Viability of infectious viral particles of HIV and BMCs in breast milk.

BACKGROUND: Infectious factors in breast milk such as viral particles and living infected cells are of prime importance in the transmission of HIV by breastfeeding. OBJECTIVES: To perform effective approaches for reducing HIV transmission via breastfeeding, we investigated the biological importance of infectious viral particles and infected BMCs in breast milk. STUDY DESIGN: Alteration of viral infectivity was monitored using a modified experimental infection assay that exploited the cytotoxicity of breast milk, and BMC viability was evaluated by flow-cytometric analysis. RESULTS: Infectious viral particles were found to decrease time-dependently after contact with breast milk, whereas BMCs showed prolonged survival in breast milk. CONCLUSIONS: The biological importance of infected BMCs in breast milk for the transmission of HIV via breastfeeding was considered.     

Neutrophil dysfunction in chronic neutrophilic leukemia without rearrangements of bcr and immunoglobulin heavy chain genes

A 67-year-old man was admitted to our hospital with abdominal distension due to hepatosplenomegaly. The peripheral blood revealed Hb content 6.5 g/dl, platelet count 4.7 x 10(4)/microliter, and WBC count 105.8 x 10(3)/microliter with 88% of mature neutrophils. The neutrophil alkaline phosphatase score was 421. Bone marrow aspiration revealed hypercellularity with increased megakaryocytes and myeloid hyperplasia. 46, XY, del 20(q 11) without Philadelphia chromosome was identified by cytogenetic study. The patient was diagnosed as having chronic neutrophilic leukemia and was successfully treated with busulfan, but he died of atypical mycobacteriosis about 20 months later. Analysis of neutrophil function demonstrated diminution of deformability, random mobility, and chemotaxis, but almost normal phagocytosis and bactericidal capacity. Southern analysis showed no rearrangements of breakpoint cluster region (bcr) gene and immunoglobulin heavy chain gene.     

High-speed imaging at 3 Tesla: a technical and clinical review with an emphasis on whole-brain 3D imaging.

Improvements to the inherently high specific-absorption rate (SAR) of high-speed imaging at 3T are necessary in order to render this method clinically feasible. Various efforts have been undertaken to improve the associated hardware and software. In this review, we focus on whole-brain isotropic 3D imaging with a turbo spin-echo sequence with variable flip-angle echo trains (3D-TSE-VFL) and present its technical and clinical features. This sequence can be used to acquire images of various contrasts including T2-weighted, fat-suppressed T2-weighted, fluid-attenuated inversion recovery (FLAIR), fat-suppressed FLAIR, and STIR (short tau inversion recovery). Various aspects of 3D-TSE-VFL are discussed, including CSF (cerebrospinal fluid) and metal artifacts, STIR contrast, small-part visualization other than brain, and the possibility of serial subtraction. Some images from clinical cases are presented.     

Effective diminution of amniotic prostaglandin production by selective inhibitors of cyclooxygenase type 2

OBJECTIVE: Cyclooxygenase inhibitors are effective tocolytic agents, but significant adverse effects limit their use. We hypothesized that selective inhibitors of the isozyme cyclooxygenase 2 would effectively diminish labor-associated prostaglandin production. STUDY DESIGN: We analyzed cyclooxygenase type 1 and 2 expression in amnion, chorion, decidua, and myometrium from laboring or nonlaboring women and tested the efficacy of selective cyclooxygenase 2 inhibition in diminishing prostaglandin production. RESULTS: The expression of cyclooxygenase 2 in amnion from women in labor, either preterm or at term, was significantly higher than in amnion before labor. In contrast, cyclooxygenase 1 expression was unchanged by labor. The enhanced expression of amniotic cyclooxygenase 2 was associated with increased prostaglandin E(2) levels in laboring women. Amniotic prostaglandin E(2) production was effectively diminished by the selective cyclooxygenase 2 inhibitors SC-236 and NS-398 but not by the cyclooxygenase 1 inhibitor SC-560. CONCLUSION: Selective inhibitors of cyclooxygenase 2 are effective in diminishing prostaglandin production in vitro and may be useful in prevention of preterm deliveries.     

Effect of Er:YAG laser irradiation on acid resistance to bovine dentin in vitro

Resin bond strength to Er:YAG laser irradiated dentin has been reported to be lower than that of unlased dentin. The reasons have been much discussed, but not clarified. One hypothetical cause has been discussed that lased dentin is acid resistant, therefore, the etching effect of acid conditions decreases. The purpose of this study was to evaluate the acid resistance of laser-irradiated dentin and compare it with the dissolved mineral of Er:YAG laser irradiated dentin and unlased dentin. This experiment was a pilot study to assess the etching effect of pre-conditioner for resin bonding to lased dentin. Bovine dentin was irradiated by Er:YAG laser and immersed in 0.1 M lactic buffer solution (pH 4.0). The dissolved Ca and P in the solution were then both measured. Dissolved Ca from lased dentin was not significantly different from that coming from unlased dentin (p > 0.05). The molar ratio of Ca/P did not differ significantly between lased and unlased dentin, either (p > 0.05). Under FE-SEM view before immersion, the dentin surface was covered with a smear layer in unlased dentin, but this layer was not clearly observed in lased dentin. These results suggested that the lased dentin had little or no resistance to lactic buffer solution.     

Delayed satiety-like actions and altered feeding microstructure by a selective type 2 corticotropin-releasing factor agonist in rats: intra-hypothalamic urocortin 3 administration reduces food intake by prolonging the post-meal interval.

Brain corticotropin-releasing factor/urocortin (CRF/Ucn) systems are hypothesized to control feeding, with central administration of 'type 2' urocortins producing delayed anorexia. The present study sought to identify the receptor subtype, brain site, and behavioral mode of action through which Ucn 3 reduces nocturnal food intake in rats. Non-food-deprived male Wistar rats (n=176) were administered Ucn 3 into the lateral (LV) or fourth ventricle, or into the ventromedial or paraventricular nuclei of the hypothalamus (VMN, PVN) or the medial amygdala (MeA), regions in which Ucn 3 is expressed in proximity to CRF(2) receptors. LV Ucn 3 suppressed ingestion during the third-fourth post-injection hours. LV Ucn 3 anorexia was reversed by cotreatment with astressin(2)-B, a selective CRF(2) antagonist and not observed following equimole subcutaneous or fourth ventricle administration. Bilateral intra-VMN and intra-PVN infusion, more potently than LV infusion, reduced the quantity (57-73%) and duration of ingestion (32-68%) during the third-fourth post-infusion hours. LV, intra-PVN and intra-VMN infusion of Ucn 3 slowed the eating rate and reduced intake by prolonging the post-meal interval. Intra-VMN Ucn 3 reduced feeding bout size, and intra-PVN Ucn 3 reduced the regularity of eating from pellet to pellet. Ucn 3 effects were behaviorally specific, because minimal effective anorectic Ucn 3 doses did not alter drinking rate or promote a conditioned taste aversion, and site-specific, because intra-MeA Ucn 3 produced a nibbling pattern of more, but smaller meals without altering total intake. The results implicate the VMN and PVN of the hypothalamus as sites for Ucn 3-CRF(2) control of food intake.     

Expression and antigenicity of virus-like particles of norovirus and their application for detection of noroviruses in stool samples

Human noroviruses (NoVs), members of the genus Norovirus in the family Caliciviridae, are the leading agents of nonbacterial acute gastroenteritis worldwide. Human NoVs are currently divided into at least two genogroups, genogroup I (GI) and genogroup II (GII), each of which contains at least 14 and 17 genotypes. To explore the genetic and antigenic relationship among NoVs, we expressed the capsid protein of four genetically distinct NoVs, the GI/3 Kashiwa645 virus, the GII/3 Sanbu809 virus, the GII/5 Ichikawa754 virus, and the GII/7 Osaka10-25 virus in baculovirus expression system. An antigen enzyme-linked immunosorbent assay (ELISA) with hyperimmune serum against the four recombinant capsid proteins and characterized previously three capsid proteins derived from GI/1, GI/4, and GII/12 was developed to detect the NoVs antigen in stools. The antigen ELISA was highly specific to the homotypic strains, allowing assignment of a strain to a Norovirus genetic cluster within a genogroup.     

Conformational changes in the antibody constant domains upon hapten-binding

Bacterial proteins A and G (SpA and SpG) are immunoglobulin receptors that can be used as probes for monitoring change in the conformation of heavy chain constant (C(H)) domains. Interaction of anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) antibody (Ab) with SpA and SpG were measured by isothermal titration calorimetry and surface plasmon resonance in order to address the question of whether hapten-binding induces a conformational change in the C(H) domain. The interactions of IgG2a or its enzymatic fragments with SpA were measured in the presence or absence of the hapten. Although binding of Fab and F(ab')2 fragments were not observed to free SpA, they did bind to immobilized SpA. In addition, the association constant (K(a)) for interaction of IgG2a with immobilized SpA was approximately 20-fold higher than that with free SpA. This was explained in terms of high avidity resulting from multivalent interaction between IgG2a and immobilized SpA on the chip. Interestingly, the hapten-binding weakened the interaction between the F(ab')2 fragment and SpA. Furthermore, approximately half of the IgG2a was incapable of binding to immobilized SpA in the presence of hapten. These results were explained using a model which assumed the formation of two kinds of SpA/IgG complexes; one through sites on F(ab')2 arms and the other through sites on the Fc region. The former type dissociated as a result of hapten-binding, as did the F(ab')2 fragment and suggested that a conformational change had occurred around the Fab arms, while the latter type did not dissociate because of the higher avidity of the Fc region. However, using a mutant SpA with a lower K(a) value for the interaction with IgG2a, it was shown that hapten-binding induced long range conformational changes in the Fc region of IgG2a. Similar evidence of conformational change upon hapten-binding was also obtained using SpG as a probe.