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The purpose of this study was to investigate the expansion of the maxillary sinus and the inclinations of posterior teeth in orthodontic patients with maxillary second premolar agenesis. A total of 30 subjects with one or two congenitally missing maxillary second premolars and retained maxillary deciduous second molars (a agenesis group) were selected and divided into a unilateral agenesis group (20 subjects with one maxillary second premolar missing) and a bilateral agenesis group (10 subjects with two maxillary second premolars missing). As controls, 30 sex- and age-matched subjects without agenesis of the maxillary second premolars were selected. Oblique cephalograms were used to investigate the association of maxillary second premolar agenesis and lower maxillary sinus size and posterior tooth inclinations. Agenesis of the maxillary second premolars induced significantly large lower maxillary sinus length, depth, area and mesial inclination of the maxillary first premolar, a significantly small anterior maxillary length, and a significantly more distal position for root apex of the maxillary first premolar. There were no significant differences in any measurements of the lower maxillary sinus and posterior teeth between the non-agenesis side in the unilateral agenesis group and the control group. Maxillary second premolar agenesis caused inferior and anterior expansion of the lower maxillary sinus and the mesial inclination of the maxillary first premolars with a distal position of root apex. Unilateral agenesis of the maxillary second premolar did not affect on the lower maxillary sinus size or posterior tooth inclinations of the unaffected antimere.

The aim of this study was to reveal the correlations between bilateral agenesis of third molars (M3s) and skeletal morphology in Japanese male and female orthodontic patients. Sixty patients (30 males, 30 females), with bilateral agenesis of maxillary M3s and without agenesis of mandibular M3s (group U), and 60 patients (30 males, 30 females), with bilateral agenesis of mandibular M3s and without agenesis of maxillary M3s (group L), were selected as agenesis groups. Additionally, 60 patients (30 males, 30 females) with all four M3s were selected as the control group (group C). Patients in these three groups had no agenesis of teeth other than M3s. Lateral cephalograms of each patient were used to evaluate skeletal morphology of the maxilla and mandible. Two-way analysis of variance was used for statistical comparisons. Groups U and L had significantly smaller maxillary length and area than group C. Group U exhibited a significantly smaller lower facial height than group C. Males showed significantly larger maxillary length; total mandibular and mandibular body length; mandibular ramus height; SNB angle; maxillary area; and mandibular symphysis, corpus and ramus areas than females. Females had significantly larger lower facial height, gonial angle and ANB angle than males. Smaller maxillary length and area and lower facial height should be considered in planning orthodontic treatment for patients with bilateral agenesis of maxillary and mandibular M3s.

The objective was to clarify whether dietary palmitic acid supplementation affects glucose-stimulated insulin secretion (GSIS) and the endoplasmic reticulum (ER) stress pathway in pancreatic islets in mice. Eight-week-old male C57BL/6J mice were randomly divided into three treatment diet groups: control diet, palmitic acid-supplemented diet (PAL) and oleic acid-supplemented diet (OLE). After 2 weeks of treatment, intraperitoneal glucose tolerance test and intraperitoneal insulin tolerance test were performed. GSIS was assessed by pancreatic perfusion in situ with basal (100 mg/dL) glucose followed by a high (300 mg/dL) glucose concentration. We measured mRNA levels of ER stress markers such as C/EBP homologous protein (CHOP), immunoglobulin heavy-chain binding protein (BIP) and X-box binding protein (XBP)-1 using real-time polymerase chain reaction (PCR) analyses in isolated islets. Immunohistochemical staining was also performed. Mice fed PAL showed significantly decreased glucose tolerance (p 0.05). In the perfusion study, GSIS was significantly suppressed in the PAL group (p 0.05). Semi-quantitative RT-PCR revealed that islet CHOP, BIP, and XBP-1 mRNA expression were significantly increased in the PAL group (p 0.05). TUNEL-positive β-cells were not detected in all groups. Dietary palmitic acid-supplementation for 2 weeks might suppress GSIS and induce ER stress in pancreatic islets in mice, in the early stage of lipotoxicity.

The effects of heteropolyoxotungstate (K7[PTi2W10O40]· 6H2O; PM-19) on the replication of herpes simplex virus type 2 (HSV-2) were examined using a semiquantitative polymerase chain reaction of intracellular viral DNA established by us and also other methods. Vero cells were infected with HSV-2 strains: either the standard strain 169, or the acyclovir-resistant strain YS-4C-1. PM-19 was added at various stages during the replication of HSV-2. PM-19 strongly inhibited the synthesis of viral genomic DNA when it was added at the time of infection. The addition of PM-19 60–90 min after viral inoculation time-dependently decreased the antiviral activity and increased the relative yield of viral DNA, and the addition of PM-19 was completely ineffective at times later than 90 min. These results suggested that PM-19 inhibited viral penetration but did not affect the synthesis of viral DNA. Furthermore, PM-19 strongly inhibited a second round of infection.

The keggin-type heteropolyoxotungstate K 7 [PTi 2 W 10 O 40 ]·6H 2 O (PM-19) is a potent polyoxometalate (PM) inhibitor of the replication of herpes simplex virus (HSV). Pretreatment of Vero cells with PM-19 prior to HSV-2 infection enhanced the antiviral potency of PM-19 almost 10-fold compared with treatment of the cells only after infection. The pretreatment effect of PM-19 is called ‘the memory effect’. The memory effect was reflected by inhibition of plaque formation and decrease of intracellular virus DNA quantity, and was strongest when PM-19 was present during the penetration stage of HSV-2 infection. The effect was maintained under conditions of fusion induced by polyethyleneglycol treatment. This suggests that PM-19 does not act at the fusion stage of infection. Using the infectious center assay method, it was clarified that a second round of infection was inhibited by about 30% in the presence of PM-19 at the penetration stage compared with the virus control in nontreated cells. The inhibition was enhanced to about 60% by PM-19 pretreatment prior to infection. This suggests that PM-19 pretreatment of the cells protects them against HSV-2 infection.

It is well known that alpha 2-adrenergic agonism inhibits insulin secretion and stimulates glucagon secretion in both animal and human studies. Recently, alpha 2-adrenergic blockers (DG-5128, MK-912, and SL 84.0418) have been studied as antihyperglycemic agents in human subjects. To clarify the action mechanism(s) of these agents, we investigated the effects of alpha 2 agonists and antagonists (10(-10) to 10(-4) mol/L) and pretreatment by pertussis toxin (PTX) on glucose-stimulated insulin secretion using the hamster insulinoma cell line HIT-T15. The imidazoline-derivative alpha 2-adrenoceptor agonists clonidine and oxymetazoline at concentrations as low as 10(-8) mol/L significantly inhibited glucose-stimulated insulin secretion by 63% and 65%, respectively (P.01 for both). These inhibitory effects were abolished by 20-hour preincubation of these cells with PTX 100 ng/mL. The imidazoline-derivative alpha 2-adrenoceptor antagonist DG-5128 at a concentration of 10(-4) mol/L doubled insulin secretion with or without pretreatment by PTX (P.01 for both). Furthermore, both clonidine and oxymetazoline at a high concentration of 10(-4) mol/L stimulated insulin secretion with pretreatment of the cells by PTX (P.05 for both). These results indicate that glucose-stimulated insulin secretion is inhibited by alpha 2-adrenoceptor agonists through PTX-sensitive G-protein in HIT-T15 cells. It is also suggested that imidazoline compounds at high concentrations directly stimulate insulin secretion.

In an IDDM model mouse which was inoculated with a diabetogenic variant, DK-27, of encephalomyocarditis (EMC) virus, the relation between a stable glycated hemoglobin A1C (St-HbA1C) level and plasma glucose level (PGL) in the progress of diabetes was studied. The St-HbA1C level and PGL were examined every 2 weeks for 10 weeks in normal male DBA/2 and IDDM-onset mice. PGLs of normal control mice did not change, but their St-HbA1C levels were slightly increased by 0.52 to 1.03%. On the other hand, in IDDM mice, their PGLs were greatly increased (> 400 mg/dl) and hyperglycemia was maintained throughout this observation period, with their St-HbA1C noticeably increased to 3.8% according to the progress of diabetes for 10 weeks. A highly significant correlation between St-HbA1C levels and averaged PGLs for the past weeks was found in IDDM mice. To examine the reflection of St-HbA1C levels to delicately varied PGLs, we also estimated both values in IDDM mice which were treated with insulin at a minimal effective dose once a day for 4 weeks. The St-HbA1C levels in insulin-injected IDDM mice were significantly lower than those in control IDDM mice. These findings suggest that the estimation of the St-HbA1C level is useful in following up blood glucose control in a long-term therapeutical study with small laboratory animals.

A-4166 is a new type of oral hypoglycemic agent that does not contain a sulfonylurea moiety. To clarify the mechanism of insulin secretion by A-4166, a specific receptor for A-4166 was investigated in a hamster pancreatic β cell line (HIT T-15), using [ 3 H]A-4166 or [ 3 H]glibenclamide as a ligand. The saturation binding of [ 3 H]A-4166 to HIT cell membranes was not observed up to 10 μM. In the displacement study, unlabeled A-4166 inhibited [ 3 H]A-4166 binding to HIT cell membranes, but glibenclamide did not. On the other hand, A-4166 inhibited [ 3 H]glibenclamide binding to the sulfonylurea receptor ( K i = 248 nM). A-4166 inhibited 86 Rb efflux from HIT cells ( ic 50 = 350 nM). The ec 50 for insulin secretion by A-4166 was 20 μM in HIT cells when they were incubated for 30 min in Krebs-Ringer bicarbonate buffer containing 16 mM HEPES supplemented with 5 mg/mL BSA in the absence of glucose. These data demonstrate the possibility of the presence of two kinds of binding sites for A-4166: one of them is the sulfonylurea receptor, and the other might be a binding site specific for A-4166.

A murine diabetes mellitus induced with a new diabetogenic variant (DK-27) which we isolated from the M variant of the encephalomyocarditis (EMC) virus was characterized. Male DBA/2 mice (9.5 weeks old) were infected with various infectious doses of DK-27 intraperitoneally. Blood glucose and insulin levels were examined in association with the viral replication. Pancreatic pathology and hormone contents and stable hemoglobin A1c (St-A1c) levels were also examined on the final day of observation (35 days of post-infection). In infected mice, blood glucose levels rapidly elevated at 72 hr, slightly decreased between 7 and 10 days and finally became sustained hyperglycemia. On the other hand, blood insulin levels elevated at 48 hr, promptly decreased, and subsequently became sustained hypoinsulinemia. Viral replication in pancreases reached the highest titers at 48 hr and rapidly disappeared with all infectious doses used. Pancreatic insulin contents in infected mice were not detectable, and glucagon contents were not affected. In pathological examination, atrophy of islets and marked diminution of B-cells were observed, and A-cells occupied the major part of an infected islet. St-A1c levels reflected lasting hyperglycemia. These findings show that DK-27 causes insulin-dependent diabetes mellitus by the specific and direct destruction of pancreatic B-cells in susceptible mice. Such a diabetic model mouse will be useful for therapeutic studies.

Using the in situ hamster pancreatic perfusion system, the stimulating action of A-4166 on insulin release was examined in comparison with that of glibenclamide. Both antidiabetic agents stimulated insulin release, but its onset by A-4166 was faster than that by glibenclamide. In the presence of a basal glucose concentration (3 mmol/l), insulin releases induced by A-4166 and glibenclamide were inhibited by preexisting diazoxide. At higher glucose concentrations (5-16.7 mmol/l), however, A-4166 was able to reverse the inhibitory effect of diazoxide on the first and second phases of insulin release, while glibenclamide did not reverse the first-phase release. On the other hand, in the presence of 16.7 mmol/l of glucose A-4166 completely reversed the inhibitory action of diazoxide added simultaneously, but glibenclamide reversed it only partially. In the presence of 8 mmol/l of glucose, the stimulating action of A-4166 and glibenclamide on insulin release was hardly affected by inhibitors of ATP production. These results indicate that the stimulating action of A-4166 on insulin release is different from glibenclamide in response to the inhibitory action of diazoxide. These results also suggest that A-4166 is an effective agent for release of insulin by acting on the KATP channel, especially under an impaired function of pancreatic B cells.

Antitumor polyoxomolybdates have been recognized in the course of study on the medical utilization of polyoxometalates, inorganic polymers of metal oxide. [NH3Pri]6[Mo7O24].3H2O (PM-8) was found as a representative of antitumor polyoxomolybdates. The growth suppressions of PM-8 against Co-4 human colon cancer xenografted under the subrenal capsule in cd-1 mice were equal or superior to that of 5-FU, MMC, ACNU, ADM and CDDP. Potent antitumor activity of PM-8 is also established against MX-1 human breast and OAT human lung cancer xenografted in athymic nude mice. Polyoxomolybdate is a new type of antitumor substance.

有機ケイ素化合物の抗腫瘍活性を培養細胞系と実験動物移植腫瘍系を用いて検討した。50種類の化合物には培養細胞系においてEhrlichがん,肉腫-180.Lewis肺がんおよびB-16メラノーマに対して細胞増殖抑制効果を示す化合物が認められ,また,マウス移植腫瘍系においても上記の各固形腫瘍に対して経口投与によって有効な抗腫瘍活性を現わす化合物が見いだされた.中でも2-(2-Trimethylsilylethy1)thiaethylamine(SDK-12A)は最も強い活性を示し,その効果は抗がん剤5-FUのそれに匹敵するものであった。さらに,SDK-12AはLewis肺がんの転移を有意に抑制し.遅延型免疫機能賦活化作用も認められた。またSDK-12Aは実験動物において毒性面では安全性の高い化合物であった。これらの結果は,SDK-12Aが多面的作用を持った特異な化合物であり,抗がん剤開発の研究領域からみて興味ある物質である。

To elucidate the mechanisms by which troglitazone, which is a direct ligand for peroxisome proliferator-activated receptor (PPAR) gamma, ameliorates insulin resistance, we have demonstrated that PPAR gamma is expressed in a pancreatic beta cell line, INS-1, using reverse transcription-polymerase chain reaction (RT-PCR). We incubated the cells with 5 μmol/l troglitazone and 1 mmol/l of each major free fatty acid (FFA; palmitic acid, oleic acid, and linoleic acid), alone or in combination, for 48 h. After that, we evaluated glucose-stimulated insulin secretion (GSIS) and 25 mmol/l KCl-induced insulin secretion in the presence of diazoxide, which clamps membrane potential. Our results showed: (1) treatment with troglitazone for 48 h caused enhancement of GSIS, although troglitazone significantly suppressed cell viability assessed by MTT assay. (2) In cells co-treated with troglitazone and FFA, troglitazone ameliorated lipotoxicity due to FFA. (3) In the presence of 300 μmol/l diazoxide and 25 mmol/l KCl, troglitazone did not affect the recovery of GSIS in INS-1 cells. These results suggest that insulin secretion from the rat insulinoma cell line, INS-1, is modulated by troglitazone, acting somewhere in the ATP-sensitive K + channel pathway, possibly through PPAR gamma.

The effects of long-term exposure of a pancreatic beta cell line, INS-1, to major free fatty acids (FFA; palmitic acid, oleic acid and linoleic acid) and leptin on insulin secretion and cell viability by C, N -diphenyl- N ′-4,5 dimethyl thiazol 2-yl tetrazolium bromide (MTT) assay were examined. The cells were incubated with 1 mmol/l of each FFA and 25 or 100 ng/ml leptin, alone or in combination, for 4, 24 or 48 h before the insulin secretion experiments. Palmitic acid (C 16:0) significantly suppressed cell viability, and suppressed insulin secretion at 24 h. Treatment with oleic acid (C 18:1) or linoleic acid (C 18:2) enhanced basal insulin secretion and diminished glucose-stimulated insulin secretion (GSIS) at 48 h. In these groups, there were no differences in cell viability as compared to cells treated without FFA. Leptin did not affect insulin secretion at 4, 24 and 48 h, and in the cells co-treated with FFA and leptin, leptin did not ameliorate lipotoxicity. These results suggest that, in INS-1 cells, different FFA have different patterns of lipotoxicity with chronic exposure, and leptin has little direct effect on insulin secretion.

A method for quantitation of the DNA of Herpes simplex virus type 2 (HSV-2)-infected Vero cells by the polymerase chain reaction (PCR) was developed. This method allowed recognition of several molecules of viral DNA among the total DNA extracted from cells. The method could be applied to a very large range (10 −0 –10 −7 ) of initial amounts of template. Products of PCR were collected after each cycle for kinetic analysis. Poducts were subjected to electrophoresis and amplified bands were stained with ethidium bromide. The intensity of fluorescence of each band was measured with a charge-coupled device (CCD) image analyzer. The time course of increases in the relative yield of viral DNA was determined. Two-fold amplification of viral DNA occurred each 6-h cycle from 7 h after infection. Using this method, the yields of viral DNA after treatment with the drug acyclovir (ACV) at 0.1 and 2 μg/ml were about 1/10 and 1/80 of those from nontreated infected cells, respectively. These results indicate that this method makes clear the inhibitory effect of ACV on the synthesis of viral DNA.

We have reported that N-[(trans-4-isopropyl-cyclohexyl)-carbonyl]-D-phenylalanine (A-4166) stimulates insulin secretion in animal studies. To further elucidate the mechanisms underlying the actions of this agent, we investigated the effects of A-4166 on insulin and glucagon secretion with or without diazoxide, an ATP-sensitive potassium channel opener, using isolated perfused rat pancreas preparations, and compared the results with those of glibenclamide. Both 30 mumol/l A-4166 and 3 mumol/l glibenclamide significantly stimulated insulin secretion and reduced glucagon secretion to similar levels at a glucose concentration of 5.6 mmol/l (p0.01 for both vs. basal levels). After infusion of A-4166 was stopped, insulin levels promptly returned to the basal values, while insulin levels increased further even after discontinuation of glibenclamide. Furthermore, 100 mumol/l diazoxide significantly inhibited the insulin-stimulatory effects of both 30 mumol/l A-4166 and 3 mumol/l glibenclamide (p0.05 and p0.01, respectively). However, the effects of diazoxide on glucagon secretion differed between the two groups; 30 mumol/l A-4166 produced a transient increase in glucagon secretion (p0.05 vs. basal levels) but 3 mumol/l glibenclamide suppressed glucagon secretion further (p0.01 vs. without diazoxide) with concomitant administration of 100 mumol/l diazoxide. These findings suggest that A-4166 directly stimulates insulin secretion, at least in part, through mechanisms resembling those of sulfonylurea, but exerts different effect on glucagon secretion in isolated perfused rat pancreas.

N-[(trans-4-isopropylcyclohexyl)-carbonyl]-D-phenylalanine (A-4166) has a structure which differs from those of other known blood glucose-lowering agents including sulfonylureas. It has been shown that oral administration of A-4166 exerts blood glucose-lowering effects in animal in vivo studies. In the present study, we investigated the effects of A-4166 on insulin and glucagon secretion at several glucose concentrations using isolated perfused rat pancreas preparations. Both 3.0 and 30 mumol/l A-4166 significantly stimulated insulin secretion as compared with basal levels at glucose concentrations of 8.0 and 11.0 mmol (p < 0.01 and p < 0.05, respectively). In contrast, glucagon secretion was not affected by administration of A-4166 up to 30 mumol/l at these glucose concentrations. At a glucose concentration of 5.6 mmol/l, neither 0.3 nor 3.0 mumol/l A-4166 produced significant changes in insulin and glucagon secretion. However, A-4166 at 30 mumol/l significantly stimulated insulin secretion and inhibited glucagon secretion as compared with basal levels (p < 0.01 and p < 0.01, respectively). We conclude that A-4166 at 3.0 and 30 mumol/l directly stimulates insulin secretion but has little effect on glucagon secretion in isolated perfused rat pancreas at glucose concentrations of 8.0 and 11.0 mmol/l. these results, taken together with previously published data, suggest that oral administration of A-4166 could be a useful strategy for stimulating endogenous insulin secretion in non-insulin-dependent diabetic patients.

A screening for inhibitors of human immunodeficiency virus type 1 (HIV-1) among various types of isopolyoxomolybdates and heteropolyoxomolybdates was carried out by using an in vitro assay system measuring the cytopathogenicity of HIV-1 in CD4+ human MT-4 cells. A novel heteropolyoxomolybdate named PM-104 with the chemical formula (NH4)12H2(Eu4(MoO4)(H2O)16(Mo7O24)4).13H2O was found to be associated with potent anti-HIV-1 activity. PM-104 interferes with virus infection at a very early step such as adsorption and/or penetration into the cells. In addition to the cytopathic effect of HIV-1 on MT-4 cells, syncytium formation between mock-infected MOLT-4 cells and MOLT-4 cells chronically infected with either HIV-1 or HIV-2 is suppressed by PM-104. PM-104 also blocks the replication of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). The antiviral properties of PM-104 could be attributed to the combined effect of europium atoms and its peculiar three-dimensional anion structure.

Polyoxotungstates with Keggin-type structure were found to demonstrate marked antiherpetic activity. K 7 [Ti 2 W 10 PO 40 ]·6H 2 O (PM-19) caused a decrease in plaque formation by several strains of herpes simplex virus (HSV) type 1, including acyclovir-resistant (thymidine kinase-negative) strains, at concentrations which were not toxic to the host cells. The 50% plaque-inhibiting concentration (EC 50 ) for the different strains was between 20 and 50 μg/ml. Single-cycle HSV growth was also inhibited by PM-19. PM-19 inhibited viral DNA synthesis in HSV-infected cells at a concentration of 5 μg/ml but did not exhibit a virucidal effect, and pretreatment of the host cells with PM-19 did not provide resistance to herpes infection. Yet, virus adsorption to the cells was markedly affected at PM-19 concentrations higher than 25 μg/ml. PM-19 was also effective against human cytomegalovirus, but not against adenoviruses and varicella-zoster virus, although it did delay the development of the cytopathic effect of these viruses.

Environmental substances were examined for their effect on interferon induction and delayed type hypersensitivity (DTH) responses in mice. Amaranth, safrole, phenacetin and nicotine suppressed the DTH response, and suppressed the serum interferon titers induced by virus infection. However, they did not affect the interferon titers which were induced by tilorone, a chemical inducer. The peak of interferon titer was 12 hours after infection with herpes simplex virus type 2 (HSV-2). Therefore, amaranth, safrole, phenacetin and nicotine were given to mice intraperitoneally 24 hours before, and 2 and 18 hours after infection with HSV-2. It was found that safrole and nicotine shortened the mean survival time of HSV-2 infected mice when they were given to mice 2 hours after virus inoculation. Amaranth and phenacetin showed similar effects. However it was not definite statistically. In biochemical and hematological tests, these four substances did not affect the functions of liver, kidney and carbohydrate metabolism in normal mice. These results suggest that substances which may often be taken into the body have the potential to affect the onset of virus infectious diseases as a result of the suppression of the host defense reaction.

N-Benzoyl-DL-phenylalanine (1) was found to possess hypoglycemic activity. A series of the analogues of compound 1 were prepared and evaluated for their blood glucose lowering activity. Both the steric effects of the phenylalanine moiety and the effects of variations in the acyl moiety were investigated. This study elucidated some of the structure-activity relationships and led to the development of N-(4-ethylbenzoyl)-D-phenylalanine (34), which was 50 times more potent than the initial compound 1.

We previously reported in this journal that the TC-83-vaccine strain of Venezuelan encephalitis (VE) virus results in a sustained diminution of glucose-stimulated insulin release in golden Syrian hamsters, persisting as long as 90 days after viral infection. This study was designed to examine the metabolic and pathologic consequences of TC-83 VE virus infection in C57 BL/Ks mice (+/+) and in genetic variants of this strain homozygous (db/db) and heterozygous (db/+) for the diabetic gene, db. Five-week-old mice of each genetic variant were inoculated subcutaneously, in groups of 18, with 100,000 plaque-forming units (PFU) of TC-83 that had not been passaged in chick embryo cells or with diluent (control mice). The clinical course in all three groups of mice following VE inoculation was mild, with a 5 to 10% mortality. By light microscopy, control +/+, TC-83 VE-infected +/+, control db/+, and TC-83 VE-infected db/+ pancreases manifested no appreciable difference in morphology. Uninfected db/db mice showed typical changes, including a definite decrease in the number of aldehyde fuchsin-staining granules in beta cells. TC-83 VE-infected db/db mice exhibited a profound decrease in pancreatic beta cell granulation on aldehyde fuchsin staining. After TC-83 VE inoculation, the most striking alterations in carbohydrate metabolism occurred in db/db mice, which showed further worsening of glucose tolerance 120 min after intraperitoneal glucose as well as significantly decreased basal and glucose-stimulated immunoreactive insulin (IRI) levels. After TC-83 VE infection, pancreatic IRI content was not decreased in +/+ or db/+ mice but was virtually absent in db/db mice. The data support the ability of TC-83 VE to inhibit glucose-stimulated IRI release in three genetic variants of C57 BL/Ks mice. In addition, this model demonstrates the heightened susceptibility of the remaining diabetic beta cells in the db/db mice to subsequent infection with a pancreatropic virus.

Benzamidine derivatives exhibited a high antiviral effect in vivo against influenza virus strains A2/Adachi and B/Lee. The inhibiting properties of anti-influenza benzamidine derivatives on the virus-induced inflammation undoubtedly plays an important role in the clarification of the mechanism of action of these drugs.

A series of analogues of N-(cyclohexylcarbonyl)-D-phenylalanine (5) have been synthesized and evaluated for their hypoglycemic activity. Relationships were studied between the activity and the three-dimensional structure of the acyl moiety, which was characterized by high-resolution 1H NMR spectroscopy and MNDO calculations. The role of the carboxyl group of the phenylalanine moiety was also studied by comparing the activities of the enantiomers, the decarboxyl derivative, the esters, and the amides of the phenylalanine derivatives. Thus, the structural requirements for possessing hypoglycemic activity was elucidated and a highly active compound, N-[(trans-4-isopropylcyclohexyl)carbonyl]-D-phenylalanine (13) was obtained, which showed a 20% blood glucose decrease at an oral dose of 1.6 mg/kg in fasted normal mice.

14C-Pipethanate hydrochlorideは，マウスに経口投与されると30分後には，血中および胆汁中放射活性は最高濃度に達し，時間推移とともに減少した．胆汁への排泄は24時間後まで続くが，48時間後には全く消失した．放射活性はほぼ全身に分布し，特に肝臓や消化管に高い放射活性が見られたが，時間とともに減少して，蓄積性や残留性を示す臓器組織は認められなかった．尿中へは，3～6時間後には，全投与量の約50%が排泄され，24～48時間までに投与された量のほとんどが体外へ排泄され，その割合は，尿に約70%，糞便中に約30%である．呼気中への排泄は，ごくわずかで0.35%であった．以上の所見は，同時に行なった凍結全身オートラジオグラフィの結果によっても裏づけられる．

(1) In order to find more effective agents than guanidine, potentiating agents were searched by means of the salt formation.(2) The potentiating effect of some amino aicd antagonists was examined, and ethionine was found as the most marked potentiation agent among the antagonists.(3) The potentiating effect of some of methionine homologs was examined, and any of the homologs was found not to have an effect more significant than ethionine.(4) The potentiating effect of some of commoner organic acids was examined, but any of these acids was found not to show any potentiating effect.(5) The mechanismus of the potentiation of guanidine with ethionine was speculated, from the hypotheses presented so far.

1) All of Mahoney, MEF1 and Saukett strains of polio virus developed guanidine resistant mutants in all of employed cell-lines, the HeLa, the FL, the KB and the Hep. No. 2 cells.2) In the assay of guanidine resistance of the reproduced progeny virus, all of employed cell-lines did not show any difference in the efficiency.3) Guanidine resistant virus could not be recovered in point of view of the resistance even after the serial passages in the culture using guanidine free medium.4) Guanidine resistant virus becomes guanidine dependent virus after the further passages in the culture using guanidine containing medium.5) The rate of the intracellular multiplication of guanidine resistant virus in the culture using guanidine containing medium was just the same to that in the culture using guanidine free medium, while guanidine sensitive virus was remarkably inhibited in the culture using guanidine containing medium.6) To develop guanidine resistant virus, the incubation-time from 24 hours to 34 hours after viral inoculation is necessary.7) When the second growth cycle of virus is stopped by the addition of anti-Mahoney serum into guanidine containing medium, guanidine sensitive Mahoney did not develope guanidine resistant Mahoney so remarkably compared with the experiments using the medium containing guanidine.8) The results described in both of 6) and 7) suggest that guanidine resistant virus is produced by selection-mechanism.9) Guanidine resistant virus does not possess t-marker character.

A new antiviral inhibitor was produced from cell-culture in the absence of Interferon inducing factors. This substance was referred to as ANTIVIRIN. This inhibitor was produced from the culture of many stable-cell-lines and primary cultured cells. The effect of this inhibitor was of rather wide spectrum. Both of RNA viruses and DNA ones were inhibited with this inhibitor. According to the experimental results, it was suggested that this inhibitor must be a complicated protein, but it is not sure at the present time whether sugar or nucleic acid participate in the active component or not.

To find chemotherapeutic agents for influenza and adenoviruses, 3-(alkoxyphenoxy)-1, 2-propanediols were synthesized. Among these derivatives, 3-(m-dodecyloxyphenoxy)-1, 2-propanediol possessed a fairly inhibitory effect on the multiplication of the PR-8 strain of influenza virus in chorioallantoic membrane culture. By the primary screening test on the type-1 strain of adenovirus, 3-(p-decyloxyphenoxy)-, 3-(p-dodecyloxyphenoxy)-, 3-(o-hexyloxyphenoxy)-, 3-(o-octyloxyphenoxy)-, 3-(o-decyloxyphenoxy)-, 3-(m-butoxyphenoxy)-, 3-(m-pentyloxyphenoxy)-, 3-(m-hexyloxyphenoxy)-, 3-(m-octyloxyphenoxy)-, 3-(m-decyloxyphenoxy)-, and 3-(m-dodecyloxyphenoxy)-1, 2-propanediols were selected. By the secondary screening test, 3-(o-hexyloxyphenoxy)-, 3-(o-octyloxyphenoxy)-, and 3-(p-dodecyloxyphenoxy)-1, 2-propanediols were found to be effective on 10×TCD50 of the type-1 of adenovirus. Among these three compounds, 3-(o-hexyloxyphenoxy)-1, 2-propanediol was effective on the type-4 and -6 strains, and 3-(o-octyloxyphenoxy)-1, 2-propanediol was effective on the type-4 strain virus, while 3-(p-dodecyloxyphenoxy)-1, 2-propanediol did not show any effect on the type-2, -3, -4, -6, and -7 strains of adenovirus. These three compounds did not possess any direct action for inactivating adenovirus and it was suggested from the experiments that the effectiveness of the compounds was due to the inhibition of viral multiplication.