Quantitative assays of pharmacologic aerosols used in studying asthma

To more nearly accurately quantitate the dose of pharmacologic agents delivered to human and animal airways via aerosols, we have developed a monodisperse aerosol containing either methacholine or histamine that permits a light scattering device (tyndallometry) to measure accurately the quantity of inspired and expired particles. These aerosols (described in previous studies) are simultaneously tagged with a radioactive label (technetium 99m) to permit the use of external gamma camera imaging. Present work focuses on the development of assay techniques to measure the quantity of methacholine delivered in these aerosols. The lack of specific radioimmune or radioenzyme assays coupled with the cross-reaction of organic contaminants with conventional chemical reagents for measuring methacholine required the development of separative techniques to isolate the methacholine from the organic aerosol contaminants. With aqueous extraction and column separation we have been able to completely isolate the methacholine from these contaminants. This allows the application of standard spectrophotometric assays for methacholine to quantitate the methacholine in the resulting solution. These separative techniques will permit the use of these aerosols in quantitative studies of airway reactivity.     

2-Mercapto-ethanol enhancement of agglutination reaction--a possible in vitro serological correlate for assessment of functional immunity in simian malaria

Conventional indirect haemagglutination test was performed in rhesus monkey sera (collected from Plasmodium knowlesi infected animals) with and without prior treatment of sera with 2-mercapto-ethanol (2-ME). Surprisingly, many sera samples showed significant enhancement of final titre with 2-ME. The 2-ME enhancement effect was more pronounced in the sera of hyperimmune monkeys on further injection of antigen or parasites. It was also noticeable in the sera during primary drug-suppressed P. knowlesi infection and appeared to have a bearing on the immune status of the animals to rechallenge. The use of a soluble antigen prepared from P. knowlesi infected erythrocytes was found to be essential in IHA test to demonstrate the 2-ME enhancement effect. Antigen prepared from freed parasites (commonly used) failed to show a similar effect in IHA. The possible role of certain T-lymphocyte products - antigen binding, non-agglutinating, 2-ME sensitive molecules - in malarial immunology has been proposed.     

Absence of chromosome heterogeneity between classical Fanconi's anemia and the Estren-Dameshek type

Chromosome investigations were carried out in 7 patients with Fanconi's anemia, type Estren-Dameshek. The frequency and types of chromosome instability found in cultured lymphocytes were in accord with those detected in individuals with classical Fanconi's anemia. The break-point distribution indicates a significant excess of breaks in chromosomes No. 1, 2, and 7 and a deficit in No. 18 and X and Y chromosomes. There was a clear clustering of breaks at certain locations in chromosomes No. 1, 2, 3, 7, 9, and 14. The location of the breaks with respect to the bands demonstrated an almost exclusive involvement of the lighter bands, regardless of the banding method used. These results suggest that most breaks take place in the interbands between the G and R bands. In all patients, chromosome instability was less frequent in direct bone marrow preparations than in lymphocyte cultures. However, cultured bone marrow cells showed a significant increase of chromosome aberrations. On the whole, the chromosome data derived from this series of patients are in agreement with those obtained in individuals with classical Fanconi's anemia and give no support to the idea of cytogenetic heterogeneity between subjects affected by these two forms of childhood aplastic anemia.     

Neurological aspects of del(1q) syndrome

We have studied three children with de novo terminal deletion of the long arm of chromosome 1 (46,XX,del(1)(q43)). They all have minor anomalies and neurological signs (severe psychomotor developmental delay, generalized hypotonia, and seizures) that have been described previously. In addition, all of these three patients have autistic-like behavior. They avoid eye contact, show no interest in people, express little emotion, and repeat stereotypic movements such as head nodding and purposeless finger manipulation. They also spend excessive time in making unusual sounds consisting of a high-pitched shrill cry with little intonation in infancy and a harsh, strained, and glottal stridency in later life. They make no labial, lingual, or nasal sounds. We suggest that these observations may be unique clinical manifestations of certain terminal 1q deletions.     

三维超声测胎儿股容积估计胎重的价值

目的通过对57例妊娠分别使用二维、三维超声测量相关参数估计胎重，并作统计学比较，从而评价三维超声对于估测胎儿体重、诊断胎儿发育迟缓及巨大胎的优势和价值。方法对2005年10月至2006年3月间在我院分娩的57例妊娠，研究组用三维超声测胎儿股容积估计胎重，对照组用二维超声同步测量相关参数估计胎重。结果对两组估重结果与胎儿实际出生体重作相关统计学分析，研究组相关系数为0．983，对照组为0．906（P〈0．05），前者与实际体重关系更密切。结论三维超声测量胎儿股容积估测胎儿体重，较传统二维超声具有更大优势，对于发现胎儿发育迟缓（FGR）及巨大胎意义重大，有利于指导临床冶疗、选择分娩方式。     

经舟骨月骨周围脱位的诊断和治疗

目的 探讨经舟骨月骨周围脱位的诊断和治疗。方法 回顾性分析17例经舟骨月骨周围脱位病例，背侧型13例，掌侧型4例；新鲜损伤组10例中8例行撬拨复位成功，仅2例因舟骨对位不良接受手术治疗；陈旧性组7例中均接受手术，4例行开放复位内固定，3例行近排腕骨切除术。结果 采用Cooney评价标准，优11例，良3例，可1例，差2例，优良率82．35％。结论 经舟骨月骨周围脱位的早期治疗简单且疗效较好，而一旦误诊，则多数需手术治疗，且残留不同程度的功能障碍。提高对本病损伤机制、腕部体征和影像学特点的认识和了解，减少误诊。是提高本病预后的关键因素之一。     

Day-night differences in estimated rates of 5-hydroxytryptamine turnover in the rat pineal gland

Summary Rates of 5-hydroxytryptamine synthesis in various brain tissues can be estimated from the linear increase in 5-hydroxytryptophan levels following inhibition of 5-hydroxytryptophan decarboxylation with RO4-4602 or NSD-1015. In addition, NSD-1015 can prevent 5-hydroxytryptamine oxidative-deamination via monoamine oxidase inhibition, leading to linear decreases in a major metabolite of this amine, 5-hydroxyindole acetic acid. In the rat pineal gland we demonstrated similar increases in 5-hydroxytryptophan levels after nocturnal or diurnal injection of RO4-4602 (100 mg · kg^–1) or NSD-1015 (200 mg · kg^–1). Similar decreases in 5-hydroxyindole acetic acid were also observed after nocturnal or diurnal injection of NSD-1015 or pargyline (an inhibitor of monoamine oxidase) (75 mg · kg^–1). 5-Hydroxytryptamine levels increased after nocturnal pargyline injection but remained constant after diurnal pargyline administration. 5-Hydroxytryptamine levels exhibited little change following nocturnal injection of NSD-1015 but decreased linearly after diurnal injection of NSD-1015. We suggest that (1) rat pineal 5-hydroxytryptamine synthesis is increased nocturnally, (2) metabolic utilization, primarily by oxidative-deamination, of 5-hydroxytryptamine is increased diurnally and (3) basal levels of pineal 5-hydroxytryptamine may be stored within adrenergic nerve endings which innervate the pinealocytes responsible for synthesizing this amine, thus protecting or otherwise making unavailable this pool of 5-hydroxytryptamine for metabolic utilization.     

NMDA receptor mediated dendritic plasticity in cortical cultures after oxygen-glucose deprivation

Dendrites and spines undergo dynamic changes in physiological and pathological conditions. Dendritic outgrowth has been observed in surviving neurons months after ischemia, which is associated with the functional compensation. It remains unclear how dendrites in surviving neurons are altered shortly after ischemia, which might reveal the mechanisms underlying neuronal survival. Using primary cortical cultures, we monitored the dendritic changes in individual neurons after oxygen-glucose deprivation (OGD). Two to four hours of OGD induced approximately 30–50% cell death in 24 h. However, the total dendritic length in surviving neurons was significantly increased after OGD with a peak at 6 h after re-oxygenation. The increase of dendritic length after OGD was mainly due to the sprouting rather than the extension of the dendrites. The dendritic outgrowth after 2 h of OGD was greater than that after 4 h of OGD. Application of NMDA receptor blocker MK-801 abolished OGD-induced dendritic outgrowth, whereas application of AMPA receptor antagonist CNQX had no significant effects. These results demonstrate a NMDA receptor-dependent dendritic plasticity shortly after OGD, which provides insights into the early response of surviving neurons after ischemia.     

Cell lines that support replication of a novel herpes simplex virus 1 UL31 deletion mutant can properly target UL34 protein to the nuclear rim in the absence of UL31

Previous results indicated that the herpes simplex virus 1 (HSV-1) U(L)31 gene is necessary and sufficient for localization of the U(L)34 protein exclusively to the nuclear membrane of infected Hep2 cells. In the current studies, a bacterial artificial chromosome containing the entire HSV-1 strain F genome was used to construct a recombinant viral genome in which a gene encoding kanamycin resistance was inserted in place of 262 codons of the 306 codon U(L)31 open reading frame. The deletion virus produced virus titers approximately 10- to 50-fold lower in rabbit skin cells, more than 2000-fold lower in Vero cells, and more than 1500-fold lower in CV1 cells, compared to a virus bearing a restored U(L)31 gene. The replication of the U(L)31 deletion virus was restored on U(L)31-complementing cell lines derived either from rabbit skin cells or CV1 cells. Confocal microscopy indicated that the majority of U(L)34 protein localized aberrantly in the cytoplasm and nucleoplasm of Vero cells and CV1 cells, whereas U(L)34 protein localized at the nuclear membrane in rabbit skin cells, and U(L)31 complementing CV1 cells infected with the U(L)31 deletion virus. We conclude that rabbit skin cells encode a function that allows proper localization of U(L)34 protein to the nuclear membrane. We speculate that this function partially complements that of U(L)31 and may explain why U(L)31 is less critical for replication in rabbit skin cells as opposed to Vero and CV1 cells.