Characteristics of paediatric patients with 2009 pandemic influenza A(H1N1) and severe, oxygen-requiring pneumonia in the Tokyo region, 1 September-31 October 2009.

Few reports describe the features of 2009 pandemic influenza A(H1N1) pneumonia in children. We retrospectively reviewed 21 consecutive children admitted to hospital from September to October 2009 in the Tokyo region. The diagnosis of 2009 pandemic influenza A(H1N1) virus infection was based on positive results of real-time RT-PCR or rapid influenza antigen test. All patients were hospitalised for pneumonia with respiratory failure and severe hypoxia. The median interval from onset of influenza symptoms to admission was 14 hours (range: 5-72 hours) and the median interval from the onset of fever (≥38 degrees C) to hospitalisation was 8.5 hours (range: 0-36 hours). All patients required oxygen inhalation. Four patients required mechanical ventilation. Chest radiography revealed patchy infiltration or atelectasis in all patients. Antiviral agents and antibiotics were administrated to all patients. Antiviral agents were administered to 20 patients within 48 hours of influenza symptom onset. No deaths occurred during the study period. Paediatric patients with this pneumonia showed rapid aggravation of dyspnoea and hypoxia after the onset of influenza symptoms.

[Therapeutic strategies for acute type A aortic dissection].

We herein present the early and mid-term outcomes of therapeutic strategies for acute type A aortic dissection in our department. Subjects were 75 patients who were admitted to our department from January 2001 to October 2006. A total of 33 patients had thrombosed dissection: emergent surgery was indicated for cases in which the maximal ascending aortic diameter was > or =50 mm or when ulcer-like projection (ULP) was observed in the ascending aorta. Only 1 case of rupture-related death was observed, in a patient who had a maximal ascending aortic diameter of 52 mm and refused surgery. Although 8 patients were converted to surgery during the chronic phase, elective surgery was recommended in all cases. Surgery consisted of entry resection using open distal anastomosis under circulatory arrest at a bladder temperature of 25 degrees C, with antegrade cerebral perfusion into the 3 cervical branches of arch aorta based on bilateral axillary artery. In-hospital mortality of the 62 patients who underwent surgery was low (4.8%) and no dissection-related deaths were reported for the midterm outcomes. In addition, a low rate of cerebral infarction was observed among cases who had residual dissection of the brachiocephalic arteries after surgery. These findings demonstrate the validity of the therapeutic strategies in our department.

Phase I trial of nedaplatin and paclitaxel for patients with non-small cell lung cancer.

A phase I study was conducted to evaluate the maximum tolerated dose and feasibility of combination with nedaplatin (NDP) and paclitaxel in patients with non-small cell lung cancer (NSCLC). Fifteen patients under 75 years old, with unresectable NSCLC who had not previously received chemotherapy or radiotherapy, with a performance status of 0-1, were enrolled. The dose escalation levels (NDP/Paclitaxel; mg/m2 day 1) were 80/150 (level 1), 80/180 (level 2), 90/180 (level 3) and repeated every 28 days. All patients receiving level 3 had dose-limiting toxicity. One patient developed grade 4 neutropenia with infection, two had incomplete recovery of neutropenia and thrombocytopenia by the 28th day after the first cycle of chemotherapy. Non-hematologic toxicities, including nephrotoxicity, nausea/vomiting, alopecia, and hypersensitivity reaction, were tolerated. Three of the 15 patients achieved partial responses. We concluded that the recommended dose was paclitaxel 180 and NDP 80 mg/m2 due to the hematologic toxicity.

Diurnal regulation of a DNA binding protein to the period repeat sequence in the SCN nuclear extract of rat brain.

We previously reported the mammalian period repeat mRNA fluctuates during circadian time in the rat suprachiasmatic nucleus (SCN) which is considered to be a clock pacemaker in mammalian brain. Presently we discovered a period repeat sequence (PR) DNA-binding protein in the rat SCN nuclear extract. In the SCN, the binding activity of PR DNA-binding protein to (ACAGGC)3 was most highest during the late day and most lowest during the late night by electro-mobility shift assay (EMSA). In the cortex nuclear extract, the binding of PR DNA-binding protein did not show a significant variation during a day. This is the first report to show the existence of diurnal regulated PR DNA-binding protein in the SCN.

A new method for preparation of an antiserum to penicillin and its application for novel enzyme immunoassay of penicillin.

A new method for the preparation of ampicillin-BSA conjugate by a three step procedure was developed. The first step is the introduction of a maleimide residue to ampicillin by a cross-linking reagent, MBS. The second step is reductive cleavage of disulfide bonds in BSA. The third step is thioether formation between the introduced maleimide residues and the reduced thiol groups. The obtained ampicillin-BSA conjugated raised an anti-ampicillin serum in rabbits. A new reagent, MPGS, was used for enzyme labelling of ampicillin to avoid immunological cross reaction. Using the enzyme labelled ampicillin and anti-ampicillin serum, enzyme immunoassay of ampicillin was successful in detecting 4 ng to 1 mug. Cross reactivities of anti-ampicillin to ampicillin analogs were studied by the enzyme immunoassay to find that the antiserum is specific to penicillin especially to ampicillin but hardly reacts with cephalosporins or the penicilloic acid derivative of ampicillin.

Circadian rhythm abnormalities of wrist activity of institutionalized dependent elderly persons with dementia.

BACKGROUND: The study objective was to clarify the descriptive characteristics of circadian rhythm abnormalities of wrist activity of the institutionalized elderly with dementia. METHODS: We studied 82 elderly persons with dementia who were institutionalized in a long-term medical care facility. The ambulatory continuous monitoring of their wrist activity was conducted for 7 days at 1-minute intervals. The time series data were analyzed using the double-plotted chronogram, spectral analysis was performed using the fast Fourier transformation and periodogram analysis was performed as well. RESULTS: The frequency of circadian rhythm abnormalities of wrist activity rhythm in elderly persons with dementia was 57.3% (47 out of 82). The abnormalities were classified into four categories: severely impaired circadian rhythm type with no boundary between day and night, free-running rhythm type, decreased circadian amplitude type, and accentuation of ultradian rhythm type. CONCLUSION: This four-category classification system provides a scientific approach for studying the mechanisms of circadian activity rhythm abnormalities of elderly persons with dementia.

Glutamate release and neuronal injury after intrathecal injection of local anesthetics.

High concentrations of local anesthetics are neurotoxic, but the mechanism for this neurotoxicity is obscure. Here, we report increased concentrations of glutamate in the cerebrospinal fluid after intrathecal injections of high concentrations of tetracaine (a local anesthetic). The peak concentrations of glutamate after administration of 1%, 2%, and 4% tetracaine were 4-fold, 6-fold, and 10-fold higher than baseline values, respectively. Animals in the 1% group were all neurologically normal one week after tetracaine injection. In the group receiving 4%, no animal was able to hop and vacuolation of the white matter and/or central chromatolysis of the motor neurons were observed. Because high concentrations of glutamate are known to be neurotoxic, our results may provide some insight into the mechanisms for neurotoxicity of intrathecal local anesthetics.

Additive effect of mPer1 and mPer2 antisense oligonucleotides on light-induced phase shift.

It is well known that light induces both mPer1 and mPer2 mRNA in the suprachiasmatic nucleus. We have reported that mPer1 antisense oligonucleotides (ODNs) inhibited the light-induced phase delays of mouse locomotor rhythm. In this study, we asked whether both or either mPer1 or mPer2 expression is necessary to induce the phase shift. We examined the effects of inhibition of mRNA expression on light-induced phase delays of mouse circadian behavior rhythm. Light-induced phase delays were moderately attenuated by microinjection of mPer1 or mPer2 antisense ODN, but not by mPer3 antisense or mPer1, mPer2 scrambled ODNs, whereas following simultaneous injection of both mPer1 and mPer2 antisense ODNs they disappeared. The present results suggest that acute induction of mPer1 and mPer2 gene play an additive effect on photic entrainment.

c-fos CRE-binding activity of CREB/ATF family in the SCN is regulated by light but not a circadian clock.

The DNA-binding activity of cAMP-responsive element binding protein (CREB) in the suprachiasmatic nucleus (SCN) was examined with Ca or cAMP-responsive element (Ca/CRE) in upstream sequence of c-fos gene using electro-mobility shift assay (EMSA). By using supershift assay, Ca/CRE-binding activity in the SCN was shown to contain not only CREB but also activating transcription factor-1 (ATF-1). Furthermore, photic-induction of CREB binding activity to Ca/CRE in the SCN occurred at all circadian times tested, indicating that CREB DNA-binding in the SCN was not gated by the endogenous clock. These results implicated the role of CREB/ATF family in photic neuronal signaling in the SCN.

Isomerization reaction of olefin using RuClH(CO)(PPh(3))(3)

When methyl 5-(tert-butyldiphenylsilyl)oxy-2-pentenoate was refluxed in toluene in the presence of RuClH(CO)(PPh(3))(3) (5 mol %), double-bond migration took place to afford methyl 5-(tert-butyldiphenylsilyl)oxy-4-pentenoate in high yield. This means that the double bond conjugated with the ester moiety migrates to a deconjugated position by a ruthenium catalyst. We planned to prepare an enol ether from alpha,beta-unsaturated compounds having an ether moiety in a tether using ruthenium-catalyzed isomerization of the double bond. As a result, silyl or benzyl enol ether was obtained from the alpha,beta-unsaturated ester having alcohol protected by the silyl or benzyl group in a tether in high yield. In this reaction, double bond migration of alpha,beta-unsaturated ketone and alpha,beta-unsaturated amide took place to produce deconjugated compounds. Moreover, the double bond of alpha, beta-unsaturated ester having a triple or double bond in a molecule migrated to produce conjugated enyne and diene. On the other hand, treatment of a bis-metalated compound having an alpha, beta-unsaturated ester moiety or the double bond in a tether with RuClH(CO)(PPh(3))(3) gave allyl bis-metalated compound in good yield. These compounds are useful units in synthetic organic chemistry.

Examination of DNA-binding activity of neuronal transcription factors by electrophoretical mobility shift assay.

Electrophoretical mobility shift assay (EMSA) is a simple, rapid, and highly sensitive technique for detection of single- or double-stranded DNA-binding proteins such as transcription factors in crude nuclear extracts (F.M. Ausubel, R. Brent, R.E. Kingston, D. D. Moore, J.G. Seidman, J.A. Smith, K. Struhl (Eds.), Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley-Interscience, 1989, pp. 12.0.1-12.2.10 [1]; J. Carey, Gel Retardation. Methods Enzymol., 208 (1991) 103-117 [2]). By using this technique, it is possible to quantify the abundance, relative affinity and binding specificity of DNA-binding proteins. Since proteins which bind specifically to radiolabeled DNA probes retard the mobility of the probe during electrophoresis (it also called gel retardation assay), discrete bands correspond to the individual DNA-protein complexes. Furthermore, EMSA allows one to determine which member(s) of a certain protein family are included in the DNA-protein complex by means of specific antibodies raised against the DNA-binding protein (supershift assay).

Neuroethological and morphological (Neo-Timm staining) correlates of limbic recruitment during the development of audiogenic kindling in seizure susceptible Wistar rats.

Acute audiogenic seizures are a model of generalized tonic-clonic seizures, induced by high intensity acoustic stimulation in genetically susceptible rodents. The neural substrate are sensory motor brainstem nuclei. Recruitment of forebrain structures takes places upon repetition of acoustically evoked seizures. The term audiogenic kindling means forebrain kindling evoked by repeated brainstem seizures and has been described in several strains of genetically epilepsy-prone rats. Thus, the present work was conducted in order to test the hypothesis that audiogenic kindling recruits the forebrain, which may be behaviorally evaluated and associated with morphological changes as well. The behavioral sequences observed during the development of audiogenic kindling were assessed by neuroethological methods (cluster analysis), with the ETHOMATIC program. Seizure severity indexes (brainstem and limbic seizures) and latencies of wild running and tonic-clonic seizures were measured to quantify seizure evolution. Densitometric analysis of Neo-Timm staining was used for assessing morphological changes associated with audiogenic kindling. In group I, II resistant (R) and 16 susceptible (S) animals were stimulated (120 dB) 21 times, and allowed a 10 day recovery period prior to retesting. In group II, 22 R and 20 S were stimulated 60 times, and allowed a 2 month recovery period prior to retesting. Repetition of the acoustic stimulation in group I and group II susceptible animals led to a progressive and statistically significant attenuation of the behaviors associated with brainstem seizures and a concomitant increased expression of the behaviors associated with limbic seizures. After either a 10 day (group I) or 2 month (group II) recovery period, acoustic stimulation preferentially evoked brainstem-associated behaviors and seizures rather than limbic ones in the audiogenic susceptible animals, although in some animals overlapped brainstem and limbic seizures were detected. Latencies for the wild running and tonic seizures after acoustic stimulation significantly increased during audiogenic kindling for both group I and group II susceptible animals. The quantitative ethological evaluation in both group I and group II, illustrated by flowcharts, showed the evolution of the kindling installation by the presence of limbic seizure clusters, competing in time with the original tonic-clonic clusters. Expression of limbic seizures by group I animals, after acoustic stimulation, was not associated with changes in the mossy fiber Neo-Timm staining pattern of these animals. In group II however, Neo-Timm staining revealed mossy fiber sprouting in the ventral hippocampus (but not in the dorsal), and a significant change in the optical density of amygdaloid nuclei and perirhinal cortex in susceptible animals as compared to resistant ones. In conclusion, audiogenic kindling effectively recruits forebrain structures, responsible for the appearance of limbic seizures. It is possible that the paradigm used in group I was subthreshold for the development of clear-cut synaptic reorganization in the hippocampal mossy fiber system, since the behavioral patterns reverted ten days after the last seizure induction. In group II, however, an increased number of evoked seizures and a more prolonged time after the last chronic seizure showed structural re-arrangements in amygdala, perirhinal cortex and hippocampus, associated with permanence in terms of behavioral data (lack of regression of limbic seizures to control values).

Reduced exploratory activity of audiogenic seizures susceptible Wistar rats.

Patients with epilepsy and animals with experimental usually show behavioral changes such as increased anxiety. Audiogenic seizures (AS) are a model of generalized tonic-clonic limbic seizures induced by sound stimulation in genetically susceptible animals. The objective of this paper was to evaluate the exploratory activity of an inbred strain derived from Wistar progenitors that has been selected in our laboratory for AS susceptibility. The exploratory activity of audiogenic seizures susceptible (S) and resistant (R) Wistar rats was measured in two situations: an open arena and the elevated plus maze, an animal model of anxiety. S animals displayed a reduced exploration in both the open arena (reduced total distance moved) and the elevated plus maze (reduced number of enclosed-arm entries). In the latter there was also a decrease in open-arm exploration, particularly of the distal part of these arms. This effect persists even when the effect of a decreased number of enclosed-arm entries is removed by analysis of covariance. Therefore, the results indicate that audiogenic seizure genetically susceptible Wistar rats display a reduced exploration of novel environments. Moreover, the results with the elevated plus maze suggest that these animals are more anxious than AS resistant rats.

A multicentre randomized controlled trial of recombinant interferon-alpha-2a in the treatment of patients with chronic hepatitis C.

Sixty-one chronic hepatitis C patients were randomly assigned to receive either 6 x 10(6) or 9 x 10(6) U of recombinant interferon-alpha-2a (IFN alpha-2a) six days a week for the first two weeks of treatment, followed in both cases by 6 x 10(6) U three days a week for the next 22 weeks. In the low dose group, 11 patients showed a complete response maintained for at least six months, 12 responded but then relapsed and nine did not respond; the corresponding figures in the high dose group were 10, 15 and five patients, respectively. The differences between groups are not statistically significant. Thus, this study provides no evidence of therapeutic benefit from increasing the initial dose of IFN alpha-2a. In both treatment groups, complete responders had significantly lower pretreatment viral titres than nonresponders and were significantly more likely to be infected by type 2a versus type 1b virus.

Parathyroid scintigraphy with 99mTc-MIBI and 123I subtraction: a comparison with magnetic resonance imaging and ultrasonography.

The aim of this study was to evaluate the efficacy of 99mTc-MIBI and 123I subtraction scintigraphy for the detection of abnormal parathyroid glands to be referred for surgical treatment. Thirty-nine consecutive patients, including 35 primary and four secondary cases of hyperparathyroidism, were evaluated. 99mTc-MIBI/123I subtraction scintigraphy (MIBI/I) was performed on all patients, and the results were compared with delayed images of 99mTc-MIBI (D-MIBI), magnetic resonance imaging (MRI) and ultrasonography (US). The overall sensitivity of MIBI/I, MRI, US and D-MIBI was 55.9%, 43.4%, 50.8% and 39.0%, respectively. In cases of single-gland disease, the sensitivity of MIBI/I, MRI, US and D-MIBI was 62.1%, 48.3%, 55.2% and 44.8%, respectively. In cases of multi-gland disease, the sensitivity of MIBI/I, MRI, US and D-MIBI was 50.0%, 37.5%, 46.7% and 36.7%, respectively. In cases of parathyroid adenoma, the sensitivity of MIBI/I, MRI, US and D-MIBI was 71.4%, 50.0%, 71.4% and 50.0%, respectively. In cases of parathyroid hyperplasia, the sensitivity of MIBI/I, MRI, US and D-MIBI was 55.2%, 42.3%, 50.0% and 39.7%, respectively. It is concluded that 99mTc-MIBI/123I subtraction is more useful than the delayed imaging of 99mTc-MIBI, MRI and US.

[Microbiological determination of netilmicin using a thin-layer chromatography scanner].

We presented a new colorimetric bioassay of aminoglycoside antibiotics, which were represented by netilmicin (NTL) in this study, based on the discoloration of thymolphthalein (TP) in paper (indicator-disc) by carbon dioxide produced by Bacillus subtilis. To evaluate the amount of the carbon dioxide, the following experiment was carried out. One milliliter of B. subtilis suspension containing 4.5 x 10(7) colony forming units/ml, 1 ml of nutrient broth, 0.9 ml of 0.1 M phosphate buffer (pH 8.0) and 0.1 ml NTL sample solution were added to an incubation container, which was then placed in a water-bath (37 degrees C) for 3 hours. The oxygen concentration in the head space of flask was determined using gas-chromatograph. The dose-response curves showed good correlation between amounts of NTL and carbon dioxide produced by B. subtilis. The indicator-disc containing TP and sodium hydroxide was placed into the Reacti-flask and then incubated in the same manner as described above. After incubation, concentration of blue colored TP was determined using a TLC scanner. The discoloration of blue color to white showed the proportionality between NTL concentrations and the degrees of discoloration of TP. The method can accurately measure NTL levels down to 2.5 micrograms/ml in water using 0.1 ml samples, and should be adequate for rapid bioassay.

[Studies on the erythrocyte membrane skeleton in a patient with chorea-acanthocytosis--theoretical speculation on the mechanism of neurological involvement].

Patients with chorea-acanthocytosis exhibit symptoms of self-biting, choreic movement, and acanthocytosis, but not dementia. The mechanism of choreic movements is still unknown. In order to clarify the etiologic mechanism underlying these movements, we evaluated the erythrocyte membrane in one patient with chorea-acanthocytosis. A 35-year-old female was admitted to Saitama Medical School Hospital because of involuntary movements. She was alert, well-oriented, and had no gross memory defects. She had slurred speech, choreic movements and lip biting. Laboratory examination showed acanthocytes in her peripheral red blood cells, normal serum lipid values, and caudate atrophy on her brain CT scan. In analyzing the acanthocytes, we initially evaluated the size of the acanthocyte population by incubating her red blood cells with plasma. The cell population approximately doubled after 2 hours incubation. Next we examined the protein composition of erythrocyte ghost by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). There was no significant difference between the patient's erythrocyte ghosts and those of a control. Then we investigated morphological changes in the patient's erythrocyte by scanning and transmission electron microscopy (SEM and TEM). SEM showed the typical acanthocyte shape. The quick-freeze, freeze-substitution method confirmed that the routine TEM section was not artifactual, and was in fact in accurate reflection of the actual features of acanthocytes. TEM of the sections prepared from erythrocyte ghosts demonstrated that spectrin tended to be accumulated in the thorn region. Furthermore, TEM of quick-freeze, deep-etched replica of the ghost revealed more clearly a spectrin network densely packed on the inner hydrophilic surface.(ABSTRACT TRUNCATED AT 250 WORDS)

[Perioperative management of endovascular stent graft placement for abdominal aortic aneurysm].

We retrospectively examined the changes in hemodynamics, oxygen index and renal function along with the complications in 25 patients who had undergone endovascular stent graft placement (ESG) surgery for abdominal aortic aneurysm. During stent graft placement, mean arterial pressure decreased to 58 +/- 8 mmHg by increasing the dose of anesthetics and/or using vasodilators. Except for this intended hypotensive period, mean arterial pressure and heart rate were relatively stable and adequately maintained during surgical manipulation. Oxygenation index was well maintained. A patient with a high preoperative creatinine level underwent prophylactic hemodialysis postoperatively. In other patients except one who died in early postoperative period, both BUN and creatinine levels were kept within normal ranges. Four patients died postoperatively and the causes of the death in two patients are related to the surgical procedure; one with multiple emboli possibly due to released atheloma from the aortic wall during procedure, the other with sepsis due to infected stent graft. Although ESG is a well tolerated procedure, embolism is the most serious complication. Careful preoperative evaluation of the ascending arch and descending aortic wall and monitoring with transcranial doppler are necessary.

[Quantitative X-ray computed tomography].

Quantitative X-ray Computed Tomography (QCT) has an advantage to be able to determine three-dimensional bone density of lumbar vertebral body compared with dual energy X-ray absorptiometry (DXA) method. We introduced the high reproductable QCT method as a simplified manual operation avoiding simultaneous imaging of both patients and bone mass phantoms. The coefficient of variation of values measured by phantom study was 0.6% for 7 years in our hospital. In measuring of 144 healthy Japanese women, the bone mineral density of lumbar spine was most likely to be decreased linealy with age.

Microbial contamination of a disinfectant-soaked unwoven cleaning cloth.

In December 2009, a 76-year-old male patient developed pneumonia due to Burkholderia cepacia whilst in an intensive care unit at a Japanese university hospital. During the subsequent environmental investigation to find the source, B. cepacia with an identical DNA type was found in his denture storage solution. Open packets of unwoven rayon cloths soaked in 0.2% alkyldiaminoethylglycine hydrochloride, used for environmental cleaning, were shown to be contaminated with B. cepacia, Alcaligenes xylosoxidans, Pseudomonas fluorescens and Pseudomonas aeruginosa. B. cepacia of a different DNA type was found in five of 42 samples from sealed packets of cloths.