Autosomal recessive cystinuria caused by genome-wide paternal uniparental isodisomy in a patient with Beckwith-Wiedemann syndrome.

Approximately 20% of Beckwith-Wiedemann syndrome (BWS) cases are caused by mosaic paternal uniparental disomy of chromosome 11 (pUPD11). Although pUPD11 is usually limited to the short arm of chromosome 11, a small minority of BWS cases show genome-wide mosaic pUPD (GWpUPD). These patients show variable clinical features depending on mosaic ratio, imprinting status of other chromosomes, and paternally inherited recessive mutations. To date, there have been no reports of a mosaic GWpUPD patient with an autosomal recessive disease caused by a paternally inherited recessive mutation. Here, we describe a patient concurrently showing the clinical features of BWS and autosomal recessive cystinuria. Genetic analyses revealed that the patient has mosaic GWpUPD and an inherited paternal homozygous mutation in SLC7A9. This is the first report indicating that a paternally inherited recessive mutation can cause an autosomal recessive disease in cases of GWpUPD mosaicism. Investigation into recessive mutations and the dysregulation of imprinting domains is critical in understanding precise clinical conditions of patients with mosaic GWpUPD.

A novel de novo point mutation of the OCT-binding site in the IGF2/H19-imprinting control region in a Beckwith-Wiedemann syndrome patient.

The IGF2/H19-imprinting control region (ICR1) functions as an insulator to methylation-sensitive binding of CTCF protein, and regulates imprinted expression of IGF2 and H19 in a parental origin-specific manner. ICR1 methylation defects cause abnormal expression of imprinted genes, leading to Beckwith-Wiedemann syndrome (BWS) or Silver-Russell syndrome (SRS). Not only ICR1 microdeletions involving the CTCF-binding site, but also point mutations and a small deletion of the OCT-binding site have been shown to trigger methylation defects in BWS. Here, mutational analysis of ICR1 in 11 BWS and 12 SRS patients with ICR1 methylation defects revealed a novel de novo point mutation of the OCT-binding site on the maternal allele in one BWS patient. In BWS, all reported mutations and the small deletion of the OCT-binding site, including our case, have occurred within repeat A2. These findings indicate that the OCT-binding site is important for maintaining an unmethylated status of maternal ICR1 in early embryogenesis.

Influenza A-associated encephalopathy with bilateral thalamic necrosis in Japan.

Two cases of acute encephalopathy in young children clearly showed evidence of influenza A virus infection and bilateral thalamic lesions. Influenza-associated encephalopathy with bilateral thalamic lesions has mostly been reported in Japan; it differs from Reye's syndrome in several respects. Other factors in addition to influenza virus infection may have contributed to the etiology of encephalopathy in our case patients.

[A case of influenza A virus associated encephalopathy with bilateral thalamic hemorrhage].

A three-year old girl was hospitalized in a semi-conscious state following a febrile convulsion. She did not recover despite treatment and died 16 days after admission. Influenza A virus (H3N2) was detected from a throat swab from the patient, and serum hemagglutinin-inhibiting antibodies to the virus elevated from less than 8 to 256. Brain CT revealed bilateral thalamic hemorrhage and peripheral low density. Subarachnoid hemorrhage was also observed thereafter. Based on clinical manifestations and neuroimaging, this patient was diagnosed as an atypical case of acute necrotizing encephalopathy associated with influenza A virus infection. Such rapid progressive encephalopathies may occur due to intracranial vascular injury including vasculitis or spasms. Although it is clear that influenza A virus triggered this case, we cannot confirm that it was a pathogen. Also, it might be advisable to consider other possible contributing factors such as drugs administered before hospitalization.

[Clinical and bacteriological studies on panipenem/betamipron in pediatrics. Kanagawa Research Group for Infectious Diseases of Children].

Following its introduction into the market, PAPM/BP (panipenem/betamipron) was clinically studied in 188 evaluable cases out of 207 cases primarily of respiratory infectious diseases treated at the pediatric departments of 15 hospitals. In the clinical evaluation, the drug proved effective in three of three cases of sepsis; three of three cases of suppurative meningitis; nine of ten cases of laryngopharyngitis, six of seven cases of tonsillitis, 56 of 63 cases of acute bronchitis, 90 of 98 cases of pneumonia, and one of one case of phyothorax, all of which are respiratory infectious diseases; one of one case of secondary infection of a chronic respiratory disease; and two of two cases of lymphadenitis, which is a disease of the soft dermal structure. The overall efficacy rate was 91.0% (171/188 cases). In the bacteriological study, Gram-positive bacteria were eliminated in five of five strains of S. aureus, 30 of 31 strains of S. pneumoniae (96.8%), and three of three strains of S. pyogenes. Gramnegative bacteria were eliminated in 15 of 17 strains of H. influenzae (88.2%), three of four strains of M. catarrhalis, and two of two strains of K. pneumoniae. The overall elimination rate was 92.1% (70/76 strains). In the 23 strains of S. pneumoniae that were examined, penicillin-resistant strains accounted for 56.5%, showing an elimination rate of 100%. No serious adverse effects were observed, and the incidence of adverse effects was 1.45%. As for abnormalities in laboratory tests, levels of GOT and GPT increased in eight cases (3.88%), LDH increased in one case (0.48%), and neutropenia occurred in one case (0.51%). These results suggest that PAMP/BP could be considered the first choice in the treatment of infectious diseases in pediatrics, due to its effectiveness and high level of safety.

The first Japanese case of Hb Santa Ana, an unstable abnormal hemoglobin, identified rapidly by electrospray ionization mass spectrometry.

Two members of a family had chronic hemolytic anemia due to unstable hemoglobin. The abnormal beta-chain with a molecular weight of 16 u smaller than the normal beta-chain was found within 5 minutes by analysis using electrospray ionization mass spectrometry. The substitution of amino acid was also determined rapidly by this new strategy. The leucine at the 88th position of the normal beta-chain was substituted by proline in the hemoglobin as in Hb Santa Ana. This is the first report of a Japanese case of Hb Santa Ana; the clinical course was similar to that in the previously reported cases.

[Low-dose cytarabine ocfosfate therapy in an elderly acute myelogenous leukemia].

A 71-year-old man was admitted for severe anemia. Bone marrow puncture revealed 48% of blast cells. A diagnosis of acute myelogenous leukemia (AML-M 4) was made. As the patient was old, we administered 300mg of cytarabine ocfosfate (SPAC) for 21 days. Blast cells in bone marrow decreased 5.6%, and SPAC was considered effective. We treated him with the same dose of SPAC for 14 days after a 21-day interval from the end of the first treatment. Although leukemic cells were still seen in bone marrow after two treatments, we considered him in partial remission, and he was discharged. After discharge, the hematological findings remain almost normal with intermittent treatment of 150 mg of SPAC for over one year. Thus, cytarabine ocfosfate might be useful in elderly AML patients.

Bone marrow-derived cultured mast cells and peritoneal mast cells as targets of a growth activity secreted by BALB/3T3 fibroblasts.

When fibroblast cell lines were cultured in contact with bone marrow-derived cultured mast cells (CMC), both NIH/3T3 and BALB/3T3 cell lines supported the proliferation of CMC. In contrast, when contact between fibroblasts and CMC was prohibited by Biopore membranes or soft agar, only BALB/3T3 fibroblasts supported CMC proliferation, suggesting that BALB/3T3 but not NIH/3T3 cells secreted a significant amount of a mast cell growth activity. Moreover, the BALB/3T3-derived growth activity induced the incorporation of [3H]thymidine by CMC and the clonal growth of peritoneal mast cells in methylcellulose. The mast cell growth activity appeared to be different from interleukin 3 (IL-3) and interleukin 4 (IL-4), because mRNAs for these interleukins were not detectable in BALB/3T3 fibroblasts. Although mast cells are genetically deficient in tissues of W/Wv mice, CMC did develop when bone marrow cells of W/Wv mice were cultured with pokeweed mitogen-stimulated spleen cell-conditioned medium. Because BALB/3T3 fibroblast-conditioned medium (BALB-FCM) did not induce the incorporation of [3H]thymidine by W/Wv CMC, the growth activity in BALB-FCM appeared to be a ligand for the receptor encoded by the W (c-kit) locus. Because CMC and peritoneal mast cells are obtained as homogeneous suspensions rather easily, these cells may be potentially useful as targets for the fibroblast-derived mast cell growth activity.

Proliferative potential of murine peritoneal mast cells after degranulation induced by compound 48/80, substance P, tetradecanoylphorbol acetate, or calcium ionophore A23187.

Proliferative potential of degranulated mast cells was investigated. Mast cells were collected from the peritoneal cavity of mice, and degranulation was induced by compound 48/80, substance P, 12-O-tetradecanoylphorbol 13-acetate (TPA), or calcium ionophore A23187. The potentiality of colony formation in methylcellulose was not reduced by treatment of various concentrations of compound 48/80, substance P and TPA. When degranulation was induced by compound 48/80, substance P or TPA, proportion of highly degranulated mast cells containing less than five granules was rather small. In contrast, considerable proportion of highly degranulated mast cells was obtained after the treatment with the low concentration (0.1 microgram/ml) of A23187. These highly degranulated mast cells, which were individually picked up by the micromanipulator, proliferated not only in methylcellulose but also in the skin of mast cell-deficient WBB6F1-W/Wv mice. Inasmuch as we have already shown the proliferation of IgE-sensitized and Ag-stimulated mast cells, degranulated mast cells appear to retain the proliferative potential in general.

An in vitro model of cell migration: evaluation of vascular endothelial cell migration.

In vivo vascular endothelial cell (VEC) migration is thought to play a central role in the development of new capillaries as well as the resurfacing of large vessels. Recently, we have developed an in vitro VEC migration assay system based on the ability of VEC to migrate off of tissue culture microcarrier beads. For these studies, bovine pulmonary artery VEC were grown to confluence on Cytodex 3 microcarrier beads (MCB). Next, the confluent VEC covered microcarrier beads were pipetted into 4-cm2 wells of a tissue culture plate and incubated at 37 degrees C/5% CO2. At various time intervals, the movement of the VEC off of the MCB onto the tissue culture surface was evaluated microscopically. Using this assay, we have studied the effect of endothelial cell growth supplement and various matrices (i.e., fibronectin, gelatin, and Matrigel) on VEC migration. These studies demonstrated that: (i) gelatin had no effect on normal or mitomycin C-pretreated VEC migration; (ii) fibronectin had no effect on normal VEC migration, but stimulated the relative migration of mitomycin pretreated VEC; and (iii) Matrigel significantly suppressed both normal and mitomycin C-pretreated VEC migration. Endothelial cell growth supplement (ECGS) stimulated both normal and mitomycin C-pretreated VEC migration on fibronectin at concentrations of 10 micrograms/ml ECGS. Pretreatment with ECGS had no effect of normal or mitomycin C VEC migration on gelatin. Finally, ECGS stimulated a statistically significant increase in the migration of normal and mitomycin C-pretreated VEC migration on Matrigel.(ABSTRACT TRUNCATED AT 250 WORDS)

Proliferative potential of degranulated murine peritoneal mast cells.

The fate of mast cells after degranulation was investigated. Purified peritoneal mast cells of WBB6F1-+/+ mice were sensitized with monoclonal anti-dinitrophenol (DNP) IgE antibodies and stimulated with DNP conjugated with human serum albumin. Mast cells were vitally stained with neutral red, and highly degranulated mast cells were identified under a phase-contrast microscope and individually picked up with the micromanipulator. When these highly degranulated mast cells were individually plated in methylcellulose, their potential to produce a cluster or a colony was comparable to that of morphologically intact mast cells. Moreover, when highly degranulated mast cells were injected into the skin of genetically mast cell-deficient WBB6F1-W/Wv mice, the proportion of injection sites at which mast cell clusters appeared was comparable to the value observed when morphologically intact mast cells were injected. The present result indicates that proliferative potential of mast cells is not reduced by their degranulation.

Pentoxifylline enhancement of defective neutrophil function and host defense in neonatal mice.

Decreased neutrophil (PMN) chemotaxis is thought to contribute to the increased morbidity and mortality from infection in newborn infants. Pentoxifylline, a methylxanthine, has previously been shown to augment PMN chemotaxis in vitro. The authors therefore investigated the effects of pentoxifylline on 1) in vitro PMN chemotaxis, 2) in vivo leukocyte accumulation, and 3) protection against Staphylococcus aureus infection in newborn mice. Using a modified Boyden chamber system, they demonstrated that pentoxifylline significantly enhanced neonatal PMN chemotaxis in a dose-dependent manner. Additionally, pentoxifylline was found to increase PMN accumulation in vivo in a proteose peptone-induced peritonitis model. Finally, the survival rate in experimentally induced S aureus infection was 51% in neonatal mice given pentoxifylline, compared with 17% in a control (nonpentoxifylline) group (P less than 0.01). These data demonstrate pentoxifylline modulation of PMN migration and enhancement of host defense against bacterial infection.

[A pharmacokinetic study in (mature and premature) neonates treated with amikacin through intravenous drip infusion].

A pharmacokinetic study was conducted in neonates (mature, premature) to which amikacin (AMK) was administered through intravenous drip infusion. The results of the study are summarized below. 1. Changes in blood concentrations of AMK obtained after intravenous drip infusion over a period of 30 or 60 minutes were comparable to those after intramuscular injection. 2. When AMK was administered to neonates (mature, premature) at a single intravenous (30 or 60 minutes) dose of 6 mg/kg, peak levels of 15.5 to 26.3 micrograms/ml were attained. These values were within the range of 15 to 30 micrograms/ml which are considered to be safe and effective peak levels. 3. In 0 day-neonates, half-lives of blood AMK levels rather long and widely varied (3 to 8 hours) but, in about 7 day-neonates, half-lives were 3 to 4 hours. 4. It is considered from the above results that the safe and effective blood concentrations of AMK in 0 to 7 day-old neonates can be obtained from intravenous administrations at each dose of 6 mg/kg repeated with intervals of 12 or 24 hours and, in 8 days or older neonates, from intravenous drip infusions over 30 or 60 minutes at each dose of 6 mg/kg repeated with intervals of 12 hours. 5. For neonates with very low birth weights, individual doses and intervals should be separately investigated.

[Experimental and clinical evaluation of cefpiramide in pediatrics].

Fundamental and clinical studies on cefpiramide (CPM), a new semisynthetic cephalosporin, were made and the following results were obtained. The antibacterial activities of CPM against clinical isolates were almost similar to those of conventional cephems except for Pseudomonas aeruginosa. The antibacterial activity of CPM against P. aeruginosa was excellent and superior than those of the others. Ten or twenty mg/kg of CPM was given intravenously at one shot to 11 cases. The mean serum levels of CPM reached 231 micrograms/ml at 15 minutes, 119 micrograms/ml at 30 minutes, 88 micrograms/ml at 1 hour, 65 micrograms/ml at 2 hours and 33 micrograms/ml at 6 hours after administration at a single dose of 10 mg/kg, respectively with the half-life of 3.42 hours. In case of 20 mg/kg, the mean serum levels attained 306 micrograms/ml at 15 minutes, 245 micrograms/ml at 30 minutes, 160 micrograms/ml at 1 hour, 118 micrograms/ml at 2 hours and 66 micrograms/ml at 6 hours respectively after administration with the half-life of 5.20 hours. CPM was given intravenously to 12 patients with various bacterial infections. The clinical effects were excellent in 5 cases, good in 6 cases and poor in 1 case and the effective rate was 92%. No side effect was observed in all cases.

[Experimental and clinical studies of an ampicillin suppository (KS-R1) in pediatrics].

Blood levels of ampicillin (ABPC) were measured in 10 childish patients with heart disease after the rectal administration of KS-R1 at doses of 125 mg and 250 mg. Average blood levels of ABPC at 15, 30 minutes, 1, 2 hours and 4 hours after the administration of KS-R1 were 6.8, 6.9, 3.1, 1.1 mcg/ml and 0.1 mcg/ml with half-life of 0.64 hours in patients of age from 1 year to 4 years 7 months old (dose level 8.9 approximately 13.9 mg/kg, average 10.5 mg/kg), and 5.2, 6.1, 3.4, 1.0 mcg/ml and 0.1 mcg/ml with half-life of 0.65 hours in patients of age from 7 years 10 months to 10 years 7 months old (dose level 8.3 approximately 13.9 mg/kg, average 9.8 mg/kg), respectively. Clinical effective rate (excellent and good) was 87% in 55 childish patients with infections. Bacteriologically, 13 strains (74%) out of 18 strains which were isolated from the patients were eradicated. No severe side effects were observed. Diarrhea was observed in 3 cases.

[Experimental and clinical evaluation of cefotetan in pediatrics].

Preclinical studies were carried out on cefotetan (CTT), together with clinical studies in the field of pediatrics. The following results were obtained. A total of 114 clinical isolates that have been stored in the authors' department was employed to determine the minimum inhibitory concentrations (MICs) of CTT against various bacterial species. Against E. coli, Salmonella, K. pneumoniae and P. mirabilis, the MICs of CTT showed a peak at 0.78 micrograms/ml, and most of the strains were inhibited by a CTT concentration of 6.25 micrograms/ml or less. The MICs for S. marcescens strains showed a peak at 25 micrograms/ml, with 25% of the strains having MICs of 3.13 micrograms/ml or less, and 67% having MICs of 25 micrograms/ml or more. All of the P. aeruginosa strains had MICs of over 100 micrograms/ml. Against all of the tested strains of S. aureus, a Gram-positive bacterium, CTT showed MICs of 12.5 micrograms/ml or more, while all of the strains of S. faecalis were found to have MICs of over 100 micrograms/ml. CTT was administered intravenously to pediatric patients as a bolus injection, and then the concentration of the antibiotic in the serum was determined as a function of time. When the dosage rate was 10 mg/kg, the mean serum levels were as follows; 58.2 micrograms/ml at 30 minutes, 45.5 micrograms/ml at 1 hour, 33.6 micrograms/ml at 2 hours, 18.0 micrograms/ml at 4 hours and 11.7 micrograms/ml at 6 hours after the injection. The half-life of CTT in the serum at this dosage was thus 2.40 hours. Similarly, at a dosage rate of 20 mg/kg, the mean values at the various times were; 98.6 micrograms/ml at 30 minutes, 75.6 micrograms/ml at 1 hour, 57.8 micrograms/ml at 2 hours, 35.5 micrograms/ml at 4 hours and 23.2 micrograms/ml at 6 hours subsequent to the injection. The half-life of CTT in the serum in these cases was 2.73 hours. CTT was drip-infused intravenously over a period of 1 hour, and then the serum concentration of the drug was monitored with the passage of time. Subsequent to the administration of 10 mg/kg, the mean serum concentrations were as follows; 48.8 micrograms/ml at 30 minutes, 81.5 micrograms/ml at 1 hour, 42.2 micrograms/ml at 2 hours, 23.6 micrograms/ml at 4 hours and 14.8 micrograms/ml at 6 hours subsequent to the injection. The half-life of CTT in the serum after this intravenous drip infusion was thus 2.13 hours.(ABSTRACT TRUNCATED AT 400 WORDS)

[Experimental investigation on various administration methods of gentamicin. Comparison of the effect of gentamicin on phagocytosis].

We compared the efficacy of intravenously, intramuscularly administered or intravenously infused over 45 minutes gentamicin (GM) by the experimental system in rabbits using diffusion chambers. Serum concentrations of GM obtained with 3 administration methods were different, while the peak values after intramuscular injection and intravenous drip infusion were similar. Similar concentration time curves of GM in the chambers were revealed after intramuscular injection and intravenous infusion. No marked difference in 3 administration methods was showed in the effects on the growth of P. aeruginosa in the chamber. Viable bacterial number in the chambers decreased when the GM concentration in the chamber was 4--5 times of MIC, and thereafter regrowth was observed after the decrease of GM concentration in the chambers to 2--3 times of MIC. About 8 hours were required for growth to the base line value. In the leucocyte containing chambers, viable bacterial number similarly decreased, but the rate of regrowth was slow. About 12 hours were required for growth to the base line value. The regrowth rate in the GM containing chamber was similar to that in the antibiotic free chamber. From this result, it is suggested that, when the antibiotic concentration in the chamber is 1--2 times of MIC, antibiotic does not show the growth suppressive effect and the activating effect on phagocytosis of leucocytes in the environment closed to the practical pathological condition such as this model. For the patients with qualitative or quantitative abnormal changes of phagocyte, short interval of drug administration might be needed. For this purpose, intravenous drip infusion under monitoring of serum concentration is more suitable than intramuscular injection, which might be accompanied with severe pain and contracture of injected muscle.

[Pharmacokinetics study on gentamicin intravenous drip infusion in children].

Pharmacokinetics of gentamicin in children after intravenous infusion over 60 minutes were compared with that after intramuscular injection. 1. Mean measured peak serum levels after intravenous infusion of 2.5 mg/kg and intramuscular injection of 2.0 mg/kg were 6.1 micrograms/ml at termination of infusion and 6.5 micrograms/ml at 30 or 60 minutes after injection, respectively. Older children showed higher serum levels. 2. There was no difference in serum half-life between both modes of administration. 3. The AUC after intravenous infusion was slightly larger than that after intramuscular injection. 4. It was suggested that the efficacy and safety of the treatment by intravenous infusion in children are comparable to that by the intramuscular injection, and optimum single dose is 1.5--2.5 mg/kg.