Brain fatty acids in perinatal asphyxia

In hypoxic or ischemic states the release of fatty acids is proposed to have several harmful effects on brain structure and function. We therefore decided to study brain FFA in a simple, clinically related animal model resembling intrauterine perinatal asphyxia (PA). Cerebral blood flow (CBF), brain fatty acids (C14:0, C16:1, C16:0, C18:1, C1 8:0, sigma C), plasma glucose, lactate, beta-hydroxybutyrate (beta-OHB), non-esterified fatty acids (NEFA) and insulin were determined in PA and compared to the normoxic state. Brain C 14:0 FFA were not significantly different from normoxic rats. Brain FFA C 16:0 were comparable between groups but significantly decreased at 20 min of PA. C 18:0 FFA showed a trend to increase with the length of PA reaching significance at 10 min of asphyxia only and were declining at 20 min, however, not significantly. Brain C 16:1 and C 18:1 FFA concentrations were comparable between groups. The parameters cerebral blood flow, glucose and lactate showed a stepwise and significant increase with the length of PA, whereas beta-HOB, NEFA and insulin showed no changes. CBF, glucose and lactate showed a strong association whereas other parameters failed to correlate with each other. Only inconsistent trends of increased brain FFA were found and the association between brain glucose and brain FFA could be ruled out. Although CBF was manifold and significantly elevated in PA, brain FFA pattern suggests that the increase of CBF is obviously not mediated by brain FFA. We conclude that FFA may not be involved in the early phase-pathogenesis of PA.     

Genes involved in deafness.

Remarkable progress has been made over the past few years in the field of hereditary deafness. To date, mutations in at least 35 genes are known to cause hearing loss. We are now beginning to understand the function of many of these genes, which affect diverse aspects of ear development and function.     

Energy metabolism in graded perinatal asphyxia of the rat

Although information on energy metabolism during hypoxemic-ischemic states is abundant, data on perinatal asphyxia (PA) are limited. As results from hypoxia-ischemia cannot be directly extrapolated to PA, a clinical entity characterized by acidosis, hypoxemia and hypercapnia, we decided to use a rat model of graded PA during delivery. Cesarean section was performed at the 21st day of gestation and the pups, still in the uterus horns, were asphyxiated from 0 to 20 minutes. In this model survival decreases with the length of asphyxia. Early changes of energy-rich phosphates in brain, heart and kidney were determined by HPLC. ATP and phosphocreatine gradually decreased with the length of asphyxia, with highest ATP depletion rate occurring in the kidney. ATP: brain 1.39 +/- 0.71 (0 min) to 0.06 microM/g wwt (20 min); heart 4.73 +/- 0.34 (0 min) to 1.08 +/- 0.47 (20 min); kidney 1.62 +/- 0.11 (0 min) to 0.02 +/- 0.02 (20 min). Phosphocreatine: brain 1.65 +/- 0.68 (0 min) to 0.51 +/- 0.45 microM/g (20 min); heart 6.98 +/- 0.38 (0 min) to 6.17 +/- 1.07 (20 min); kidney 8.23 +/- 0.86 (0 min) to 3.76 +/- 0.54 (20 min). We present data on energy derangement in a rat model of PA, closely resembling the clinical situation, showing that energy depletion precedes cell damage and death.     

Myelination deficits in brain of rats following perinatal asphyxia

Perinatal asphyxia remains a major cause of acute mortality and of permanent neurodevelopmental disability in infants and children. However, the pathophysiologic features of hypoxic-ischemic encephalopathy are still incompletely understood. Animal studies have been focussing on grey matter pathology but information on white matter lesions is limited. The aim of the study was to investigate white matter lesions after three months following graded perinatal asphyxia in the rat using a well-documented, reproducible, clinically relevant and simple animal model of perinatal asphyxia. Brains of rat pups (n=10 per group) exposed to asphyctic periods of 10 and 20 minutes were examined histologically and compared to normoxic brain using Kluever-Barrera myelin staining, immunohistochemically with antibodies against myelin basic protein, 2',3'-cyclic-nucleotide'-phosphodiesterase as markers for myelination, antibodies against neurofilaments for the evaluation of axonal density and antibodies against glial fibrillary acidic protein as a marker for astrocytic gliosis. Morphometry three months after perinatal asphyxia showed significant reduction of corpus callosum in asphyctic brains. Patchy myelination deficits were found in hippocampal fimbriae and cerebellum, lobulus L 8, accompanied by reduced axonal density. Hypothalamus and striatum did not show any myelination deficit. Up to now only short term effects of perinatal asphyxia on myelination have been reported and this communication reveals long-term myelination deficit in three brain regions after three months following perinatal asphyxia. As myelination deficit was regularly accompanied by reduction of neurofilament immunoreactivity, we suggest that white matter lesions are paralleling grey matter damage, a subject still controversial in pathophysiology of brain damage in perinatal asphyxia.     

Histological changes and neurotransmitter levels three months following perinatal asphyxia in the rat.

The involvement of excitatory amino acids (EAA) in the pathogenesis of hypoxic-ischemic states is well-documented. Information on the role of overexcitation by EAA in perinatalasphyxia (PA), however, is limited and data from adult models cannot be directly extrapolated to immature systems. Moreover, most adult models of ischemia are representing stroke rather than PA. We decided to study long term effects in a non-invasive rat model of PA resembling the clinical situation three months following the asphyctic insult. Morphometry on Nissl - stained sections was used to determine neuronal death in frontal cortex, striatum, hippocampus CA1, hypothalamus and cerebellum L1, and the amino acids glutamate, glutamine, aspartate, GABA, taurine, arginine as well as histamine, serotonin and 5-hydroxy-indoleacetic acid were determined in several brain regions and areas. Morphometry revealed that neuronal loss was present in the hippocampal area CA1 in all groups with PA and that morphological alterations were significantly higher in the cerebellar granular layer. The prominent light microscopical finding in all areas of asphyctic rats studied was decreased Nissl-staining, suggesting decreased cellular RNA levels. Glutamate, aspartate and glutamine were significantly elevated in the hypothalamus of asphyctic rats probably indicating overstimulation by EAA. Excitotoxicity in this area would be compatible with findings of emotional / behavioral deficits observed in a parallel study in our model of PA. Our observations point to and may help to explain behavioral and emotional deficits in Man with a history of perinatal asphyxia.     

Physical and neurological state of the newborn after perinatal asphyxia

Asphyxia neonatorum is the most prevalent birth trauma. Usually it leads to the birth of weak and nonvigorous child. Perinatal asphyxia is the reason of not only postasphyxic hypoxic-ischemic encephalopathy, but also multiple organ dysfunctions. Neurological sequelae and consequences of multisystem dysfunction affect on newborn status in first life months and later on by the muscle tone disturbances, physical and mental retardations. In this paper it is presented modern conceptions about asphyxia origin, risk factors, neurological and systemic complications for child nervous system and organism.     

The role of genetic determinant in the development of severe perinatal asphyxia

The frequency of GSTT1 and GSTM1 gene deletion polymorphism was determined in a case-control study of full-term Ukrainian newborns including patients with perinatal asphyxia. Multiplex polymerase chain reaction was used for genotyping 245 full-term newborns. The investigated full-term newborns with perinatal asphyxia were subdivided in the subgroups depending of severity of perinatal asphyxia and neonatal outcome. No significant differences in allele frequencies of homorygous null genotypes of GSTT1 and GSTM1 gene were detected among newborns with moderate perinatal asphyxia and healthy control. However, association with the development of severe perinatal asphyxia was detected for the deletion polymorphism in GSTT1 gene and the combination of the GSTT1 absent/GSTM1 absent in the newborns. The study shows that severe perinatal asphyxia may develop in the consequence of genetic predisposition to this condition as compare with moderate.     

精子发生过程中的相关基因

在哺乳动物精子发生过程中, 原生殖细胞发育成为精原细胞, 再发育为精母细胞, 精母细胞经过两次减数分裂成为圆形精细胞, 这些圆形精细胞经过细胞变态形成精子。精子发生过程经历了复杂的细胞分化阶段, 这一阶段受许多因素的调控作用, 其中生精细胞内的基因调节起着决定作用。精子发生中的重要基因与一系列精子发生过程中阶段性的细胞事件密切相关, 例如减数分裂重组、联会丝复合物的形成、姊妹染色体的结合、减数分裂后精子的变态以及减数分裂周期中的关键点和必需因子等。生精细胞许多特异基因的阶段特异性表达, 参与了精子发生这一特殊的细胞分化过程。近年来随着基因克隆、表达和功能研究技术的发展和应用, 发现了许多与精子发生相关的基因, 而且有的被证明在精子发生过程中具有重要作用。文章较全面综述了这一研究领域的一些进展, 着重讨论了与精子发生相关的周期蛋白基因、原癌基因、无精子因子基因、细胞骨架基因、热休克基因、核蛋白转型基因、中心体蛋白基因和细胞凋亡相关基因等。     

Perinatal asphyxia pathophysiology in pig and human: a review

In utero fetuses are evidently exposed to several factors that cause an interruption of the oxygen flow through the umbilical cord causing asphyxia leading to hypoxia and metabolic acidosis. These conditions are important causes of intra-partum and neonatal mortality. The main objective of this review is to provide current information regarding the pathophysiology of asphyxia in piglets around parturition; the physiological mechanisms invoked by affected piglets to compensate perinatal hypoxemia are discussed. This review also addresses some similarities and differences of asphyxia between piglets and other mammals, including human neonates. Metabolic acidosis and hypoxia are sequela to asphyxia and can cause profound health effects in postnatal performance because of an abnormal suckling, a reduced absorption of colostrum and inadequate passive transfer of neonatal immunity. Acidosis also cause hypothermia, increased mortality and reduced survival in neonates. One of the first deleterious effects of intrauterine hypoxia is the expulsion of meconium into the amniotic sac leading to meconium staining of the skin, and in severe cases, meconium aspiration into the lungs. Even though there have been technological changes and improvements in husbandry, piglet mortality due to asphyxia remains a major problem. One potential alternative to reduce neonatal mortality in pigs is the monitoring of fetal stress during birth and the implemention of strategies such as the Apgar score, that is often used in human pediatrics. It is also important to consider the physiological, behavioral and biochemical changes that take place during parturition which subsequently impact the vitality, maturity and development of neonatal pigs. Understanding the pathophysiology of fetal hypoxia should help practitioners and farmers implement more effective delivery techniques aimed at reducing neonatal mortality and improving postnatal performance.     

Genes involved in androgen biosynthesis and the male phenotype.

A series of enzymatic steps in the testis lead to the conversion of cholesterol to the male sex steroid hormones, testosterone and 5 alpha-dihydrotestosterone. Mutations in any one of these steps are presumed to alter or block the development of the male phenotype. Most of the genes encoding the enzymes involved in this pathway have now been cloned, and mutations within the coding regions of these genes do, in fact, block development of the male phenotype.     

Thyroid hormone concentrations in foals affected by perinatal asphyxia syndrome

The hypothalamus-pituitary-thyroid axis has specific functions, mostly related to metabolic activities, cell differentiation, and development. To the authors’ knowledge, there are no studies about thyroid hormone (TH) concentrations in foals affected by perinatal asphyxia syndrome (PAS). Hence, the aims of the study are (1) to evaluate plasma TH concentrations (T₃ and T₄) in healthy foals during the first 7 days of life; (2) to evaluate plasma TH concentration (T₃ and T₄) in critically ill foals affected by PAS during the first 7 days of hospitalization; and (3) to compare TH concentrations between surviving and nonsurviving critically ill foals. Forty-five Standardbred foals were enrolled in this prospective observational study: 21 healthy foals (group 1) and 24 foals affected by PAS (group 2). Jugular blood samples were collected within 10 minutes from birth/admission and every 24 hours for 7 days (t0–t7). TH concentrations were analyzed by RIA. In both groups, T₃ concentration was significantly lower at t4, t5, t6, and t7 compared with t1 (P < 0.05), and T₄ concentration was significantly higher at birth than at all other time points (P < 0.01). No differences were found in TH concentrations at admission between surviving (n = 20) and nonsurviving (n = 4) foals. Statistical comparison between healthy and PAS foals divided into age groups showed significantly lower TH concentrations at t0 in PAS foals <12 hours old at admission (P < 0.01). In conclusion, PAS may cause lower T₃ and T₄ concentrations in affected foals than in age-matched healthy foals, as reported for other systemic illnesses, such as sepsis and prematurity. TH concentrations showed no prognostic value, which maybe due to the small number of nonsurviving foals in this study. Further studies are needed to find out if thyroid replacement therapy could be useful in the treatment of critically ill foals affected by PAS.     

Long term neurological and behavioral effects of graded perinatal asphyxia in the rat

Perinatal hypoxic-ischemic states can cause irreversible damage to the brain, ranging from minimal brain dysfunction to death. Only few studies have been reported describing neurological, cognitive and behavioral deficits following perinatal asphyxia. We therefore decided to study long term effects of perinatal asphyxia in a well-documented animal model resembling the clinical situation. Caeserean section in rats was performed and the pups, still in the uterus horns, were placed into a water bath at 37 degrees C for periods of 5-20 min; pups were then given to surrogate mothers and examined at three month of age. Examinations consisted of a battery of motor and reflex tests, Morris water maze, multiple T-maze, elevated plus maze and open field studies. No abnormalities were found in rats even with long periods of perinatal asphyxia by neurological examination, in the open field and in mazes. Interestingly, in the elevated plus maze rats with long lasting exposure to hypoxia (15 and 20 min of asphyxia) showed reduced anxiety-related behavior. This finding may be relevant for the explanation of anxiety related disorders in adulthood with a tentative history in the perinatal period.     

Genes potentially involved in plaque rupture

PURPOSE OF REVIEW: Rupture of an atherosclerotic plaque is the predominant underlying event in the pathogenesis of acute coronary syndromes and stroke. While ruptured plaques are morphologically well described, the precise molecular mechanisms involved in plaque rupture are still incompletely understood. Over the last few years, techniques like microarray, suppression subtractive hybridization and differential display enabled us to study complex gene expression profiles that occur during the process of atherogenesis. In this review we focus on recent large-scale gene expression profiles performed on whole mount vascular specimens. RECENT FINDINGS: The gene expression profiles on whole mount vascular tissue confirmed that at least three mechanisms are involved in plaque rupture: (1) a disturbed balance in extracellular matrix turnover, (2) disturbed regulation of cell turnover and (3) processes involved in lipid metabolism. Animal models exhibiting features of plaque rupture reflect the involvement of these three mechanisms. The most dramatic mouse phenotypes were observed after interventions in at least two of these mechanisms. SUMMARY: The observation of plaque rupture in recent mice models is indicative of the multifactorial process of plaque rupture. This multifactorial character of plaque rupture suggests that interventions may be most effective when they influence more than one mechanisms at a time.     

Genes involved in cytokinin signal transduction

Although cytokinin plays a central role in plant development, our knowledge about the signal transduction pathway initiated by this plant hormone is fragmentary. By randomly introducing enhancer elements into theArabidopsis genome throughAgrobacterium-mediated transformation, 5 cytokinin independent mutant calli (cki1-1, −2, −3, −4 andcki2) were obtained. These mutants exhibit typical cytokinin responses, including rapid proliferation, chloroplast differentiation, shoot induction and inhibition of root formation, in the absence of cytokinin. TheCKl1 gene encodes a product similar to the sensor histidine kinases of two-component systems, and its overexpression in plants induces typical cytokinin responses (Kakimoto 1996). Here I report that overexpression of this gene did not alter the auxin reqirement ofArabidopsis. Another mutant,many shoots, which was also identified on the same screening, produced many adventitious shoots on cotyledons, petioles and true leaves. The extended abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International Prize for Biology “Frontier of Plant Biology”     

Genes involved in senescence and immortalization

Senescence is now understood to be the final phenotypic state adopted by a cell in response to several distinct cell physiological processes, including proliferation, oncogene activation and oxygen free radical toxicity. The role of telomere maintenance in immortalization and the roles of p16(INK4A), p19(ARF), p53 and other genes in senescence are being further elucidated. Significant progress continues to be made in our understanding of cellular senescence and immortalization.     

Genes involved in the uptake and catabolism of gluconate by Escherichia coli.

The isolation and properties of a mutant of Escherichia coli K12 that is totally unable to take up and utilize gluconate are described. Genetical analysis shows this phenotype to be associated with two lesions. One phenotype, designated GntM-, is the result of a mutation in a gene co-transducible with malA; the other, designated GNTS-, is the result of a mutation in a gene (GntS) co-transducible with fdp. The GntS--phenotype differs little from that of wild-type cells, but GntM- GntS+ organisms grow on gluconate only after a prolonged lag and form a gluconate uptake system that is strongly repressed by pyruvate. Moreover, such GntM- mutants readily give rise to further mutants that form a gluconate uptake system, gluconate kinase and 6-phosphogluconate dehydratase consititutively; in partial diploids, this constitutivity is recessive to the inducible character. It is postulated that the GntM- phenotype is due to malfunction of a negative control gene gntR, and that gntS+ specifies the activity of a gluconate uptake system.     

Genes involved in the biosynthesis of PQQ fromAcinetobacter calcoaceticus

From a gene bank of theAcinetobacter calcoaceticus genome a plasmid was isolated that complements four different classes of PQQ^- mutants. Subclones of this plasmid revealed that the four corresponding PQQ genes are located on a fragment of 5 kilobases. The nucleotide sequence of this 5 kb fragment was determined and by means of Tn5 insertion mutants the reading frames of the PQQ genes could be identified. Three of the PQQ genes code for proteins of M_r 29700 (gene I), M_r 10800 (gene II) and M_r 43600 (gene III) respectively. In the DNA region where gene IV was mapped however the largest possible reading frame encodes for a polypeptide of only 24 amino acids. A possible role for this small polypeptide will be discussed. Finally we show that expression of the four PQQ genes inAcinetobacter lwoffi andEscherichia coli lead to the synthesis of the coenzyme in these organisms.