bluuebunny/crossref_metadata_2025 huggingface dataset:
Hormonal Control of Rat Fetal Pancreas Development
Authors: Morisset Jean , Lainé Jean , Mimeau-Worthington Tracy
Published: April 2003

Rat fetal pancreas development and maturation were investigated in vitro and in vivo, and the informations available on their controls do not agree. Our main objective was to reinvestigate fetal pancreas growth in vivo through treatments of the dams during their entire pregnancy. Pregnant rats were thus implanted subcutaneously with Alzet minipumps and received cerulein (0.25 μg kg–1 h–1), gastrin-releasing peptide (GRP; 0.18 μg kg–1 h–1), GRP antagonist (12 μg kg–1 h–1), pentagastrin (2.38 μg kg–1 h–1), L-365,260, a cholecystokinin B (CCKB) receptor antagonist (120 μg kg–1 h–1), and hydrocortisone (417 or 833 μg kg–1 h–1). After sacrifice at the end of pregnancy, the pancreata of the dams and those of their fetuses were excised for weight, protein, RNA, DNA, and digestive enzyme determinations. In the fetus, pancreas growth defined as hyperplasia was observed only in response to hydrocortisone, while aplasia occurred in response to cerulein. Gastrin and the GRP antagonist were the most effective hypertrophic agents, and the effect of the CCKB receptor antagonist was atrophic. In conclusion, hydrocortisone caused proliferation of the fetal rat pancreas, whereas gastrin induced its differentiation and maturation probably through CCKB receptor occupation.
Cross-Desensitization to Furosemide and Salbutamol in Isolated Neonatal Guinea Pig Airways
Authors: Gelber Rebecca P. , Christ Michael J. , Iwamoto Lynn M. , Marinkovich G. Andre , Fujiwara Naomi , Nakamura Kenneth T.
Published: April 2003

Airway hyperresponsiveness in neonatal chronic lung disease is treated with both furosemide, a diurectic that inhibits the Na-K-2Cl cotrasporter, and salbutamol, a β2-adrenoceptor agonist. Tachyphylaxis to both drugs in vitro has been described. This study was conducted to determine if the relaxation response in newborn guinea pig airways to furosemide and salbutamol can be cross-desensitized in vitro. Tracheal ring segments from 4- to 7-day-old guinea pigs were suspended in HEPES buffer for measurement of isometric tension. Segments were pre-treated with either furosemide (300 μM, 1 h) or salbutamol (10 μM, 30 min). After constriction with 3 μM acetylcholine, relaxation response to salbutamol or furosemide, respectively, was measured. Pretreatment with furosemide diminished relaxation response to salbutamol [87 ± 3% (n = 11) vs. 117 ± 8% (n = 10), p < 0.05], as compared to saline-treated controls. In addition, pretreatment with salbutamol diminished relaxation response to furosemide [53 ± 2% (n = 11) vs. saline-treated (83 ± 7%, n = 7, p < 0.05) and DMSO-treated controls (69 ± 5%, n = 5, p < 0.05)]. Measurements of 86Rb uptake, cyclic AMP levels and responses in the presence of charybdotoxin make it unlikely that Na-K-2Cl cotransporter activity, stimulation of cAMP, or opening of maxi-K+ channels are mechanisms involved in the cross-desensitization to furosemide and salbutamol in vitro.
Is the CRIB Score (Clinical Risk Index for Babies) a Valid Tool in Predicting Neurodevelopmental Outcome inExtremely Low Birth Wei ght Infants?
Authors: Lago P. , Freato F. , Bettiol T. , Chiandetti L. , Vianello A. , Zaramella P.
Published: April 2003

The study aimed to assess how well the severity of clinical conditions in extremely low birth weight infants in the first 12 h of life, as measured by the CRIB (clinical risk index for babies), relates to hospital outcome and subsequent disability at 18 months of age. The CRIB was confirmed as a valid index of initial neonatal risk, even in extremely low birth weight infants, predicting hospital outcome (death or major brain lesions) more accurately than birth weight or gestational age. However, an adjustment of the CRIB score for gestational age might enhance its positive predictive value in relation to short-term developmental outcome in this particular population.
Indomethacin Treatment Decreases Renal Blood Flow Velocity in Human Neonates
Authors: Kang Nam Soo , Yoo Kee Hwan , Cheon Haewon , Choi Byung Min , Hong Young Sook , Lee Joo Won , Kim Soon Kyum
Published: April 2003

This study was designed to evaluate the effect of indomethacin (ID) on renal perfusion in 13 neonates with symptomatic patent ductus arteriosus (PDA). Serial blood flow velocity in the left renal artery was measured just before and at 10, 30, 45, 60, 75 and 90 min after ID administration. Serum creatinine (Cr), sodium (Na), and osmolarity were measured just before, at 12 and 24 h, and at 3 days after ID administration. Timed urine also was collected for measurement of amount, fractional excretion of Na (FENa), and creatinine clearance (CCr). ID decreased end-diastolic flow velocity of renal artery and increased Pourcelot’s index, starting at 10 min and lasting for 75 min (p < 0.05). Serum Cr significantly increased at 12 h, and hourly urine output and CCr decreased for 24 h. Serum Na and osmolarity decreased for a period of at least 3 days (p < 0.05). FENa decreased at 12–24 h (p < 0.05). We conclude that ID treatment can induce significant renal dysfunction due to diminution of renal perfusion in human neonates.
Effects of Milk and Colostrum on Superoxide Generation of Oral Polymorphonuclear Neutrophils and the Changes That Occur during Storage at Low Temperature
Authors: Kanematsu Hiroshi , Kanno Tomoko , Nakahara Hiroko , Ishisaka Rumi , Akiyama Jitsuo , Horton Alan A. , Yasuda Tatsuji , Yoshioka Tamotsu
Published: April 2003

Human oral polymorphonuclear neutrophils (OPMN) generate reactive oxygen species even in the absence of stimulants. Because OPMN from newborn babies are exposed to colostrum and mature milk, the biological properties of these cells including the generation of reactive oxygen species might possibly be affected by the constituents of colostrum and milk. The present work reports the effects of colostrum and mature milk, including the effects of storage at low temperature, on superoxide generation by OPMN. Fresh colostrum and mature milk did not affect either endogenous or formyl-methionyl-leucyl-phenylalanine-induced generation of superoxide by OPMN. However, superoxide generation stimulated by phorbol myristate acetate or arachidonic acid was inhibited by colostrum and mature milk presumably due to binding of the ligands to milk proteins. During the storage of milk at 4°C, free forms of unsaturated long-chain fatty acids increased, and there was concomitant increase in the ability of milk to generate superoxide radicals in OPMN. Kinetic analysis suggested that colostrum and mature milk regulate superoxide generation by OPMN, thereby modulating the bactericidal activity of these cells in the oral cavity.
Effect of SOD-Mimetic Fe-Chlorine e6-Na on the Level of Brain Lipid Peroxide of Rat Fetal Brains Exposed to Reactive Oxygen Species Leading to Intrauterine Growth Retardation
Authors: Saito Katsu , Maeda Munenori , Yoshihara Hajime , Amano Kan , Nishijima Masahiro , Nakamura Kunie
Published: April 2003

The influence of oxidative stress in rat brain and liver on intrauterine growth retardation (IUGR) in rat fetuses was examined. Twenty pregnant Wistar rats were used. On the 15th day of pregnancy, uterine artery and vein were ligated bilaterally using a modified Wigglesworth method. On the 21st day of pregnancy, the fetuses were delivered by hysterotomy. Fetal blood was collected by cardiac puncture. Fetal brain and liver were removed for the analysis of lipid peroxide. Sham surgical operations were performed in the control rats. Within the ligated group, a superoxide dismutase mimicking substance, Fe-chlorine e6-Na (FeCNa), was injected intraperitoneally once a day from day 15 of gestation to day 20. Fetal blood, brain, and liver were stored at –70°C until analysis. Control rats received an equivalent volume of saline. In growth-retarded fetuses, both superoxide released from erythrocytes and brain lipid peroxide showed significantly higher levels, but not superoxide dismutase in erythrocytes and liver lipid peroxide. These changes were alleviated by injection of superoxide dismutase-mimicking substance, FeCNa. Rat fetuses with intrauterine growth retardation suffered from a significant oxidative stress in utero. The increase in reactive oxygen species was alleviated by an injection of FeCNa.
Decreased mRNA Expression of G-CSF Receptor in Cord Blood Neutrophils of Term Newborns
Authors: Gessler Peter , Neu Stefan , Brockmann Yvonne , Speer Christian P.
Published: April 2003

Granulocyte colony-stimulating factor (G-CSF) promotes neutrophil production and enhances neutrophil function. The effects of G-CSF are mediated by binding to its receptor. Since neutrophils are an essential part of the neonatal host defense system, we studied G-CSF receptor expression in neonatal neutrophils. We determined protein and mRNA expression of G-CSF receptor in freshly isolated neutrophils from cord blood of healthy term newborns (n = 16) and of adults (n = 6) as well as the in vitro effect of supplemented recombinant human G-CSF (rhG-CSF) and tumor necrosis factor-α (TNF-α) on G-CSF receptor expression of neutrophils. Expression of G-CSF receptor on the surface of neutrophils of cord blood was significantly lower compared to adults (61 ± 6 vs. 89 ± 2%). G-CSF receptor mRNA transcripts of neutrophils from newborns compared to adults was lower, too (77 ± 14 vs. 152 ± 33%). Neutrophils isolated from cord blood showed a decrease of G-CSF receptor expression within 24 h of culture. Moreover, we were able to show that supplemented rhG-CSF is necessary for maintenance of G-CSF receptor expression. TNF-α, however, down-regulated G-CSF receptor expression. We conclude that low protein and mRNA expression of G-CSF receptor in neutrophils of neonates compared to adults may adversely affect granulopoiesis and neutrophil functions by decreased responsiveness to G-CSF. Furthermore, G-CSF receptor expression on neutrophils was modified not only by G-CSF itself, but also by TNF-α.
Spectral Analysis of Heart Variability in the Newborn Infant
Authors: Rosen H. , Craelius W. , Curcie D. , Hiatt M. , Hegyi T.
Published: April 2003

We investigated the relationship between spectral power and both mean heart rate (HR) and heart rate variability (HRV). Spectral power was calculated using digital heart rate recordings from term infants. Regression analysis revealed a positive correlation between low-frequency (LF) sympathetic power and HR, and a negative correlation between high-frequency (HF) parasympathetic power and HR. HRV correlated positively in all regions of the power spectrum. In awake infants, the contribution of HF power to total power (HF/TP) was significantly decreased. LF power tended to be greater, however, this trend was not statistically significant. By following expected autonomic patterns, the findings of this study confirm that spectral analysis provides a noninvasive method for the assessment of autonomic activity influencing the newborn heart. The correlation between spectral power and HRV can serve as an additional tool in the study of autonomic dysfunction.
A Longitudinal Study of Fetal Growth Variability
Authors: Guihard-Costa A.M. , Droullé P. , Thiebaugeorges O. , Hascoet J.M.
Published: April 2003

The aim of this study was to assess the variability in individual fetal growth rhythms in comparison to averaged standard curves obtained from cross-sectional data. Biparietal diameter (BDP), abdominal transverse diameter (ATD) and femur length (FL) were measured by ultrasonography in 24 normal subjects, and the variance in growth rates determined for four time intervals: 12–26, 26–34, 34–37 and 37–39 weeks gestation. BPD, ATD and FL growths were always linear until 26 weeks with low variances in growth rates. Growth rates decreased thereafter whereas related variances increased significantly with a great diversity in individual growth trajectories. This study questions the relevance of mathematically smoothed curves which lead to an erroneous impression of growth trajectory uniformity when ultrasonography does not seem to be able to predict accurately newborn biometrical characteristics by the end of gestation.
Cortisol and 17-Hydroxyprogesterone Levels in Saliva of Healthy Neonates
Authors: Klug Ingo , Dressendörfer Regina , Strasburger Christian , Kühl Gonne Peter , Reiter Hans-Ludwig , Reich Andrea , Müller Grit , Meyer Kerstin , Kratzsch Jürgen , Kiess Wieland
Published: April 2003

The measurement of cortisol and 17-hydroxyprogesterone (17-OHP) in saliva has become a reliable tool for both the scientist and the clinician for studying adrenal cortical function in the adult and the older child. We have now established in parallel normative data for salivary cortisol and 17-OHP levels in healthy neonates. We have asked whether or not there is a circadian rhythm of cortisol and 17-OHP saliva levels in neonates. Furthermore, we have asked whether salivary hormone levels correlated with auxologic and clinical data and time of sampling. Cortisol and 17-OHP levels in saliva samples from 119 healthy neonates (55 girls, 64 boys) were measured using in-house time-resolved fluorescent immunoassays. Saliva samples were obtained using a saliva collecting tube three times a day on the first or second day of life. Gender and gestational age did not influence salivary cortisol and 17-OHP levels. No significant circadian rhythm of salivary hormone levels was detected in this group of newborns. However, body mass index, arterial cord blood pH and time of saliva sampling significantly influenced salivary hormone levels. In conclusion, measurement of cortisol and 17-OHP in saliva is feasible in healthy neonates. The existence of normative data forms the basis for future studies on pathophysiologic states in the newborn period.
Brain Hemodynamic Changes in Preterm Infants after Maintenance Dose Caffeine and Aminophylline Treatment
Authors: Dani C. , Bertini Giovanna , Reali Maria F. , Tronchin M. , Wiechmann Lisa , Martelli Elena , Rubaltelli F.F.
Published: April 2003

Objective: To investigate the acute effects of low-dose caffeine and aminophylline on cerebral blood flow in preterm infants, using both near-infrared spectroscopy (NIRS) and cerebral Doppler ultrasonography. Methods: Preterm infants with a gestational age of <32 weeks and birth weight of <1,500 g were randomized to receive either caffeine or aminophylline treatment for apnea of prematurity. The study period went from 30 min before to 60 min after the administration of the maintenance dose of pure caffeine (2.5 mg/kg once a day) or aminophylline (1.25 mg/kg twice a day). NIRS was used to measure changes in oxygenated hemoglobin (O2Hb), deoxygenated hemoglobin (HHb), oxidized-reduced cytochrome aa3 (CtOx), and mean cerebral oxygen saturation (SmO2 = O2Hb/total Hb). Changes in cerebral blood volume (ΔCBV) after caffeine or aminophylline administration were calculated. Cerebral blood flow velocity (CBV) in the pericallosal artery was evaluated by cerebral Doppler ultrasounds. Results: Data collected by NIRS and cerebral Doppler ultrasounds did not show significant differences before and after caffeine treatment. We observed a significant increase in O2Hb and HHb concentration and in CBV at 30 min after the infusion of aminophylline, which tended to return to baseline at the end of the study period. Conclusion: Caffeine does not significantly affect brain hemodynamics, while aminophylline induces a significant transient increase in O2Hb and HHb concentration and CBV.
Effects of Oxytocin Treatment in Early Life on Body Weight and Corticosterone in Adult Offspring from ad libitum-Fed and Food-Restricted Rats
Authors: Sohlström Annica , Carlsson Christin , Uvnäs-Moberg Kerstin
Published: April 2003

The aims of this study were: (1) to assess the effects of maternal undernutrition during pregnancy on adult offspring with regard to growth, body composition and plasma levels of glucose, insulin and corticosterone, and (2) to investigate whether oxytocin treatment early in life could ameliorate the adverse effects of food restriction in utero. Pups from ad libitum-fed and food-restricted (60% of ad libitum intake during pregnancy) rats were injected subcutaneously once a day with oxytocin or saline on days 1–14 after birth. At adult age (62 days), male offspring from food-restricted dams had lower body weight, less adipose tissue, lower plasma glucose but higher corticosterone levels, compared to offspring from ad libitum-fed dams. However, oxytocin-treated food-restricted males had higher body weight, higher glucose and lower corticosterone levels compared to their saline-treated counterparts. In conclusion, oxytocin treatment early in life seems to ameliorate some of the adverse effects of food restriction in utero.
Influence of Inhibition of Nitric Oxide Synthesis on Cardiac Function in the Newborn Lamb after Hypoxic-Ischemic Injury
Authors: Dorrepaal Caroline A. , van Bel Frank , Steendijk Paul , Shadid Majidah , van de Velde Enno T. , Baan Jan
Published: April 2003

The aim of the present study was to investigate the effect of immediate post-hypoxic-ischemic (HI) inhibition of nitric oxide synthesis by Nω-nitro-L-arginine (NLA) on cardiac function and reactive oxygen species production. Fifteen newborn lambs were subjected to severe HI. Upon resuscitation 5 received 10 mg NLA/kg, 4 40 mg NLA/kg and 6 a placebo. Left ventricular (LV) contractility, cardiac output (CO), non-protein-bound iron (NPBI), ratio of reduced/oxidized ascorbic acid, α-tocopherol, sulfhydryl groups and malondialdehyde were measured before and 15, 60 and 120 min after resuscitation. There was a significant decrease in CO in all 3 groups at 60 min post-HI (p < 0.05). Reactive oxygen species production was also highest at 60 min post-HI (significantly increased NPBI and decrease in sulfhydryl groups in control lambs; p < 0.05). These results suggest neither a positive nor a negative effect of nitric oxide synthesis inhibition on post-HI myocardial performance but may suggest a positive effect of NLA on reactive oxygen species-mediated post-HI damage.
Surfactant Tracheobronchial Lavage for the Management of a Rabbit Model of Meconium Aspiration Syndrome
Authors: Lam Barbara C.C. , Yeung C.Y. , Fu K.H. , Wong K.Y. , Chan F.L. , Tsoi N.S.
Published: April 2003

We evaluated the effect of tracheobronchial lavage with diluted surfactant solution (bovine lipid extract surfactant, bLES) in a rabbit model of meconium aspiration. All animals were anaesthetized, tracheotomized and given 3–4 ml/kg of 25% slurry of human meconium into the endotracheal tube and mechanically ventilated for 1 h. The animals were then randomly assigned to surfactant lavage (n = 12) with 15 ml/kg of diluted surfactant at a concentration of 5.4 mg phospholipid/ml administered in aliquots of 2 ml; or simple endotracheal suction (control n = 12) when the oxygenation index (OI) was ≥15. Changes in the arterial blood gases and the histomorphological and radiological appearances of the lungs were recorded. The OI and arterial/alveolar oxygen tension (a/A PO2) of the surfactant lavage group improved significantly at 5 min post-treatment, and these improvements were observed throughout the ensuing 4 h of ventilation. There was significantly more solid content recovered by surfactant lavage compared with the control group (p = 0.0001). Radiologically, the post-treatment air space opacification scores of the lavage group were significantly lower compared with the control (p = 0.002). The post-treatment radiographs of the lavage-treated rabbits were rated by the radiologist, who was blinded to the treatment groups, as much improved in 5 and improved in 4, whereas the control rabbits were rated as much worse in 3 and worse in 4. Histological examination showed the lungs of the lavaged rabbits had significantly more normal airway (p < 0.0001), more fields showing completely normal airspace (p = 0.0001) and less fields showing severe overdistension with meconium (p = 0.0005). We concluded that lavage with diluted surfactant solution effectively washed out the meconium, improved gases exchanges, and improved the histological and radiological appearances in the rabbit model of MAS.
Spontaneous Motility in Preterm Infants Treated with Indomethacin
Authors: Bos Arend F. , Venema Ida M.J. , Bergervoet Mariska , Zweens Mar J. , Pratl Barbara , van Eykern Leo A.
Published: April 2003

Objective: To determine in preterm infants with a patent ductus arteriosus (PDA) the effect of indomethacin treatment on spontaneous motor activity. Study Design: Motor activity was assessed from repeated videotape recordings in 32 preterm infants (≤33 weeks gestation). Sixteen infants required indomethacin therapy for treatment of PDA, 16 were control infants, matched for gestational age. Indomethacin (0.2 mg/kg i.v. in 5 min) was given thrice, with an interval of 12 h. One-hour recordings were made immediately before and immediately after the first dose of indomethacin and 24 h later before and after the third dose. The same recording schedule was used for the control infants. The effects of indomethacin on quantity and quality of spontaneous motor activity were examined. Results: A significant reduction in the quantity of several spontaneous movement patterns and an increase in the occurrence of rest periods were found following the first indomethacin administration (p < 0.01). Concerning the quality of general movements, a reduction in the speed was found (p < 0.05). Both effects were not found after the third indomethacin administration. Conclusion: In preterm infants with a PDA, treatment with indomethacin leads to a transient reduction in the quantity of spontaneous movement patterns and to a decrease in the speed of general movements. We recommend a cautious use of bolus indomethacin for the treatment of PDA.
Sublethal Endotoxemia Promotes Pulmonary Cytokine-Induced Neutrophil Chemoattractant Expression and Neutrophil Recruitment but Not Overt Lung Injury in Neonatal Rats
Authors: Tillema Mary S. , Lorenz Kathryn L. , Weiss Marc G. , Dries David J.
Published: April 2003

Gram-negative sepsis and septic shock remain major causes of morbidity and mortality in the newborn. Respiratory failure is a common feature in neonatal sepsis regardless of the presence or absence of associated pneumonia. In adult animal models, cytokine-induced neutrophil chemoattractant (CINC) is a potent chemoattractant for neutrophils and believed to play a role in endotoxin-induced lung injury. We examined this in a neonatal model. Ten-day-old Sprague-Dawley rats were injected with Salmonella enteritidis endotoxin (ETX) 0.03 mg/kg i.p. and sacrificed at baseline, 30 min, 1, 2, 4, 8 and 16 h post-ETX. Blood was collected by cardiac puncture. After bronchoalveolar lavage, lung tissue was collected and evaluated for neutrophil (polymorphonuclear leukocyte) recruitment by myeloperoxidase assay (MPO). Lung CINC expression was measured by Northern blot and ELISA. Peripheral blood leukocytosis was noted at 1 h (p < 0.001) with counts below baseline at 2 and 4 h. Differential counts revealed neutrophilia at 8 h (p < 0.001). MPO revealed pulmonary PMN recruitment peaking at 1 h (p < 0.05) and CINC RNA and protein expression peaked slightly later at 2 h (p < 0.001). No overt lung injury was noted by bronchoalveolar lavage cell counts or by histology. Therefore, pulmonary CINC expression and neutrophil recruitment follows LPS exposure in neonatal rats. This may represent priming of the lung tissue and a secondary event may be necessary for injury to occur
A zinc finger domain gene in the lizard, Calotes versicolor, shows extensive homology with the mammalian ZFX and is expressed embryonically
Authors: Ganesh S. , Choudhary B. , Raman R.
Published: April 2003

A 590-bp long zinc finger domain DNA fragment has been isolated by polymerase chain reaction from the lizard, Calotes versicolor, employing the primers used for amplifying the zinc finger domain of the human Y-chromosomal gene, ZFY. Cloned in pUC18, the fragment, called CvZfa, was sequenced and its expression during development was studied. At the nucleotide and amino acid level CvZfa shows respectively 83% and 90% identity with the human ZFY, but its extent of homology is greater with the ZFX of human (86% at nucleotide and 92% at amino acid level) and the ZFY-like genes of turtle and chick. Similarly its homology with the mouse Zfx and Zfa is much greater than that with Zfy-1 and Zfy-2. It appears that the mammalian ZFX (Zfx) evolved from reptilian ancestors with a considerable degree of conservation, but the ZFX to ZFY divergence within the class mammalia was more rapid. The CvZfa transcripts were seen in all the embryonic stages from which RNA was analysed. The whole mount in situ hybridization with the posteriorly placed mesonephros and the gonadal primordia of 10 to 25 day old embryos showed signal selectively in mesonephros of the 20 and 25 day embryos. There was no signal in the genital ridge. Thus CvZfa may not have a direct role in gonadogenesis of C. versicolor, but the possibility of its inductive role in the formation of adreno-gonadal axis through mesonephros cannot be discounted.
Mobile genetic elements, chiasmata, and the unique organization of beta-heterochromatin
Authors: Holmquist G.P. , Kapitonov V.V. , Jurka J.
Published: April 2003

Beta-heterochromatin in Drosophila and the Syrian hamster share a similar DNA organization, few unique sequences, and scrambled repeats of mobile elements without tandem repetition. DNA in α-heterochromatin is tandemly repetitious, and we now show that the repeat unit can either contain or lack a mobile element. The tandem repeat organization of α-heterochromatin is presumably due to a concertina-like mechanism of unequal exchange between repeat units. Although both heterochromatin types are late replicating and can incorporate mobile retroposons, the sequence distinction between the two heterochromatins appears to be due to a property conferred by chiasmata upon the process of homologous recombination in β-heterochromatin but not in α-heterochromatin. Chiasmata seem to suppress the concertina mechanism of unequal exchange and impart to β-heterochromatin its nontandem, scrambled repeat organization.
Unusual chromosomal organization of telomeric sequences and expeditious karyotypic differentiation in the recently evolved Mus terricolor complex
Authors: Sharma G.G. , Sharma T.
Published: April 2003

The Mus terricolor complex displays a stable homozygous arrangement of autosomal heterochromatin variations in the form of accretion of definitive autosomal short arms among three nonoverlapping populations, in concert with an expeditious evolutionary differentiation into three chromosomal species: M. terricolor I, II, and III. In contrast to the highly conservative M. musculus-like chromosomes in the coexisting sibling species, M. booduga, reshuffling and differentiation of centric heterochromatin has occurred in harmony with a revision of centric configurations, resulting in acrocentric and submetacentric autosomes. The chromosomal distribution of the prevalent vertebrate telomeric sequence (TTAGGG)n was examined by fluorescence in situ hybridization to metaphase cells of M. terricolor I, II, and III. An unusual centric organization of internal telomeric sequences was detected in all the submetacentric and acrocentric autosomes. An auxiliary role of these presumably fragile, recombinogenic telomeric sequences in the evolutionary revision of centric configurations in the terricolor complex is hypothesized.
X-linked sex-reversing genes
Authors: Wachtel S.S.
Published: April 2003

Rearrangement of the X chromosome generates fertile XY females in the wood lemming, and a duplication has been discovered in Xp21 in some XY females in the human. This has enabled identification and mapping of a novel sex-reversing gene in the human, double dosage of which blocks development of the testis. Whether or not the human gene is related to the gene that causes sex reversal in the wood lemming remains to be seen. Yet similar genes must exist in all eutherian mammals, according to Ohno’s Law.
A novel testis-specific gene, SPAG4, whose product interacts specifically with outer dense fiber protein ODF27, maps to human chromosome 20q11.2
Authors: Tarnasky H. , Gill D. , Murthy S. , Shao X. , Demetrick D.J. , van der Hoorn F.A.
Published: April 2003

The product of the spermatid-specific rat SPAG4 gene interacts specifically with the major 27-kDa outer dense fiber protein ODF27 in elongating spermatids. In this study we isolated the human homolog of the spermatid-specific rat SPAG4 gene and mapped it by fluorescence in situ hybridization to chromosome 20q11.2.
FHIT gene transcript alterations occur frequently in myeloproliferative and myelodysplastic diseases
Authors: Luan X. , Ramesh K.H. , Cannizzaro L.A.
Published: April 2003

Twenty-five primary biopsy samples, obtained from patients diagnosed with chronic/acute myeloproliferative disorders, myelodysplastic disorders, in addition to seven cell lines established from patients with leukemias arrested at different stages of myeloid differentiation, were examined for alterations in an alternatively spliced form of the FHIT gene. Transcript alterations of this gene were detected in 80% of the primary biopsies and in two of the cell lines. Reverse transcription PCR (RT-PCR) detected deletions of one or more specific exons in the translated or untranslated portion of the FHIT gene. Point mutations in exons 3, 4, and 5 of the FHIT gene were also detected in 4 patients by single stranded conformational PCR analysis. Transcript alterations were detected in all primary hematopoietic samples which contained chromosome abnormalities, as well as, in hematopoietic disorders which did not show alterations of the 3p14 region. These findings suggest FHIT gene involvement in the transformation of hematopoietic stem cells to leukemia.
Physical mapping of the t(12;22) translocation breakpoints in a family with a complex type of 3/3′/4 synpolydactyly
Authors: Debeer P. , Schoenmakers E.F.P.M. , Thoelen R. , Fryns, J.-P. , van de Ven W.J.M.
Published: April 2003

We previously reported clinical and radiological findings in a Belgian family with a complex type of synpolydactyly associated with metacarpal and metatarsal synostoses, cosegregating with a balanced t(12;22). Recently, expansions of a polyalanine stretch within the first exon of the HOXD13 gene, which resides on chromosome 2q31, have been shown to cause synpolydactyly (SPD). Using exon amplification followed by direct sequencing, we were able to exclude the direct involvement of the HOXD13 gene in this family. As a first step toward the positional cloning of a candidate disease gene on chromosome 12 and/or 22 responsible for the type of complex synpolydactyly observed in this family, we report here the construction of a somatic cell hybrid retaining only the der(22) of the t(12;22)(p11.3;q13.3). STS content mapping and FISH experiments allowed us to position the chromosomal breakpoints between markers D12S1596 and D12S1034 on chromosome 12 and markers N73F4 and D22S158 on chromosome 22.
The cytogenetic and molecular characterization of benign and malignant soft tissue tumors
Authors: Sreekantaiah C.
Published: April 2003

Cytogenetic analyses of benign and malignant soft tissue tumors have led to the description of recurrent, specific, and even pathognomonic chromosomal translocations and/or other rearrangements in most types of soft tissue tumors. The consistent karyotypic rearrangements have provided critical diagnostic information in this group of neoplasms that often presents significant diagnostic challenges to the clinician and the pathologist. These findings have also been instrumental in the characterization of the abnormalities at the molecular level. Novel genes have been isolated from the translocation junctions and the mechanisms of their deregulation identified. This has increased our understanding of the histogenesis of these tumors, paved the way for the molecular diagnosis of many sarcomas, aided in directing therapy, and also provided important prognostic information.
Genomic organization of the bovine aromatase encoding gene and a homologous pseudogene as revealed by DNA fiber FISH
Authors: Brunner R.M. , Goldammer T. , Fürbass R. , Vanselow J. , Schwerin M.
Published: April 2003

In cattle, the CYP19 locus comprises the aromatase cytochrome P450-encoding gene (CYP19) and a homologous pseudogene (CYP19P1). It has been assigned to chromosome region 10q26. Cloning of genomic DNA revealed that the CYP19 gene covers more than 56 kb. Its precise extent is still unknown because the DNA spanning the untranslated first exon 1.1 and the coding region (exons 2 to 10) have not been isolated. Furthermore, the chromosome arrangement of closely linked CYP19 and CYP19P1 was also not clear. To establish a high resolution physical map of the entire CYP19 locus, fluorescence in situ hybridization to extended bovine genomic DNA fibers (fiber FISH) was performed. The results demonstrate (1) that the clone containing exon 1.1 is located about 19 kb upstream from the CYP19 coding region. (2) Within the chromosome region 10q26 CYP19 and CYP19P1 are arranged “tail-to-head”, being separated by a distance of about 24 kb between the labeled clones. (3) The physical size of the bovine CYP19 locus amounts to a minimum of 130 kb.
Cloning of TCFL5 encoding a novel human basic helix-loop-helix motif protein that is specifically expressed in primary spermatocytes at the pachytene stage
Authors: Maruyama O. , Nishimori H. , Katagiri T. , Miki Y. , Ueno A. , Nakamura Y.
Published: April 2003

We have isolated a novel human gene that is expressed specifically in primary spermatocytes in the testis. The cDNA contains an open reading frame of 1356 bp, encoding a 452-amino-acid protein that includes a basic Helix-Loop-Helix (bHLH) motif. The gene, which was mapped to chromosome region 20q13.3→qter by fluorescence in situ hybridization, consists of six exons and spans approximately 24 kb of genomic DNA. Immunohistochemical staining located the gene product exclusively in cell nuclei of primary spermatocytes at the pachytene stage, but not in those at the leptonema stage. We named this gene TCFL5 (transcription factor-like 5, basic helix-loop-helix). The cell-type and stage-specific expression of TCFL5 indicates that this protein may function in a crucial role in spermatogenesis as a transcription factor by regulating cell proliferation or differentiation of cells through binding to a specific DNA sequence like other bHLH molecules.
Linkage mapping of Lims1l, the murine homolog of the human LIM domain gene PINCH, to mouse Chromosome 10
Authors: Abu-Hayyeh, S. , Eddleston J. , Murdoch J.N. , Tham M. , Copp A.J. , Stanier P.
Published: April 2003

The human LIM domain gene LIMS1 was used to identify a mouse homolog. The resulting mouse sequence was used to identify a polymorphism by SSCP analysis. Linkage studies performed in the EUCIB backcross placed Lims1l on the proximal portion of mouse Chromosome 10. This localisation makes it an interesting candidate for the deafness mutant, waltzer (v).
Structure and refinement of the physical mapping of the γ-glutamylcysteine ligase regulatory subunit (GLCLR) gene to chromosome 1p22.1 within the critically deleted region of human malignant mesothelioma
Authors: Rozet J.-M. , Gerber S. , Perrault I. , Calvas P. , Souied E. , Châtelin S. , Viegas-Péquignot E. , Molina-Gomez D. , Munnich A. , Kaplan J.
Published: April 2003

Abstract. Glutathione is a ubiquitous antioxidant in mammalian tissues. The first step of its synthesis is catalyzed by the glutamate-cysteine ligase (GLCL) which consists of a heavy, catalytic subunit and a light, regulatory subunit (GLCLR). Previous genetic analyses have revealed frequent losses of chromosome 1p22→p21 in human malignant mesothelioma and the shortest region of overlapping deletions has been narrowed between the two loci D1S435 and D1S236. An expressed sequence tag of the GLCLR gene was found within a YAC contig encompassing the same interval and was therefore considered as a good candidate gene for predisposition to human mesothelioma. We report here the characterization of the genomic structure of the GLCLR gene and refine its physical mapping to chromosome 1p22.1.
ZOO-FISH and R-banding reveal extensive conservation of human chromosome regions in euchromatic regions of river buffalo chromosomes
Authors: Iannuzzi L. , Di Meo G.P. , Perucatti A. , Bardaro T.
Published: April 2003

Commercially available human chromosome (HSA) painting probes were hybridized on river buffalo (Bubalus bubalis, 2n = 50) chromosomes by using FISH and R-banding techniques. Clear hybridization FITC-signals revealed extensive conservation of human chromosome regions in this species and demonstrated that human chromosome probes primarily paint euchromatic regions (R-bands). The present results are discussed in the light of previous gene mapping data obtained in river buffalo and ZOO-FISH data in cattle, and in relation to the standard bovine chromosome nomenclatures. In particular, HSA 8, HSA 10, HSA 11, and HSA 16+7 paint, respectively, BBU 1p, BBU 4p, BBU 5p, and BBU 24, which are homoeologous, respectively, to cattle chromosomes 25, 28, 29 and 27. Thus, these river buffalo chromosome arms can serve as markers to resolve discrepancies in the nomenclature of cattle and related species.
A serpin gene cluster on human chromosome 6p25 contains PI6, PI9 and ELANH2 which have a common structure almost identical to the 18q21 ovalbumin serpin genes
Authors: Sun J. , Stephens R. , Mirza G. , Kanai H. , Ragoussis J. , Bird P.I.
Published: April 2003

The human genes encoding the “ovalbumin” subgroup of closely related serine proteinase inhibitors (serpins) are located at 18q21.3 and 6p25. Those at 6p25 include proteinase inhibitor 6 (PI-6; gene symbol PI6), proteinase inhibitor 9 (PI-9; gene symbol PI9) and monocyte neutrophil elastase inhibitor (M/NEI; gene symbol ELANH2). Here we describe the fine mapping of these genes to a 200-kb region of chromosome 6 that includes the markers WI-8835 and D6S1338, and the establishment of the gene order: tel–PI6–PI9–ELANH2–cen. PI6 and ELANH2 are transcribed towards the telomere, and structural analysis shows that PI6 and PI9 are organized identically, having seven exons and six introns. PI6 and PI9 are almost identical in structure to the ovalbumin serpin genes at 18q21.3. The 18q21.3 genes have an extra exon and intron, otherwise all the other exon/intron boundaries are conserved between the two groups. These results represent the first detailed map of the chromosome 6 serpin gene cluster, and demonstrate that although they are very closely related, the 6p25 and 18q21→q23 ovalbumin serpin genes form two structurally distinct groups. These findings do not support a previously proposed model for evolution of the clusters which invoked an inter-chromosomal duplication of the entire 6p25 group to 18q21.3.
Identification, genomic organization, and alternative splicing of KNSL3, a novel human gene encoding a kinesin-like protein
Authors: Okamoto S. , Matsushima M. , Nakamura Y.
Published: April 2003

Proteins of the kinesin superfamily are microtubule-dependent molecular motors that play important roles in organelle transport and cell division. Through genomic sequencing and use of the RT-PCR technique, we have identified and characterized KNSL3 (kinesin-like 3), a novel member of the kinesin-like protein family in humans. We determined its genomic organization and detected four alternatively spliced transcripts. KNSL3 was expressed ubiquitously, but sizes and relative amounts of the major products were different in each of the tissues examined. Alternative splicing, along with the multiplicity of genes in the molecular family that includes KNSL3, produce diversity among the C-terminal ends of kinesins. These observations may contribute to an understanding of the specificity of different kinesins with respect to organelle binding.
Two extended arrays of a satellite DNA sequence at the centromere and at the short-arm telomere of Chinese hamster chromosome 5
Authors: Faravelli M. , Moralli D. , Bertoni L. , Attolini C. , Chernova O. , Raimondi E. , Giulotto E.
Published: April 2003

We have cloned a Chinese hamster chromosome-specific repeated sequence (SatCH5). This satellite is composed of a 33-bp unit organized in two extended tandem arrays. It is localized at the centromere and at the short-arm subtelomere of chromosome 5. Altogether, SatCH5 covers about 1–2 Mb per diploid genome and is not present in other species, including the Syrian hamster and mouse. Since it is known in the Chinese hamster and numerous other vertebrate species that telomeric (TTAGGG)n repeats are localized at the centromeres of several chromosomes, we studied the localization of SatCH5 relative to (TTAGGG)n sequences. Using two-color fluorescence in situ hybridization on stretched chromosomes and on DNA fibers, we have shown that at the centromere of chromosome 5 SatCH5 and the (TTAGGG)n arrays are contiguous. SatCH5 is the first chromosome-specific repetitive sequence located at both the pericentromeric and subtelomeric regions of the same chromosome.
Genomic structure, sequence, and refined mapping of the human intersectin gene (ITSN), which encompasses 250 kb on chromosome 21q22.1→q22.2
Authors: Guipponi M. , Scott H.S. , Hattori M. , Ishii K. , Sakaki Y. , Antonarakis S.E.
Published: April 2003

The ubiquitously expressed and brain-specific human intersectin (ITSN) isoforms are scaffold proteins probably involved in general endocytosis and synaptic vesicle recycling, respectively. Here, analysis of 21q22.1→q22.2 genomic sequence revealed that ITSN consists of 41 exons spanning ∼250 kb and maps between GART and D21S325. The probable function of the ITSN isoforms and mapping position of ITSN suggest that disproportionate expression of this gene may be implicated in the phenotypic characteristics of Down syndrome.
Cloning of a novel gene (ING1L) homologous to ING1, a candidate tumor suppressor
Authors: Shimada Y. , Saito A. , Suzuki M. , Takahashi E. , Horie M.
Published: April 2003

The ING1 gene encodes p33ING1, a putative tumor suppressor for neuroblastomas and breast cancers, which has been shown to cooperate with p53 in controlling cell proliferation. We have isolated a novel human gene, ING1L, that potentially encodes a PHD-type zinc-finger protein highly homologous to p33ING1. Fluorescence in situ hybridization and radiation-hybrid analyses assigned ING1L to human chromosome 4. Both ING1 and ING1L are expressed in a variety of human tissues, but we found ING1L expression to be significantly more pronounced in tumors from several colon-cancer patients than in normal colon tissues excised at the same surgical sites. Although the significance of this observation with respect to carcinogenesis remains to be established, the data suggest that ING1L might be involved in colon cancers through interference with signal(s) transmitted through p53 and p33ING1.
Sperm analysis in a subfertile male with a Y;16 translocation, using four-color FISH
Authors: Giltay J.C. , Kastrop P.M.M. , Tiemessen C.H.J. , van Inzen W.G. , Scheres J.M.J.C. , Pearson P.L.
Published: April 2003

Sperm analysis was performed in a male with oligoasthenoteratozoospermia (OAT) and a reciprocal t(Y;16) (q11.21;q24), using four-color FISH. Intracytoplasmic sperm injection (ICSI) treatment in this patient had resulted in the birth of one chromosomally balanced and two chromosomally normal children. To assess the risk of having a chromosomally unbalanced conception after ICSI, morphologically normal spermatozoa were studied with a set of probes allowing detection of all segregation variants. There were 51% normal or balanced sperm cells. The fraction of sperm products resulting from alternate and adjacent I segregation was 87%, 12% were products of 3:1 disjunction, and the other 1% had other types of aneuploidy. If morphologically abnormal cells were also included in the FISH analysis, nearly 90% of all the spermatozoa were unbalanced. We conclude that although the majority of males with a Y/autosome translocation are infertile due to azoospermia, our patient produces sufficient morphologically and chromosomally normal spermatozoa to have chromosomally normal or balanced offspring after ICSI. Assuming that ICSI with an unbalanced spermatozoon from this patient would result in a nonviable embryo in many cases, the combination of in vitro and subsequent in vivo selection probably results in a risk of unbalanced offspring of much less than 50%. Hence, FISH studies on the sperm of translocation carriers are useful for estimating the risk of having unbalanced offspring after ICSI and in understanding the mechanisms underlying infertility in such carriers.
A complete set of repeat-depleted, PCR-amplifiable, human chromosome-specific painting probes
Authors: Bolzer A. , Craig J.M. , Cremer T. , Speicher M.R.
Published: April 2003

We describe the generation of a complete set of human chromosome-specific painting probes depleted in repetitive sequences. These probes yield highly specific signals when hybridized without the addition of a blocking agent, such as Cot-1 DNA, and without probe preannealing prior to hybridization. Fluorescent intensities and signal-to-background ratios for these probes are comparable to those of untreated probes hybridized with Cot-1 DNA. We demonstrate the suitability of these probes for applications with very complex probe sets, such as multiplex-FISH.
Chromosome-type aberrations induced in chromosome 9 after treatment of human peripheral blood lymphocytes with mitomycin C at the G₀ phase
Authors: Kusakabe H. , Takahashi T. , Tanaka N.
Published: April 2003

To determine the fate of chromosome aberrations induced primarily by clastogenic chemicals, aberrations of chromosome 9 in cultured human peripheral blood lymphocytes were analyzed after exposure to mitomycin C (MMC) at G₀ phase. Chromosome 9 painting by fluorescence in situ hybridization revealed that the translocation of 9p or 9q to another chromosome and the centric fragment representing the entire length of 9p were characteristically generated from chromatid-type aberrations involving the centromeric region of chromosome 9. These changes were not observed at 48 h after culture initiation, but persistently appeared at later stages (72–120 h postinitiation). Induction of centric fragments of 9p and micronuclei without the α satellite DNA of chromosome 9 suggested that most of the breaks were induced near the α satellite DNA locus on 9q. Modified patterns of chromosome 9 aberrations were also observed, being related to the copy number of the short or long arm of the chromosome. Such unbalanced karyotypes could remain in the lymphocyte genome over further cell divisions for at least 120 h after culture initiation, indicating that these aberrant cells can survive and that they could pose a health risk.
Mapping and complex expression pattern of the human NPAP60L nucleoporin gene
Authors: Trichet V. , Shkolny D. , Dunham I. , Beare D. , McDermid H.E.
Published: April 2003

From a clone mapping to human chromosome 22q13.3, we have identified NPAP60L, the human homolog of the rat nuclear pore-associated protein gene, Npap60. The expression pattern of the human copy is much more complex that that of the rat, although conservation of the potential specific function of NPAP60L in male germ cells can be seen for one of the five transcripts. The exon-intron organization of the NPAP60L gene shows the presence of at least three alternate 3′ ends, and Northern analysis indicates the possible presence of alternate 5′ ends. Somatic cell hybrid mapping revealed additional related copies of NPAP60L on human chromosomes 5, 6, and 14, although it is not known if these are functional genes.