Homogenous,real-time duplex loop-mediated isothermal amplification using a single fluorophore-labeled primer and an intercalator dye: Its application to the simultaneous detection of Shiga toxin genes 1 and 2 in Shiga toxigenic Escherichia coli isolates

We developed a completely homogeneous duplex loop-mediated isothermal amplification (LAMP) method. The present LAMP method employed a combination of a 6-carboxyfluorescein (FAM)-labeled primer (donor) for one target gene, a non-labeled primer for the other, and an intercalator ethidium bromide (EtBr) dye (acceptor) on the basis of fluorescence resonance energy transfer (FRET) between the FAM donor and EtBr acceptor. Measuring changes in fluorescence of FAM enabled the LAMP method to detect two different genes simultaneously. This method was used to detect Shiga toxin genes in Shiga toxigenic Escherichia coli isolates, demonstrating simultaneous detection of two different genes with rapidity and accuracy.     

Surgically-induced astigmatism following single-site phacotrabeculectomy,phacotrabeculotomy and advanced non-penetrating phacotrabeculectomy

We study surgically-induced astigmatism following three kinds of glaucoma surgeries combined with cataract surgery with 6 months of follow-up. The mean surgically-induced astigmatism at 6 months postoperatively was 1.92 ± 1.87 diopters in phacotrabeculectomy (n = 45), 0.72 ± 0.40 diopters in advanced non-penetrating phacotrabeculectomy (n = 40) and 0.76 ± 0.47 diopters in phacotrabeculotomy (n = 49), which appeared to be stabilized by 3 months. Each group revealed a tendency of “with the wound” induced astigmatism. Phacotrabeculectomy could induce greater surgically-induced astigmatism than advanced non-penetrating phacotrabeculectomy and phacotrabeculotomy.     

Fixation behavior in advanced stage glaucoma assessed by the MicroPerimeter MP-1

Purpose  

To investigate fixation behavior in eyes with advanced glaucoma using the MicroPerimeter MP-1.     

Management of neonatal cow's milk allergy in high-risk neonates

Background:  We conducted a multicenter clinical survey to clarify the current attitudes to diagnosis or treatment of neonatal milk allergy (NMA) in institutions providing medical care for high-risk neonates in Japan.
Methods:  Questionnaires were distributed to 263 institutions that provide medical care for high-risk infants. Information was requested on the number of hospitalized neonates between January 2004 and December 2005, the number of neonates diagnosed with milk allergy, frequent clinical symptoms, and clinical tests performed routinely when NMA was suspected.
Results:  Responses were received from 145 institutions (55.1%). Of 69 796 hospitalized neonates, a diagnosis of cow's milk allergy was made in 0.21%. The incidence in infants with birthweight <1000 g was 0.35%. Gastrointestinal symptoms were identified as the most frequent symptoms that suggested NMA by 80% of institutions. A challenge test in each suspected case was performed in only 15% of institutions, even though it was considered to be the most significant test for diagnosis. Most institutions considered a specific immunoglobulin E test in cases of suspected NMA, but only one-third agreed on its diagnostic significance. A lymphocyte stimulation test was performed in only 5.5% of institutions.
Conclusions:  This study is the first to show the incidence of NMA in institutions providing medical care for high-risk neonates in Japan. Current clinical tests may be insufficient for diagnosis of NMA in which non-immunoglobulin-E-mediated delayed allergic reactions are involved. Therefore, awareness of the clinical features of this disorder is required among neonatologists and allergists.     

Activation of c-Jun N-Terminal Kinase Cascades Is Involved in Part of the Neuronal Degeneration Induced by Trimethyltin in Cortical Neurons of Mice

The organotin trimethyltin (TMT) is known to cause neuronal degeneration in the central nervous system. A systemic injection of TMT produced neuronal damage in the cerebral frontal cortex of mice. To elucidate the mechanism(s) underlying the toxicity of TMT toward neurons, we prepared primary cultures of neurons from the cerebral cortex of mouse embryos for use in this study. Microscopic observations revealed that a continuous exposure to TMT produced neuronal damage with nuclear condensation in an incubation time–dependent manner up to 48 h. The neuronal damage induced by TMT was not blocked by N-methyl-D-aspartate receptor channel–blocker MK-801. The exposure to TMT produced an elevation of the phosphorylation level of c-Jun N-terminal kinase (JNK)^p46, but not JNK^p54, prior to neuronal death. Under the same conditions, a significant elevation was seen in the phosphorylation level of stress-activated protein kinase 1, which activates JNKs. Furthermore, TMT enhanced the expression and phosphorylation of c-Jun during a continuous exposure. The JNK inhibitor SP600125 was effective in significantly but only partially attenuating the TMT-induced nuclear condensation and accumulation of lactate dehydrogenase in the culture medium. Taken together, our data suggest that the neuronal damage induced by TMT was independent of excitotoxicity but that at least some of it was dependent on the JNK cascades in primary cultures of cortical neurons.     

Preoperative peripheral na?ve / memory ratio and prognosis of nonsmall-cell lung cancer patients

PURPOSE: This study focused on the relationship between preoperative peripheral blood CD4(+) na?ve/memory ratio and prognosis following surgery for patients with nonsmall-cell lung cancer. METHODS: After measuring CD3, CD4, CD8, CD45RA, CD45RO, CD25, and intracellular CTLA4 levels, CD4(+) na?ve/memory ratios were determined for 13 controls and 38 patients. RESULTS: Although we observed no significant difference in the ratios, the patients exhibited a wider range of values than the controls. Of the 38 patients, 24 subsequently underwent surgery and were divided into three groups based on their preoperative CD4(+) na?ve/memory ratio: Group I: <0.3; Group II: 0.3-0.8; and Group III: >0.8. Survival times were then evaluated. Group II survival was significantly better than Groups I and III. CONCLUSION: Our data suggest that the preoperative CD4(+) na?ve/memory ratio may serve as a prognostic factor for nonsmall-cell lung cancer patients.     

Surgical approaches to pineal region tumors

Direct surgical resection of pineal region tumors has become safer, more effective, and now plays an essential role in their management. Tissue diagnosis allows for the initiation of appropriate therapies and resection can be curative or improve the efficacy of adjuvant therapies. Several approaches have been reported. Based on our operative experience with 57 patients over a 20-year period, we conclude that the Infratentorial Supracerebellar and Parieto-Occipital Paramedian Transtentorial approaches provide excellent exposure while allowing minimally invasive, relatively low risk access to the majority of pineal region tumors. Indications, positioning, techniques, advantages, and disadvantages are discussed. A review of other approaches, pertinent historical remarks, and a discussion of the role of surgery in the contemporary management of pineal region tumors are presented.