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Biomedical subjects

A Kobayashi

Publications and source records attributed to A Kobayashi.

At least 199 records · Page 11Linked to original sources

pICln predominantly localizes at luminal surface membranes of distal tubules and Henle's ascending limbs.

We produced a highly specific antibody to the C-terminal peptide sequence of pICln. It recognized pICln with a 38-kDa molecular mass on SDS-polyacrylamide gel electrophoresis, coinciding with that previously reported. During native polyacrylamide gel electrophoresis, three immunoreactive bands (38, 70, and 130 kDa) were detected. The isoelectric point of pICln was calculated to be 4.0. Subcellular localization study showed the presence of pICln in the soluble and microsomal fraction. pICln can be easily solubilized from the membrane fraction with Triton X-100. From immunohistochemical observations, we found pICln to be obviously located on the luminal surface membranes of the distal tubules and Henle's loop ascending limbs, and it can also be found inside proximal tubular cells. The present results suggested that pICln functions as a "cytosolic anchor = membrane insertion" model, and it plays important roles in the "urine dilution segment" cells of nephrons.

Animals↗

Cys2/His2 zinc-finger protein family of petunia: evolution and general mechanism of target-sequence recognition.

The EPF family is a group of Cys2/His2zinc-finger proteins in petunia. In these proteins, characteristically long spacer regions have been found to separate the zinc fingers. Our previous DNA-binding studies demonstrated that two-fingered proteins (ZPT2-1 and ZPT2-2), which have spacers of different lengths, bind to two separate AGT core motifs in a spacing specific manner. To investigate the possibility that these proteins might distinguish between the target sequences on the basis of spacing between the core motifs, we screened petunia cDNA library for other proteins belonging to this family. Initial screening by PCR and subsequent cloning of full-length cDNAs allowed us to identify the genes for 10 new proteins that had two, three or four zinc fingers. Among the two-fingered proteins the spacing between zinc fingers varied from 19 to 65 amino acids. The variation in the length of spacers was even more extensive in three- and four-fingered proteins. The presence of such proteins is consistent with our hypothesis that the spacing between the core motifs might be important for target sequence recognition. Furthermore, comparison of diverse protein structures suggests that three- and two-fingered proteins might have resulted due to successive loss of fingers from a four-fingered protein during molecular evolution. We also demonstrate that a highly conserved motif (QALGGH) among the members of EPF family and other Cys2/His2 zinc-finger proteins in plants is critical for the DNA-binding activity.

Amino Acid Sequence↗

[Two surgical cases of right parasternal minimal incision for aortic valve replacement].

Although median sternotomy has been used as a good approach to all cardiac valves and coronary arteries, advantages of the minimal invasive cardiac operation have been reported recently. We employed the right parasternal minial incision, reported by Cosgrove et al. for two cases of aortic valve replacement. In the first case, we were able to get a good operation field and easily implanted a mechanical prosthesis. In contrast, we had some difficulties with the second case where in addition to the third and fourth cartilages, a second cartilage resection was necessary because the aortic root lay at a cephalic level. Moreover, the operation field was restricted because we left several rib cartilages to preserve the right internal thoracic artery. The cannula had to be inserted via the right atrium as we failed to insert it in the right femoral vein, and the aortic root deviated more medially than usual. From these experiences, it is important to check the position of aortic root, and if the cannula cannot be inserted in the femoral vein, cannulation via the right atrium can ve utilized in the minimal invasive cardiac operation.

Aortic Valve↗

Temperature- and capsaicin-sensitive nerve fibers in brown adipose tissue attenuate thermogenesis in the rat.

We examined the function of putative sensory fibers that are contained in intercostal nerves and innervate interscapular brown adipose tissue (IBAT) in urethane-anesthetized rats. Warming the IBAT to 40-44 degrees C with two small heaters placed bilaterally on the skin above it attenuated the subsequent noradrenaline-induced thermogenesis (NIT) of the IBAT. In this range of warming, higher IBAT temperatures resulted in more attenuation. Denervation of IBAT blocked the effect of thermal stimulation on the NIT. Thus, activation of nerve fibers in IBAT that are sensitive to warmth or to the nociceptive effects of heat probably attenuated the NIT. Since the putative sensory fibers in the IBAT contain calcitonin gene-related peptide (CGRP) and substance P, which are thought to act in peripheral tissues, we tested the effects of injection of these neuropeptides into the IBAT. Administration of 5.2 nmol CGRP but not substance P or vehicle saline mimicked the effect of thermal stimulation of IBAT. As the neuropeptide-containing primary sensory neurons are characterized by their sensitivity to capsaicin, we also tested its effects (1 mg/kg, s.c.) and found that it also attenuated the NIT. Denervation of the IBAT or pretreatment with capsazepine, a capsaicin receptor antagonist, blocked the effect of capsaicin. We propose that temperature- and capsaicin-sensitive nerve fibers release CGRP to attenuate the NIT of brown adipocytes.

Adipose Tissue, Brown↗

Sulfur-containing compounds from Scorodocarpus borneensis and their antimicrobial activity.

Three novel sulfur-containing compounds were isolated from Scorodocarpus borneensis. Their chemical structures were determined to be 2,4,5,7-tetrathiaoctane 4,4-dioxide (CH3SCH2SO2SCH2SCH3, 1) and 5-thioxo-2,4,6-trithiaheptane 2,2-dioxide (CH3SO2CH2SCSSCH3, 2), while the other compound was assumed to be O-ethyl S-methylthiomethyl thiosulfite (CH3SCH2SS(O)OCH2CH3, 3) on the basis of spectroscopic data. This first isolation of compound 1 strongly suggests that S. borneensis possesses a polysulfide formation pathway similar to that of other Allium species. Compounds 1 and 2 exhibited antimicrobial activity against some bacteria and fungi.

Anti-Bacterial Agents↗

Infective endocarditis complicated by mycotic cerebral aneurysm: two case reports of women in the peripartum period.

Mycotic cerebral aneurysm is a relatively rare but very serious complication of infective endocarditis. Infective endocarditis is a rare but a potentially fatal complication of pregnancy. We report here two very rare cases of infective endocarditis associated with mycotic cerebral aneurysm in peripartum women. In one case, cardiac surgery was performed prior to cerebral surgery and after delivery. In the other case, emergency cerebral surgery was performed due to rupture on the day cardiac surgery had been scheduled, 45 days after delivery. The surgical management of a patient with infective endocarditis and mycotic cerebral aneurysm is reviewed. The surgical strategy for a pregnant patient is also reviewed.

Adult↗

Molecular cloning and chromosomal localization of a prophenoloxidase cDNA from the malaria vector Anopheles gambiae.

A cDNA clone for prophenoloxidase was isolated from the most important human malaria vector, Anopheles gambiae. The clone encoded a polypeptide of 79341 Da that contains the two copper binding domains common to all invertebrate prophenoloxidases and haemocyanins. Expression of the prophenoloxidase gene was detected throughout all life stages from egg to imago in two strains of A. gambiae; however, the strongest expression was observed in developing embryos in eggs. The prophenoloxidase gene was mapped to the inversion rich region of the right arm of chromosome-2 in region 13B.

Amino Acid Sequence↗

Seven zinc-finger transcription factors are expressed sequentially during the development of anthers in petunia.

The development of anthers and pollen involves several stages. In an attempt to identify the transcription factors that are involved in the regulation of gene expression associated with the development of anthers, seven genes for new zinc-finger proteins that are expressed in anthers were detected using RT-PCR. Subsequent cloning and sequence analysis of the full-length cDNA clones revealed that they all encoded zinc-finger proteins of the EPF type. However, the encoded proteins were very different from each other in terms of size, number of zinc fingers (two, three and four) and the spacings between the fingers. Northern blot analysis revealed that the genes were expressed preferentially in anthers. Moreover, it was found that the seven genes were expressed transiently for short periods that corresponded to different developmental stages. It appeared that the genes were activated sequentially during the development of the anther. Considering these observations, a possible mechanism that could account for the autonomous progression of the development of the anther via a regulatory cascade of these transcription factors is discussed.

Amino Acid Sequence↗

The role of ansa-to-recurrent-laryngeal nerve anastomosis in operations for thyroid cancer.

OBJECTIVE: To study the recovery in phonation after reconstruction of the recurrent laryngeal nerve (RLN) in patients whose thyroid cancer was invading the nerve, and to evaluate the role of ansa cervicalis to RLN anastomosis (ARA) in operations for thyroid cancer. DESIGN: Retrospective study. SETTING: University hospital and private thyroid clinic hospital, Japan. SUBJECTS: 34 patients with thyroid cancer who underwent reconstruction of unilateral RLN and 331 consecutive patients operated on for thyroid cancer. INTERVENTIONS: Reconstruction was direct anastomosis (DA), free nerve grafting (FNG), vagus-RLN anastomosis (VRA) or ARA, including anastomosis behind the thyroid cartilage. MAIN OUTCOME MEASURES: Maximum phonation time (34 normal subjects and 26 patients with vocal cord paralysis served as controls), laryngoscopic examination, and the ratio of reconstruction in patients who needed resection of the RLN. RESULTS: The maximum phonation time started to increase rapidly 2-5 months postoperatively in most cases as the patients' voices recovered, and 12 months after reconstruction was significantly longer than in those patients with vocal cord paralysis (P < 0.0001). It was comparable to that of the normal subjects, although the reinnervated cords were fixed in the median. The number of reconstructions in the series of 331 patients increased from 18% to 82% after we started doing ARA with the meticulous technique of anastomosis inside the thyroid cartilage. CONCLUSIONS: ARA is as effective as DA or FNG in improving phonation in patients who need resection of a unilateral RLN. As ARA has several advantages over FNG it has a definite place in operations for thyroid cancer.

Adult↗

Functional outcomes of "floating elbow" injuries in adult patients.

OBJECTIVES: To assess elbow function, complications, and problems of floating elbow fractures in adults receiving surgical treatment. DESIGN: Retrospective clinical review. SETTING: Level I trauma center in Kanagawa, Japan. PATIENTS: Fourteen patients with fifteen floating elbow injuries, excluding one immediate amputation, seen at the Kitasato University Hospital from January 1, 1984, to April 30, 1995. INTERVENTION: All fractures were managed surgically by various methods. In ten cases, the humeral and forearm fractures were treated simultaneously with immediate fixation. In three cases, both the humeral and forearm fractures were treated with delayed fixation on Day 1, 4, or 7. In the remaining two cases, the open forearm fracture was managed with immediate fixation and the humerus fracture with delayed fixation on Day 10 or 25. MAIN OUTCOME MEASUREMENTS: All subjects underwent standardized elbow evaluations, and results were compared with an elbow score based on a 100-point scale. The parameters evaluated were pain, motion, elbow and grip strength, and function during daily activities. Complications such as infections, nonunions, malunions, and refractures were investigated. RESULTS: Mean follow-up was forty-three months (range 13 to 112 months). At final follow-up, the mean elbow function score was 79 points, with 67 percent (ten of fifteen) of the subjects having good or excellent results. The functional outcome did not correlate with the Injury Severity Score of the individual patients, the existence of open injuries or neurovascular injuries, or the timing of surgery. There were one deep infection, two nonunions of the humerus, two nonunions of the forearm, one varus deformity of the humerus, and one forearm refracture. CONCLUSION: Based on the present data, we could not clarify the factors influencing the final functional outcome after floating elbow injury. These injuries, however, potentially have many complications, such as infection or nonunion, especially when there is associated brachial plexus injury. We consider that floating elbow injuries are severe injuries and that surgical stabilization is needed; beyond that, there are no specific forms of surgical treatment to reliably guarantee excellent results.

Activities of Daily Living↗

An image analyzing system using an artificial neural network for evaluating the parenchymal echo pattern of cirrhotic liver and chronic hepatitis.

To objectively evaluate the parenchymal echo pattern of cirrhotic liver and chronic hepatitis, we applied an image analyzing system (IAS) using a neural network. Autopsy specimens in a water tank (n = 13) were used to examine the relationship between the diameter of the regenerative nodule and the coarse score (CS) calculated by IAS. CS was significantly correlated with the diameter of the regenerative nodule (p < 0.0001, r = 0.966). CS is considered to be useful for evaluating the coarseness of the parenchymal echo pattern.

Hepatitis, Chronic↗

Fine structural and histochemical study of equine Paneth cells.

Ultrastructure, lysozyme and glycoconjugate activity in duodenal Paneth cells were observed concurrently in the horse. Paneth cells were seen to uniformly line the base of the equine intestinal glands. The round secretory granules have centrally located electron densities with peripherally located electron lucent halos. Histochemically, the peripheral halo layer was positively stained for carbohydrates by the periodic acid-thiocarbohydrazide-silver protein-physical development (PA-TCH-SP-PD) method and the entire granules reacted positively to the WGA. The central core area reacted with anti-lysozyme. We identified a young (Type I) and an old (Type II) cell population in the same crypt, but we suggest that the observed populations are variations of the same cell type with the varied appearance due to aging of the secretory granules.

Animals↗

[Two cases of Achromobacter xylosoxidans sepsis].

Achromobacter xylosoxidans is a gram-negative bacterium whose natural habitat has not been clearly defined. It has been isolated from ear discharge and the large intestine of humans and from various hospital or environmental water sources. Infection with A. xylosoxidans in humans has been documented, and resulting illnesses include meningitis, pneumonia, cholecystitis, peritonitis and urinary tract infection. Bacteremia due to A. xylosoxidans is rare, and little information on treatment is available. Two cases of bacteremia due to A. xylosoxidans in patients with hemapoietic malignancies are reported herein. Case 1 involved a 70-yr. male whose clinical diagnosis was IgA lambda-type plasmacytoma. Case 2 involved 72-yr. male whose clinical diagnosis was acute lymphatic leukemia (L2). Both patients had been catheterized. Neutropenia was noted and the white blood cell counts were 20/microliter in case 1 and 35/microliter in case 2 when A. xylosoxidans was isolated from the blood culture. We suggest that bacteremia due to A. xylosoxidans may have been related to the presence of the catheter and neutropenia.

Aged↗

Role of alveolar macrophages in initiation and regulation of inflammation in Pseudomonas aeruginosa pneumonia.

To evaluate the role of alveolar macrophages (AMs) in acute Pseudomonas aeruginosa pneumonia in mice, AMs were depleted by aerosol inhalation of liposomes containing clodronate disodium. AM-depleted mice were then intratracheally infected with 5 x 10(5) CFU of P. aeruginosa. In addition to monitoring neutrophil recruitment and chemokine releases, lung injury was evaluated soon after infection (8 h) and at a later time (48 h). At 8 h, depletion of AMs reduced neutrophil recruitment, chemokine release, and lung injury. At 48 h, however, depletion of AMs decreased bacterial clearance and resulted in delayed movement of neutrophils from the site of inflammation with aggravated lung injury. With instillation of 5 x 10(7) CFU of bacteria, AM-depleted mice showed low mortality within 24 h of infection but high mortality at a later time, in contrast to non-AM-depleted mice. These results demonstrate that depletion of AMs has beneficial early effects but deleterious late effects on lung injury and survival in cases of P. aeruginosa pneumonia.

Acute Disease↗

Structural organization of virulence-associated plasmids of Yersinia pestis.

The complete nucleotide sequence and gene organization of the three virulence plasmids from Yersinia pestis KIM5 were determined. Plasmid pPCP1 (9,610 bp) has a GC content of 45.3% and encodes two previously known virulence factors, an associated protein, and a single copy of IS100. Plasmid pCD1 (70,504 bp) has a GC content of 44.8%. It is known to encode a number of essential virulence determinants, regulatory functions, and a multiprotein secretory system comprising the low-calcium response stimulation that is shared with the other two Yersinia species pathogenic for humans (Y. pseudotuberculosis and Y. enterocolitica). A new pseudogene, which occurs as an intact gene in the Y. enterocolitica and Y. pseudotuberculosis-derived analogues, was found in pCD1. It corresponds to that encoding the lipoprotein YlpA. Several intact and partial insertion sequences and/or transposons were also found in pCD1, as well as six putative structural genes with high homology to proteins of unknown function in other yersiniae. The sequences of the genes involved in the replication of pCD1 are highly homologous to those of the cognate plasmids in Y. pseudotuberculosis and Y. enterocolitica, but their localization within the plasmid differs markedly from those of the latter. Plasmid pMT1 (100,984 bp) has a GC content of 50.2%. It possesses two copies of IS100, which are located 25 kb apart and in opposite orientations. Adjacent to one of these IS100 inserts is a partial copy of IS285. A single copy of an IS200-like element (recently named IS1541) was also located in pMT1. In addition to 5 previously described genes, such as murine toxin, capsule antigen, capsule anchoring protein, etc., 30 homologues to genes of several bacterial species were found in this plasmid, and another 44 open reading frames without homology to any known or hypothetical protein in the databases were predicted.

Base Composition↗

The second-largest subunit of the mouse DNA polymerase alpha-primase complex facilitates both production and nuclear translocation of the catalytic subunit of DNA polymerase alpha.

DNA polymerase alpha-primase is a replication enzyme necessary for DNA replication in all eukaryotes examined so far. Mouse DNA polymerase alpha is made up of four subunits, the largest of which is the catalytic subunit with a molecular mass of 180 kDa (p180). This subunit exists as a tight complex with the second-largest subunit (p68), whose physiological role has remained unclear up until now. We set out to characterize these subunits individually or in combination by using a cDNA expression system in cultured mammalian cells. Coexpression of p68 markedly increased the protein level of p180, with the result that ectopically generated DNA polymerase activity was dramatically increased. Immunofluorescence analysis showed that while either singly expressed p180 or p68 was localized in the cytoplasm, cotransfection of both subunits resulted in colocalization in the nucleus. We identified a putative nuclear localization signal for p180 (residues 1419 to 1437) and found that interaction with p68 is essential for p180 to translocate into the nucleus. These results indicate that association of p180 with p68 is important for both protein synthesis of p180 and translocation into the nucleus, implying that p68 plays a pivotal role in the newly synthesized DNA polymerase alpha complex.

3T3 Cells↗