Biomedical subjects

N Kobayashi

Publications and source records attributed to N Kobayashi.

At least 235 records · Page 13Linked to original sources

PubMed2000

[Anaplastic thyroid carcinoma with lung metastasis producing CA 19-9 and GM-CSF].

A 68-year-old Japanese woman was admitted to our hospital because of hoarseness, dysphagia and a mass on the right side of her neck. Chest radiographs showed multiple nodular shadows in both lung fields. Detailed investigations resulted in a diagnosis of multiple lung metastasis of anaplastic thyroid carcinoma transformed from papillary adenocarcinoma. Both serum CA 19-9 and GM-CSF levels were elevated, to 70.5 U/ml (normal range: 0-37 U/ml) and 343.4 pg/ml (normal range: 0-8 pg/ml), respectively. Immunostaining disclosed that the primary and metastatic tumors were positive for CA 19-9, but not for GM-CSF antigens. Serum levels of these two parameters slowly decreased after chemo-radiotherapy, suggesting that the tumor may have produced GM-CSF as well as CA 19-9. Recent studies have indicated that the prognosis is poor for non-small cell lung cancers that produce G-CSF or CA 19-9. To our knowledge, this is the first case report of anaplastic thyroid carcinoma characterized by high serum levels of both CA 19-9 and GM-CSF, with metastasis to the lung and other organs.

Adenocarcinoma, Papillary↗

PubMed2000

[Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (1998)].

The bacteria isolated from the patients with lower respiratory tract infections were collected by institutions located throughout Japan, since 1981. Ikemoto et al. have been investigating susceptibilities of these isolates to various antibacterial agents and antibiotics, and analyzed some characteristics of the patients and isolates from them each year. Results obtained from these investigations are discussed. In these 18 institutions around the entire Japan, 532 strains of presumably etiological bacteria were isolated mainly from the sputa of 438 patients with lower respiratory tract infections during the period from October in 1998 to September in 1999. MICs of various antibacterial agents and antibiotics were determined against 85 strains of Staphylococcus aureus, 100 strains of Streptococcus pneumoniae, 96 strains of Haemophilus influenzae, 75 strains of Pseudomonas aeruginosa (non-mucoid strains), 6 strains of Pseudomonas aeruginosa (mucoid strains), 38 strains of Moraxella subgenus Branhamella catarrhalis, 26 strains of Klebsiella pneumoniae etc., and the susceptibilities of 517 strains were assessed except for those strains that died during transportation. S. aureus strains for which MICs of oxacillin (MPIPC) were higher than 4 micrograms/ml (methicillin-resistant S. aureus: MRSA) accounted for 60.0%. Vancomycin (VCM) and arbekacin (ABK) showed the most potent activities against MRSA. But one of MRSA showed resistance to ABK with the MIC of 64 micrograms/ml. The sensitive strains of MRSA to VCM have decreased. The frequency of penicillin (PC)-intermediate S. pneumoniae (PISP) + PC-resistant S. pneumoniae (PRSP) have increased in 46.0% for 1998 comparatively from 30.9% of 1997's. But PRSP decreased, and PISP increased into 39.0% of 1998 years from 19.8% of 1997's. Panipenem (PAPM), imipenem (IPM) and faropenem (FRPM) showed the most potent activities against S. pneumoniae with MIC80s of 0.125 microgram/ml or below. Against H. influenzae and M. (B.) catarrhalis, almost all the drugs showed good activities. The sensitive strains of them against ceftazidime (CAZ) decreased in 1997, but those have increased in 1998. Inversely, the susceptibility of them against cefotiam (CTM) had been higher in 1997, but those have been lower in 1998. Tobramycin (TOB) showed the most potent activity against P. aeruginosa (both mucoid and nonmucoid strains). All drugs except ampicillin (ABPC) were active against K. pneumoniae. A quite few of K. pneumoniae showed low susceptibilities. Also, we investigated year to year changes in the characteristics of patients, their respiratory infectious diseases, and the etiology. The examination of age distribution indicated that the proportion of patients with ages over 70 years was 48.6% of all the patients showing a slight increase in every year. About the proportion of diagnosed diseases as follows: Bacterial pneumonia was the most frequent with 40.2%. The ratio of it has increased slightly, and the increased rate was 10% in patients with ages over 70 years compared with the results in 1997. Chronic bronchitis have decreased slightly with 27.6% in 1998. Number of strains isolated from patients before administration of antibiotics were more than those after administration of them in chronic bronchitis, but these were almost same number in bacterial pneumonia. Administration of antibiotics has changed the results of the frequency of isolation of bacterial species. Bacterial isolations before administration of antibiotics were as follows: S. pneumoniae 26.7%, H. influenzae 23.8%, S. aureus 13.3% and M. (B.) catarrhalis 10.8%. The frequencies of S. aureus decreased after antibiotics administration over 15 days, but the frequencies of P. aeruginosa (both mucoid and non-mucoid) was not affected. The frequencies of P. aeruginosa was 45.5% after administration over 15 days. The frequencies of S. pneumoniae decreased upon administration of antibiotics, these were only 4.5% over 15 days. The frequencies of H. (

Anti-Bacterial Agents↗

PubMed2000

[Evaluation of coronary flow reserve in patients with vasospastic angina].

OBJECTIVES: The presence of microvascluar impairment was evaluated in 154 patients with vasospastic angina identified by the acetylcholine provocation test. METHODS: Coronary flow reserve was evaluated with a Doppler flow guidewire in 128 vessels of 72 patients with chest pain, but no significant coronary stenosis(less than 50% stenosis) and no clinical factors that affect coronary flow reserve. Coronary flow reserve was obtained from the ratio of adenosine triphosphate-induced maximum/baseline averaged peak velocity. These vessels were classified into 2 categories according to whether acetylcholine-induced vasospasm was positive or negative. Vasospasm positive was defined as more than 90% stenosis provoked with chest pain and/or ischemic ST change. Positive vessels were subdivided according to focal or diffuse vasospasm. These vessels were also classified into 2 other categories according to whether vasospasm in the distal artery was positive or negative. RESULTS: Coronary flow reserve was significantly lower in vessels with vasospasm than in vessels without vasospasm in patients without vasospasm(2.9 +/- 0.8 vs 3.6 +/- 1.0, p = 0.0005). Coronary flow reserve was significantly lower in vessels without vasospasm in patients with vasospasm than in vessels without vasospasm in patients without vasospasm(3.0 +/- 0.8 vs 3.6 +/- 1.0, p = 0.03). There was no significant difference in coronary flow reserve between vessels with vasospasm and vessels without vasospasm in patients with vasospasm(2.9 +/- 0.8 vs 3.0 +/- 0.8, p = 0.8). There was no significant difference in coronary flow reserve between focal and diffuse vasospasm(3.2 +/- 0.8 vs 2.9 +/- 0.8, p = 0.3). Coronary flow reserve was significantly lower in vessels with vasospasm in the distal artery than in vessels without vasospasm in the distal artery (2.8 +/- 0.8 vs 3.4 +/- 1.0, p = 0.004). CONCLUSIONS: Patients with vasospastic angina have microvascular impairment in both vessels with vasospasm, and vessels without vasospasm. Microvascular impairment is prominent in vessels with vasospasm in the distal artery.

Acetylcholine↗

PubMed2000

[The effects of aerobic training on the emotional response in patients who underwent cardiac surgery].

It has been reported that cardiac surgery and aerobic training influence the patient's emotional response. We investigated the changes in emotional response before and after cardiac surgery and during aerobic exercise training as a cardiac rehabilitation using profile of mood states (POMS). Subjects were thirty-five patients (25 men and 10 women, average 57 years) who underwent cardiac surgery. All patients participated in the rehabilitation program which included aerobic exercise training after cardiac surgery. Aerobic training consisted of cycle ergometer or treadmill. Emotional states were evaluated by POMS score at the preoperative phase, early postoperative phase (about 10 days after surgery) and aerobic training (about 20 days after surgery). Most of the emotional state (tension, anxiety, anger and hostility) significantly improved after cardiac surgery. In particular, aerobic training has an additional effect for improving one of the emotional state which is tension and anxiety. However, several factors such as deconditioning, postoperative complications and high age delayed the improving of emotional response.

Adolescent↗

PubMed2000

Establishment of a tightly regulated human cell line for the development of hepatocyte transplantation.

Hepatocyte transplantation (HTX) could be an attractive treatment for patients with liver failure and liver-based metabolic disease. Human primary hepatocytes are ideal in this modality, but the shortage of human livers available for hepatocyte isolation severely limits the use of this form of therapy. A tightly regulated human hepatocyte cell line that grows economically in culture and exhibits differentiated liver functions would be an attractive alternative to the primary human hepatocytes. To test the feasibility, human hepatocytes were immortalized by a retroviral vector expressing simian virus 40 large T antigen and herpes simplex virus-thymidine kinase. A highly differentiated immortal hepatocyte line NKNT-3 was established. NKNT-3 cells grew in chemically defined serum-free medium, retained highly differentiated liver functions, and were sensitivity to ganciclovir as a prodrug. Essentially unlimited availability of NKNT-3 cells may be clinically useful for HTX and bioartificial liver.

Antigens, Viral↗

PubMed2000

Effects of theophylline on human eosinophil functions: comparative study with neutrophil functions.

The understanding of theophylline as a bronchodilator has been reconsidered in recent years. We undertook to determine its immunomodulatory actions in granulocytes and elucidate their mechanism. Preincubation of neutrophils with theophylline (10(-5) to 5 x 10(-3) M) had a biphasic effect on O2(-) production stimulated with N-formyl-methionyl-leucyl-phenylalanine or C5a. Theophylline potentiates O2(-) production via adenosine A(2A) receptor antagonism induced by receptor-linked agonists from neutrophils, but not from eosinophils. The addition of theophylline caused a significant decline in neutrophil chemotaxis at lower concentrations than those for eosinophil motility. Theophylline reduces neutrophil chemotaxis via adenosine A1 receptor antagonism. At high concentrations, with an intracellular cAMP accumulation as a result of phosphodiesterase (PDE) inhibition, theophylline also exerts an inhibitory effect on the O2(-) production and chemotaxis of both types of cells. The difference in theophylline's effect on neutrophils and eosinophils appears to depend on the existence of specific adenosine receptors. Theophylline thus modulates granulocyte functions in association with specific adenosine receptor antagonism and cAMP-PDE inhibition.

Bronchodilator Agents↗

PubMed2000

Margin clearance and HPV infection do not influence the cure rates of early neoplasia of the uterine cervix by laser conization.

A lesion existing in the endocervical and/or ectocervical conized margin and HPV-DNA existing in a conized specimen are reported to be at risk of persistence or recurrence of early neoplasia of the cervix when treated by conization. The aim of this study was to investigate whether margin clearance and HPV infection influenced the outcome in our series of laser conization. Excisional conization with the KTP/YAG Surgical Laser System or Nd-YAG laser was performed in this study. Eighty patients with cervical neoplasias were included: 47 with dysplasia, 25 with carcinoma in situ (CIS) and eight with microinvasive carcinoma. The endocervical and ectocervical conized margins were examined microscopically. HPV-DNA was analyzed with the primer for types 16, 18, 31, 33, 35, 52b and 58 amplified by the PCR method. The margins of the conized specimens were confirmed histopathologically to be clear in 58 cases (73%), whereas in 22 cases (27%) they were involved by neoplasia. HPV-DNA was positive in 38% of dysplasias, 40% of CISs and 50% of microinvasive carcinomas. The overall rate of the initial cure at 10 weeks after treatment appeared to be 100% in all 80 cases. Primary cure rates were 100% for 47 cases with dysplasia, 96% for 24 cases with CIS and 100% for four cases with microinvasive carcinoma regardless of margin positivity and HPV-DNA status. Involved margins and HPV infection did not influence the cure of early neoplasia of the uterine cervix achieved by our laser conization procedure. The favorable results may be due to the procedure of vaporizing the cut surface forming a dome-shaped tissue defect.

Conization↗

PubMed2000

[A case of neurocysticercosis presenting as focal seizure].

In Japan neurocysticercosis is a rare disease. We report a case of neurocysticercosis with a review of the literature. A 29-year-old man presented focal seizure on the left side. Physical and neurological findings on admission were unremarkable. CT and MRI showed a mass with ring-like enhancement and perifocal edema in the right frontal lobe, as well as multiple small masses with calcification. The main mass measuring 8 mm in diameter, was suspected as the eliptogenic focus, and was totally removed via the transcortical approach. Postsurgically seizures did not recur, and histological examination of the mass confirmed a cysticercus. A therapeutic course of albendazole and praziquantel administered postsurgically was effective for the multiple intraparenchymal masses.

Adult↗

PubMed2000

[Pituitary cystic mass with spontaneous disappearance: three cases of equivocal Rathke's cleft cyst].

Three cases with a pituitary cystic mass that spontaneously disappeared were discussed. Common characteristics of these 3 cases were presence of headache, cystic mass, spontaneous disappearance, and no recurrence. Based on clinical course, MRI findings such as cystic mass with ring enhancement, no calcification, and displacement posteriorly of the normal pituitary gland, and previous reports, these 3 cases were diagnosed as an equivocal Rathke's cleft cyst. We suggested that operative indication in an equivocal Rathke's cleft cyst, even though it was symptomatic, should be decided after sufficient follow-up, because there may be a possibility of spontaneous reduction or disappearance like our cases.

Adult↗

PubMed2000

Fas antigen expression and outcome of oral squamous cell carcinoma.

Paraffin sections of biopsy specimens obtained from 46 patients with oral squamous cell carcinomas were stained with both anti-peptide antibody against human Fas antigen and monoclonal mouse antibody against human proliferating cell nuclear antigen (PCNA). The patients received chemotherapy with a combination of carboplatin and peplomycin sulfate or mitomycin C and peplomycin sulfate before surgery. The relation between the expression of Fas antigen and the clinical features of each case was examined. The correlation between PCNA and Fas antigen expression was also studied. The mean PCNA labeling index of the 22 Fas-negative cases was 46.9%, which was significantly higher than that of the 24 Fas-positive cases (39.5%). Strong correlations were found between the expression of Fas antigen and the response to chemotherapy, tumor recurrence, and survival. The Fas-negative group had only a minor response to chemotherapy and a poor outcome, whereas the Fas-positive group had a better response to chemotherapy and a good outcome. Although lymph node metastasis was significantly related to survival, there was no correlation between Fas antigen expression and lymph node metastasis. The Kaplan-Meier survival curve of patients positive for Fas antigen was significantly better than that of patients negative for Fas antigen. Our results suggest that Fas antigen expression is an independent predictor of outcome whose usefulness should be evaluated in prospective studies.

Actuarial Analysis↗

PubMed2000

[Does quinapril improve coronary vasoconstriction in vasospastic angina?].

OBJECTIVES: Endothelial dysfunction is one of the important mechanisms of coronary spasm. Recently, the angiotensin converting enzyme (ACE) inhibitor quinapril, which has a high affinity for vascular ACE, has improved endothelial dysfunction in coronary artery disease. This study investigated whether quinapril improves coronary vasoconstriction induced by acetylcholine in vasospastic angina. METHODS: Twenty-four patients with vasospastic angina without significant organic stenosis diagnosed by the acetylcholine provocation test (vessel with spasm defined as > or = 90% stenosis provoked with chest pain and/or ischemic ST change) were enrolled in this study. Patients were randomly assigned to 2 groups: the quinapril group (receiving quinapril 20 mg/day, n = 12), and the non-quinapril group (not receiving quinapril, n = 12). All patients received calcium antagonist. Six months later, coronary angiography was repeated and changes in coronary spasm were compared between the groups. Seven patients were withdrawn from this study, so 17 patients were evaluated (the quinapril group: 8 patients, the non-quinapril group: 9). RESULTS: Angina symptoms were completely or almost suppressed in all patients during the study period. Angiographically, the number of patients with improvement of spasm, patients with deterioration of spasm and patients with stability of spasm were 1, 0, 7 in the quinapril group versus 0, 1, 8 in the non-quinapril group, respectively. There was no significant change between the 2 groups. Quantitative angiographic analysis showed that the coronary spasm rate (percentage of minimal lumen diameter at the site of spasm after administration of acetylcholine to minimal lumen diameter after administration of isosorbide dinitrate) at the first and second angiography were 71 +/- 10% and 62 +/- 21% in the quinapril group versus 73 +/- 14% and 67 +/- 15% in the non-quinapril group, respectively. There were no significant interval changes between the 2 groups (p = 0.60). CONCLUSIONS: Our results did not indicate that quinapril had improved coronary vasoconstriction induced by acetylcholine in patients with vasospastic angina.

Acetylcholine↗

PubMed2000

[Down's syndrome].

Explore the source record for details and available documents.

Diagnosis, Differential↗

PubMed2000

A new approach to develop a biohybrid artificial liver using a tightly regulated human hepatocyte cell line.

Currently patients with liver failure have been treated with a various liver support systems including a whole liver perfusion, a non-biological artificial liver, and a biohybrid artificial liver. In a hepatocyte-based bioreactor, porcine hepatocytes or transformed human liver tumor cells have been utilized because of the ease of preparation. According to the clinical data reported as of now, satisfactory results have not been obtained from the use of currently available liver support devices. One of the problems is limited availability of primary human liver cells for developing live support systems because of the shortage of human liver. To resolve this issue, human hepatocytes were immortalized with a retroviral vector SSR#69 which contained the genes of simian virus 40 large T antigen (SV40Tag) and herpes simplex virus-thymidine kinase (HSV-TK). One of the immortal cell lines, NKNT-3, showed the gene expression of differentiated liver functions, grew steadily in chemically defined serum-free CS-C medium, and doubled in number in about 48 hours. Essentially unlimited availability of NKNT-3 cells supports their clinical use for liver support devices. To realize the high density culture of NKNT-3 cells in a bioartificial liver device, we have developed cellulose microspheres (CMS) which contain cell adhesive GRGDS (Gly-Arg-Gly-Asp-Ser) peptides. Within 24 hours after starting a stirring suspension culture, GRGDS-CMS efficiently immobilized NKNT-3 cells. An electron microscopic examination demonstrated that NKNT-3 cells attached on GRGDS-CMS had well-developed mitochondria, rough reticulums, and villous extensions. In this article, we review the history of extracorporeal liver support systems and describe an attractive strategy for developing a novel extracorporeal liver assist device using NKNT-3 cells and GRGDS-coated cellulose microspheres.

Animals↗

PubMed1999

(-)-Epigallocatechin gallate inhibits mos activation-mediated xenopus oocyte maturation induced by progesterone.

(-)-Epigallocatechin gallate (EGCG), one of the constituents of green tea known to have a tumor preventing effect, inhibited maturation of Xenopus laevis oocytes induced by progesterone when this polyphenol was microinjected into oocytes at a final concentration of about 1 mM. Western blot and activity measurement analyses showed that Mos translation and the subsequent activations of mitogen-activated protein kinase and p90(rsk), probably by protein phosphorylation, seemed to have been inhibited by the microinjection of EGCG. These results suggest that EGCG may have the ability to control Xenopus oocyte maturation at least during the stage of Mos activation.

Animals↗

PubMed1999

An immunomodulatory arabinomannan extracted from Mycobacterium tuberculosis, Z-100, restores the balance of Th1/Th2 cell responses in tumor bearing mice.

The regulatory effect of Z-100 on the balance of Th1/Th2 cell responses in BALB/c mice bearing Meth-A fibrosarcoma was investigated. In tumor bearing mice, Th1 cytokine production (IL-2, IFN-gamma) are suppressed and Th2 cytokine production (IL-4, IL-10) are increased, as compared with those of normal mice. The administration of Z-100 (10 mg/kg) to tumor bearing mice restored the balance of Th1/Th2 cell responses from Th2 dominant state to the normal state. This regulatory effect of Z-100 was eliminated by depletion of adherent cells from splenocytes derived from tumor bearing mice, and by the treatment with 2-ClAdo (a macrophage inhibitor). Similarly, this regulatory effect was diminished by the treatment with anti-IL-12 mAb and anti-IFN-gamma mAb. In addition, the IL-12 p40 mRNA expression in splenic adherent cells and IFN-gamma mRNA expression in CD4+ T cells were increased by the administration of Z-100 to tumor bearing mice. These results suggested that Z-100 restored the balance of Th1/Th2 cell responses to the normal one in tumor bearing mice through the activation of macrophages and up-regulation of IL-12 production from macrophages and IFN-gamma production from CD4+ T cells.

Adjuvants, Immunologic↗

PubMed1999

NMR analysis of the binding of a rhodanese peptide to a minichaperone in solution.

A detailed structural analysis of interactions between denatured proteins and GroEL is essential for an understanding of its mechanism. Minichaperones constitute an excellent paradigm for obtaining high-resolution structural information about the binding site and conformation of substrates bound to GroEL, and are particularly suitable for NMR studies. Here, we used transferred nuclear Overhauser effects to study the interaction in solution between minichaperone GroEL(193-335) and a synthetic peptide (Rho), corresponding to the N-terminal alpha-helix (residues 11 to 23) of the mitochondrial rhodanese, a protein whose in vitro refolding is mediated by minichaperones. Using a 60 kDa maltose-binding protein (MBP)-GroEL(193-335) fusion protein to increase the sensitivity of the transferred NOEs, we observed characteristic sequential and mid-range transferred nuclear Overhauser effects. The peptide adopts an alpha-helical conformation upon binding to the minichaperone. Thus the binding site of GroEL is compatible with binding of alpha-helices as well as extended beta-strands. To locate the peptide-binding site on GroEL(193-335), we analysed changes in its chemical shifts on adding an excess of Rho peptide. All residues with significant chemical shift differences are localised in helices H8 and H9. Non-specific interactions were not observed. This indicates that the peptide Rho binds specifically to minichaperone GroEL(193-335). The binding region identified by NMR in solution agrees with crystallographic studies with small peptides and with fluorescence quenching studies with denatured proteins.

ATP-Binding Cassette Transporters↗

PubMed1999

Comparison of immediate and follow-up results of the short and long NIR stent with the Palmaz-Schatz stent.

The intrinsic characteristics of a stent including stent length may affect both procedural success and long-term outcome. The present study evaluated the immediate and follow-up results after implantation of the short and long NIR stent and compared these results with the Palmaz-Schatz stent. Between July 1995 and December 1996, stenting with a 16-mm NIR stent (NIR-16), a 32-mm NIR stent (NIR-32), or a Palmaz-Schatz stent (PS) was performed in 68, 57, and 155 lesions, respectively. There were no significant differences in the incidences of delivery failure (PS, 2.6%: NIR-16, 4.4%; NIR-32, 5.3%; p = NS) and procedural success (PS, 92%; NIR-16, 93%; NIR-32, 93%; p = NS) among the 3 groups. The reference vessel diameter was smaller in lesions with a 32-mm NIR stent than in those with a Palmaz-Schatz stent (PS, 3.14+/-0.58, NIR-16, 3.00+/-0.50; NIR-32, 2.90+/-0.47 mm; p <0.05). The lesion length was longer in lesions with a 32-mm NIR stent than in those with a Palmaz-Schatz or a 16-mm NIR stent (PS, 8.9+/-5.0; NIR-16, 11.0+/-4.1; NIR-32, 26.1+/-9.7 mm; p <0.01). After the procedure, the lesions with a 32-mm NIR stent had a smaller minimal lumen diameter than those with a Palmaz-Schatz stent (PS, 3.17+/-0.61; NIR-16, 2.99+/-0.51; NIR-32, 2.89+/-0.49 mm; p <0.01). At follow-up, a smaller minimal lumen diameter was observed in lesions with a 32-mm NIR stent than in those with a Palmaz-Schatz or a 16-mm NIR stent (PS, 2.32+/-0.98; NIR-16, 2.25+/-0.80; NIR-32, 1.68+/-0.79 mm; p <0.01). Restenosis rates were 16.5% in lesions with a Palmaz-Schatz stent, 13.3% in those with a 16-mm NIR stent, and 47.4% in those with a 32-mm NIR stent (p <0.01). Although stent delivery and procedural success of a long NIR stent were acceptable, the restenosis rate of a long NIR stent was high compared with a short NIR stent or a Palmaz-Schatz stent.

Adult↗

PubMed1999

D-configuration of serine is crucial in maintaining the phalloidin-like conformation of viroisin.

NMR studies have revealed that the conformation of the monocyclic viroisin is dissimilar to that of the corresponding monocyclic derivative of phalloidin, dethiophalloidin, but has much similarity with the conformation of the bicyclic phalloidin. Obviously, one of three structural features found exclusively in the virotoxins is able to compensate for the conformational strain that in the bicyclic phallotoxins maintains the toxic conformation. Synthetic work on virotoxin analogues has shown that both the additional hydroxy group in allo-hydroxyproline and the methylsulfonyl moiety in the 2'-position of tryptophan are unlikely to represent the structural element in question, leaving the D-serine moiety as the supposed key element. In this study we asked whether it is the hydroxy group of this amino acid or its D-configuration that is responsible for the effect. We synthesized four viroisin analogues and submitted them to conformational analysis by NMR as well as to an actin binding assay. While the rotating-frame nuclear Overhauser effect (ROESY) spectra of the analogues with L-configured amino acids showed several sets of signals, indicating the existence of conformers interconverting more slowly than the NMR time scale, the spectra of the analogues with D-configured amino acids showed only one set of signals. Remarkably, the two viroisin analogues with D-serine and D-alanine also had distinctly higher affinities for filamentous actin than their L-configured counterparts, suggesting that the high biological activity may be correlated with the absence of multiple and slowly interconverting conformers. Anyhow, D-configuration of serine is the structural element that maintains the phalloidin-like structure, while the hydroxy group does not contribute to conformational stability but is likely to be in contact with the actin surface.

Actins↗