bluuebunny/crossref_metadata_2025 huggingface dataset:
Lectin Binding in the Human Umbilical Cord from Normally Grown Pregnancies and from Pregnancies Complicated by Intra-Uterine Growth Retardation with Absent or Reversed Diastolic Flow
Authors: Sgambati E. , Brizzi E. , Marini M. , Gheri Bryk S. , Gheri G.
Published: August 2003

The content and the distribution of the glycoconjugate sugar residues were investigated in the human umbilical cord from normally grown pregnancies and from pregnancies complicated by intra-uterine growth retardation with absent or reversed flow in the umbilical artery. The gestational age range was 29–41 weeks. For this purpose, a battery of eight horseradish peroxidase conjugated lectins was used (ConA, WGA, PNA, SBA, DBA, LTA, UEA I, and OOA; see text). Our data showed that in both study groups ConA, WGA and PNA (after neuraminidase digestion) and LTA, UEA I, and OOA reacted with Wharton’s jelly and vessels of the umbilical cord from the first weeks of gestation considered. SBA reactivity was observed only in the amniotic epithelial cells of the jelly and in the endothelial cells of the vessels. The reactivity with the most part of the lectins decreased or was lacking in the various components of the umbilical cord between 39 and 41 weeks of gestation in the normal group and between 36 and 37 weeks of gestation in the pathological one. Moreover, the intensity of reactivity with ConA, WGA and PNA (after neuraminidase digestion) and LTA, UEA I, and OOA in the normal group and with PNA (before and after neuraminidase digestion) and LTA and OOA in the pathological one was stronger in the peripheral portion of the jelly with respect to the adventitial one. The reactivity with ConA and UEA I was stronger in the vein than in the arteries in both study groups. Some hypotheses are reported on the role played by the sugar residues in the normal and pathological umbilical cords.
Analysis of DNA Damage Using the Comet Assay in Infants Fed Cow’s Milk
Authors: Dündaröz Ruşen , Ulucan Hakan , Aydın Halil İbrahim , Güngör Tayfun , Baltacı Volkan , Denli Metin , Sanisoğlu Yavuz
Published: August 2003

It has been hypothesized that non-human milk feeding may increase the risk for cancer or for a specific cancer or group of cancers as well as the risk for diseases such as type-1 diabetes mellitus and Crohn’s disease. Regarding DNA damage leading to cancer development in the absence of human milk protection, a comparison between infants fed human milk and cow’s milk has been performed. Each group consisted of 35 infants, whose ages ranged from 9 to 12 months. The level of DNA damage in the peripheral blood lymphocytes of infants has been studied by the comet assay. A significant increase has been found in the number of limited DNA-damaged (p < 0.001) and extensive DNA-damaged (p < 0.001) cells of infants fed cow’s milk. To our knowledge, this is the first study using the comet assay on infants not breast-fed. Supporting our previous SCE study, these results suggest that there is some level of DNA damage in the lymphocytes of infants not breast-fed and this may lead to malignancy in childhood or later in life.
Sequential Postnatal Changes in Erythrocyte Glutathione and Sulfhydryl Content: A Possible Adaptational Response to the Extrauterine Environment
Authors: Jean-Baptiste Dominique , Rudolph Nathan
Published: August 2003

Glutathione is a key factor in intracellular antioxidant defense mechanisms. We postulated that it might modulate the adaptational response of the fetus to a higher oxygen environment after birth. Therefore, we determined total glutathione (GSH + GSSG) and non-protein sulfhydryl (–SH) content in cord blood RBC hemolysates, and sequential values in blood from the same infants for up to 72 h after birth. We noted a consistent rapid decline in total glutathione content by 24 h in both term (–18%) and preterm (–24%) infants, with subsequent stabilization of levels in term infants. However, erythrocyte (–SH) content (measured only in term babies) remained stable and unchanged from levels in cord blood, indicating the presence of (–SH) in one or more other moieties in addition to GSH. We speculate on the possibility of a role for GSH as a (–SH) donor in modulating postnatal fetal adaptation.
Long-Term Effects of Antepartum Bed Rest on Offspring
Authors: Bellieni Carlo Valerio , Bagnoli Franco , Perrone Serafina , Caparelli Ninetta , Cordelli Duccio Maria , Melissa Bernardino , Buonocore Giuseppe
Published: August 2003

We studied the children of mothers who were confined to bed during pregnancy for more than 15 consecutive days (mean 3.69 months) in the years 1986–1990 (bed rest offspring; BRO). We studied 86 children: 43 BRO and 43 control children. Data were obtained by means of a 20-item questionnaire filled in by the mothers. The BRO group differed from the control group in incidence of allergies (p = 0.005), motion sickness (p = 0.03), and need to be rocked to fall asleep (p = 0.01). More BRO born at term than controls played musical instruments later in life. Two possible explanations for more allergies among the BRO group are the use of beta-stimulating drugs against premature delivery and the effects of prolonged stress on the developing immune system. Understimulation of the developing vestibular system during gestation may affect some aspects of its development and may explain the higher incidence of motion sickness and need for vigorous rocking in BRO.
Enteral Crude Red Kidney Bean (Phaseolus vulgaris) Lectin – Phytohemagglutinin – Induces Maturational Changes in the Enterocyte Membrane Proteins of Suckling Rats
Authors: Kruszewska Danuta , Kiela Pawel , Ljungh Åsa , Erlwanger Kennedy H. , Weström Björn R. , Linderoth Ann , Pierzynowski Stefan G.
Published: August 2003

This study aimed to investigate the effect of enterally administered crude red kidney bean (Phaseolus vulgaris) lectin, PHA, on the expression of brush-border membrane vesicle (BBMV) proteins, in particular Na+/H+ exchangers (NHEs), in the small intestine of suckling rats. Gavage of PHA to 14-day-old rats for 3 days resulted in altered protein/glycoprotein patterns as analyzed by SDS-PAGE. Immunoblots demonstrated the appearance of two 71- and 27-kD protein bands indicative for NHE3 – one of the NHE isoforms – and PHA, respectively. PHA treatment also resulted in an augmented uptake of 22Na+ by the BBMV indicating an increase in NHE activity. Overall, the data suggests that enteral PHA exposure may induce maturational changes in enterocyte membrane proteins in young rats. In view of these findings, an investigation into the addition of PHA to infant formulas and weaning diets is warranted.
Alterations in Cerebral Mitochondria during Acute Hypoglycemia
Authors: Ballesteros Juan R. , Mishra Om P. , McGowan Jane E.
Published: August 2003

To test the hypothesis that acute hypoglycemia leads to free radical induced alterations in cerebral mitochondria, newborn piglets were subjected to 2 h of insulin-induced hypoglycemia (blood glucose 1 mmol/l). The effects of free radicals were determined in cerebral cortical synaptosomes, mitochondria, and neuronal nuclei by measuring membrane lipid peroxidation. Fragmentation of nuclear and mitochondrial DNA was also examined. Lipid peroxidation was significantly increased in hypoglycemic mitochondrial membranes as compared to controls, but no increase in peroxidation in hypoglycemic synaptosomal or nuclear membranes was observed. An increase in low molecular weight DNA fragments was observed only in mitochondrial DNA from hypoglycemic piglets. We speculate that alteration of cerebral mitochondria due to increased free radical production is one of the early events in the pathogenesis of hypoglycemic brain injury.
Postischemic Hyperthermia Induced Caspase-3 Activation in the Newborn Rat Brain after Hypoxia-Ischemia and Exacerbated the Brain Damage
Authors: Fukuda Hirotsugu , Tomimatsu Takuji , Kanagawa Takeshi , Mu Junwu , Kohzuki Masatomo , Shimoya Koichiro , Hosono Takayoshi , Kanzaki Toru , Murata Yuji
Published: August 2003

The effects of postischemic hyperthermia were investigated in the newborn rat brain after hypoxia-ischemia (HI). Seven-day-old rats were subjected to left carotid artery ligation followed by 8% oxygen for 30 min, and divided into a hyperthermia group (rectal temperature at 39°C for 6 h) and a normothermia group. Hyperthermia resulted in an approximately 5-fold increase in activated caspase-3 24 h after HI when compared with the normothermia group, and gross loss of brain tissue was observed only in the hyperthermia group at 7 and 30 days after HI. Our results show that postischemic hyperthermia exacerbates HI injury in immature brains, and that the mechanism is strongly associated with activation of an apoptotic pathway.
Vascular Reactivity of Pulmonary Arteries from Premature Lambs Subjected to Liquid Ventilation
Authors: Villamor Eduardo , Degraeuwe Pieter L.J. , De Mey Jo G.R. , Blanco Carlos E.
Published: August 2003

This study aimed to evaluate the effect of tidal liquid ventilation (TLV) and liquid assisted high frequency oscillatory ventilation (LA-HFOV) on the contractile and relaxing properties of fetal ovine pulmonary arteries. 0.85 term lambs were subjected, after surfactant instillation, to 5 h of TLV or LA-HFOV (5 ml·kg–1 perfluorocarbon liquid). After euthanasia of the animals, intrapulmonary arteries (fourth branch) were dissected and mounted in a myograph for isometric tension recording. Arteries from unventilated age-matched fetuses were also studied. The contractions induced by K+, and the thromboxane A2 mimetic U46619 were not significantly different in the TLV and the LA-HFOV groups. Acetylcholine-induced relaxation was absent after TLV and LA-HFOV but present in unventilated animals. Sodium nitroprusside-induced relaxation was also similar after TLV and LA-HFOV, but reduced when the two groups were compared with unventilated fetuses. We conclude that after TLV and LA-HFOV the pulmonary arterial responses to receptor- and non receptor-mediated contraction and to endothelium-dependent and -independent relaxation were similar.
Increased Wave Latency in Auditory Brainstem Response to Anemia in Newborn and Adult Sheep
Authors: Quiñónez Rafael E. , Widness John A.
Published: August 2003

The objective of the present study was to determine whether phlebotomy-induced anemia in newborn lambs and adult sheep results in prolonged auditory brainstem response (ABR) wave latencies, and if found, whether developmental differences exist between the two groups. Among newborn lambs, hemoglobin (Hb) levels were reduced from 100–120 to 30–50 g/l over a 3-day period. Over the next 2 days, serial red blood cell transfusions were administered to restore Hb to pre-phlebotomy levels. In adult sheep, comparable levels of phlebotomy-induced anemia were achieved within a 24-hour period, after which Hb levels returned to pre-phlebotomy levels. During the induction of anemia and its reversal, a significant (p < 0.05) increase in ABR latency followed by a return to pre-anemia ABR latencies that was most prominent in the peripheral auditory pathway was observed in both newborn and adult sheep as Hb levels fell below 50 g/l (p < 0.05). There was no difference between adults and newborns in ABR latency relative to the Hb level at which ABR wave latencies increased. We speculate that ABR wave latency prolongation might serve as an indicator of the need for red blood cell transfusion.
Neurogenesis in the Hippocampus of an Adult Marsupial
Authors: Harman Alison , Meyer Peter , Ahmat Alicia
Published: August 2003

In the adult eutherian brain, stem cells in the dentate gyrus continually divide throughout adult life and into old age producing new granule cells. However, it was not known whether this is also the case for marsupials. Previously, in fact, it was thought that marsupials did not have continued neurogenesis in the mature brain. Here we examined neurogenesis in the adult brain of a small marsupial, the fat-tailed dunnart, using 3H-thymidine to label newly generated cells. We showed that neurogenesis takes place in the adult dentate gyrus along its inner margin, as seen in eutherian mammals. Control animals had similar numbers of labeled cells 24 h and 1 month after 3H-thymidine injection. An enriched environment resulted in similar numbers of cells being generated as controls. However, there was a significant decline in the number of labeled cells one month later. Stress and old age resulted in significantly lower numbers of new cells being generated. In immunohistochemically treated control brains, 3H-thymidine-labeled cells at the early stage were sometimes GFAP positive, were not calbindin positive at either stage examined and at the later stage were PSA-NCAM positive. We hypothesize that, as seen in eutherian mammals, the new cells progressed from being GFAP positive at stem cell stage to PSA-NCAM positive during outgrowth of mossy fibers 1 month later, to calbindin positive when mature. It is possible that maturity of these cells was not reached by 1 month as marsupials have a slower metabolic rate and this species also undergoes daily periods of torpor.
Brain-Body Size Relations in Grasshopper Mice
Authors: Mann Michael D. , Towe Arnold L.
Published: August 2003

Cranial volumes were measured on museum specimens of two species of grasshopper mice, Onychomys leucogaster brevicaudus and Onychomys torridus pulcher. These were compared with body weights and body lengths, as recorded on the museum tags. On average, females were heavier and slightly longer than males, but they had smaller cranial volumes by about 3%. Statistically, the differences were marginally significant (with no difference in body length for one species), but the pattern of differences was the same in both species, and therefore probably real. This casts suspicion on the common notion that a larger body size requires a larger brain. It also questions the real meaning of encephalization quotients.
Parvalbumin-, Calbindin-, and Calretinin-Immunoreactive Neurons in the Prefrontal Cortex of the Owl Monkey (Aotus trivirgatus): A Standardized Quantitative Comparison with Sensory and Motor Areas
Authors: Elston Guy N. , González-Albo Maria C.
Published: August 2003

Recent studies have revealed regional variation in the density and distribution of inhibitory neurons in different cortical areas, which are thought to reflect area-specific specializations in cortical circuitry. However, there are as yet few standardized quantitative data regarding how the inhibitory circuitry in prefrontal cortex (PFC), which is thought to be involved in executive functions such as cognition, emotion and decision making, compares to that in other cortical areas. Here we used immunohistochemical techniques to determine the density and distribution of parvalbumin (PV)-, calbindin (CB)-, and calretinin (CR)-immunoreactive (ir) neurons and axon terminals in the dorsolateral and orbital PFC of the owl monkey (Aotus trivirgatus), and compared them directly with data obtained using the same techniques in 11 different visual, somatosensory and motor areas. We found marked differences in the density of PV-ir, CB-ir, and CR-ir interneurons in several cortical areas. One hundred and twenty eight of all 234 possible between-area pair-wise comparisons were significantly different. The density of specific subpopulations of these cells also varied among cortical areas, as did the density of axon terminals. Comparison of PFC with other cortical areas revealed that 40 of all 66 possible statistical comparisons of the density of PV-ir, CB-ir, and CR-ir cells were significantly different. We also found evidence for heterogeneity in the pattern of labeling of PV-ir, CB-ir, and CR-ir cells and axon terminals between the dorsolateral and orbital subdivisions of PFC. These data are likely to reflect basic differences in interneuron circuitry, which are likely to influence inhibitory function in the cortex.
Spatio-Temporal Distribution of Microglia/Macrophages during Regeneration in the Cerebellum of Adult Teleost Fish, Apteronotus leptorhynchus: A Quantitative Analysis
Authors: Zupanc Günther K.H. , Clint Sorcha C. , Takimoto Noriko , Hughes Alun T.L. , Wellbrock Ursula M. , Meissner Daniela
Published: August 2003

In contrast to mammals, adult teleost fish exhibit an enormous capacity to replace damaged neurons with newly generated ones after injuries in the central nervous system. In the present study, the role of microglia/macrophages, identified by tomato lectin binding, was examined in this process of neuronal regeneration in the corpus cerebelli of the teleost fish Apteronotus leptorhynchus. In the intact corpus cerebelli, or after short survival times following application of a mechanical lesion to this cerebellar subdivision, microglia/macrophages were virtually absent. Conversely, approximately 3 days after application of the lesion, the areal density of microglia/macrophages started to increase at and near the lesion site in the ipsilateral hemisphere, as well as in the contralateral hemisphere, and reached maximum levels at approximately 10 days post lesion. The density remained elevated until it reached background levels approximately one month after the injury. By comparing the time course of the appearance of microglia/macrophages with that of other regenerative events occurring within the first few weeks of wound healing in this model system, we hypothesize that one possible function of microglia/macrophages might be to remove debris of cells that have undergone apoptotic cell death at the lesion site.
Expression of Nitric Oxide Synthase in the Preoptic-Hypothalamo-Hypophyseal System of the Teleost Oreochromis niloticus
Authors: Bordieri Loredana , Persichini Tiziana , Venturini Giorgio , Cioni Carla
Published: August 2003

In the present study, we have analyzed the expression of nitric oxide synthase (NOS) in the preoptic-hypothalamo-hypophyseal system of the teleost Oreochromis niloticus. The assay for enzyme activity demonstrated that a constitutive NOS activity is present both in soluble and particulate fractions of the homogenates of diencephalons. Western blot analysis using an antibody against the N-terminus of human nNOS revealed two bands both in the supernatant and in the pellet. One band co-migrates at approximately 150 kDa with that detected in the rat cerebellum homogenates and presumably corresponds to neuronal NOS (nNOS) of mammals. The additional band, which migrates at approximately 180 kDa, might be attributed to an alternatively spliced nNOS isoform. Using NADPH diaphorase (NADPHd) histochemistry in combination with NOS immunohistochemistry, nNOS expression has been detected in preoptic nuclei, hypophysiotrophic nuclei of the ventral hypothalamus, and the pituitary gland. Various degrees of dissociation of NADPHd activity and nNOS immunoreactivity have been detected that could be attributed to the expression of different subtypes of nNOS in the preoptic/hypothalamo/hypophysial system of tilapia. In this paper, we also investigated the colocalization of nNOS with arginine-vasotocin (AVT) by means of immunolabeling of consecutive sections. Results suggest that NO may be colocalized with AVT in a subpopulation of neurosecretory neurons. Present findings suggest that nitric oxide (NO) is implicated in the modulation of hormone release in teleosts in a similar way to mammals.
Queen Discrimination Ability of Ant Workers (Camponotus japonicus) Coincides with Brain Maturation
Authors: Hara Kenji
Published: August 2003

Discrimination behavior is a fundamental feature of social insects, and the discrimination ability allows the individual to be integrated into the colony. To address the brain functions associated with this task, which underlies intraspecific communication, I examined the ability to discriminate queens in young workers of the ant Camponotus japonicus together with a histological analysis of their brains. Workers raised in a foster colony were tested with respect to their ability to discriminate the foster queen from an unfamiliar queen. During the first 3 days after eclosing, some young workers made erroneous decisions whereas after 3–4 days the decisions were always correct. To assess brain maturation, mitotic activity in the brains of workers was individually analyzed using BrdU injection immediately after the behavioral test. Neurogenesis appeared to be complete at eclosion, but in 3 of 8 young workers that showed incorrect behavioral decisions, mitotic activity was observed in the antennal lobe. In contrast, no mitotic activity was observed in the brains of young workers who correctly discriminated their foster queens from alien queens. These results suggest that in newly emerged ant workers the ability to discriminate queens coincides with the completion of cell proliferation in the brain.
Estradiol Control of Expression and Levels of Estradiol-Binding Proteins in the Medial Preoptic Area, Medial Hypothalamus and Pituitary
Authors: Gao Guang , Herbert Zsöfia , Kong Jing , Gabrielson Nathan , Mautz Alan , Wu Dongming , Jirikowski Gustav F. , Caldwell Jack D.
Published: August 2003

The brains of mammals have at least three estradiol-binding proteins: estradiol receptor-α (ERα), ERβ, and sex hormone-binding globulin (SHBG). In this study we compare the effects of estradiol treatment on the expression of mRNA for these three estradiol-binding proteins in two reproductively important brain areas, the medial preoptic area-anterior hypothalamus (MPOA-AH) and medial hypothalamus (MH) as well as in the hippocampus in ovariectomized rats, using the reverse transcriptase-polymerase chain reaction (RT-PCR). We also used surface-enhanced laser desorption ionization time of flight (SELDI-TOF) mass spectrometry (MS) to analyze the effects of estradiol in ovariectomized rats on SHBG levels in the MPOA-MH as well as the neurohypophysis. In vivo estradiol treatment in ovariectomized rats eliminated or significantly reduced expression of all three estradiol-binding proteins in both the MPOA-AH and MH. This change in ERα, ERβ, and SHBG expression did not occur in the hippocampus. Both Northern blot and DNA sequence analysis confirmed the results of the RT-PCR for SHBG. SELDI-TOF MS analysis demonstrated that in vivo estradiol treatments resulted in dramatically decreased levels of SHBG in the hypothalamus and that a reduction in SHBG mRNA by estradiol treatment also resulted in a reduction in SHBG protein levels. Estradiol treatment also eliminated detectable SHBG from the neurohypophysis, suggesting that estradiol controls SHBG levels in this release site. That in vivo estradiol treatments had the same inhibitory effects on mRNA levels for SHBG and both ERs suggests similar translational control mechanisms for all three steroid-binding proteins in the brain. That estradiol treatments also reduced pituitary SHBG suggests that such treatment releases SHBG from the neurohypophysis.
Estrogen Receptor Expression in Laryngeal Muscle in Relation to Estrogen-Dependent Increases in Synapse Strength
Authors: Wu Kwok H. , Tobias Martha L. , Kelley Darcy B.
Published: August 2003

In Xenopus laevis, the laryngeal neuromuscular synapse is the final effector for sexually differentiated song production. Females have stronger laryngeal synapses than males, and synapse strength is estrogen dependent. Estrogen-induced increases in synaptic strength require at least 3 weeks of exposure, suggesting that the hormone acts via a classical genomic mechanism involving the estrogen receptor (ER). The locus of the sex difference in synapse strength, determined using quantal analysis, is presynaptic, leading to the prediction that estrogen acts directly on vocal motor neurons. However, laryngeal motor neurons do not accumulate estrogen. Estrogen might instead affect motor neuron transmitter release via a retrograde signal from its target muscle. To test this hypothesis, we determined whether laryngeal muscle expresses ER. With RT-PCR using primers that recognize highly conserved domains of the ERα, mRNA products of the predicted size were amplified from laryngeal muscle as well as from other classical target tissues (forebrain and oviduct). Northern blots using a portion of the PCR product as primer revealed the same-sized band in oviduct and laryngeal muscle. Immunocytochemistry and Western blots confirmed the presence of ER protein in laryngeal muscle fibers and revealed several proteins in laryngeal muscle, brain and liver; among these was an approximately 66-kD protein – presumed to be full-length ER – that was the only one found in oviduct. Estrogen treatment of juveniles resulted in an upregulation of the 66-kD ER protein concomitant with an increase in quantal content. Taken together, these experiments strongly suggest that the ER is expressed by laryngeal muscle; this receptor could mediate estrogen-dependent changes in synaptic strength via retrograde signaling.
Effects of Sex Steroid Hormones on the Expression of Somatostatin Receptors sst1 and sst5 in Goldfish Pituitary and Forebrain
Authors: Canosa Luis Fabián , Lin Xinwei , Peter Richard E.
Published: August 2003

In the present paper the effects of estradiol and testosterone on the expression of the types 1 and 5 somatostatin receptors (sst1 and sst5) in the goldfish forebrain and pituitary were investigated. Estradiol increased the sst1 expression in both the forebrain and pituitary in a dose- and time-dependent manner. In addition, estradiol also increased the pituitary expression of sst5. On the other hand, testosterone had no effects on the expression of these receptor subtypes. Mature female goldfish were found to have higher sst1 and sst5 expression in the pituitary, as well as a higher expression of sst1 in the forebrain compared to sexually regressed animals. As estradiol treatment increases serum growth hormone levels in goldfish, these data suggest that sst1 and sst5 receptors are likely not directly involved in this aspect of growth hormone release.
Central Insulin Administration Maintains Reproductive Behavior in Diabetic Female Rats
Authors: Kovacs Peter , Morales Jose C. , Karkanias George B.
Published: August 2003

Diabetic female rats have decreased ovulation, reproductive behavior and luteinizing hormone surges. Peripheral insulin treatment restores the phenotype to normal and also corrects many of the metabolic changes. To further evaluate the role of insulin in the specific maintenance of reproductive behavior, rather than administering it peripherally, we gave insulin intracerebroventricularly (ICV) at doses that did not correct the peripheral metabolic changes associated with diabetes such as hyperglycemia. During the experiment, we assessed the general activity and reproductive behavior of gonadectomized, estrogen- and progesterone-treated diabetic and control, nondiabetic female rats. Some of the animals received ICV saline, the rest received ICV insulin. General activity (ambulation, rearing and corner sniffing) was not affected by any of the treatments. Streptozotocin-induced diabetes resulted in a significantly decreased lordosis quotient, and ICV insulin was able to completely prevent this reduction. Furthermore, ICV insulin was able to cause a modest increase in the lordosis quotient among nondiabetic rats as well. Moreover, ICV insulin also increased the quality of lordosis among diabetic rats. Our results support a central role of insulin in the control of reproductive functions, beyond maintaining the homeostatic and metabolic balance.
α-Melanocyte-Stimulating Hormone Stimulates Prolactin Secretion through Melanocortin-3 Receptors Expressed in Mammotropes in the Mouse Pituitary
Authors: Matsumura Ryusei , Takagi Chika , Kakeya Tomoshi , Okuda Kiyoshi , Takeuchi Sakae , Takahashi Sumio
Published: August 2003

The intermediate lobe of rodent pituitaries is involved in the regulation of prolactin (PRL) secretion from the anterior lobe. In a previous study, we demonstrated the stimulatory effect of α-melanocyte-stimulating hormone (α-MSH) on PRL release and the expression of melanocortin-3 receptors (MC3-Rs) in cultured mouse pituitary cells. The aim of the present study was to clarify whether α-MSH directly stimulates PRL release through the MC3-Rs by determining the cell type of MC3-R-expressing cells in the mouse pituitary anterior lobe. Northern blot analysis revealed a 2.7-kb transcript for MC3-R mRNA in the anterior and neurointermediate lobes of pituitary glands of adult male and female mice. Dual cellular localization of MC3-R mRNA and PRL or growth hormone (GH) in the mouse pituitary glands was performed by in situ hybridization analysis of MC3-R mRNA followed by immunocytochemical detection of PRL or GH. MC3-R mRNA was detected in most mammotropes and some somatotropes. α-MSH increased PRL release and stimulated DNA replication in mammotropes, and these effects were blocked by SHU9119, an antagonist of MC3-R and MC4-R. These results indicate that α-MSH stimulates PRL release and proliferation of mammotropes through MC3-Rs, and suggest that α-MSH from intermediate lobes can regulate mammotrope functions in the mouse pituitary.
Direct Inhibitory Action of Galanin on Hypothalamic Arcuate Nucleus Neurones Expressing Galanin Receptor Gal-R1 mRNA
Authors: Poulain Pierre , Decrocq Noëlle , Mitchell Valérie
Published: August 2003

Galanin may act in the regulation of reproduction and feeding behaviour through a direct action on arcuate nucleus (ARC) neurones but evidence for a postsynaptic action is absent. The effects of rat galanin(1–29) on rat ARC neurones were examined during recordings with biocytin-filled sharp microelectrodes from brain slices. Neurones were subsequently visualised and hybridized for the mRNA of the galanin receptor Gal-R1. Current-clamp recordings revealed that ARC neurones displayed a functional heterogeneity in their firing properties. A great majority of neurones showed low-threshold spikes, instantaneous and time-dependent inward rectifications and an outward rectification. Two types of bursting behaviours were recorded, depending on the level of the membrane potential. Galanin (500 nM) elicited a reversible hyperpolarisation with a decrease in the resistance or a decrease in resistance only in 9 of 34 tested neurones, independent of the electrophysiological phenotype. The reversal potential for the galanin response was about –94 mV, near the equilibrium potential for K+. Galantide (10 nM) antagonised the effects which were not blocked by tetrodotoxin, indicating a postsynaptic action. In situ hybridisation revealed that Gal-R1 mRNA occurred in the neurones that responded to galanin, whereas the unresponsive neurones never exhibited the hybridisation signal. This suggests that the responsive neurones possess functional postsynaptic Gal-R1. The study demonstrates for the first time a direct inhibitory action of galanin on ARC neurones, probably mediated by activation of Gal-R1. This action may have important implications for the functioning of neurones involved in the regulation of reproduction and feeding behaviour, such as proopiomelanocortin neurones.
Circadian Determinations of Cortisol, Prolactin and Melatonin in Chronic Methyl-Phenyl-Tetrahydropyridine-Treated Monkeys
Authors: Barcia Carlos , Bautista Víctor , Sánchez-Bahillo Angel , Fernández-Villalba Emiliano , Navarro-Ruis Juana-María , Barreiro Andrés Fernández , Poza y Poza Máximo , Herrero María-Trinidad
Published: August 2003

The aim of this study was to investigate whether prolactin, melatonin and cortisol are altered in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys and if so, whether MPTP may alter the availability of these hormones in chronic experimental parkinsonism. Furthermore, vegetative and sleep disorders have been described in both parkinsonian patients and in MPTP chronic monkeys; these may result indirectly from concomitant hormonal variations. Seven adult male cynomolgus monkeys were used for this experiment. Five were treated with systemic doses of intravenous MPTP but not with L-DOPA or dopaminergic agonists. In their 3rd year of parkinsonism, plasma samples were obtained day and night at 3-hour intervals. Sample collection was repeated three times for each animal. Prolactin, melatonin and cortisol concentrations were determined by enzyme immunoassay and compared with samples taken from the control group. Both MPTP-treated monkeys and the control group displayed a similar secretion pattern for the three hormones, except at several specific times when prolactin and melatonin showed significant differences. No changes were found for cortisol. The results suggest a possible alteration of hormonal metabolism in chronic MPTP parkinsonian monkeys.
Local Anesthetics Inhibit Uterine Activity in vitro
Authors: Fauza Dario O. , Kohane Daniel S. , Beeuwkes Elizabeth B. , Clayton Nicholas , Maher Timothy J.
Published: August 2003

Objective: Analysis of the effects of local anesthetics upon the gravid uterus. Methods: Isolated strips of pregnant-rat myometrium were placed within tissue baths. Muscular activity was then recorded by a force transducer, at cumulative concentrations of either procaine, lidocaine, or ropivacaine. Results: The effects of myometrial activity were non-monotonic. At lower concentrations, an increase in contraction frequency was noted, concomitant with a depression in amplitude. As the concentrations increased, both amplitude and frequency were depressed until, eventually, activity was abolished. Conclusions: Local anesthetics administered in situ consistently inhibit or completely arrest overall myometrial activity. These agents may prove clinically useful in preterm labor management, if administered locally to the myometrium via suitable slow release drug delivery systems, warranting further trials in vivo.
Intrauterine Repair of Gastroschisis in Fetal Rabbits
Authors: Till H. , Muensterer O. , Mueller M. , Klis V. , Klotz S. , Metzger R. , Joppich I.
Published: August 2003

Objective: Infants with gastroschisis (GS) still face severe morbidity. Prenatal closure may prevent gastrointestinal organ damage, but intrauterine GS repair (GSR) has not been established yet. Methods: In New Zealand White rabbits we developed and compared GS versus GSR: creation of GS was achieved by hysterotomy, right-sided laparotomy of the fetus and pressure on the abdominal wall to provoke evisceration. GSR was accomplished by careful reposition of eviscerated organs and a running suture of the fetal abdominal wall. For study purposes, 18 animals were divided equally into 3 groups: GS, GS with GSR after 2 h, and unmanipulated controls (C). Vitality was assessed by echocardiography. After 5 h all animals were sacrificed. Results: GSR inflicted no increased mortality, because all fetuses survived GS or GS with GSR. All fetuses with GS demonstrated significant evisceration of abdominal organs. In contrast, the abdominal wall of the fetuses from GSR was intact. Conclusion: The present animal model demonstrated the technical feasibility and success of an intrauterine repair of GS for the first time. However, further long-term studies (leaving GS and GSR in utero for several days) will be necessary to compare survival rates and intestinal injury, motility or absorption. The clinical application of GSR in utero remains a vision so far.
Discordant Fetal Growth Patterns in Monochorionic Twin Pregnancies Described by Simple Algebraic Relations
Authors: Umur Asli , van Gemert Martin J.C. , Schaap Arty H.P. , Ross Michael G.
Published: August 2003

Objective: To test whether twin-twin transfusion syndrome (TTTS) and non-TTTS cases that were suspect but not confirmatory for developing the syndrome (non-TTTS-with-symptoms) have discordant fetal growth patterns that correlate with our previously derived algebraic relations. Methods: In 25 monochorionic twin pregnancies, fetal growth was determined by standard ultrasonography. The difference between estimated fetal weights (dEFW) as well as the difference divided by the average of the two weights, the difference average ratio (DAR), were fitted to the predicted trends of discordant fetal growth for TTTS and non-TTTS pregnancies. The best fits were compared with the clinical data. Results: Out of 13 TTTS cases, dEFW analysis correctly correlated with 8 (62%) and DAR analysis correctly with 10 (77%). Out of 12 non-TTTS-with-symptoms cases, dEFW analysis correctly correlated with 7 (58%) and DAR analysis correctly with 9 (75%). If TTTS correlated best, dEFW analysis was correct in 8/12 (67%), and DAR analysis in 10/11 (91%) cases. If non-TTTS correlated best, dEFW analysis was correct in 7/9 (78%), and DAR analysis in 9/12 (75%). The likelihood ratios of TTTS and non-TTTS were 1.9 and 0.26, respectively, with dEFW analysis, and 9.2 and 0.31 with DAR analysis. Conclusion: The simple algebraic relations derived to identify trends of fetal discordant growth show evidence that clinical TTTS and non-TTTS manifestations are quantifiable. The relations may contribute to future risk stratification in monochorionic twin pregnancies, e.g., by prospectively distinguishing between cases that will develop TTTS and those that will not, despite presenting with symptoms.
Identification of Fetal Nucleated Red Blood Cells in the Maternal Circulation during Pregnancy Using Anti-Hemoglobin-ε Antibody
Authors: Mavrou A. , Kolialexi A. , Antsaklis A. , Korantzis A. , Metaxotou C.
Published: August 2003

Aim: To investigate the use of anti-hemoglobin-Ε antibody in order to identify fetal cells in the maternal circulation during pregnancy. Materials and Methods: 48 blood samples were obtained from pregnant women, 26 in the 1st trimester and 22 in the 2nd trimester. Magnetic activated cell sorting was used for fetal cell enrichment followed by immunophenotyping with a monoclonal antibody against hemoglobin-Ε. FISH with X, Y and 21 chromosome-specific probes was performed in 29 cases. Results: The mean number of Ε-positive cells was 9.2 (range 2–23) in the 1st trimester, 4.8 (range 3–13) in the 2nd trimester and 22 (range 15–28) in pregnancies with Down syndrome. No significant difference was noted in the number of Ε-positive nucleated red blood cells (NRBCs) isolated from carriers and noncarriers of β-thalassemia. FISH analysis was successful in 24 cases. In 4 cases with known male fetuses, an average of 4.7 Ε-positive cells showed a Y signal. In 4 cases with Down syndrome, all Ε-positive cells showed 3 signals for chromosome 21. Conclusion: Anti-hemoglobin-Ε antibody has increased specificity for fetal NRBCs and should be preferentially used to improve noninvasive prenatal diagnosis of chromosome abnormalities from fetal cells in maternal blood.
Chorioamniotic Membrane Separation following Open Fetal Surgery: Pregnancy Outcome
Authors: Wilson R. Douglas , Johnson Mark P. , Crombleholme Timothy M. , Flake Alan W. , Hedrick Holly L. , King Mary , Howell Lori J. , Adzick N. Scott
Published: August 2003

Objective: To review the incidence of posthysterotomy chorioamniotic membrane separation and delivery outcome following open fetal surgery [myelomeningocele (MMC); cystic adenomatoid malformation (CCAM); congenital diaphragmatic hernia (CDH); sacrococcygeal teratoma (SCT)]. Study Design: Retrospective review of a maternal population undergoing open fetal surgery at a single tertiary level program (1998–2001) following the initiation of close postoperative ultrasound follow-up for membrane separation. Onset of membrane separation was coded as not present (NP), immediate (<2 weeks) or delayed (>2 weeks) from day of surgery. Results: Fifty-three charts were reviewed: MMC 43, CCAM 7, CDH 1, and SCT 2. In the MMC group there were 26 NP, 8 immediate, and 9 delayed. Preterm labor occurred in 4 patients with only 2 having had membrane separation. Risk of membrane separation is increased for surgery done at less than 23 weeks gestation (p < 0.005). Delay from MMC surgery to delivery was 11.0, 9.8, 12.0 weeks for NP, immediate, and delay, respectively. In the MMC group, there were 3 neonatal deaths (NND) at 9, 9, and 21 days post surgery (PROM/PTL; chorioamnionitis, PROM/PTL, respectively). No membrane separation was present in the CCAM, CDH, and SCT cases. Conclusions: (1) Membrane separation was significantly more likely to occur if surgery was performed prior to 23 weeks. (2) Membrane separation post hysterotomy (17/50 = 34%) may be associated with an increased risk of PROM but not delivery before 30 weeks gestation. (3) Delivery prior to 33 weeks gestation for MMC groups was 12/43 (28%) with 3 NND (7%). (4) Elective delivery at 36–37 weeks gestation was possible for 43% of the fetal surgery population.
Maternal Serum and Amniotic Fluid Hydroxyproline Levels in Neural Tube Defects
Authors: Özyurt Hüseyin , Totan Aylin , Şahin Şemsettin , Kılınç Cumhur , Söğüt Sadık , Akyol Ömer
Published: August 2003

Objective: The present study was aimed to evaluate the alterations of maternal serum and amniotic fluid OH- proline levels in neural tube defects (NTD). Methods: 38 patients and 23 control subjects were selected from the patients followed up by the Departments of Genetics and Antenatal Care, Zekai Tahir Burak Women’s Health, Education and Research Hospital. Amnion fluid and blood samples were synchronously obtained from NTD and control subjects. OH-proline levels were detected spectrophotometrically. Results: Amniotic fluid OH-proline levels were found to be increased in NTD patients compared to control subjects (p < 0.05). There was no statistically significant difference in serum OH-proline levels between the groups. Conclusion: The results showed an increase in amniotic fluid OH-proline levels in NTD compared to the control group. It may be suggested that the increased OH-proline levels reflect increased collagen turnover in NTD patients. Indeed, amniotic fluid OH-proline levels may be a candidate as an additional diagnostic parameter in NTD.
Prenatal Diagnosis of Fetal Intracranial Hemorrhage at 25 Weeks of Gestation
Authors: Zanders Erik H.J. , Buist Fayette C.D. , van Vugt John M.G.
Published: August 2003

Objective: To determine whether magnetic resonance imaging (MRI) can give further information for the prenatal diagnosis of intracranial hemorrhage, when ultrasound (US) is not conclusive. Methods: We report on a fetus in the 26th week of gestation with a grade-III hemorrhage in the right cerebral hemisphere, which was suspected on US examination and specified by MRI examination in a patient with pre-eclampsia. Results: Doppler measurement of the umbilical artery, which showed the absence of end diastolic velocity, was indicative of placental insufficiency. The MRI specified the findings found by US examination and confirmed the poor outcome of the fetus. Labor was induced by prostaglandins in the 27th week of gestation. Postmortem examination showed a female fetus with a grade-III intracranial hemorrhage in the center of the right hemisphere with a diameter of 4 cm. A hemorrhage was also found in the subarachnoid space and in the occipital region of the left hemisphere. Conclusion: We suggest that fetuses at high risk of developing hypoxia should be sonographically evaluated and MRI should be considered when intracranial echogenicities are seen.
Prenatal Diagnosis in Beta-Thalassemia: An Indian Experience
Authors: Agarwal S. , Gupta A. , Gupta U.R. , Sarwai S. , Phadke S. , Agarwal S.S.
Published: August 2003

Objectives: Thalassemia is the most common single gene disorder and is widely distributed in Asian Indians with an average prevalence rate of 4%, with a high prevalence among Sindhis, Punjabis, Gujratis and Bengalis. Prevention and control of β-thalassemia disease require the accurate diagnosis of carriers and proper genetic counseling. Method: Prenatal diagnosis can be performed in the first or second trimester of pregnancy by DNA analysis using polymerase chain reaction. Since there are 17 mutations as well as rare ones causing β-thalassemia in Asian Indians, the point mutation detection by reverse dot blot (RDB) allele-specific oligonucleotide hybridization for common mutations along with the amplification refractory mutation system (ARMS) technique was developed for prenatal diagnosis. Maternal contamination of fetal DNA was ruled out by the variable number of tandem repeat analysis using apolipoprotein B site. Results: Using both techniques (RDB and ARMS) we were able to offer complete diagnosis in 53 pregnancies. On molecular analysis 23% were found to be normal, 48.0% were carriers, and 29.0% were affected with β-thalassemia. Parents were counseled to continue the pregnancy when the fetuses were either normal or had traits whereas in the case of an affected fetus, the parents opted for termination of the pregnancy. Conclusion: Prenatal diagnosis of β-thalassemia by the RDB or ARMS technique can prevent the birth of an affected child in developing countries in which β-thalassemia is quite prevalent.
Epinephrine Inhibits Tracheal Occlusion Induced Lung Growth in Fetal Sheep
Authors: Kitano Yoshihiro , von Allmen Daniel , Kanai Masaki , Adzick N. Scott , Flake Alan W.
Published: August 2003

Objective: Although the mechanisms responsible for lung growth following tracheal occlusion (TO) are not fully understood, lung fluid accumulation is a requirement for growth to occur. It is known that betamimetics such as epinephrine (Epi), terbutaline, and ritodrine inhibit fetal lung fluid production late in gestation. We hypothesized that continuous infusion of Epi would have a detrimental effect on lung growth following TO in the fetal sheep model. Methods: Twenty fetal lambs were divided into four groups. Group 1: no TO without Epi (n = 4); group 2: TO without Epi (n = 4); group 3: no TO with Epi (n = 5), and group 4: TO with Epi (n = 7). TO was performed on days 130–134 of gestation (full term = 145 days). In groups 3 and 4, Epi was infused into the fetus at a rate of 2 µg/min using a miniosmotic pump implanted subcutaneously at the time of TO. The fetuses were sacrificed after 4 days, and lung volume (LV, ml/kg), drained lung fluid (LF, ml/kg), wet lung weight/body weight ratio (LW/BW, %), whole right-lung dry weight per body weight ratio (dRLW/BW, g/kg), volume density of lung parenchyma (vVp), and right-lung DNA and protein contents were compared among the four groups by one-way Anova. Results: LW/BW and dRLW/BW of group 4 (3.91 ± 0.52 and 2.10 ± 0.28, mean ± SD) were significantly lower than those of group 2 (5.18 ± 0.57 and 2.67 ± 0.15) and were not statistically significantly different from those of group 1 or 3. LV, LF, VVp, and DNA and protein contents all showed a similar trend. Conclusions: Continuous infusion of Epi to the fetus results in less fluid accumulation within the TO lungs and abolishes significant lung growth after TO in the late-gestation fetal lamb. Drugs affecting lung fluid secretion may have a major impact on TO-induced lung growth.
Antenatal Evaluation of an Encephalocele in a Dizygotic Twin Pregnancy Using Fast Magnetic Resonance Imaging
Authors: Kojima Kazuhisa , Suzuki Yoshikatsu , Miyajima Shinichi , Tanemura Mitsuyo , Yamaguchi Nobuyuki , Ueda Yukihiko , Suzumori Kaoru
Published: August 2003

We report a case of an encephalocele in a dizygotic twin pregnancy, following ovulatory induction. In the involved fetus, an abnormal shadow like an encapsulated-solid tumor located on the occiput was found by routine maternal transabdominal ultrasonography at 17 weeks of gestation. The parents did not accept induced abortion because of the presence of another fetus with no abnormality on ultrasonography. At 35 weeks of gestation, transabdominal ultrasound examination showed a large occipital cyst, composed of protrusive fetal brain and cerebrospinal fluid. Fast-scanning magnetic resonance imaging delineated more clearly the inside of the abnormal lesion and thus allowed confirmation of the putative diagnosis of fetal encephalocele during pregnancy. Surgical report was possible in this case, and the patient had no severe physical or neurological abnormalities 10 months after birth. Since the prognosis appears to depend primarily on how prominent the brain tissue is inside the herniated sac, this approach had benefit for clinical decision making.