used as a stopgap. After confirmation of its performance and safety, the automated ESAAS cleaning system was introduced. To find out whether the decrease in bacteria secondarily caused a decrease in the Et concentration or whether the ESAAS directly inactivated the Et, an in vitro experiment was carried out. Highly concentrated Et, which had been left in the reverse osmosis (RO) drainage pipeline, was used as a sample to investigate the effects of ESAAS on Et at various concentrations and temperatures and on the recovery test. The results showed that ESAAS directly inactivated Et. This paper reports the results of the crossover test. The results of parallel tests carried out over an approximately 4 year period have already been reported. No significant problems occurred in the dialysis. The automated ESAAS cleaning system that was developed proved to be more economical than the conventional disinfecting method.

The effect of electrolyzed strong acid aqueous solution on hemodialysis equipment.

We reported the high effectiveness of electrolyzed strong acid aqueous solution (ESAAS) in cleaning hemodialysis lines. Although ESAAS has a strong bactericidal action, one concern is its strong acidity. It has a pH of 2.3-2.7, more than 1,000 mV in oxidation-reduction potential (ORP), and 10-50 ppm of available chlorine. The possibility of metal corrosion, degradation of synthetic resins, chorine gas emission, or dissolving calcium carbonate (CaCO3) deposits due to ESAAS's acidity was tested using in vitro experiments. The bactericidal and antiviral effects of various ESAAS's were also tested. Metal corrosion and synthetic resin degradation, although they occurred, were not serious. There were no problems with chlorine gas emission and dissolving of CaCO3 deposits. Each type of ESAAS showed almost the same bactericidal and antiviral effect, but in some cases differences were observed.

Immunohistochemical localization of thrombomodulin in the stratified epithelium of the rat is restricted to the keratinizing epidermis.

The expression and function of thrombomodulin (TM), an endothelial cofactor protein for thrombin-mediated protein C activation, in the epithelium are not fully characterized. This report describes the distribution and localization of TM in the various types of epithelia in the rat by light and electron microscopic immunocytochemistry. TM showed a limited distribution and was expressed by the keratinizing stratified epithelia of the skin, tongue, and esophagus, but was not present on the non-keratinizing epithelia of the vagina, ureter, trachea, stomach, or gut. An identical pattern of TM expression was seen in mucocutaneous junctions, transitional zones from a non-keratinizing stratified epithelium to a keratinizing epithelium at the edge of the eyelid and in the anal canal. As the keratinization of the stratified epithelia proceeded, the staining intensity increased in the transitional zones. Within the keratinizing stratified epithelia, TM staining was limited to the keratinocytes of the spinous layer, the spinous cells. The subcellular localization of TM on the spinous cells was restricted to the plasma membrane facing the intercellular spaces. TM was not detectable on the desmosomes or the two membranes making up the junction, presumably the nexus. The functional significance of TM in keratinizing epithelia is discussed.

Immunocytochemical localization of thrombomodulin in the aqueous humor passage of the rat eye.

This report describes the distribution and localization of thrombomodulin (TM) in the rat eye by light and electron microscopic immunocytochemistry. In addition to the endothelium of the entire vasculature, TM was found on the non-vascular structures lining the cavities of the posterior and anterior chambers and the limbus. TM was localized on the basal, lateral, and apical plasma membranes of the inner and outer ciliary epithelium, and the posterior iris epithelium in which there was no polarized expression of TM. In the anterior chamber, TM was localized on the luminal surface of the corneal endothelium, but was negative on the anterior border layer of the iris, which is composed of a discontinuous layer of fibroblasts and collagen fibers. Thus, TM was present at sites of cell-to-cell contact. TM was also present on the endothelia of the trabecular meshwork and the Schlemm's canal in the limbus. TM was localized not only on the luminal plasma membrane, but also on the cytoplasmic giant vacuoles in the endothelial cells of the Schlemm's canal. These findings extend the importance of anticoagulant mechanisms to the systems of secretion, circulation, and drainage of the aqueous humor.

Granulation in amine-storage organelles of mouse megakaryocytes: X-ray microprobe analysis and radioautography.

The mechanisms and the processes of the storage of bivalent cations, ATP and 5-hydroxytryptamine (5HT) in the precursors of the amine-storage organelles of megakaryocytes were studied at the electron microscopic level. Although the precursors of the amine-storage organelles in the megakaryocytes fixed with glutaraldehyde and osmium tetroxide were empty, the electron opaque granules were observed in these organelles of the freeze-substituted megakaryocytes cut onto ethylene glycol. X-ray microprobe analysis demonstrated that they contained P, Mg and Ca. Quantitative differences in bivalent cations in the granules were not observed between megakaryocytes and blood platelets. Electron opaque uranaffin-reaction products were observed in the precursors of the amine-storage organelles of the megakaryocytes after treatment with the uranaffin reaction for ATP. However, few chromaffin positive granules were observed in the precursors of the amine-storage organelles after the chromaffin reaction for monoamines. Radioautographic analysis demonstrated that blood platelets avidly took up 3H-5HT but megakaryocytes were not able to accumulate 3H-5HT in vivo. These results indicate that megakaryocytes do not yet acquire the well developed uptake system of 5HT in vivo, while they readily accumulate cations and ATP in the precursors of the amine-storage organelles.