Efficacy and safety of live varicella zoster vaccine in diabetes: a randomized, double-blind, placebo-controlled trial.

AIMS: To elucidate varicella zoster virus (VZV)-specific cell-mediated immunity and humoral immunogenicity against live attenuated Oka varicella zoster vaccine concurrently vaccinated with 23-valent pneumococcal polysaccharide vaccine (PPSV23) in elderly people with diabetes mellitus. METHODS: This double-blind randomized controlled single-centre study of 60-70-year-old people with diabetes compared immunity and safety profiles 3 months after one dose of varicella zoster vaccine or placebo. PPSV23 was immunized simultaneously. Primary analysis evaluated cell-mediated immunity using the VZV skin test. Secondary analyses were a VZV interferon-γ enzyme-linked immunospot (ELISPOT) assay and immunoadherence haemagglutination test. Adverse experiences were recorded using diary questionnaires. RESULTS: By intent-to-treat analysis, 27 participants with diabetes who had been administered the vaccine were compared with 27 participants who were given a placebo. Changes in skin test scores were 0.41 ± 0.80 and 0.11 ± 0.93 (P = 0.2155), and geometric mean fold rises of the ELISPOT counts were 1.2 [95% confidence interval (CI) 0.2, 7.9] and 1.2 (95% CI 0.2, 7.3) (P = 0.989) in the vaccine and placebo groups, respectively. The geometric mean titre did not increase 3 months after vaccination in either group. No vaccination-related severe adverse experience was reported and no participant developed herpes zoster. DISCUSSION: Our previous results demonstrated that varicella zoster vaccine safely enhanced VZV-specific immunity in elderly people with or without diabetes. The results of this study showed that varicella zoster vaccine can be used safely, but it cannot boost virus-specific immunity in elderly people with diabetes when administered with concurrent PPSV23. Alternative strategies are needed to prevent VZV-associated diseases in this population.

Monodomain Blue Phase Liquid Crystal Layers for Phase Modulation.

Liquid crystal "Blue Phases" (BP) have evolved, in the last years, from a scientific curiosity to emerging materials for new photonic and display applications. They possess attractive features over standard nematic liquid crystals, like submillisecond switching times and polarization- independent optical response. However, BPs still present a number of technical issues that prevent their use in practical applications: their phases are only found in limited temperature ranges, thus requiring stabilization of the layers; stabilized BP layers are inhomogeneous and not uniformly oriented, which worsen the optical performance of the devices. It would be essential for practical uses to obtain perfectly aligned and oriented monodomain BP layers, where the alignment and orientation of the cubic lattice are organized in a single 3D structure. In this work we have obtained virtually perfect monodomain BP layers and used them in devices for polarization independent phase modulation. We demonstrate that, under applied voltage, well aligned and oriented layers generate smoother and higher values of the phase shift than inhomogeneous layers, while preserving polarization independency. All BP devices were successfully stabilized in BPI phase, maintaining the layer monodomain homogeneity at room temperature, covering the entire area of the devices with a unique BP phase.

Various costimulatory pathways are essential for induction of regulatory cells by intratracheal delivery of alloantigen.

BACKGROUND: We previously reported that intratracheal delivery of alloantigen induced regulatory cells in a mouse heart transplantation model. We investigated the roles of costimulatory pathways in the induction of regulatory cells by intratracheal delivery of alloantigen. METHODS: CBA (H-2k) mice were pretreated with intratracheal delivery of splenocytes (1 x 10(7)) from C57BL/10 (H-2b) mice and administration of monoclonal antibodies (mAb) specific for programmed death (PD)-1 and its ligands, programmed death-ligand (PD-L)1 and PD-L2, CD70, CD134 ligand (CD134L), CD153, CD137L, or receptor activator of nuclear factor-kappaB (NF-kappaB) (RANK). Seven days later, naive CBA mice underwent adoptive transfer of splenocytes (5 x 10(7)) from the pretreated CBA mice and transplantation of C57BL/10 heart. RESULTS: Adoptive transfer of splenocytes from CBA mice that had been pretreated with intratracheal delivery of C57BL/10 splenocytes significantly prolonged the survival of C57BL/10 allograft (median survival time [MST], 68 days) as compared with adoptive transfer from untreated CBA mice (MST, 12 days). Concomitant administration of control immunoglobulin (Ig)G, anti-PD-L2 mAb, or anti-CD137L along with intratracheal delivery did not significantly affect the prolongation (MST, 72, 68, and 65 days, respectively). In contrast, anti-PD-1, anti-PD-L1, anti-CD70, anti-CD134L, anti-CD153, or anti-RANK mAb abrogated the prolongation induced by adoptive transfer from the pretreated mice with intratracheal delivery (MST, 18, 17, 16, 14, 10, and 18 days, respectively). CONCLUSION: The PD-1/PD-L1, CD27/CD70, CD134/CD134L, CD30/CD153, and tumor necrosis factor (TNF)-related activation-induced cytokine (TRANCE)/RANK interactions are independently required for generation of regulatory cells by intratracheal delivery of alloantigen.

Purification and some properties of two kinds of keratin-hydrolyzing enzymes of cow snout epithelium.

Two kinds of keratin-hydrolyzing enzymes (KHEs) from cow snout epithelium were highly purified by affinity chromatography using soybean trypsin inhibitor-bound Sepharose. On gel filtration chromatography, the KHEs were eluted at a volume corresponding to a relative molecular mass (Mr) of 21,000. They were separated from each other by ion exchange chromatography. One of the enzymes had the same characteristics as urea extracted alkaline proteinase, of which optimal pH was at 8.5 to 9.0. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed a single band with Mr of 21,500 in the presence or absence of a reducing agent. The other enzyme was a neutral proteinase, with an optimal pH of 7.5. Both enzymes were inhibited by phenylmethylsulfonyl fluoride and soybean trypsin inhibitor. Among the fluorogenic peptides that were hydrolyzed most effectively by the alkaline proteinase were peptidyl MCAs (4-methyl-coumaryl-7-amides) with extended sequences, Boc-Leu-Ser-Thr-Arg-MCA, and then Boc-Val-Pro-ARg-MCA. The neutral proteinase hydrolyzed the latter most effectively. They hydrolyzed preferentially high Mr keratins of cow snout and of newborn mouse epidermis, and showed a limited proteolysis toward 68,000 polypeptide, giving rise to distinct products. The high substrate specificity and extended subsites of the KHEs suggest their role on the metabolism of the high Mr keratins.

Water sorption-desorption test of the skin in vivo for functional assessment of the stratum corneum.

Based on the evidence from our previous studies that we can evaluate the hydration state of the skin surface quickly and quantitatively in terms of conductance to the high frequency electric current of 3.5 MHz, we have established a simple in vivo function test that furnishes information on the hygroscopic property and water-holding capacity of the stratum corneum in a few minutes. The test procedure consists of electromeasurements before and after application of a droplet of water on the skin for 10 seconds to obtain data on the hygroscopic property of the skin surface and later serial measurements at an interval of 30 seconds for 2 min to evaluate the water-holding capacity. Under usual ambient conditions normal skin surface showed a high rise in conductance just after application of water, which was followed by a rapid fall-off within 30 seconds, thereafter by gradual return to the prehydration levels by 2 min. By this method we have demonstrated that (i) the superficial horny layer of normal skin is much less hygroscopic and less capable of holding water than the corresponding deeper portions and that (ii) scaly skin shows functional defects in both hygroscopicity and water-holding capacity, between which the former normalizes much faster than the latter.

Folate supplements and phenytoin-salicylate interaction.

We found biphasic fluctuations of serum phenytoin level when aspirin was added to chronic phenytoin therapy for an epileptic patient. The total serum phenytoin level was lowered initially by addition of aspirin. However, after 4 months on phenytoin-aspirin combination therapy, he showed elevated serum phenytoin levels, mild nystagmus, and serum folate deficiency.

Self-limited neonatal familial hyperparathyroidism associated with hypercalciuria and renal tubular acidosis in three siblings.

Three siblings with neonatal familial hyperparathyroidism diagnosed at age 4 months, 2 months, and 5 days, respectively, were treated. Hypercalciuria, nephrocalcinosis, and renal tubular acidosis were present in each child. In all three, there were higher responses of serum parathyroid hormone to serum calcium and higher elevation of serum calcium with oral calcium loading. The metabolism of vitamin D and calcitonin seemed to be intact. Hypercalcemia associated with the abnormal response of parathyroid hormone secretion disappeared when the children passed the age of approximately 2 years, although renal tubular acidosis and nephrocalcinosis remained. An autosomal recessive inheritance seems likely.

Establishment of feeder-independent cloned caprine trophoblast cell line which expresses placental lactogen and interferon tau.

A feeder-independent cloned trophoblast cell line, HTS-1, was established from a mature placenta of Shiba goat (Capra hircus). During the growth phase, single HTS-1 cells exhibited ruffled membranes or lamellipodia often accompanied by elongated cell shape, indicating highly motile nature of the cells. At or near confluence, HTS-1 cells formed monolayers with few sign of cellular overlapping. Binucleate cells were found at a high frequency especially in the peripheral regions of monolayers. In small colonies and the monolayers, majority of HTS-1 cells assumed polygonally shaped cobble-stone like morphology characteristic to epithelial cells, although considerable variations in cellular morphology were observed despite of repeated cloning. Time-lapse video recordings of HTS-1 cells during culture revealed that not only the small colonies but also the monolayers near or at confluence were remarkably motile, often causing extreme elongation of the cells within them. The extremely plastic nature of HTS-1 cells in vitro is likely to be the reflection of the extraordinary capacity of caprine trophoblast cells to be stretched to extreme thinness in vivo as shown by electron microscopy. HTS-1 cells cultured on matrigel are highly invasive, and express MT1-MMP which, in the mouse, has been known to be expressed at the invasive edge of trophoblast both in vitro and in vivo. HTS-1 cells express placental lactogen (PL) and interferon-tau (IFNtau), as confirmed by immunocytochemistry, Western blotting and RT-PCR analysis. Both PL and IFNtau expression in the cells appeared to be down-regulated by cell-cell contact. In the medium conditioned by HTS-1 cells, the presence of secretory form of PL and IFNtau was confirmed by Western blotting. The HTS-1 cell line will serve as a useful in vitro model for the analysis of the molecular and/or cellular mechanisms underlying synepitheliochorial placentation in bovidae animals.

Interaction of fibrinogen with detergent.

Both cationic and anionic detergents were found to precipitate fibrinogen by forming fibrinogen-detergent complexes. These complexes were soluble in distilled water, but the aqueous solutions were very unstable and the complexes precipitated in the presence of salt. In the interaction of fibrinogen with the cationic detergent, stearyltrimethyl-ammonium chloride, approximately 160 molecules of detergent were found to bind to one molecule of fibrinogen. In distilled water, the fibrinogen-stearyltrimethylammonium complex (FG-STA(Cl)) remained soluble in the presence of thrombin [ED 3.4.21.5] although the same peptides were released as those released from fibrinogen. Precipitation of FG-STA(Cl) by salt was found to be closely related to adsorption of the anion of the salt by the complex. Futher addition of salt resulted in solubilization of the precipitate, and the solubilization was also due to further adsorption of the anion onto the precipitate.

Chemical modification of carboxyl groups of fibrinogen and its effect on the binding of cationic detergent.

Carboxyl groups of native human fibrinogen were modified with glycine methyl ester and 1-ethyl-3(3-dimethylaminopropyl)carbodiimide. It seemed likely that the modification occurred stepwise. Approximately 26% of the carboxyl groups of fibrinogen was modified finally. The modified fibrinogen had no interaction with cationic detergent, and did not form any complex with the detergent. In dilute acid, fibrinogen was observed to show only a slight interaction with cationic detergent. It is probable that the exposed and ionized carboxyl groups are essential for the formation of a complex between fibrinogen and cationic detergent.

Purification of fibrinogen using cationic detergent.

Stearyltrimethylammonium chloride was used to isolate human fibrinogen, and purified protein was obtained by removing the detergent bound to it. Medium consisting of 0.015--0.03 mM fibrinogen-detergent complex, 0.85 M NaCl, 0.03 M sodium caprylate, and 30 per cent ethanol was found to be effective for renaturation of fibrinogen from the complex. The purified fibrinogen did not form any fibrils on incubation for 15 days with Ca-2+ at pH 7.2, and 37 degrees. The clottability of the purified fibrinogen was over 99 per cent. Immunochemical studies showed that the purified fibrinogen produced one precipitation line with a mixture of anti-human fibrinogen and anti-human serum. Although highly purified, the fibrinogen preparation still contained a trace of plasminogen.

Inverse relation of serum Helicobacter pylori antibody titres and extent of intestinal metaplasia.

AIMS: To clarify the relation between the serum titre of anti-Helicobacter pylori (H pylori) antibody and the extent of intestinal metaplasia of the gastric mucosa. METHODS: The serum anti-H pylori IgG titres of 95 asymptomatic individuals (mean age 65 years) undergoing an annual health examination were measured and compared with the extent of intestinal metaplasia (absent, moderate, or extensive), determined by examination of multiple endoscopic mucosal biopsy specimens. Serum pepsinogen I (PGI) levels, as a marker for gastric atrophy, were also measured. RESULTS: The prevalence of seropositivity for H pylori antibody was high (> 80%), regardless of the extent of metaplasia. However, there was a negative association between the extent of metaplasia and the anti-H pylori titre: 75% of the subjects in the group without metaplasia had high (3+) antibody levels, as did 43% with moderate, and 37% with extensive metaplasia (absent v extensive). The inverse relation between the titre and the extent of metaplasia was evident when examined in those with normal PGI (> 30 ng/ml), whereas no such relation was apparent in subjects with low PGI (< or = 30 ng/ml). CONCLUSIONS: The anti-H pylori titre correlates inversely with the extent of intestinal metaplasia, particularly in subjects with less marked gastric atrophy.

Mature and immature myeloid cells decrease the granulocyte colony-stimulating factor level by absorption of granulocyte colony-stimulating factor.

We studied the effects of polymorphonuclear neutrophils (PMN) and immature myeloid cells on the granulocyte colony-stimulating factor (G-CSF) level in vitro to better understand the regulatory mechanisms of neutropoiesis. Intact normal PMN decreased the G-CSF level after incubation with recombinant human (rh) G-CSF in a time- and dose-dependent manner. The percent reduction decreased as the concentration of rhG-CSF increased. However, the cell-free PMN-conditioned medium (PMN-CM) did not decrease the G-CSF level. The intact PMN also decreased the granulocyte-macrophage (GM)-CSF level after culture with rhGM-CSF, but did not affect the monocyte (M)-CSF level after culture with rhM-CSF. Normal bone marrow (BM) immature neutrophilic cells and G-CSF-dependent acute myeloid leukemic cells (OCI/AML la) also decreased the G-CSF level, whereas K-562 cells, which have no detectable G-CSF receptors, did not affect it. Phenylarsine oxide (PhAsO), an inhibitor of endocytosis of ligand receptor complex, abrogated this decreasing effect of intact PMN and OCI/AML la cells. These findings suggest that mature and immature myeloid cells negatively regulate neutropoiesis by, at least in part, decreasing the G-CSF level probably through receptor-mediated continual absorption and metabolism of G-CSF.

Glucocorticoid-induced apoptotic pathways in eosinophils: comparison with glucocorticoid-sensitive leukemia cells.

Glucocorticoids are known to promote apoptosis of eosinophils, normal and neoplastic lymphoid cells, and blastic cells in some patients with acute myeloid leukemia. We investigated the biochemical signal transduction pathways, in particular, the generation of reactive oxygen species (ROS) and activation of caspases in dexamethasone (DEX)-induced apoptosis of eosinophils, and we compared them with those in DEX-sensitive myeloid and lymphoid leukemia cell lines. The GC-receptor antagonist completely abolished DEX-induced apoptosis of eosinophils and leukemia cells. Among inhibitors related to the ROS system, diphenylene iodonium (DPI), a nicotinamide adenine dinucleotide diphosphate (NADPH) oxidase inhibitor, strongly inhibited both spontaneous and DEX-induced apoptosis of eosinophils at concentrations as low as 0.2 to 2 mumol/L, while promoting apoptosis of leukemia cells in a dose-dependent manner. Apocynin, another NADPH oxidase inhibitor, and antioxidants did not affect the apoptosis of eosinophils or leukemia cells. DEX treatment did not change intracellular production of O2- and H2O2, and it decreased the extracellular release of O2- in both cells. These results suggest little or no involvement of ROS generation in DEX-induced apoptosis of both cells. Although among peptide-based caspase inhibitors, only z-VAD-FMK, a broad caspase inhibitor, partially inhibited the apoptosis of eosinophils and leukemia cells, DEX treatment increased the activities of caspases 2-, 3-, 6-, and 8-like proteases assessed by colorimetry in both cells, suggesting the involvement of a similar caspase activation pathway in DEX-induced apoptosis in both cells. DPI markedly reduced caspase 3-like activity in eosinophils, while augmenting the activity in leukemia cells, indicating that DPI acts upstream of caspase 3 activation opposingly in both cells. Thus, the action of DPI in eosinophils seems peculiar in respect to apoptosis induction, and DPI appears to exert an influence on unknown targets rather than those involved in NADPH oxidase inhibition.

Low-dose cytarabine and aclarubicin in combination with granulocyte colony-stimulating factor (CAG regimen) for previously treated patients with relapsed or primary resistant acute myelogenous leukemia (AML) and previously untreated elderly patients with AML, secondary AML, and refractory anemia with excess blasts in transformation.

We used the CAG regimen (low-dose cytarabine [10 mg/m2 per 12 hours, days 1-14], aclarubicin [14 mg/m2 per day, days 1-4], and granulocyte colony-stimulating factor [200 micrograms/m2 per day, days 1-14]) for the treatment of patients with primary resistant acute myelogenous leukemia (AML) and previously untreated elderly patients with AML, secondary AML, and refractory anemia with excess blasts in transformation (RAEB-T) in addition to relapsed AML. Forty-three of 69 (62%) patients achieved complete remission (CR), including 29 of 35 (83%) patients with relapsed AML, 1 of 8 patients with primary resistant AML, 5 of 8 elderly patients with previously untreated AML, and 8 of 18 patients with previously untreated secondary AML or RAEB-T. Ten of 22 (45%) patients > or = 65 years old achieved CR. The patients who achieved CR received at least 1 course of modified CAG therapy as the first consolidation therapy, followed by various second consolidation and intensification therapies. The median disease-free survival and overall survival were 8 and 15 months, respectively, for relapsed AML; 11 and 8 months for the elderly patients; and 8 and 17 months for secondary AML and RAEB-T. Myelosuppression was mild to moderate, and other than fever, severe nonhematologic toxicity was rare. CAG as the induction therapy seems promising for the treatment of various categories of poor-prognosis AML.

L-2-hydroxyglutaric aciduria: two Japanese adult cases in one family.

We report two adult Japanese sisters with L-2-hydroxy-glutaric aciduria (acidemia), both of whom were much older (aged 57, 47 years old) than previously reported patients (from neonate to 44 years old), and who presented with differing severity. Magnetic resonance imaging revealed typical subcortical white matter lesions in both cases and showed brainstem atrophy and thickness of the calvarium in the elder sister. L-2-Hydroxyglutaric acid levels were increased in urine, plasma, and cerebrospinal fluid. These cases suggest that organic acid analysis is necessary even in elderly patients who seem to have neurodegenerative disorders.

Bimodal pattern of killing of Chinese hamster V79 variant cells by hydrogen peroxide.

To elucidate the mechanism of cytotoxicity of H2O2, we selected H2O2-resistant Chinese hamster V79 cells by single-step selection from a pool of spontaneous variants. The resistant cells showed bimodal sensitivity to H2O2 without exhibiting a significantly higher level of the detoxicating enzymes, catalase, glutathione peroxidase and superoxide dismutase. Mode-one and mode-two killing were observed at lower (< 300 microM) and higher (> 2 mM) H2O2 concentrations, respectively. Mode-one but not mode-two killing was prevented by iron chelators. Pretreatment with low concentrations of ascorbic acid preferentially enhanced the killing at higher H2O2 concentrations. These resistant cells were cross-resistant to t-butyl hydroperoxide and cumene hydroperoxide.

Mitochondrial biosynthesis controls the sensitivity of Chinese hamster cells to hydrogen peroxide.

The mechanism of H2O2-resistance of Hpr-4, a variant of Chinese hamster V79 cells, was investigated. The effect of H2O2 on the mitochondria of the parental and Hpr-4 cells was compared. First, both biochemical and ultrastructural results showed that mitochondria in the parental cells were damaged by exposure to H2O2, while those in Hpr-4 cells recovered from the damage. Second, the H2O2-resistance of Hpr-4 cells was reversibly reduced or recovered by the addition or removal of inhibitors of mitochondrial biosynthesis, respectively. Third, the parental cells were auxotrophic to pyruvate after exposure to H2O2. Fourth, H2O2-sensitivity of the parental cells was also enhanced by the inhibition of mitochondrial biosynthesis. From these results, it was concluded that the mitochondria of Hpr-4 cells apparently had a greater resistance to H2O2 than those of the parental cells and that functional mitochondria were involved in the recovery of Chinese hamster V79 cells from H2O2-induced damage.

Immunoblot analysis of antibodies in human strongyloidiasis.

An electrophoretic transfer technique was used to investigate qualitatively the production of antibodies to Strongyloides stercoralis larvae in 56 patients with strongyloidiasis. SDS-PAGE analysis of the larval extract revealed the presence of at least 33-39 polypeptide bands under either reducing or non-reducing condition. In the immunoblot analysis, almost all patients showed positive reactivity to the polypeptide bands. The reactivity, however, revealed significant variation among the patients, ranging in number of bands from only one to more than 18. Among the bands, 4 with molecular sizes of 97, 66, 41 and 26 kDa were frequently recognized by the patients' sera, indicating that these antigenic components may form an available antigen for immunological testing for strongyloidiasis. On the other hand, the reactivities were very faint in cases of overwhelming strongyloidiasis.

Quantification of myonecrosis and comparison of necrotic activity of snake venoms by determination of creatine phosphokinase activity in mice sera.

Serum creatine phosphokinase (CK) values were measured in venom-injected mice. The increase of CK levels was proportional to both the amount of venom injected and the myonecrotic actions determined microscopically. The CK isoenzymes were regarded as due to the predominant increase of CK-MM, which was considered to be of skeletal muscle origin in the venom-injected area. No CK isozyme variants were found by electrophoretic examination. For a comparative study on necrotic intensity, nine snake venoms were examined. The elapid venoms induced higher values of serum CK with a stronger necrotic effect than the crotalid venoms, and the data agreed with the histological findings. On application of the antivenom, an initially elevated CK value was significantly decreased and no severe damage was observed under light microscopy. Since the changes of serum CK activities paralleled necrotic intensity, the CK value is useful as an indicator of myonecrosis due to snake venom.