Optimizing BM harvesting from normal adult donors.

The experience at a single institution of BM harvesting (BMH) in general anesthetic for allogeneic transplantation from 49 healthy adult donors since March 2002 is presented in detail, together with an analysis of all the donor complications. In this study, we analyzed the advantages through the change from an aspiration needle with one hole (group A, n=18) to a system with additional five side holes (group B, n=31) in April 2005 for faster aspiration of large volumes of BM. In group B, the operation time was reduced by 50%, which is 12 min to date (1006 ml BM). Furthermore, the collection rate (volume BM/time) was significantly increased, namely to 81.9 ml/min in group B. The yields of total nucleated cells and CD34+ cells are nearly identical and adequate in both systems. The proportion of donors treated as day cases--that is, able to be discharged on the same day as the procedure--was 56% in group A and 81% in group B. There was no significant operative site morbidity. BMH accomplished by trained personal is a safe procedure for healthy adult donors on an outpatient basis as standard in our collection center.

Long-term abnormalities in brain glucose/energy metabolism after inhibition of the neuronal insulin receptor: implication of tau-protein.

The triplicate intracerebroventricular (icv) application of the diabetogenic compound streptozotocin (STZ) in low dosage was used in 1-year-old male Wistar rats to induce a damage of the neuronal insulin signal transduction (IST) system and to investigate the activities of hexokinase (HK), phosphofructokinase (PFK), glyceraldehyde-3-phosphate dehydrogenase (GDH), pyruvate kinase (PK), lactate dehydrogenase (LDH) and alpha-ketoglutarate dehydrogenase (alpha-KGDH) in frontoparietotemporal brain cortex (ct) and hippocampus (h) 9 weeks after damage. In parallel, the concentrations of adenosine triphosphate (ATP), adenosine diphosphate (ADP), guanosine triphosphate (GTP) and creatine phosphate (CrP) were determined. We found reductions of HK to 53% (ct) and 60% (h) of control, PFK to 63/64% (ct/h); GDH to 56/61% (ct/h), PFK to 57/59% (ct/h), alpha-KGDH to 37/35% (ct/h) and an increase of LDH to 300/240% (ct/h). ATP decreased to 82/87% (ct/h) of control, GTP to 69/81% (ct/h), CrP to 82/81% (ct/h), approximately P to 82/82% (ct/h), whereas ADP increased to 189/154% (ct/h). The fall of the activities of the glycolytic enzymes HK, PFK, GDH and PK was found to be more marked after 9 weeks of damage when compared with 3- and 6-week damage whereas the diminution in the concentration of energy rich compound was stably reduced by between 20 and 10% relative to control. The abnormalities in glucose/energy metabolism were discussed in relation to tau-protein mismetabolism of experimental animals, and of sporadic AD.

Relationship between cerebral energy metabolism in parietotemporal cortex and hippocampus and mental activity during aging in rats.

The present investigation demonstrates differences in both formation and utilization of the energy-rich compounds adenosine triphosphate ATP and creatine phosphate (CrP) in behaviorally well and poorly performing inbred male Wistar rats in parietotemporal cerebral cortex and hippocampus with aging from 1 to 2ys. Also, differences in learning and memory capacities (behavior) became obvious. By holeboard testing, good (GP) and poor performers (PP) have been discriminated. The pools of energy-rich phosphates as determined in parietotemporal cerebral cortex and in hippocampus under resting conditions were found to be reduced by nearly 10% for both creatine phosphate and the whole available energy pools in PP as compared to GP. Aging from 1 to 2y diminished the concentrations of energy-rich phosphates in the cerebral areas studied and in both GP and PP under resting conditions. Additionally, an age-related aggravation of the energy deficit became obvious between GP and PP. Repeated mental activation from 1y to 2y resulted in the maintenance of improvement as registered for the mean run time and the number of visited/revisited holes in GP. In contrast, PP deteriorated (mean run time), and could not maintain improvement (number of visited/revisited holes) over time. Repeated mental activation normalized the energy pool by increased formation of the energy-rich compounds ATP and CrP in both cerebral areas studied in GP and PP. However, differences became obvious between GP and PP. The energy-turnover in the latter group was found to be significantly reduced for both cerebral areas studied. GP could meet the enhanced energy demand of mental activation during aging by increasing formation and utilization of energy. PP could increase energy formation but were unable to sufficiently adapt energy utilization under the same conditions. This disturbance in energy metabolism may have impacts on energy-consuming processes in PP which may contribute to the markedly reduced cognitive reserve in PP. In human beings, PP approximately poorly educated people found to be prone to sporadic Alzheimer disease.

Nematic liquid crystals with a tetrafluoroethylene bridge in the mesogenic core structure.

A dramatic increase of the clearing temperatures of liquid crystals based on bis(cyclohexyl)ethane 1 by 50 to 70 K can be achieved by the perfluorination of the central ethylene link. Conformational analysis indicates that this effect is due to the increased rigidity of the mesogenic core structure and to the suppression of conformers with a bent shape. Materials based on bis(cylohexyl)tetrafluoroethane 2 might play a crucial role as materials for the next generation of active matrix LCDs with reduced power consumption.

Renal failure in multiple myeloma "the myeloma kidney":state of the art.

Renal failure is present in about 20% of patients with multiple myeloma (MM) at diagnosis. Renal function impairment is usually caused by the so-called "myeloma kidney" and is associated with shortened survival in patients treated with conventional therapy. Renal failure is reversible in up to 50% of patients, particularly when its degree is moderate and it is related to precipitating factors such as hypercalcemia. In our experience, approximately 10% of newly diagnosed patients with MM have renal failure severe enough to require dialysis. Despite its frequency, there are few reports dealing with MM and renal failure.

Chronic exposure to low-level lead impairs learning ability during aging and energy metabolism in aged rat brain.

The neurotoxic effect of chronic exposure to low-level lead (Pb) with advancing age is becoming an important social issue of public health. To examine the effects of low-level Pb treatment on behavior, cognition and brain energy metabolism in aging, we administered 200 ppm Pb acetate to adult (10-month-old) male Wistar rats for 12.5 months. After 12.5 months' exposure, the mean Pb levels in blood and brain had increased to 17.5 µg/dl and 0.07 µg/g, respectively, and the rats showed impaired learning and memory functions in a holeboard spatial memory test. No significant difference was found between experimental and control groups in locomotor activity and passive avoidance tests. By HPLC analysis of energy-rich phosphate concentrations, mild abnormalities were found in parietotemporal cortex and hippocampus, but only the 4.4% decrease of ATP in the parietotemporal cortex was statistically significant. These results suggest that chronic exposure to Pb during aging stage may selectively impair learning and memory functions and may cause slight cerebral energy impairment.

Damaged neuronal energy metabolism and behavior are improved by Ginkgo biloba extract (EGb 761).

The standardized extract EGb 761 from the dried green leaves of Ginkgo biloba is a complex mixture of ingredients with an uniquely broad spectrum of pharmacological activities on the central nervous system e.g. in memory enhancing properties and in the regulation of cerebral glucose/energy metabolism. To test these effects on both behavioral and metabolic brain parameters, the animal model of intracerebroventricular (i.c.v.) streptozotocin (STZ) treatment was used. To quantify the experimental data more precisely, animals that were good performers were separated from poor performers by means of the holeboard test before i.c.v. administration of STZ. Good performers only were considered for the study. After a 1-week training period on the holeboard improvement was seen in all animals in learning, memory and cognition, and the improvement was maintained over the investigation period of 12 weeks in the control group. In this group, the energy pool in the cerebral parietotemporal cortex was found to be large and the energy turnover high. After triplicate i.c.v. STZ injection, working memory (WM), reference memory (RM), and passive avoidance (PA) behavior fell off and continued to deteriorate throughout the investigation period. Otherwise there were no significant differences in locomotor activity, excluding the possibility that activity per se might have contributed to the behavioral abnormalities. These were accompanied by a permanent deficit in cerebral energy metabolism. The ongoing deterioration in behavior and the maintained deficit in cerebral energy metabolism occurring after a triplicate i.c.v. STZ injection were significantly slowed down by EGb761. The deficits in learning, memory and cognition were partially compensated, and the disturbances in cerebral energy metabolism returned to almost completely normal values. These findings underscore the beneficial effect of EGb761 that had been reported in dementia disorders.

Effects of Estradiol (-17beta) on learning, memory and cerebral energy metabolism in male rats after intracerebroventricular administration of streptozotocin.

Treatment of adult rats with intracerebroventricularly (i.c.v.) injected streptozotocin (STZ) may provide a relevant animal model of chronic neuronal dysfunction that is characterized by a decrease in both the neuronal metabolism of glucose and the formation of energy. The present study was designed to evaluate whether or not rats treated with triplicate i.c.v. STZ injection would show long-term effects in learning and memory behavior as measured by means of a holeboard test, closed field activity, and passive avoidance behavior over a period of 6 weeks. For this purpose, animals with good performance were discriminated from those with poor performance by the holeboard test before i.c.v. administration of STZ. After a 1-week training period with the holeboard all animals improved their abilities in learning and memory, and the improvement was maintained over the investigation period of 6 weeks in the control group. After i.c.v. STZ working memory (WM), reference memory (RM), as well passive avoidance (PA) behavior decreased, deteriorating progressively during the investigation period. The latter were accompanied by a permanent deficit in cerebral energy metabolism. The ongoing deterioration in behavior and the sustained deficit in cerebral energy metabolism occurring after a triplicate i.c.v. STZ administration lead us to assume that this animal model may be appropriate for the investigation of mechanisms characteristic for sporadic Alzheimer disease. In this context, the effect of Estradiol-17beta (E2) on behavior and energy metabolism was studied. We found that E2 slowed down the i.c.v. STZ-induced deterioration in memory functions, partially compensated the learning deficit, and improved the disturbances in cerebral energy metabolism to the extent that it was almost completely normal again. These findings underscore the beneficial effect of E2 in dementia disorders.

Intracerebroventricular administration of streptozotocin causes long-term diminutions in learning and memory abilities and in cerebral energy metabolism in adult rats.

Drastic abnormalities have been demonstrated to occur in cerebral glucose and energy metabolism in sporadic Alzheimer's disease, pointing to a primary disturbance in neuronal insulin and insulin receptor signal transduction and contributing to the causation of dementia. The compound streptozotocin (STZ) is known to inhibit insulin receptor function. The study was designed to investigate whether intracerebroventricularly (icv) applied STZ would inhibit neuronal insulin receptor function and would induce changes in both behavior and neuronal energy metabolism. Adult rats with icv-injected STZ developed long-term and progressive deficits in learning, memory, and cognitive behavior, indicated by decreases in working and reference memory in the holeboard task and the passive avoidance paradigm, along with a permanent and ongoing cerebral energy deficit. This animal model may be appropriate for investigations related to sporadic Alzheimer's dementia.

Functional organization of the Golgi apparatus in glycosphingolipid biosynthesis. Lactosylceramide and subsequent glycosphingolipids are formed in the lumen of the late Golgi.

Biosynthesis of plasma membrane sphingolipids involves the coordinate action of enzymes localized to individual compartments of the biosynthetic secretory pathway of proteins. These stations include the endoplasmic reticulum and the Golgi apparatus. Although a precise localization of all the enzymes that synthesize glycosphingolipids has not been achieved to date, it is assumed that the sequence of events in glycosphingolipid biosynthesis resembles that in glycoprotein biosynthesis, i.e. that early reactions occur in early stations (endoplasmic reticulum and cis/medial Golgi) of the pathway, and late reactions occur in late stations (trans Golgi/trans Golgi network). Using truncated analogues of ceramide and glucosylceramide that allow measurement of enzyme activities in intact membrane fractions, we have reinvestigated the localization of individual enzymes involved in glycosphingolipid biosynthesis and for the first time studied the localization of lactosylceramide synthase after partial separation of Golgi membranes as previously described (Trinchera, M., and Ghidoni, R. (1989) J. Biol. Chem. 264, 15766-15769). Here, we show that the reactions involved in higher glycosphingolipid biosynthesis, including lactosylceramide synthesis, all reside in the lumen of the late Golgi compartments from rat liver.

Brain glucose metabolism is controlled by amplification and desensitization of the neuronal insulin receptor.

Glucose metabolism is essential for brain function and structure. Glucose contributes to the formation of neurotransmitters and is normally the only source for energy formation. There is increasing evidence that brain glucose metabolism is under control of the neuronal insulin/insulin receptor signal transduction. The present data clearly show that intracerebroventricularly administered insulin exerts anabolic effects on cerebral glucose/energy metabolism (amplification of the neuronal insulin receptor complex) whereas cortisol (corticosterone) acts antagonistically (desensitization of the neuronal insulin receptor complex). It is also shown that short-term cortisol (corticosterone) enhanced energy turnover in temporoparietal cortex and hippocampus. In contrast, long-term cortisol (corticosterone) reduced energy turnover in both brain structures studied. This metabolic pattern is reminiscent of that found in very old age. Therefore, it is assumed that long-term cortisol accelerates the aging process in the brain and thus the risk for age-related disorders such as dementia.

Lactosylceramide is synthesized in the lumen of the Golgi apparatus.

Recently, synthesis of lactosylceramide has been described to occur on the cytosolic face of the Golgi [(1991) J. Biol. Chem. 266, 20907-20912]. The reactions following in the biosynthesis of higher glycosphingolipids are known to take place in the lumen of the Golgi. For our understanding of the functional organization of the multiglycosyltransferase system of glycosphingolipid synthesis in the Golgi, the knowledge of the topology of individual reactions is a prerequisite. We have developed a simple and quick assay system for sphingolipid biosynthesis and have obtained evidence that lactosylceramide is synthesized in the lumen of the Golgi. Because lactosylceramide is generated by galactosylation of glucosylceramide which, in turn, is synthesized from ceramide and UDP-Glc on the cytosolic surface of the Golgi apparatus, further efforts will be directed to the characterization of a glucosylceramide-translator in the Golgi membranes rather than a lactosylceramide-translocator.