Wall shear stress distribution inside growing cerebral aneurysm.

BACKGROUND AND PURPOSE: Hemodynamic stimulation has been suggested to affect the growth of cerebral aneurysms. The present study examined the effects of intra-aneurysmal hemodynamics on aneurysm growth. MATERIALS AND METHODS: Velocity profiles were measured for 2 cases of AcomA aneurysms. Realistically shaped models of these aneurysms were constructed, based on CT angiograms. Flow fields and WSS in the models were measured by using particle image velocimetry and LDV. In 1 case, hemodynamic changes were observed in 4 stages of growth over a 27-month period, whereas no development was observed in the other case. RESULTS: The growing model had a smaller and more stagnant recirculation area than that in the nongrowth model. The WSS was markedly reduced in the enlarging region in the growing models, whereas extremely low WSS was not found in the nongrowth model. In addition, a higher WSSG was consistently observed adjacent to the enlarging region during aneurysm growth. CONCLUSIONS: The results indicated that the flow structure of recirculation itself does not necessarily lead to high likelihood of cerebral aneurysm. However, WSSG and WSS were distinctly different between the 2 cases. Higher WSSG was found surrounding the growing region, and extremely low WSS was found at the growing region of the growing cerebral aneurysm.

Regulation of Th17 cell differentiation by intestinal commensal bacteria.

Interleukin (IL)-17-producing CD4 T cells ('Th17 cells') are most abundant at the intestinal mucosa, and play a critical role in the maintenance of mucosal barrier function. Recent studies indicate that accumulation of intestinal Th17 cells depends on stimulation by intestinal commensal bacteria - particularly by segmented filamentous bacterium. In this review, we summarise recent advances in our understanding of the mechanisms of intestinal Th17 synthesis in mice, and discuss their relevance to infectious and inflammatory diseases.

Gene arrangement at the Rhesus blood group locus of chimpanzees detected by fiber-FISH.

The Rhesus (Rh) blood group system in humans is encoded by two genes with high sequence homology. These two genes, namely, RHCE and RHD, have been implied to be duplicated during evolution. However, the genomic organization of Rh genes in chimpanzees and other nonhuman primates has not been precisely studied. We analyzed the arrangement of the Rh genes of chimpanzees (Pan troglodytes) by two-color fluorescence in situ hybridization on chromatin DNA fibers (fiber-FISH) using two genomic DNA probes that respectively contain introns 3 and 7 of human RH genes. Among the five chimpanzees studied, three were found to be homozygous for the two-Rh-gene type, in an arrangement of Rh (5'-->3') - Rh (3'<--5'). Although a similar gene arrangement can be detected in the RH gene locus of typical Rh-positive humans, the distance between the two genes in chimpanzees was about 50 kb longer than that in humans. The remaining two chimpanzees were homozygous for a four-Rh-gene type, in an arrangement of Rh (5'-->3') - Rh (3'<--5') - Rh (3'<--5') - Rh (3'<--5') within a region spanning about 300 kb. This four-Rh-gene type has not been detected in humans. Further analysis of other great apes showed different gene arrangements: a bonobo was homozygous for the three-Rh-gene type; a gorilla was heterozygous for the one-Rh- and two-Rh-gene types; an orangutan was homozygous for the one-Rh-gene type. Our findings on the intra- and interspecific genomic variations in the Rh gene locus in Hominoids would shed further light on reconstructing the genomic pathways of Rh gene duplication during evolution.

New chimpanzee MHC class I alleles cloned by polymerase chain reaction.

We describe polymerase chain reaction (PCR) cloning of full-length Patr-A, -B and -Cw locus alleles from a chimpanzee using one sense/antisense primer pair. Of the six alleles cloned here, two have not previously been described. Comparison of nucleotide sequences between the two novel Patr alleles and various HLA alleles suggests that the new Patr-A allele belongs to the A1/A3/A11 family, whereas the new Patr-Cw allele is not closely related to any known HLA-Cw allele.

A Novel MAPK phosphatase MKP-7 acts preferentially on JNK/SAPK and p38 alpha and beta MAPKs.

Mitogen-activated protein kinases (MAPKs) are inactivated via dephosphorylation of either the threonine or tyrosine residue or both in the P-loop catalyzed by protein phosphatases which include serine/threonine phosphatases, tyrosine phosphatases, and dual specificity phosphatases. Nine members of the dual specificity phosphatases specific for MAPKs, termed MKPs, have been reported. Each member has its own substrate specificity, tissue distribution, and subcellular localization. In this study, we have cloned and characterized a novel MKP, designated MKP-7. MKP-7 is most similar to hVH5, a member of previously known MKPs, in the primary structure. MKP-7 is predominantly localized in the cytoplasm when expressed in cultured cells, whereas hVH5 is both in the nucleus and the cytoplasm. MKP-7 binds to and inactivates p38 MAPK and JNK/SAPK, but not ERK. Furthermore, we have found that MKPs have the substrate specificity toward the isoforms of the p38 family (alpha, beta, gamma, and delta). MKP-7 binds to and inactivates p38 alpha and -beta, but not gamma or delta. MKP-5 and CL100/MKP-1 also bind to p38 alpha and -beta, but not gamma or delta. Finally, we propose a tentative classification of MKPs based on the sequence characteristics of their MAPK-docking site.

Identification of a docking groove on ERK and p38 MAP kinases that regulates the specificity of docking interactions.

MAP kinases (MAPKs) form a complex with MAPK kinases (MAPKKs), MAPK-specific phosphatases (MKPs) and various targets including MAPKAPKs. These docking interactions contribute to regulation of the specificity and efficiency of the enzymatic reactions. We have previously identified a docking site on MAPKs, termed the CD (common docking) domain, which is utilized commonly for docking interactions with MAPKKs, MKPs and MAPKAPKs. However, the CD domain alone does not determine the docking specificity. Here we have identified a novel site on p38 and ERK2 MAPKs that regulates the docking specificity towards MAPKAPKs. Remarkably, exchange of two amino acids in this site of ERK2 for corresponding residues of p38 converted the docking specificity for MAPKAPK-3/3pk, which is a dominant target of p38, from the ERK2 type to the p38 type, and vice versa. Furthermore, our detailed analyses with a number of MAPKAPKs and MKPs suggest that a groove in the steric structure of MAPKs, which comprises the CD domain and the site identified here, serves as a common docking region for various MAPK-interacting molecules.

Mitochondrial 16S rRNA sequence diversity of hominoids.

We determined nucleotide sequences of the 16S rRNA gene of mitochondrial DNA (mtDNA) (about 1.6 kb) for 35 chimpanzee, 13 bonobo, 10 gorilla, 16 orangutan, and 23 gibbon individuals. We compared those data with published sequences and estimated nucleotide diversity for each species. All the ape species showed higher diversity than human. We also constructed phylogenetic trees and networks. The two orangutan subspecies were clearly separated from each other, and Sumatran orangutans showed much higher nucleotide diversity than Bornean orangutans. Some gibbon species did not form monophyletic clusters, and variation within species was not much different from that among species in the subgenus Hylobates.

A conserved docking motif in MAP kinases common to substrates, activators and regulators.

Mitogen-activated protein kinases (MAPKs) are specifically phosphorylated and activated by the MAPK kinases, phosphorylate various targets such as MAPK-activated protein kinases and transcription factors, and are inactivated by specific phosphatases. Recently, docking interactions via the non-catalytic regions of MAPKs have been suggested to be important in regulating these reactions. Here we identify docking sites in MAPKs and in MAPK-interacting enzymes. A docking domain in extracellular-signal-regulated kinase (ERK), a MAPK, serves as a common site for binding to the MAPK kinase MEK1, the MAPK-activated protein kinase MNK1 and the MAPK phosphatase MKP3. Two aspartic acids in this domain are essential for docking, one of which is mutated in the sevenmaker mutant of Drosophila ERK/Rolled. A corresponding domain in the MAPKs p38 and JNK/SAPK also serves as a common docking site for their MEKs, MAPK-activated protein kinases and MKPs. These docking interactions increase the efficiency of the enzymatic reactions. These findings reveal a hitherto unidentified docking motif in MAPKs that is used in common for recognition of their activators, substrates and regulators.

Molecular cloning and characterization of a novel dual specificity phosphatase, MKP-5.

A group of dual specificity protein phosphatases negatively regulates members of the mitogen-activated protein kinase (MAPK) superfamily, which consists of three major subfamilies, MAPK/extracellular signal-regulated kinase (ERK), stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), and p38. Nine members of this group of dual specificity phosphatases have previously been cloned. They show distinct substrate specificities for MAPKs, different tissue distribution and subcellular localization, and different modes of inducibility of their expression by extracellular stimuli. Here we have cloned and characterized a novel dual specificity phosphatase, which we have designated MKP-5. MKP-5 is a protein of 482 amino acids with a calculated molecular mass of 52.6 kDa and consists of 150 N-terminal amino acids of unknown function, two Cdc25 homology 2 regions in the middle, and a C-terminal catalytic domain. MKP-5 binds to p38 and SAPK/JNK, but not to MAPK/ERK, and inactivates p38 and SAPK/JNK, but not MAPK/ERK. p38 is a preferred substrate. The subcellular localization of MKP-5 is unique; it is present evenly in both the cytoplasm and the nucleus. MKP-5 mRNA is widely expressed in various tissues and organs, and its expression in cultured cells is elevated by stress stimuli. These results suggest that MKP-5 is a novel type of dual specificity phosphatase specific for p38 and SAPK/JNK.

Improved electroporation of Rhodococcus equi.

The condition of an electroporation method was re-evaluated for the introduction of foreign plasmid DNA into Rhodococcus equi. The method is based on an electroporation of the bacteria made competent by culturing in a broth containing glycine and by heat shock at 50 degrees C. Transformation of R. equi could be achieved with a chloramphenicol-resistant shuttle vector originating from Rhodococcus fascians at an efficiency of about 10(4) transformants/microgram DNA. The bacteria were also shown to become competent when they were incubated with a chemical transformation buffer prior to washing with an electroporation buffer.

[Severe laryngeal edema immediately after extubation in a 93 year old female].

A 93-year-old female, who had post-operative respiratory insufficiency, was treated with artificial ventilation for 8 days. Immediately after extubation, dyspnea, cyanosis and unconsciousness occurred. Severe laryngeal edema was found by bronchofiberscopy and reintubation seemed impossible. She underwent emergency tracheotomy and barely survived the critical state. It should be kept in mind that severe laryngeal edema might develop immediately after long term intubation in supergeriatric patients associated with hyponutrition.

[Pharmacological studies on human urinary kallidinogenase (SK-827): cerebral protective effects].

We investigated the cerebral protective effects of human urinary kallidinogenase (SK-827) in rabbits. The following results were obtained: 1) At a dose of 5.0 x 10(-3) to 1.25 x 10(-2) PNA U/kg, SK-827 significantly inhibited the decrease of focal cerebral blood flow and the impairment of electrocorticogram (ECoG) activity in infarcted rabbits. 2) SK-827 at a dose of 2.5 x 10(-3) PNA U/kg inhibited the development of infarction area induced by internal carotid arterial injection of glass beads. 3) SK-827 at a dose of 2.5 x 10(-3) to 5.0 x 10(-3) PNA U/kg inhibited the decrease of cerebral cortical pO2 induced by 5% oxygen inhalation. 4) On subacute phase 7 days after the destruction of internal capsule, SK-827 at a dose of 1.25 x 10(-2) PNA U/kg improved the abnormality of spontaneous ECoG. These findings suggest that SK-827 minimizes the reduction of cerebral function induced by ischemia.

Absence of nonpercutaneous transmission of hepatitis C virus in a colony of chimpanzees.

Transmission of hepatitis C virus (HCV) was studied in a colony of 85 chimpanzees using assays for anti-HCV and HCV-RNA. Thirteen of the 85 sera were positive for anti-HCV, and 12 of the 13 were also positive for HCV-RNA. All of the anti-HCV positive sera except one were obtained from chimpanzees which had been inoculated with non-A, non-B hepatitis virus. On the other hand, only one of 63 sera of chimpanzees without history of experimental infection of the virus was positive for anti-HCV. Transmission to this chimpanzee was thought to be a needle contaminated with HCV. All 39 samples of chimpanzees born in the center were negative for both anti-HCV and HCV-RNA. Sixteen of their mothers had undergone experimental infection, and 6 of them were positive for both anti-HCV and HCV-RNA. These results suggest that nonpercutaneous transmission, including sexual and mother-to-infant transmissions, is not an important mode of transmission. If these findings apply to humans, definition of inapparent sources of the infection is needed.

[123I-N-isopropyl-p-iodoamphetamine scintigraphy in patients with malignant melanoma].

N-Isopropyl-p-[123I]iodoamphetamine (123I-IMP) has been widely used in patients with cerebral vascular damage and epilepsy. The potential of 123I-IMP accumulation in the melanin producing cells has been reported since early stage of its development. Some authors reported the accumulation of 123I-IMP in the malignant melanoma of human and animals. We evaluated the 123I-IMP scintigraphy in 9 patients with malignant melanoma. Patients were classified into two groups: A, 4 patients with 8 lesions; B, 5 post-operative patients without lesions. In group A, the 123I-IMP uptake was seen in 4 of 8 lesions. The smallest true positive lesion was located at skin and its size was 15 mm in diameter. Two of the visualized 4 lesions were amelanotic malignant melanomas. This fact suggests that uptake of 123I-IMP in malignant melanoma may be related to the processes of melanin synthesis. In group B, two abnormal deposits had been seen in the right thigh of a female patient. However no abnormality was seen in the following 67Ga scintigraphy, TCT, MRI, and repeated 123I-IMP scintigraphy. Therefore the abnormal deposits were considered to be the false positive lesions due to urinary contamination.

[Differentiation of infiltrates between lung cancer and noncancerous lung disease by 123I-IMP lung imaging].

Lung imaging with N-isopropyl-p-123I-iodoamphetamine (123I-IMP) was performed to estimate the pulmonary lesion imaging findings in 3 patients with bronchogenic carcinoma (2: bronchioloalveolar carcinoma and 1: adenocarcinoma) and 18 with noncancerous lung diseases (10: bacterial pneumonia, 1: viral pneumonia, 1: aspiration pneumonia, 1: radiation pneumonitis, 4: pulmonary tuberculosis and 1: obstructive pneumonitis due to an endobronchial lipoma) at 30 min and 4 hr after i.v. injection of 111 MBq of 123I-IMP. These patients all exhibited infiltrates only in the chest radiograms. Decreased uptake of 123I-IMP was observed in the cancerous infiltrating lesions in 3 patients with bronchogenic carcinoma at 30 min and 4 hr, while the uptake of 123I-IMP was normal or increased at 30 min and intense at 4 hr in all 18 noncancerous infiltrating lesions. Therefore 123I-IMP lung imaging can be used to differentiate bronchogenic carcinoma from noncancerous lung disease in patients who exhibit infiltrates only in the chest radiograms.

Resuscitation by extracorporeal lung assist of a patient suffocating after inhalation of sawdust particles.

A 17-yr-old male accidentally inhaled a massive amount of sawdust particles. Severe respiratory distress developed, resulting in subcutaneous emphysema and left pneumothorax. Therefore, sufficient positive-pressure ventilation was inapplicable. Under veno-venous extracorporeal lung assist (ECLA) with an artificial lung, the sawdust particles were removed by broncho-fibroscopy and lung lavage. After 36 h, the patient was weaned from ECLA without further complications.

A case of parathyroid carcinoma visualized by 67Ga citrate and 201Tl chloride scintigraphy.

Both 67Ga citrate and 201Tl chloride accumulated in a parathyroid tumor of a 62 year-old woman with primary hyperparathyroidism. Histological studies disclosed that the tumor was a parathyroid carcinoma. The use of both tumor scanning agents, 67Ga citrate and 201Tl chloride to visualize parathyroid carcinoma in a patient with primary hyperparathyroidism has not been reported as far as we could determine.

[Clinical evaluation of I-131 metaiodobenzylguanidine (MIBG) imaging in suspected neuroblastoma].

Twenty neuroblastoma and 4 nonneuroblastoma patients were studied by 131I-MIBG imaging. The primary tumor was detected in 89% of patients (8/9) before therapy. Bone marrow metastasis was also visualized in 4 of the 8 patients with primary positive scan. True negative results were obtained in 4 nonneuroblastoma patients. After therapy, of 10 tumor-bearing patients, eight showed positive scans and 9 of 12 lesions (75%) were visualized. The accuracies of presence or absence of neuroblastoma were compared between 131I-MIBG imaging and several tumor markers. The accuracies before and after therapy were as follows: 131I-MIBG imaging; 92% (12/13), 88% (15/17), serum NSE; 80% (4/5), 93% (13/14), serum LDH; 92% (11/12), 76% (13/17), urinary VMA; 54% (7/13), 56% (9/16), and urinary HVA; 77% (10/13), 56% (9/16). It appears that 131I-MIBG imaging is useful for both locating and excluding neuroblastoma. In addition, 131I-MIBG imaging appears to be the most efficient diagnostic and follow up study for neuroblastoma when it is combined with measurements of serum NSE.

[A case of acute airway obstruction with sharp sawdust particles, successfully treated by extracorporeal lung assist].

A 17 year-old male accidentally fell into a loading shute filled with sharp particles of sawdust. He aspirated a large amount of sawdust and suffered from acute airway obstruction accompanied by barotrauma, probably due to severe cough and tissue damage from the needle sharp sawdust particles. Ordinary mechanical ventilation was not enough to keep normal gas exchange. To sustain life, a venovenous extracorporeal lung assist, ECLA, with two Kolobow membrane lungs was performed. Under ECLA, the sawdust particles were removed from the airway one by one with a bronchofiber scope taking almost 10 hours. Then the left lung lavage was performed with normal saline to remove finer sawdust. The patient survived these procedures and was weaned from ECLA after 36 hrs. His recovery was uneventful.