bluuebunny/crossref_metadata_2025 huggingface dataset:
Spectrum of Gastritis in Celiac Disease in Childhood
Authors: Jevon G.P. , Dimmick J.E. , Dohil R. , Hassall E.G.
Published: July 2002

Celiac disease (CD) may cause changes throughout the gastrointestinal tract. The pathology is best described in the distal duodenum and jejunum. It is also associated with lymphocytic gastritis (LG) and varioliform gastritis in adults and children, but the histologic spectrum in the gastric biopsy and the clinical implications are undefined. In this report we relate our experience with the clinical, endoscopic, and histologic changes in gastric biopsies in CD in childhood.

Slides (hematoxylin and eosin stained) were reviewed from 33 celiac children, 5 having had more than 1 gastric biopsy during a 7-year period. Gastric intraepithelial lymphocyte (IEL) counts were compared with those of 10 histologically normal controls (normal range, 1–7 IEL/100 antral or body epithelial cells) and 10 nonceliac chronic gastritis (CG) biopsies without H. pylori (normal range, 1–19 IEL/100 antral cells), noting changes in the epithelium and lamina propria (LP). LG was present in 29/33 initial biopsy sets. Fifteen of 29 showed LG/CG. The IEL number was greater in LG/CG than in LG only (27.2 ± 9.3, n = 14 vs. 18.6 ± 13.4, n = 15 in the antrum; 23.5 ± 2.8, n = 4 vs. 13.0 ± 8.4 in the body). In CD the difference between these mean values and those of normal and nonceliac CG controls was statistically significant. In CG/LG the inflammatory infiltrate was predominantly diffuse/superficial in the LP; mucin depletion was noted in 11/15. The IELs were in the LG/CG range in two CG controls. The IELs were normal at follow-up in five cases. There were no statistically significant differences between the groups with respect to clinical parameters or gastric endoscopic findings. No child had varioliform gastritis.

We conclude that in CD children, the stomach is endoscopically unremarkable but may show LG, or LG/CG with or without mucin depletion, or occasionally appear normal. Gastric histology returned to normal with gluten withdrawal. Normal gastric histology is not typical, but does not exclude CD.
Respiratory Syncytial Virus Infects the Bonnet Monkey, Macaca radiata
Authors: Simoes Eric A.F. , Hayward Anthony R. , Ponnuraj Esther M. , Straumanis John P. , Stenmark Kurt R. , Wilson Harry L. , Babu P. George
Published: July 2002

The Bonnet monkey model of respiratory syncytial virus (RSV) infection may be a useful nonhuman primate model for studying RSV disease in humans because Bonnet monkeys can predictably be infected to obtain an orderly sequence of morphologic, cytologic, virologic, serologic, and inflammatory changes related to time of infection. Young feral Bonnet monkeys, Macaca radiata, were infected endotracheally with 106 plaque-forming units (pfu) of the Long strain of RSV. RSV was recovered from the animals' lungs at necropsy on days 3, 5, and 7 with the highest viral titer obtained on day 3 (1.1 and 5.2 X 103 pfu/g of tissue in the upper and lower lobes, respectively). RSV antigen and F protein mRNA were detected 3–5 days after infection in alveolar macrophages and in the epithelium of bronchi, terminal bronchioles, and alveoli. Histologic analysis of RSV-infected lungs at necropsy revealed progressive bronchiolar mucosal and submucosal inflammation, periarterial mononuclear interstitial inflammation, and focal alveolitis, with a maximal response at 7 days after infection. Cell counts in bronchoalveolar lavage (BAL) increased with time with neutrophils and macrophages predominating on day 3 (6.47 and 5.85 x 105/mm3, respectively) and lymphocytes predominating on day 9 (4.18 X 105/mm3). Serum-neutralizing antibody appeared on day 5 and IgG antibody to RSV was detected on day 9. This sequence of morphologic, cytologic, virologic, serologic, and inflammatory change following RSV infection creates a useful model in the study of experimentally induced RSV disease with a potential for testing future vaccine-induced alterations in RSV disease response.
Fatal Hepatic Short-Chain l-3-Hydroxyacyl-Coenzyme a Dehydrogenase Deficiency: Clinical, Biochemical, and Pathological Studies on Three Subjects with This Recently Identified Disorder of Mitochondrial β-Oxidation
Authors: Bennett Michael J. , Spotswood Sheila D. , Ross Karen F. , Comfort Susan , Koonce Robert , Boriack Richard L. , Ijlst Lodewijk , Wanders Ronald J.A.
Published: July 2002

This report describes the clinical, biochemical, and pathological findings in three infants with hepatic short-chain l-3-hydroxyacyl-coenzyme A dehydrogenase (SCHAD) deficiency, a recently recognized disorder of the mitochondrial oxidation of straight-chain fatty acids. Candidate subjects were identified from an ongoing study of infant deaths. SCHAD analysis was performed on previously frozen liver and skeletal muscle on subjects with a characteristic urine organic acid profile. Autopsy findings were correlated with the biochemical abnormalities. Enzyme analysis in liver revealed marked deficiency in SCHAD with residual activities of 3–11%. All subjects had normal activity in skeletal muscle. However, Western blot analysis of SCHAD revealed an identical truncated protein in both liver and muscle from one patient, suggesting that SCHAD is similar in liver and muscle and that the normal activity in muscle may be due to other enzymes with C4 activity. Autopsy findings revealed marked steatosis and a muscle pattern consistent with spinal muscular atrophy in one patient. Lipid storage was less pronounced in one patient and not detected in the third patient who had a well-documented history of recurrent hypoglycemia. This is the initial pathological characterization of this enzyme defect, and our observations suggest that SCHAD deficiency is a very severe disorder contributing to early infant death.
Chemistry Test Ordering Patterns after Elimination of Predefined Multitest Chemistry Panels in a Children's Hospital
Authors: Pysher Theodore J. , Bach Phillip R. , Lowichik Amy , Petersen Mark D. , Shields Linda H.
Published: July 2002

Predefined multitest chemistry panels (PMCPs) have constituted a large proportion of laboratory tests and patient charges, even in pediatric settings, despite the absence of documented clinical utility for PMCPs and the general availability of random access analyzers that do not require predefined test combinations. We eliminated PMCPs in our tertiary children's hospital but placed no other restrictions on ordering, and observed a 32.7% reduction in the number of automated chemistry tests ordered. All 23 tests in the previous PMCPs showed a decline in utilization, >50% for 8 of the tests and 20–50% for 13 others, and this change was sustained throughout an 8-month follow-up period. The total number of orders for one or more tests increased by 8.2%, but the variety of combinations that were ordered increased by 280%. The most substantial changes included a decrease in the number of orders for combinations of >15 tests, and increases in the number of orders for single tests and combinations of 2 to 5 tests. Orders for combinations identical to all of the former PMCPs declined, with the exception of the 4-test electrolyte panel. There was a marked decline in orders for a 7-test panel identical to the recently defined HCFA-AMA Basic Metabolic Panel, and orders for combinations identical to the HCFA-AMA Liver Function and Extended Metabolic panels were vanishingly rare and nonexistent, respectively. The calculated reduction in patient charges was much greater than actual cost savings, but the reduction in total tests and increase in the variety of test combinations suggest that significant savings can be realized if clinicians are encouraged to order only the tests or combinations they need without imposing procedural, financial, and regulatory burdens.
Anomalies in Cephalothoracopagus Synotus Twins and Their Implications for Morphogenesis in Conjoined Twins
Authors: Bates A.W. , Dodd S.M.
Published: July 2002

The anatomical features of a pair of female conjoined twins of the cephalothoracopagus type are described: the anterior face was well formed but the posterior face was cyclopean with a single ear canal. Visceral abnormalities included two shared hearts linked by an anastomotic arterial ring. The smaller of the hearts was univentricular with atrial situs ambiguus and anomalous pulmonary venous return, and was associated with two trilobed lungs. This, and the presence of a single spleen, suggested asplenia syndrome in one twin. The brains, orientated with respect to the notochordal axes, were joined only at the frontal poles. The abnormalities described in previous reports of twins of this type are reviewed and compared. In addition to alterations in body pattern due to conjoining, endodermal developmental defects and abnormal laterality appear to have been involved in the morphogenesis of this case.
Fetal Membrane Histology in Preterm Premature Rupture of Membranes: Comparison to Controls, and between Antibiotic and Placebo Treatment
Authors: Bendon R.W. , Faye-Petersen O. , Pavlova Z. , Qureshi F. , Mercer B. , Miodovnik M. , Das A.F. , Meis P.J. , Moawad A.H. , Iams J.D. , McNellis D.
Published: July 2002

The objectives of this study were to test the hypotheses that antibiotic therapy will alter the histologic appearance of fetal membranes in preterm premature rupture of membranes (pPROM), and that the membrane histology will demonstrate distinct differences between term and preterm rupture of membranes. We also wished to test interobserver variability of pathologists. Placental membranes were sampled from 268 women participating in a randomized placebo-controlled trial of antibiotic therapy for pPROM at 24–32 weeks of gestation (cases) and from 4 control groups who were not in the randomized trial: (1) preterm labor without pPROM ( n = 21), ( 2) term labor ( n = 65), (3) term PROM ( n = 21), and ( 4) term cesarean section ( n = 27). The cases and controls were scored for 40 histologic features by pathologists blinded to the identity of each sample (case or control). pPROM histology of samples from patients receiving antibiotics and those receiving placebo was compared using a chi-squared test and with control groups using logistic regression. There were no histological differences between pPROM cases treated with antibiotic and those receiving placebo, nor with respect to duration of membrane rupture greater or less than 48 h. Concordance among pathologists was low for features other than acute inflammation. Logistic regression analysis controlled for race and pathologist, and demonstrated that all of the control groups had significantly fewer common markers of acute inflammation when compared with the pPROM cases. This study suggests that histopathologic evidence of infection is seen more frequently with pPROM than in preterm or term controls. The histologic features used in this study cannot be used to determine the effectiveness of antibiotic therapy.
Diagnosis of Hirschsprung Disease Using the Rectal Biopsy: Multi-institutional Survey
Authors: Qualman Stephen J. , Jaffe Ronald , Bove Kevin E. , Monforte-Muñoz Hector
Published: July 2002

Diagnostic pathologists remain uncomfortable with the diagnosis of Hirschsprung disease (HD) via rectal (mucosal/submucosal) biopsy and with performance and interpretation of the associated acetylcholinesterase (AChE) assay. This report details the different diagnostic approaches taken by four major pediatric institutions— Children's Hospital, Columbus, OH; Children's Hospital Medical Center, Cincinnati, OH; Children's Hospital, Pittsburgh, PA; Children's Hospital, Los Angeles, CA—in confirming or excluding the presence of HD. The Columbus approach emphasizes serial morphologic examination of rectal biopsies, while Cincinnati emphasizes the primary diagnostic utility of the AChE stain. Pittsburgh and Los Angeles emphasize a detailed gross and microscopic analysis of rectal biopsies to detect both conventional HD and its more rare subtypes.

The diagnostic approaches of these four institutions can be used on a complementary basis to the advantage of the general diagnostic pathologist, especially in HD cases with subtle clinical presentations. The need for careful and continual communication between the clinician and pathologist in diagnosing or excluding the presence of HD is imperative.
Circumferential Distribution of Ganglion Cells in the Transition Zone of Children with Hirschsprung Disease
Authors: White Frances V. , Langer Jacob C.
Published: July 2002

We prospectively studied the circumferential distribution of ganglion cells in the transition zone from a study population of 21 patients with Hirschsprung disease (HD) undergoing a pull-through procedure over a 26-month period. Twelve cases were satisfactory for examination, in that the transition zone was contained within a single surgical specimen and specimen distortion was minimal. Ganglion cells in the submucosa were counted in all 12 cases. In seven of the cases, the transition zone was proximal to the rectal sleeve and myenteric plexus ganglion cells were also counted.

We found an uneven circumferential distribution of ganglion cells in both myenteric plexus and submucosa of the transition zone, resulting in a “leading edge” of ganglion cells extending into aganglionic distal bowel. The maximum length of this leading edge was 2.4 cm and 2.1 cm in the myenteric plexus and submucosa, respectively. Ganglion cells at the tip of the leading edge were present in clusters of up to six ganglion cells, in marked contrast to an absence of ganglion cells for most of the remainder of the circumference. Closely spaced myenteric plexus ganglia were seen at the tip of the leading edge in some cases. The leading edge was more frequently observed along the antimesenteric side, but this was not statistically significant.

Our findings have relevance in the interpretation of intraoperative biopsies at the time of pull-through surgery and subsequent biopsies of neorectum in patients with surgically corrected HD.
Fetal Obstructive Uropathy: Patterns of Renal Pathology
Authors: Poucell-Hatton Siria , Huang Min , Bannykh Sergei , Benirschke Kurt , Masliah Eliezer
Published: July 2002

Fetal obstructive uropathy (FOU) is characterized by obstruction of the urethra, renal anomalies, ureterovesical dilatation, oligohydramnios, cryptorchidism, and abdominal muscle wall changes. The main objective of the present study was to better understand the relationship between FOU and renal pathology using a series of 15 male autopsy cases. A total of 11 cases with patent anus and 4 with imperforate anus were analyzed. Of the first group, most cases showed obstruction at the level of prostatic urethra. Seven cases showed obstruction at the level of the prostatic urethra and histologic study revealed scarring and partial or complete absence of the prostate, while in the remaining four cases the prostate was present. Of the cases with imperforate anus, two showed obstruction at the level of prostatic urethra, one showed posterior urethral valves, and one was obstructed at the proximal urethra. In all cases the kidneys showed mixed (dysplastic and cystic) changes with no significant differences between the two groups. An inverse correlation was observed between degree of renal dysplasia and gestational age, whereas the opposite was true for cystic changes. Distal and collecting tubules were more intensely immunoreactive to the anti-cytokeratin antibody when compared to proximal tubules. Moreover, anti-cytokeratin immunoreactivity was more prominent in tubules displaying cystic dilatation. DNA fragmentation analysis of renal tissue revealed a higher apoptosis of mesenchymal and tubular cells in the FOU cases, compared to gestational aged–matched controls. These results suggest that renal anomalies in FOU might be related to the gestational age at which the injury occurred and to the duration of the obstruction.
Escherichia coli 0157:H7-induced Hemolytic-uremic Syndrome: Histopathologic Changes in the Colon over Time
Authors: Murray Karen F. , Patterson Kathleen
Published: July 2002

The 1993 E. coli 0157:H7 epidemic in the Western United States has provided a unique opportunity to evaluate the histopathologic temporal progression of disease in the colon in children who developed hemolytic uremic syndrome (HUS). In this report we briefly summarize the clinical courses of eight patients and then discuss the colonic pathology observed in specimens obtained at surgery or at the time of autopsy. The patients were divided into two groups: group 1 consisted of six subjects whose colonic samples were obtained during the acute phase of disease, and group 2 consisted of two subjects whose samples were obtained late in their disease. Both the gross and microscopic findings showed that the most severely affected as well as the earliest affected regions of the colon were the left and transverse portions. Only later in the disease progression was there right-sided colon involvement. These findings are in contrast to the distribution described in E. coli 0157:H7 hemorrhagic colitis without HUS, thus suggesting a different mechanism of injury.
Detection of Trisomy 18 on Formalin-fixed and Paraffin-embedded Material by Fluorescence in Situ Hybridization
Authors: Isaksen Christina Vogt , Ytterhus Borgny , Skarsvøg Sølve
Published: July 2002

Formalin-fixed and paraffin-embedded autopsy material from 10 fetuses and infants with unknown karyotype and anomalies suggestive of trisomy 18 were subjected to fluorescence in situ hybridization (FISH). Nuclei were extracted from the tissues and hybridized with a chromosome 18–specific centromere probe. The hybridization was successful in 9 of 10 cases. Two cases showed three hybridization signals in most of the nuclei (74% and 85%). These had anomalies frequently occurring with trisomy 18 (congenital heart defect, omphalocele, and horseshoe kidney). Two cases showed a mixture of two and three signals (47%/49% and 59%/36%), suggesting the possibility of mosaicism. One of these cases had anomalies consistent with a trisomy 18 phenotype. In the other case intrauterine growth retardation and syndactylies suggested triploidy. Hybridization with a chromosome 8–specific probe gave a distribution of two and three signals (34% and 62%, respectively). This result strengthened the suspicion of a possible triploid mosaicism. In five of the cases most of the nuclei showed two signals (85% to 88%). However, as only one type of tissue was examined for enumeration of chromosome 18, the possibility of organ mosaicism or other chromosome aberrations cannot be excluded. The FISH technique is applicable on macerated and autolysed formalin-fixed tissue, making it possible to retrospectively analyze autopsy material from aborted and stillborn fetuses and infants. This analysis contributes to a better quality of perinatal autopsies and is helpful in parental counseling.
Congenital Misalignment of Pulmonary Veins with Alveolar Capillary Dysplasia Causing Persistent Neonatal Pulmonary Hypertension: Report of Two Affected Siblings
Authors: Gutierrez Carmen , Rodriguez Angeles , Palenzuela Sylvia , Forteza Carolina , Rossello Jose L. Diaz
Published: July 2002

Misalignment of pulmonary vessels with alveolar capillary dysplasia is a rare cause of persistent pulmonary hypertension of the newborn. Most of the reported cases have been sporadic. We present two consecutive affected siblings with this disorder. This is the fifth reported family occurrence of this condition. In addition to the pulmonary abnormality, one of our cases had duodenal atresia.
New Case of Beemer-Langer Syndrome
Authors: Vujanić Gordan M. , Hunt Nicholas C.A.
Published: July 2002

We present the case of a male infant born at 37 weeks gestation with multiple congenital anomalies, including hydrops fetalis, facial and visceral abnormalities, short ribs, and short limbs without polydactyly. We believe that this represents a further case of the Beemer-Langer syndrome, a relatively recently described form of lethal osteochondrodysplasia with an autosomal recessive mode of inheritance. This case also showed some less frequently described anomalies, including arachnoid cysts of the brain and short intestines.
Pediatric Neuroblastic Brain Tumors Containing Abundant Neuropil and True Rosettes
Authors: Eberhart Charles G. , Brat Daniel J. , Cohen Kenneth J. , Burger Peter C.
Published: July 2002

We have encountered a series of seven unusual neuroblastic pediatric central nervous system (CNS) neoplasms with a unique constellation of histologic, immunohistochemical, and ultrastructural features. The tumors presented in five girls and two boys, ages 1 to 3 years. In six cases the lesions involved the frontoparietal region, in one case the tectal plate. The tumors consisted of small to medium-sized, round to oval, hyperchromatic cells with poorly defined cytoplasmic borders. Cells were found in clusters and cords set in a paucicellular fibrillar neuropil matrix. Distinctive, virtually anuclear regions of neuropil were scattered throughout the lesions. True rosettes with well-formed central lumens often filled with granular debris were present, along with perivascular pseudorosettes and occasional Homer-Wright rosettes. Mitoses and apoptosis were frequent, but large regions of confluent necrosis were absent. Immunohistochemically, the neuropil-like areas as well as the perinuclear cytoplasm of many embryonal tumor cells were positive for synaptophysin and neurofilament protein. Ultrastructurally, the tumor cells showed microtubule-containing neuronal processes, some with neurosecretory granules. While the lesions were largely glial fibrillary acidic protein (GFAP) negative, there was focal GFAP positivity consistent with divergent differentiation in one case. The clinical outcome was poor, with five patients dead from their disease 5 to 14 months after initial presentation and one patient with recurrent disease 7 months after resection and chemotherapy. The final patient is alive without recurrent disease 30 months after initial presentation. These lesions present distinctive histological features within the group of primitive neuroectodermal tumors.
Absent Pulmonary Valve with Tricuspid Atresia or Severe Tricuspid Stenosis: Report of Three Cases and Review of the Literature
Authors: Litovsky Silvio , Choy Michael , Park Jeanny , Parrish Mark , Waters Brenda , Nagashima Mitsugi , Van Praagh Richard , Van Praagh Stella
Published: July 2002

Absence of the pulmonary valve occurs usually in association with tetralogy of Fallot and occasionally with an atrial septal defect or as an isolated lesion. Very rarely it occurs with tricuspid atresia, intact ventricular septum, and dysplasia of the right ventricular free wall and of the ventricular septum. We present the clinical, anatomic, and histologic findings of a new case, and for the first time, the data from two patients with absent pulmonary valve and severe tricuspid stenosis, who exhibited similar histologic findings. We also reviewed the clinical and anatomic data of 24 previously published cases and compared them with the new cases. In all three new cases, the myocardium of the right ventricle was very abnormal. In the two cases with tricuspid stenosis, large segments of myocardium were replaced with sinusoids and fibrous tissue. In the case with tricuspid atresia, the right ventricular free wall contained only fibroelastic tissue. The ventricular septum in all three patients showed asymmetric hypertrophy and in two of the three patients, multiple sinusoids had replaced large segments of myocardial cells. The left ventricular free wall myocardium and the walls of the great arteries were unremarkable. Our data indicate that myocardial depletion involving the right ventricular free wall and the ventricular septum and its replacement by sinusoids and fibroelastic tissue occur not only in cases of absent pulmonary valve with tricuspid atresia but also in cases of absent pulmonary valve with tricuspid stenosis. The degree of myocardial depletion varies and is more severe when the tricuspid valve is atretic.
Work-up and Diagnosis of Pediatric Non-Hodgkin's Lymphomas
Authors: Perkins Sherrie L.
Published: July 2002

Non-Hodgkin's lymphomas (NHL) in children and adolescents represent about 10% of childhood cancers. Although the types of NHL commonly seen in this population are relatively limited to lymphoblastic lymphomas, Burkitt's and Burkitt-like lymphomas, and large cell lymphomas, correct diagnosis and classification are essential for optimal therapy. Careful handling of pathologic specimens, along with collection of proper materials for ancillary studies such as immunophenotyping, cytogenetics, or molecular studies, will aid the pathologist in reaching a correct diagnosis. Specific morphologic, immunophenotypic, and genetic features of the commonly seen types of pediatric NHL are described.
Evidence for erythrocyte membrane glycoproteins being carriers of blood‐group P1 determinants
Authors: Haselberger Christian G. , Schenkel-Brunner Helmut
Published: July 2002

The contribution of different membrane constituents to the bloodgroup P1 activity of human erythrocytes was investigated. Pronase digestion of native red cell stroma or partition between butanol and water had no serologically detectable effect, whereas pronase‐treatment of previously butanol‐extracted membranes liberated virtually all blood‐group P1 determinants from the ghosts. On Laemmli gels, all P1 activity was found in the band 4.5 region. Thus it is concluded that, in addition to the well‐documented P1 glycolipid, also membrane glycoproteins are carriers of blood‐group P1 determinants.
Hydroxylamine as an inhibitor between Z and P680 in photosystem II
Authors: Ghanotakis Demetrios F. , Babcock Gerald T.
Published: July 2002

Upon addition of hydroxylamine to chloroplasts or photosystem II preparations, the EPR signal of Z⨥ disappears and a new signal is observed. From its shape and g‐value this signal is identified with the oxidized reaction center chlorophyll, P680+. The decay of P680+ occurs with a halftime of ⪅ 200 μs and apparently is the result of a back reaction with the reduced form of the primary acceptor, QA. This mode of hydroxylamine inhibition is reversible. These observations indicate that hydroxylamine, in addition to its well known inhibitory action on the oxygen evolving complex, is also able to disrupt physiological electron flow to P680 itself.
Long‐chain fatty acids and their acyl‐CoA esters cause the translocation of phosphatidate phosphohydrolase from the cytosolic to the microsomal fraction of rat liver
Authors: Martin-Sanz Paloma , Hopewell Roger , Brindley David N.
Published: July 2002

A translocation of phosphatidate phosphohydrolase from the cytosolic to the microsomal fraction was promoted in cell‐free extracts of rat liver by oleate and palmitate and their CoA esters. Oleate was more potent in this respect than palmitate and the CoA esters were more effective than the unesterified acids. Octanoate, octanoyl‐CoA and CoA did not cause the translocation. It is proposed that the interaction of phosphatidate phosphohydrolase with the membranes that synthesize glycerolipids causes it to become metabolically active. This enables the liver to increase its capacity for triacylglycerol synthesis in response to an increased supply of fatty acids.
Temperature‐dependent cleavage of chromatin by micrococcal nuclease near the nucleosome center
Authors: Huang Sue-Ying , Garrard William T.
Published: July 2002

Digestion of nuclei at 4°C with micrococcal nuclease results in significant intranucleosomal cleavage compared to digestion conducted at 37°C. Employing nucleoprotein gel electrophoresis in one dimension followed by DNA electrophoresis in a second dimension, we demonstrate that such temperature‐sensitive, internal cleavage predominantly occurs about 20 bp from the nucleosome center. We suggest that lower temperatures reduce the stability of hydrophobic interactions within the histone octamer and lead to a conformational alteration in nucleosomes that is detected by micrococcal nuclease.
Identification of novel GTP‐binding proteins in the human neutrophil
Authors: Bokoch Gary M. , Parkos Charles A.
Published: July 2002

We describe here the existence of previously undescribed GTP‐binding proteins within the human neutrophil. These proteins specifically bind guanine nucleotides under conditions in which the previously characterized G‐proteins are unable to bind. We have partially purified these proteins and present both functional and immunologic data which indicate that they are unrelated to Gn, the major neutrophil pertussis toxin substrate. An additional protein, of apparent molecular mass 22 kDa, may be related to the ras G‐protein family. Analysis of the structural and functional characteristics of these novel proteins will promote a better understanding of the process of neutrophil activation.
Electrogenesis associated with proton transfer in the reaction center protein of the purple bacterium Rhodobacter sphaeroides
Authors: Drachev L.A. , Mamedov M.D. , Mulkidjanian A.Ya. , Semenov A.Yu. , Shinkarev V.P. , Verkhovsky M.I.
Published: July 2002

Electrogenic events in the photosynthetic reaction center complex (RC), accompanying single‐ and two‐electron reduction of the secondar quinone acceptor Qb, were investigated. In the presence of inhibitors of electron transfer via the bc1‐complex, the kinetics of formation of the transmembrane electric potential difference induced by two successive light flashes exhibit a few phases. Besides the fast phase A which is due to the charge separation between the bacteriochlorophyll dimer P and primary quinone acceptor qa, two slower atrazine‐sensitive phases, BI and BII, were observed. Phase BI is suggested to be due to proton transfer between the amino acid residues of the reaction center protein, and phase BII due to proton uptake during the second flash‐induced formation of ubiquinol. A possible model of electrogenesis in the acceptor moiety of the RC is discussed.
Purification and characterization of a chymotrypsin Kunitz inhibitor type of polypeptide fro the venom of cobra (Naja naja naja)
Authors: Shafqat Jawed , Zaidi Zafar H. , Jörnvall Hans
Published: July 2002

A chymotrypsin Kunitz inhibitor type of polypeptide has been isolated from the venom of Naja naja naja by reverse phase HPLC and cation exchange FPLC. It is present in a considerably lower amount than that of the corresponding trypsin inhibitor. The primary structure, determined by sequence analysis of the whole molecule and its tryptic peptides, has 57 residues with an apparent molecular mass of 6.2 kDa. The main contact site with the protease (P1) has a Phe, showing the specificity of the inhibitor. Of residues considered functionally important in Kunitz‐type inhibitors, Gly‐36 is replaced by Ser in a segment of weak contacts with the protease.
Cytotoxicity of, and DNA damage by, active oxygen species produced by xanthine oxidase
Authors: Chiricolo Mariella , Tazzari Pier Luigi , Abbondanza Ada , Dinota Angelo , Battelli Maria Giulia
Published: July 2002

Toxicity to Raji cells of the xanthine oxidase/hypoxanthine system is related to the formation of single‐strand DNA breaks. DNA damage was proportional to the concentration of xanthine oxidase and to the time of exposure. It was prevented by the absence of hypoxanthine, or by the presence of allopurinol, or both superoxide dismutase and catalase. The release of 51Cr from damaged cells was detectable 12h after the inhibition of cloning efficiency and the production of DNA breakage. These data suggest that DNA damage induced by the oxygen products precedes the severe lesion to the cellular membrane.
Hepatocyte swelling leads to rapid decrease of the G‐/total actin ratio and increases actin mRNA levels
Authors: Theodoropoulos Panayotis A. , Stournaras Christos , Stoll Barbara , Markogiannakis Emmanuel , Lang Florian , Gravanis Achilles , Häussinger Dieter
Published: July 2002

Exposure of isolated rat hepatocytes to hypotonic (190 mosinol/1) incubation media lowered the cellular G‐actin level without affecting the total actin content: here the G‐/total actin ratio decreased by 15.5 ± 1.4% (n=7). Similar effects were observed following isotonic cell swelling by either addition of glutamine (10 mM) or insulin (100 nM), resulting in a decrease of the G‐/total actin ratios by 13.5 ± 2.1 % (n = 5) and 14.1 ± 1.1% (n = 11), respectively. The effects of hypotonic exposure, glutamine and insulin on the G‐/total actin ratio largely occurred within 1 min and persisted for at least 2 h in presence of the respective effectors. After a 120 min exposure to hypotonic media, glutamine or insulin the actin mRNA levels were increased 2.4‐, 2.0‐ and 3.6‐fold, respectively. Hypertonic exposure lowered the G‐/total actin ratio by only 4.9 ± 2.5% (n = 4) and increased actin mRNA levels only 1.2‐fold. There was a close relationship between glutamine‐ and hypotonicity‐induced cell swelling and the decrease of G‐/total actin ratios. The data suggest that cell swelling exerts rapid and marked effects on the state of actin polymerization and increases actin mRNA levels. Thus, cytoskeletal alterations in response to cell swelling may be involved in the regulation of hepatic metabolism by cell volume.
Receptor‐like kinase activity in membranes of Arabidopsis thaliana
Authors: Schaller G.Eric , Bleecker Anthony B.
Published: July 2002

A class of protein kinases in the plant Arabidopsis thaliana has been identified which has biochemical characteristics similar to those of the animal receptor protein kinases. These plant protein kinases are membrane‐associated, glycosylated, exhibit a preference for Mn2+ relative to Mg2+, and range from 115 to 135 kDa when identified by renaturation after protein blotting. Evidence is presented that the TMK1 protein, a receptor‐like protein kinase identified in Arabidopsis from its cloned gene, is also glycosylated in its native form, and shows a preference for Mn2+ over Mg2+ when assayed for kinase activity. All of the plant receptor‐like protein kinases identified phosphorylate only serine and threonine residues.
Regulation of the H+/e− stoichiometry of cytochrome c oxidase from bovine heart by intramitochondrial ATP/ADP ratios
Authors: Frank Viola , Kadenbach Bernhard
Published: July 2002

This paper describes the effect of intramitochondrial ATP/ADP ratios on the H+/e− stoichiometry of reconstituted cytochrome c oxidase (COX) from bovine heart. At 100% intraliposomal ATP the H+/e− stoichiometry of the reconstituted enzyme is decreased to half of the value measured below 98% intraliposomal ATP (above 2% ADP), while it remains constant up to 100% ADP. The decrease is obtained with different COX preparations, independent of the absolute value of the H+/e− stoichiometry. Decrease of H+/e− stoichiometry is prevented by preincubation of the enzyme with a tissue‐specific monoclonal antibody to subunit VIa‐H (heart‐type). Tissue‐specific regulation of the efficiency of energy transduction in COX of muscle mitochondria could have a physilogical function in maintaining the body temperature at rest or sleep, i.e. at low ATP expenditure.
Functional conservation of mouse Notch receptor family members
Authors: Kato Hisamune , Sakai Takashi , Tamura Kumiko , Minoguchi Shigeru , Shirayoshi Yasuaki , Hamada Yoshio , Tsujimoto Yoshihide , Honjo Tasuku
Published: July 2002

All the known members of the mouse Notch receptor family were examined for their biochemical function by interaction with a DNA binding protein RBP‐Jκ. mNotch2, mNotch3 and int3 (= mNotch4) were shown to interact with RBP‐Jκ by the GST‐fusion pull down assay and dominant negative competition with Epstein Barr virus nuclear antigen 2. Furthermore the intracellular region of int3 was shown to transactivate the Epstein Barr virus TP1 promoter. These results indicate that all mouse Notch family members have biochemical functions similar to mNotch1, which transduces proliferative signal by direct interaction with the DNA binding protein RBP‐Jκ.
Genotypes of cytochrome P450 and clinical response to clomipramine in patients with major depression
Authors: Bouchez J , Dumur V , Lhermitte M , Goudemand M
Published: July 2002

The genetic cytochrome P450 polymorphism is reported in factors affecting the individual response to drugs. The interindividual variation at steady-state levels or also in elimination of drugs, finds an explanation in genetic differences in the metabolism. In particular, activities of the P450-IID6 isoenzyme are related to the sparteine/debrisoquine oxidation polymorphism. Phenotyping such a system has been proposed to analyse variability in the tricyclic antidepressant level. To analyse clinical relevance of a pharmacogenetic approach, we studied the cytochrome P450 CYP2D6 genotypes and the clinical responses to clomipramine in 21 hospitalised patients who met DSM-III-R criteria for major depression. Three patients were predicted as poor metabolizers. We suggested a limitation of clomipramine (CMI) hydroxylation in poor metaboliser (PM) patients which is balanced by a desmethylation. The clinical efficacy pattern does not differ in poor metaboliser and early metaboliser patients. Firstly, there is no significant differences in the evolution of scores on MADRS and specific retardation scale into the two groups. Secondly, outcome of side effects does not occur more frequently in PM patients. Clinical relevance of such an approach needs further study.
Prothymosin α fragmentation in apoptosis
Authors: Evstafieva Alexandra G. , Belov George A. , Kalkum Markus , Chichkova Nina V. , Bogdanov Alexey A. , Agol Vadim I. , Vartapetian Andrey B.
Published: July 2002

We observed fragmentation of an essential proliferation‐related human nuclear protein prothymosin α in the course of apoptosis induced by various stimuli. Prothymosin α cleavage occurred at the DDVD99 motif. In vitro, prothymosin α could be cleaved at D99 by caspase‐3 and ‐7. Caspase hydrolysis disrupted the nuclear localization signal of prothymosin α and abrogated the ability of the truncated protein to accumulate inside the nucleus. Prothymosin α fragmentation may therefore be proposed to disable intranuclear proliferation‐related function of prothymosin α in two ways: by cleaving off a short peptide containing important determinants, and by preventing active nuclear uptake of the truncated protein.
Expression of fatty acid‐CoA ligase 4 during development and in brain
Authors: Cao Yang , Murphy Kelley J. , McIntyre Thomas M. , Zimmerman Guy A. , Prescott Stephen M.
Published: July 2002

Fatty acid utilization is initiated by fatty acid‐CoA ligase, which converts free fatty acids into fatty acyl‐CoA esters. We have cloned previously the human long‐chain fatty acid‐CoA ligase 4 (FACL4), which is a central enzyme in controlling the free arachidonic acid level in cells and thereby regulating eicosanoid production. We report here the expression of this gene in tissues, particularly in different parts of the brain. We found that FACL4 encoded a 75 kDa enzyme and that there was a modified translation product expressed in the brain. FACL4 was expressed in early stages of development with a significant amount of FACL4 mRNA detected in an E7 mouse embryo. In addition, FACL4 was highly expressed in both adult and newborn mouse brain especially in the granule cells of the dentate gyrus and the pyramidal cell layer of CA1 in hippocampus, and the granular cell layer and Purkinje cells of the cerebellum.
Preferential induction of apoptosis of leukaemic cells by farnesol
Authors: Rioja Alphonso , Pizzey Arnold R , Marson Charles M , Thomas N.Shaun B
Published: July 2002

Farnesol preferentially inhibits proliferation and induces apoptosis of tumour‐derived but not non‐transformed cell lines. We investigated whether farnesol induces apoptosis of blasts from patients with acute myeloid leukaemia (AML) and leukaemic cell lines, as compared with normal, human primary haemopoietic cells. We show that 30 μM farnesol causes apoptosis of leukaemic cell lines of T‐ and B‐lymphocyte, myeloid or erythroid lineages and primary blasts obtained from patients with AML. However, the same concentration did not kill primary monocytes, or quiescent or proliferating T‐lymphocytes. We conclude that farnesol selectively kills AML blasts and leukaemic cell lines in preference to primary haemopoietic cells.
Regulated but not constitutive human respiratory syncytial virus (HRSV) P protein phosphorylation is essential for oligomerization
Authors: Asenjo Ana , Villanueva Nieves
Published: July 2002

Purified human respiratory syncytial virus (HRSV) P phosphoprotein from transfected HEp‐2 cells is able to oligomerize forming tetramers. The bulk of constitutive P protein phosphorylation (99.8%) (serine residues 116, 117, 119, 232 and 237) can be removed without affecting protein oligomerization. However, dephosphorylated P protein, produced in bacteria, is unable to oligomerize. This difference can be explained by a transient P protein phosphorylation, detected in HEp‐2 cells, that could be essential for P protein oligomerization.
The high molecular weight isoforms of basic fibroblast growth factor (FGF‐2): an insight into an intracrine mechanism
Authors: Delrieu Isabelle
Published: July 2002

Basic fibroblast growth factor (FGF‐2) is an important modulator of cell growth and differentiation under both physiological and pathological conditions. Until recently, most investigations into the FGF‐2 signalling pathway were concerned with its interaction with specific membrane receptors. Nevertheless, while a 18 kDa protein of FGF‐2 is cytosolic, there are also co‐translated high molecular weight (HMW) isoforms that are predominantly located in the cell nucleus. An increasing amount of data strongly argue in favour of distinct biological functions depending on the subcellular location of the FGF‐2 species. This review describes the evidence concerning the strictly intracellular mode of action of the HMW isoforms of FGF‐2.
Intra‐ and intermolecular interactions of the catalytic domains of human CD45 protein tyrosine phosphatase
Authors: Hayami-Noumi Keiko , Tsuchiya Tomofusa , Moriyama Yoshinori , Noumi Takato
Published: July 2002

We have investigated protein‐protein interaction between distinct domains of the human CD45 cytoplasmic region using a yeast two‐hybrid system. Consequently, we have found that the spacer region between two tandem PTP domains specifically interacts with the membrane‐distal PTP domain (D2). This interaction is mediated by a stretch of amino acid residues in the carboxyl‐terminal half of the spacer region. Although the membrane proximal region does not directly interact with either of the two PTP domains, it appears to function in stabilizing the interaction between the spacer region and D2. We also demonstrate that the interaction between the spacer region and D2 might take place intramolecularly.
Preparation and characterization of ‘heparinocytes’: erythrocytes with covalently bound low molecular weight heparin
Authors: Müller Michele , Büchi Linda , Woodtli Karin , Haeberli André , Beer Jürg H
Published: July 2002

In an attempt to create the possibility of stable, long acting, intravascular anticoagulation, low molecular weight heparin was modified by introducing a sulfhydryl group into the molecule (LMWH‐SH). Human erythrocytes were covalently grafted with LMWH‐SH by the use of a heterobifunctional coupling reagent which reacts with the SH group of LMWH‐SH and surface exposed amino groups of erythrocytes now called ‘heparinocytes’ (HC). HC were morphologically indistinguishable from untreated erythrocytes and displayed identical osmotic resistance. The functionality of HC was analyzed by classical coagulation tests in which they dose dependently inhibited clot formation. HC were also functional in recalcified whole blood inhibiting thrombin formation as assessed by the cleavage of the chromogenic substrate S‐2238. The system appears applicable as a potential autologous, long‐term anticoagulant treatment or prophylaxis.
Characterization of the interactions between the glycine transporters GLYT1 and GLYT2 and the SNARE protein syntaxin 1A
Authors: Geerlings Arjan , López-Corcuera Beatriz , Aragón Carmen
Published: July 2002

In this study we have examined the effect of the SNARE protein syntaxin 1A on the glycine transporters GLYT1 and GLYT2. Our results demonstrate a functional and physical interaction between both glycine transporters and syntaxin 1A. Co‐transfection of syntaxin 1A with GLYT1 or GLYT2 in COS cells resulted in approximately 40% inhibition in glycine transport. This inhibition was reversed by the syntaxin 1A‐binding protein, Munc18. Furthermore, immunoprecipitation studies showed a physical interaction between syntaxin 1A and both transporters in COS cells and in rat brain tissue. Finally, we conclude that this physical interaction resulted in a partial removal of the glycine transporters from the plasma membrane as demonstrated by biotinylation studies.
Cytosolic Hsp70s are involved in the transport of aminopeptidase 1 from the cytoplasm into the vacuole
Authors: Satyanarayana Chitkala , Schröder-Köhne Stephan , Craig Elizabeth A. , Schu Peter V. , Horst Martin
Published: July 2002

Eukaryotic 70 kDa heat shock proteins (Hsp70s) are localized in various cellular compartments and exhibit functions such as protein translocation across membranes, protein folding and assembly. Here we demonstrate that the constitutively expressed members of the yeast cytoplasmic Ssa subfamily, Ssa1/2p, are involved in the transport of the vacuolar hydrolase aminopeptidase 1 from the cytoplasm into the vacuole. The Ssap family members displayed overlapping functions in the transport of aminopeptidase 1. In SSAI and SSAII deletion mutants the precursor of aminopeptidase 1 accumulated in a dodecameric complex that is packaged in prevacuolar transport vesicles. Ssa1/2p was prominently localized to the vacuolar membrane, consistent with the role we propose for Ssa proteins in the fusion of transport vesicles with the vacuolar membrane.
Crystal structure of rat heme oxygenase‐1 in complex with heme
Authors: Sugishima Masakazu , Omata Yoshiaki , Kakuta Yoshimitsu , Sakamoto Hiroshi , Noguchi Masato , Fukuyama Keiichi
Published: July 2002

Heme oxygenase catalyzes the oxidative cleavage of protoheme to biliverdin, the first step of heme metabolism utilizing O2 and NADPH. We determined the crystal structures of rat heme oxygenase‐1 (HO‐1)–heme and selenomethionyl HO‐1–heme complexes. Heme is sandwiched between two helices with the δ‐meso edge of the heme being exposed to the surface. Gly143N forms a hydrogen bond to the distal ligand of heme, OH−. The distance between Gly143N and the ligand is shorter than that in the human HO‐1–heme complex. This difference may be related to a pH‐dependent change of the distal ligand of heme. Flexibility of the distal helix may control the stability of the coordination of the distal ligand to heme iron. The possible role of Gly143 in the heme oxygenase reaction is discussed.
Involvement of the platelet‐derived growth factor receptor in angiotensin II‐induced activation of extracellular regulated kinases 1 and 2 in human mesangial cells
Authors: Mondorf Ulrich F. , Geiger Helmut , Herrero Martina , Zeuzem Stefan , Piiper Albrecht
Published: July 2002

In mesangial cells angiotensin II (Ang II) has been shown to activate extracellular regulated kinases 1 and 2 (ERK1/2). Here, we studied the role of the epidermal growth factor receptor (EGFR) and platelet‐derived growth factor receptor (PDGFR) in Ang II‐induced ERK1/2 activation in human mesangial cells. Ang II induced activation of ERK1/2 via the AT1 receptor, and this response was blocked by the PDGFR‐selective tyrosine kinase inhibitor AG1295, but not by AG1478, an EGFR‐selective tyrosine kinase inhibitor, indicating participation of the PDGFR, but not of the EGFR in Ang II‐induced ERK1/2 activation. In agreement with this assumption, Ang II caused tyrosine phosphorylation of the PDGFR and the adapter protein Shc in an AG1295‐sensitive fashion. In conclusion, our data show that Ang II‐induced activation of mitogenic signalling cascade in human mesangial cells involves ligand‐independent activation of the PDGFR, but not of the coexpressed EGFR.