Sulfogalactosylceramides in motor and psycho-cognitive adult metachromatic leukodystrophy: relations between clinical, biochemical analysis and molecular aspects

Metachromatic leukodystrophy (MLD) is a human autosomal recessive lysosomal neurodegenerative disorder that results from the accumulation of sulfatides in the central and peripheral nervous system. It is due to the enzyme deficiency of the sulfatide sulfatase i.e. arylsulfatase A (ASA). During adolescence and/or adulthood, there are 2 clinical presentations. It may be that of a degenerative disease of the central nervous system with mainly spastic manifestations or a spino-cerebellar ataxia, or that of a psychosis. As several lines of evidence indicate that the psychotic form of MLD could be a model of psychosis, we decided to do a pluridisciplinary study on 11 psycho-cognitive cases involving mental and psychiatric testing, in comparison with 5 adult motor cases, a biochemical study with enzyme assays and quantitative mass spectrometry of urinary sulfatides, so as to determine whether there were biochemical particularities related to the psychotic forms. For quantitative mass spectrometry (MS), a non physiological sulfatide with C17:0 fatty acid was synthesized. The major sulfatide isoforms were present in the 2 clinical forms with the following fatty acids and sphingoid bases: C22:1/d18:1, and /or C22:0/d18:2 (m/z 862.5), C22:0 (OH)/d18:1 (m/z 878,5), C24:0/d18:1 and / or C24:0/C23:1(OH)/d18:2 (m/z 890,3), C24:0 (OH)/d18:1(m/z 906.5). We had shown previously that there were different ASA mutations in the psychiatric adult form (heterozygous I179S) versus the adult motor form (homozygous P426L). We show here that there were no relations with the level of ASA and with the mass spectrometric study of the sulfatide isoforms which were identical in the 2 clinical forms.     

苦瓜采后某些生理生化变化与其衰败的关系

对采后常温贮藏的蓝山大白苦瓜的呼吸速率、乙烯释放速率及SOD、POD、CAT活性和AsA、GSH的含量进行了测定。结果表明,适期采收的食用苦瓜呼吸速率呈下降趋势,第8天叶绿素消失、瓜皮转黄前后转为上升,一直延续至苦瓜衰败,其呼吸类型属晚峰型;苦瓜采后乙烯释放呈上升趋势,初期乙烯释放微弱,增加缓慢,从第6天开始急剧增加。乙烯释放速率上升早于呼吸上升,二者在采收6d后迅速同步上升。苦瓜采后1～4d,保护酶系统的SOD、POD、CAT活性出现适应性上升,非保护酶系统的AsA、GSH含量也上升;在4～8d,除POD活性迅速下降后又急剧上升及AsA含量在6d后下降外,SOD、CAT活性和GSH含量均稳中有降,保持相对稳定;在叶绿素消失、瓜皮转黄的第8天后,AsA、GSH含量急剧减少,而SOD、POD、CAT活性急剧增强,视为衰老迅速恶化的结果。     

Lactic acidosis and ketoacidosis: biochemical and clinical implications.

A case of lactic acidosis presented the opportunity for review of the association between lactic acidosis and ketoacidosis. The diagnosis of lactic acidosis or the combination of lactic acidosis and ketoacidosis is established clinically by the detection of a metabolic acidosis of the "unmeasured anion gap" type in the absence of significant renal failure, poison intake or a strongly positive clinical test for ketones. Before treatment can be planned the biochemical basis of lactic acidosis and ketoacidosis must be understood -- especially the fact that lactic acidosis is not a single disease entity but has many possible causes. Among important considerations is the relation between the blood concentrations of bicarbonate and organic acid anions. After recovery from metabolic acidosis of the unmeasured anion gap type, metabolic alkalosis is common. Decreased bicarbonate excretion plays an important role in the pathogenesis of the latter and may be the result of potassium or chloride loss, or both. The deficits, if present, should be corrected with appropriate therapy.     

Microbial population and biochemical changes during production of protein-enriched fufu

Summary Candida utilis strain BKT4 and Saccharomyces cerevisiae strain BKT7 isolated from burukutu (a local wine brewed from sorghum) were used to enrich fufu. During the fermentation process, there were changes in the microbiological and biochemical characteristics of the cassava. The total viable counts increased with increasing fermentation time while the counts of the lactics and fungi increased at the later stages of the fermentation due to the acidity of the medium. Various bacteria (Bacillus, Staphylococcus, Klebsiella, Escherichia, Streptococcus, Lactobacillus, Leuconostoc, Corynebacterium), moulds (Penicillium, Aspergillus, Fusarium, Mucor, Rhizopus) and yeasts (Candida, Saccharomyces, Hansenula, Rhodotorula) were found to be associated with the fermentation process. The pH of the fermenting cassava increased from 4.2 to 5.7 after 72 h while the cyanide level decreased from 2.2 mg/kg to 0.7 mg/kg over the same period of fermentation. Fufu (prepared by crushing and sieving fermenting cassava roots) enriched with 0.5 g of C. utilis strain BKT4, S. cerevisiae BKT7 and a mixed culture of the two organisms revealed a crude protein of 7.90, 6.34 and 10.0% respectively as compared to 2% protein content of the enriched fufu. There was a corresponding increase in protein content of the product as the quantity of the enrichment yeast was increased from 0.5 to 3.0 g. The aroma of the enriched fufu was preferred to that of the commercial fufu. Generally, good acceptability and organoleptic qualities (colour, taste, texture and aroma) of the protein enriched fufu was best achieved within 48 h of enrichment. The results of this study suggest that fufu can be made more nutritious with yeasts particularly Candida utilis strain BKT4 and Saccharomyces cerevisiae strain BKT7.     

Pulmonary vascular smooth muscle: biochemical and mechanical developmental changes.

To evaluate the developmental changes in pulmonary vascular smooth muscle contractile protein content, mechanical properties, and their contribution to the high resistance characteristic of the fetal and immediate neonatal period, we studied pulmonary vessels of fetal, newborn, and adult sheep, as well as newborn and adult pigs. Strips of the second- through fifth-generation vessels were dissected, and their content of tissue total smooth muscle cell protein, myosin, and actin-to-myosin ratio were measured; the mechanical properties of the second-generation vascular strips were also studied. For all ages the smooth muscle protein and myosin content of the second-generation vessels were significantly greater than for the lower pulmonary vascular orders (P less than 0.05). The myosin content in fetal sheep (0.77 +/- 0.03 micrograms/mg wet tissue) was similar to that of the newborn (0.79 +/- 0.04) and adult (0.86 +/- 0.05). However, the smooth muscle protein content (7.94 +/- 0.21 micrograms/mg wet tissue) and the actin-to-myosin ratio of the pulmonary vascular tissue of the fetus (1.00 +/- 0.04) were lower (P less than 0.01) in the fetal than in the newborn (9.16 +/- 0.26 and 1.60 +/- 0.12) and adult (9.38 +/- 0.3 and 1.60 +/- 0.11, respectively). No differences were observed for these parameters between the newborn and adult pig. Stress (16.5 +/- 1.7 mN/mm2) and the maximum shortening capacity (13.0 +/- 1.5% of optimal length) in the newborn pulmonary vascular strips were significantly greater than for the fetus (6.8 +/- 1.4 and 5.9 +/- 1.0, respectively) but similar to those of the adult sheep.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphological and biochemical changes of LLC-PK1 cells during adaptation to glucose-free culture conditions.

The established renal epithelial cell line LLC-PK1 retained in tissue culture several differentiated properties of renal proximal tubular cells. By adapting LLC-PK1 cells to glucose-free culture conditions, we recently succeeded in isolating a gluconeogenic strain of LLC-PK1 cells capable of growing in the absence of hexoses. In contrast to the parental wild type, the isolated strain expressed fructose-1,6-bisphosphatase activity and was, therefore, designated LLC-PK1-FBPase+. Besides the differences in glucose metabolism, the isolated gluconeogenic substrain differs form the parental wild type with respect to morphological appearance and the expression of apical membrane marker enzymes. LLC-PK1-FBPase+ cells display a drastic accumulation of autophagic vacuoles, disappearance of apical membrane alkaline phosphatase activity, and increased gamma-glutamyltranspeptidase activity. In order to find out whether or not a low alkaline phosphatase activity in combination with the enhanced formation of autophagic vacuoles is related to a change in apical membrane surface, we utilized a combined light and electron microscopic morphometric procedure to determine the absolute amount of organelle volumes and membrane surface areas. This stereologic approach shows that LLC-PK1-FBPase+ cells display a tenfold increase in the volume of autophagic vacuoles and the lysosomal compartment. Analysis of lysosomal enzyme activities, however, revealed no changes as compared to wild-type cells. The apical membrane surface of gluconeogenic cells was found to be increased by 80%. Karyotype analysis revealed that LLC-PK1 wild-type cells were diploid, whereas FBPase+ cells exhibited polyploidy with a high percentage of tetraploid nuclei. Culturing LLC-PK1-FBPase+ cells in the presence of 5 mM glucose does not abolish the morphological and biochemical changes described, indicating the stability of the FBPase+ strain.     

植物衰老期间生理生化变化的研究进展

植物衰老是受内外因素控制的细胞有序降解并最终导致死亡的过程,衰老期间会出现与正常生长阶段不同的生理生化变化。植物衰老引起的各种功能的下降极大地限制了作物产量潜力的发挥,种子贮存过程中的衰变、逆境条件下植株的早衰、果蔬采后贮藏衰老导致货架寿命的缩短等均会造成极大的经济损失。研究植物衰老的生理机制及其调控具有十分重要的意义。综述了有关植物衰老时生理生化变化方面的近期研究进展,以利于人们对植物衰老生理的更深入的了解。     

Ageing of the human corneal stroma: structural and biochemical changes.

High and low angle X-ray diffraction patterns from the corneal stroma give information about the mean intermolecular spacing of the collagen molecules and the mean interfibrillar spacing of the collagen fibrils, respectively. X-ray data were collected, using a high intensity synchrotron source, from human corneas and sclera at approximately physiological hydration. The spacings were measured as a function of tissue age. Between birth and 90 years there is an increase in the cross-sectional area associated with each molecule in corneal collagen from approx. 3.04 nm2 to 3.46 nm2, and an increase in scleral collagen from approx. 2.65 nm2 to 3.19 nm2. These changes may be due to an increase in the extent of non-enzymic cross-linking between collagen molecules over the age range. We have investigated this possibility by measuring collagen glycation using the thiobarbituric acid assay and the subsequent advanced glycation end-products (AGEs) using fluorescence emission. The results obtained have shown an age-related increase in glycation and AGEs in both tissues. We have also demonstrated a decrease in the interfibrillar spacing of corneal collagen with increasing age which may be related to changes in the proteoglycan composition of the interfibrillar matrix.     

Physiological and biochemical changes in frog sciatic nerve during anoxia and recovery

Frog (Rana pipiens) sciatic nerve was incubated, with and without stimulation, in an oil bath. The correlation between changes in the magnitude of the compound action potential (α and β) and changes in metabolites, particularly energy reserves, during anoxia and recovery from anoxia was studied. The time to extinction of the action potential in anoxia was frequency-dependent. The action potential could not be restored, nor its extinction delayed, by washing the nerve in O₂-free Ringer's solution. Therefore, in this system extracellular K⁺ accumulation was not a significant factor in blocking impulse conduction. At the time of complete nerve block resulting from anoxia (90 min at rest), ATP, P-creatine and glucose were 30, 10 and 10 per cent, respectively, of initial levels. Glycogen did not fall below 42 per cent of control levels even after 5 h of anoxia. Changes in the levels of energy reserves during anoxia were used to calculate the metabolic rate of nerves at rest and during stimulation. In one series of experiments, the resting metabolic rate was 0·12 mequiv. of ‘high-energy phosphate’ (～P)/kg/min. Stimulation increased the metabolic rate to 0·22 mequiv. of ～P/kg/min at 30 Hz and to 0·29 mequiv. of ～P/kg/min at 100 Hz. The change in metabolic rate when the nerve passed from the resting to the stimulated state was quite abrupt, an observation suggesting that the slow transition observed with methods monitoring O₂, consumption was largely instrumental. In nerve stimulated to exhaustion in the absence of O₂, neither ATP nor P-creatine had fully recovered within 60 min after O₂, was readmitted, although the action potential reached supranormal levels 15 min after return to O₂. The ratio of lactate: pyruvate, which increased as expected during anoxia, paradoxically increased even further after O₂, was readmitted. The rate of energy utilization during recovery was 0·30 mequiv. of ～P/kg/min. Nerves stimulated at 100–200 Hz in O₂, exhibited no changes in levels of P-creatine, ATP or lactate, an observation implying that the nerve could not be made to use ～P faster than oxidation of glucose could provide it. This meant that the maximal metabolic rate was not limited by the rate of supply of chemical energy. Instead, the limitation may have arisen as a result of a limited rate at which ionic imbalance can result from stimulation or a limited pump capacity of the axonal membrane. Nerves stimulated at 200 Hz in O₂ for 20 min and then transferred to an O₂-free environment without further stimulation exhibited an increase in the rate of energy utilization (nearly two-fold) over the resting rate, a finding that suggested a metabolic (ionic?) debt as a result of activity which could not be met even though the energy supply was adequate. Therefore, restriction of energy expenditure by a limiting pumping rate seemed to be the most likely explanation. The resting metabolic rate of frog sciatic nerve was only one-quarter to one-third of the rate for rat sciatic nerve, when compared at the same temperature (25°C).     

Physiological, biochemical and molecular changes occurring during olive development and ripening

Since ancient times the olive tree (Olea europaea), an evergreen drought- and moderately salt-tolerant species, has been cultivated for its oil and fruit in the Mediterranean basin. Olive is unique among the commercial important oil crops for many reasons. Today, it ranks sixth in the world's production of vegetable oils. Due to its nutritional quality, olive oil has a high commercial value compared with most other plant oils. Olive oil has a well-balanced composition of fatty acids, with small amounts of palmitate, and it is highly enriched in the moneonic acid oleate. This makes it both fairly stable against auto-oxidation and suitable for human health. Nevertheless, it is the presence of minor components, in particular phenolics, contributing for oil's high oxidative stability, color and flavor, that makes olive oil unique among other oils. Moreover, as a result of their demonstrated roles in the prevention of cancer and cardiovascular diseases, olive phenolics have gained much attention during the past years. Also unique to virgin olive oil is its characteristic aroma. This results from the formation of volatile compounds, namely, aldehydes and alcohols of six carbon atoms, which is triggered when olives are crushed during the process of oil extraction. The biochemistry of the olive tree is also singular. O. europaea is one of the few species able to synthesize both polyols (mannitol) and oligosaccharides (raffinose and stachyose) as the final products of the photosynthetic CO(2) fixation in the leaf. These carbohydrates, together with sucrose, can be exported from leaves to fruits to fulfill cellular metabolic requirements and act as precursors to oil synthesis. Additionally, developing olives contain active chloroplasts capable of fixing CO(2) and thus contributing to the carbon economy of the fruit. The overall quality of table olives and olive oil is influenced by the fruit ripening stage. Olive fruit ripening is a combination of physiological and biochemical changes influenced by several environmental and cultural conditions, even if most events are under strict genetic control.