Double-blind placebo-controlled study of loratadine, mequitazine, and placebo in the symptomatic treatment of seasonal allergic rhinitis

Loratadine is a long-acting H1 antagonist devoid of anticholinergic and sedative effects. A double-blind, placebo-controlled, parallel-group study was performed in 69 patients to compare efficacy and safety of loratadine and mequitazine. Patients allergic to grass pollens were randomly assigned to one of the three treatment groups and followed up to 2 weeks during the peak of the pollen season. Symptoms of allergic rhinitis were evaluated at baseline and after 3, 7, and 14 days of treatment by the physician with patients rating their response daily on diary cards. Both loratadine and mequitazine induced a significant relief of nasal symptoms when these were compared to placebo. Loratadine was found to be significantly superior to placebo after 3 days of treatment, whereas a significant improvement was only observed after 7 days in patients treated with mequitazine. For nonnasal symptoms, none of the two anti-H1 antagonist induced a significant improvement, and this lack of effect may be related to low symptoms at baseline. Loratadine did not induce more side effects than placebo. Loratadine can be considered to be an effective and safe anti H1 histamine with a rapid onset of action.     

Polarized ion transport during migration of transformed Madin-Darby canine kidney cells

Epithelial cells lose their usual polarization during carcinogenesis. Although most malignant tumours are of epithelial origin little is known about ion channels in carcinoma cells. Previously, we observed that migration of transformed Madin-Darby canine kidney (MDCK-F) cells depended on oscillating K⁺ channel activity. In the present study we examined whether periodic K⁺ channel activity may cause changes of cell volume, and whether K⁺ channel activity is distributed in a uniform way in MDCK-F cells. After determining the average volume of MDCK-F cells (2013±270 m³; n=8) by means of atomic force microscopy we deduced volume changes by calculating the K⁺ efflux during bursts of K⁺ channel activity. Therefore, we measured the membrane conductance of MDCK-F cells which periodically rose by 22.3±2.5 nS from a resting level of 6.5±1.4 nS (n=12), and we measured the membrane potential which hyperpolarized in parallel from –35.4±1.2 mV to –71.6±1.8 mV (n=11). The distribution of K⁺ channel activity was assessed by locally superfusing the front or rear end of migrating MDCK-F cells with the K⁺ channel blocker charybdotoxin (CTX). Only exposure of the rear end to CTX inhibited migration providing evidence for horizontal polarization of K⁺ channel activity in transformed MDCK-F cells. This is in contrast to the vertical polarization in parent MDCK cells. We propose that the asymmetrical distribution of K⁺ channel activity is a prerequisite for migration of MDCK-F cells.     

Detection of Norwalk Virus and Other Genogroup 1 Human Caliciviruses by a Monoclonal Antibody, RecombinantAntigen-Based Immunoglobulin M Capture Enzyme Immunoassay

Sera obtained from two groups of adult volunteers infected with Norwalk virus (NV) and two groups of patients involved in two natural outbreaks were tested for NV-reactive immunoglobulin M (IgM) by use of a monoclonal antibody, recombinant-antigen-based IgM capture enzyme immunoassay (EIA). No NV-reactive IgM was detected in the preinoculation sera of 15 volunteers, and 14 of 15 showed NV-reactive antibodies postinfection with NV. All of the volunteers showed IgG seroconversion to NV. In the outbreak studies, all 9 persons in one outbreak and 19 of 24 in another outbreak had NV-reactive IgM. In the first outbreak, only three of nine seroconverted to NV, which was likely due to late collection of acute-phase sera. In the second outbreak, 21 of 24 showed IgG seroconversion to NV. Sequencing of viruses isolated from five stool samples selected from those in the second outbreak showed that they were human calicivirus (HuCV) genogroup 1 viruses related, but not identical, to NV. In the volunteer studies, NV-reactive IgM was first detected 8 days postinoculation. The time of development of NV-reactive IgM antibodies in natural outbreaks was estimated to be similar to that found in the volunteer studies. Sera from three Hawaii virus-infected volunteers, four Snow Mountain virus patients, and 80 healthy individuals were negative for NV-reactive IgM, indicating test specificity for HuCV genogroup I infections. This capture IgM EIA is suitable for diagnosis of NV and other HuCV genogroup I infections and is especially useful when sera and fecal samples have not been collected early in the course of an outbreak.     

Measurement of streptococcal cell wall in tissues of rats resistant or susceptible to cell wall-induced chronic erosive arthritis.

The quantity of streptococcal cell wall localized in the joints of rats of strains which are either susceptible (Sprague-Dawley, LEW/N, M520/N) or resistant (Buffalo, WKY/N, F344/N) to cell wall-induced chronic erosive arthritis was measured after intraperitoneal injection of group A streptococcal cell wall fragments. Susceptibility or resistance was not associated with a difference in the amount of cell wall localized in limbs or other tissues. It is concluded that although localization of cell wall in joint tissue is essential for development of arthritis, the relative resistance of certain rat strains reflects genetic regulation of inflammatory response rather than a quantitative difference in localization of cell wall in joints.     

CYP2D6 genotyping strategy based on gene copy number determination by TaqMan real-time PCR

The genetic polymorphism of the cytochrome P450 monooxygenase, CYP2D6, comprises at least 43 alleles giving rise to distinct drug metabolism phenotypes termed ultrarapid, extensive, intermediate, and poor metabolizers. As a consequence, drug side effects or lack of drug effect may occur if standard doses are applied. Genetic prediction of drug oxidation phenotype as a basis for dose selection requires analysis of single nucleotide polymorphisms and of alleles with duplicated or deleted genes. Here we developed a novel method to determine the CYP2D6 gene dose per genome. A TaqMan real-time PCR assay to specifically amplify genomic CYP2D6 was established by using a specific set of amplification primers and probe, located in exon 9, which effectively prevent amplification of CYP2D7 and CYP2D8 pseudogenes. Quantitative CYP2D6 amplification data were normalized to albumin as an internal reference gene which was coamplified simultaneously in a single-tube biplex assay. The assay was validated with a selection of previously genotyped DNA samples containing none, one, two, or three CYP2D6 gene copies. The results were highly reproducible and closely matched the number of genes with no overlap between the groups. Analysis of DNA samples comprising all major alleles and genotypes revealed high sensitivity and specificity of the assay, as demonstrated by agreement of the determined gene dose with the presence of CYP2D6(*)2 x 2 (gene duplication) and CYP2D6(*) 5 (gene deletion) alleles. The predictability of the new strategy was systematically evaluated. The semiautomatic TaqMan assay allows high sample throughput and will be useful for pharmacogenetic studies and in the clinical setting.     

Lymphocyte proliferation induced by autologous cells. XV. Relationships between the human autologous mixed lymphocyte reaction stimulated by non-T and activated T cells

The human {T:T act} AMLR was characterized in its relationship to the {T:Non-T} AMLR and its validity as a nonxenogeneic antigen induced response was extended. Human T cell lines, established from responding T cells in an autologous mixed lymphocyte reaction (MLR), were maintained in medium containing human serum and interleukin-2 (IL-2). These cells stimulated ³H-thymidine incorporation by autologous T cells and by autologous unfractionated blood mononuclear cells. Freshly activated T cells isolated from an autologous MLR stimulated autologous T cells to a lesser extent could be enhanced by adding IL-2. Twenty-five to 50% of T cells stimulated by activated T cells express the T8 determinant. In contrast, we have previously shown that less than 10% of T cells activated after 6 days in culture with non-T cells express the T8 determinant. The number of T8 bearing cells were increased significantly after 10 days in culture with non-T cells. This suggested that two types of reactions, the {T:Non-T} and {T:T act} AMLR, might occur in sequence when T cells and autologous non-T cells are cocultured: first, the activation of T4 cells by non-T cells, then by the activation of T8 cells by activated T4 cells. Finally, activated T cells can stimulate unfractionated autologous mononuclear cells without prior exposure to sheep erythrocytes or fetal calf serum.     

A second locus (GLC3B) for primary congenital glaucoma (Buphthalmos) maps to the 1p36 region

Primary congenital glaucoma (gene symbol: GLC3) is an ocular disorder that occurs for 0.01-0.04% of blind people. In the majority of familial cases reported so far, this condition is inherited as an autosomal recessive trait. We have recently used a group of 17 GLC3 families with a minimum of two affected offspring and consanguinity in most of the parental generation and mapped the first GLC3 locus (GLC3A) to the 2p21 region. Six families did not show any linkage to the GLC3A locus and thus provided evidence for genetic heterogeneity of this disorder. A total of eight families unlinked to the 2p21 region were used to search for the chromosomal location of the second GLC3 locus. Herein, we describe mapping of a new locus (designated GLC3B) for primary congenital glaucoma to the short arm of chromosome 1 (1p36.2-36.1) that is situated centromeric to the neuroblastoma and Charcot-Marie-Tooth type 2A (CMT2A) loci. A total of 17 DNA markers were genotyped from this region of chromosome 1. Four families showed no recombination with the two markers D1S2834 and D1S402 with a maximum lod score of 4.510 and 4.157 respectively. Pairwise and multipoint linkage analysis and inspection of the haplotypes revealed that the remaining four families are not linked to this part of chromosome 1, thus providing further evidence that at least one more locus for the autosomal recessive form of GLC3 must exist in the genome. Based on the recombination events, the overall linkage map of this region is: tel-D1S1192-D1S1635-D1S1193 - (D1S1597/-D1S489/D1S228)- [GLC3B/D1S2834/D1S402] - (D1S1176/D1S507/D1S407) - D1S2728-(MFAP2/D1S170) - D1S1368 - D1S436- D1S1592-cen.      

Striated intramural gallbladder lucencies on US studies: predictors of acute cholecystitis

Ultrasound scans of 51 consecutive patients with gallbladder wall thickening were reviewed, and specific sonographic features were correlated with surgical and clinical follow-up. Two patterns of thickening were identified as specific indicators of the presence or absence of acute cholecystitis. "Striated" wall thickening, consisting of several alternating, irregular, discontinuous, lucent and echogenic bands, was seen in eight of 13 patients (62%) with acute cholecystitis. This pattern was not encountered in any of the patients who did not have acute cholecystitis. Conversely, "three-layer" thickening, consisting of a single circumferential lucent zone between two relatively uniform echogenic layers, was seen in only one of 13 patients (8%) with acute cholecystitis but in 11 of 38 patients (29%) with other diagnoses. Other abnormalities, including the presence of intramural echogenic foci and wall irregularities, were more frequently seen in patients with acute cholecystitis but were not as helpful. Use of these features may suggest or help exclude a diagnosis of acute cholecystitis in those patients in whom the cause of gallbladder wall thickening is otherwise not apparent.     

Emergency department thoracotomy (EDT). A 26-month experience using an "agonal" protocol

EDT can be successfully performed with the proper system in place. This includes an established thoracotomy protocol, a well-integrated EMS system, and an in-house team. Time seems to be critical, and the time between injury and EDT may be the single most important factor affecting survival other than the mechanism of injury. Cardiac penetrations, especially stab wounds, were found to have a 93 per cent survival while subdiaphragmatic penetrations had only one survivor from a group of 18 patients (5.5%). The high rate of salvage in the heart wound group probably reflects the speed of prehospital transport, though all other major series have found this group to gain the maximum benefit. No patient was successfully resuscitated from blunt injury with EDT. Three additional patients had "signs of life" restored (one pediatric blunt; two subdiaphragmatic gunshot wounds) but died of coagulopathies shortly thereafter. The experience with air ambulance patients was far too small to allow any conclusions or observations. It is felt that as the use and application of helicopters to EMS situations becomes widespread, more patients will be arriving at trauma centers with no vital signs and massive blunt injury but only moments from the accident. This special group of "dying" patients will require intense scrutiny and possibly new and inventive approaches for any hopeful salvage. Emergency thoracotomy will, no doubt, have a place as part of this. The development of a simple working protocol is of extreme importance. The protocol should be one that will allow maximum selection of patients who can benefit and elimination of those patients where EDT would be useless. The primary benefactor for EDT remains the patient sustaining a stab wound to the heart who arrives at the center shortly after injury. In other areas, such as abdominal exsanguination or severe blunt injury, further study is needed to determine what factors, prehospital and resuscitative, will improve outcome.