Metabolic regulation of stem cell function.

Stem cell function is regulated by intrinsic mechanisms, such as transcriptional and epigenetic regulators, as well as extrinsic mechanisms, such as short-range signals from the niche and long-range humoral signals. Interactions between these regulatory mechanisms and cellular metabolism are just beginning to be identified. In multiple systems, differentiation is accompanied by changes in glycolysis, oxidative phosphorylation and the levels of reactive oxygen species. Indeed, metabolic pathways regulate proliferation and differentiation by regulating energy production and the generation of substrates for biosynthetic pathways. Some metabolic pathways appear to function differently in stem cells as compared with restricted progenitors and differentiated cells. They also appear to influence stem cell function by regulating signal transduction, epigenetic marks and oxidative stress. Studies to date illustrate the importance of metabolism in the regulation of stem cell function and suggest complex cross-regulation likely exists between metabolism and other stem cell regulatory mechanisms.

Multi-segment trunk kinematics during a loaded lifting task for elderly and young subjects.

The trunk is frequently modelled as one fixed segment ignoring possible multi-segmental contributions during manual handling. This study compared segmental trunk motion in a young and older population during a lifting task. Twelve elderly and 19 young subjects repeatedly lifted a 5 kg box from bench to shelf under two stance conditions. Displacement and angular trunk segment kinematics were recorded with an electromagnetic tracker system and then analysed. The elderly subjects displayed significantly increased pelvic and trunk displacement and significantly reduced pelvic and lower thorax (T10-L1) range of motion in both stance conditions. Upper thorax (C7-T10) motion was at times greater than lumbar motion and opposite to the lower segments and was related to the task while the lower segments contributed to both equilibrium and task requirements. Decreased segmental trunk angular kinematics may contribute to increased displacement kinematics and place the elderly at increased risk of injury and falling. The pelvis, lumbar spine, low thorax (T10-L1), upper thorax (C7-10) contributed uniquely and synchronously to trunk (C7-S2) mechanics during a lifting task. Reduced angular kinematics of the pelvis and low thorax contributed to increased displacement kinematics and hence increased the risk of falling in the elderly compared to the young. Investigations of trunk mechanics should include multi-segment analysis.

A case of head and neck kaposiform hemangioendothelioma simulating a malignancy on imaging.

Kaposiform hemangioendothelioma (KH) is an endothelial-derived spindle cell neoplasm often associated with Kasabach-Merritt syndrome. Most cases arise in infancy and childhood and are soft-tissue tumors. The tumor displays an appearance between capillary hemangioma and Kaposi's sarcoma. We report a case of KH in a 1-year-old girl involving a mass that showed abnormal enhancement of soft tissue superficial to the right temporal bone with partial destruction of the temporal bone, the temporomandibular joint, mandibular condyle, and occipital bone. The physical finding of a discolored mass led clinicians to consider a hemangiomatous lesion, whereas the radiological picture suggested a more aggressive diagnosis of rhabdomyosarcoma and aggressive fibromatosis.

Partition coefficients of low-molecular-weight volatile chemicals in various liquids and tissues.

Partition coefficients are required for developing physiologically based pharmacokinetic models used to assess the uptake, distribution, tabolism, and elimination of volatile chemicals in mammals. A gas-phase vial equilibration technique is presented for determining the liquid:air and tissue:air partition coefficients for low-molecular-weight volatile chemicals. This technique was developed from two previously described medium:air methods, relied solely on measurement of chemical concentration in the gas phase, and, compared to earlier work, extends the range of chemicals and tissues examined. Partition coefficients were determined with 0.9% saline, olive oil, and blood, liver, muscle, and fat tissues from rats for 55 compounds. Human blood:air coefficients were determined for 36 compounds and several blood:air values were also determined in the mouse and for one compound in the hamster. An approach is described for predicting the tissue solubilities of untested compounds based on oil:air and saline:air coefficients using regression analyses. A similar approach is used to model fat:air coefficients in terms of oil:air values and to model human blood: air coefficients in terms of rat blood:air coefficients.

Long-latency auditory event-related potentials in epilepsy.

Long latency auditory event-related potentials have been shown to change in patients with cerebral dysfunction. Some seizure patients with no evidence of brain damage or mental retardation show altered interictal cognitive and memory function. Long-latency auditory event-related potentials to tone stimulation were recorded in nineteen control subjects and seventeen patients with complex partial or partial and secondarily generalized seizures who had no evidence of brain damage, retardation, or drug intoxication, and whose seizures were controlled when studied. The latencies of N2 and P3 components were significantly longer in seizure patients than control subjects, and the P3 waveform was significantly greater in amplitude in epileptics. These findings suggest that cognitive event-related potentials are affected by partial epilepsy. The changes may be related to the recently reported involvement of the hippocampus in ERP generation, or to loss or alteration of modulatory functions, possibly cholinergic in nature, in the temporal lobe consequent upon epileptogenesis.

The folding and mutual interaction of the domains of yeast 3-phosphoglycerate kinase.

Analysis of the reversible unfolding of yeast phosphoglycerate kinase leads to the conclusion that the two lobes are capable of folding independently, consistent with the presence of intermediates on the folding pathway with a single domain folded. The domains have different free energies of stabilisation. Immunological cross-reactivity, circular dichroism and thiol reactivity provide evidence for cyanogen bromide peptide 1-173, which comprises five-sixths of the N-terminal domain, containing sufficient information to refold into a native-like structure which dimerises.

Interictal EEG prediction of response to antiepileptic drug monotherapy.

Forty-eight patients had sleep-deprived EEGs prior to antiepileptic drug monotherapy. The majority were seizure-free after one year, or had more than 50% reduction in seizure frequency. Among those with normal EEGs 50% were seizure-free, while 75% with diffuse slowing, 44% with focal abnormality, and 83% with generalized epileptiform discharges were fully controlled. Freedom from seizures was achieved in 13% taking phenobarbital, 50% taking phenytoin, 63% taking carbamazepine, and 100% taking valproate. The sleep-deprived interictal EEG should be an integral part of initial assessment and drug selection in patients with clinical histories of convulsive seizure.

Computerized spectral analysis of the interictal EEG in epilepsy.

Eight seizure patients without encephalopathy, frequent seizures, brain lesion, or medication intoxication had significant slowing of alpha-frequency activity, as compared to nonepileptic controls, that was evident in compressed spectral analysis but not in standard EEG. Those patients with cognitive or behavioral problems or taking more than one antiepileptic medication had a greater degree of slowing. Differences between seizure patients with and without cognitive or behavioral symptoms, and between specific antiepileptic drugs, were suggested but could not be assigned significance due to small numbers. The findings suggest that interictal changes in brain function that are not revealed by standard EEG may relate to observed changes in interictal behavior and cognition, and that computerized spectral analysis of the interictal EEG may be of value in the assessment of seizure patients before and during therapy.

Protein degradation in rat liver. Evidence for populations of protein degradation rates in cellular organelles.

Protein degradation in liver subcellular and submitochondrial fractions from adult rats has been measured by a double isotope technique. Protein subunits have been resolved by two-dimensional polyacrylamide gel electrophoresis. No consistent relationship between subunit size or isoelectric point and degradation rate has been found in any fraction. Frequency analysis of isotope ratios for protein subunits in subcellular and submitochondrial fractions has been carried out. The analyses demonstrate that degradation rates of resolved protein subunits in organelles or organelle subcompartments can be grouped into populations. Resolved protein subunits in subcellular and submitochondrial fractions can be grouped into one, two or three populations. The results suggest limited heterogeneity of protein degradation rates within each subcellular organelle and support the notion that morphological subcompartment rather than molecular property may be the prime determinant of rates of degradation of individual proteins.

Protein degradation in rat liver during post-natal development.

Protein degradation rates for liver subcellular and submitochondrial fractions from neonatal (8-day), weanling (25-day) and adult rats were estimated by the double-isotope method with NaH14CO3 and [3H] arginine as the radiolabelled precursors [Dice, Walker, Byrne & Cardiel (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 2093-2097]. Decreased protein degradation rates were found during post-natal development for homogenate, nuclear, mitochondrial, lysosomal and microsomal proteins. A decrease in degradation rates for the immunoisolated subunits of monoamine oxidase and pyruvate dehydrogenase was also observed in neonatal and weanling rats respectively. The results suggest coordinate degradation of the subunits of the multi-subunit enzyme pyruvate dehydrogenase. Pyruvate dehydrogenase has a faster rate of degradation in adult rat liver than does cytochrome oxidase. Data analysis suggests heterogeneity of protein degradation rates in the mitochondrial outer membrane and intermembrane space fractions at each developmental stage but not in the mitochondrial inner membrane or matrix fractions. Results obtained for protein degradation rates in adult rat liver by the method of Burgess, Walker & Mayer [(1978) Biochem. J. 176, 919-926] in general confirmed the results obtained for the adult rat liver by the above method. No evidence of a subunit-size relationship for protein degradation was found for proteins in any subcellular or submitochondrial fraction.

Coordination of protein synthesis and degradation.

The degree of coordination between protein synthesis and protein degradation in developing and mature cels is considered. Studies on specific enzyme and general protein turnover in developing liver and differentiating mammary gland are presented. In the mature liver mitochondrion average protein degradation rates are higher for outer membrane and intermembrane space proteins than for matrix and inner membrane proteins. Significant heterogeneity of protein degradation rates was observed only in the outer mitochondrial membrane. During postnatal development the rates of degradation of proteins in many liver cellular fractions are increased. In the mitochondrion only the average rates of degradation of proteins in the outer membrane and intermembrane space fractions increase during development. Evidence for hormonally regulated changes in both protein synthesis and degradation during mammary cell differentiation is given. The data indicate that a transitory decrease in protein degradation accompanies the increase in protein synthesis on hormonal stimulation of the tissue. The results from the two model systems are collated and used to formulate a phenomenological hypothesis of protein degradation and its integration with protein synthesis in steady-state and non-steady-state conditions.

Choice of precursors for the measurement of protein turnover by the double-isotope method. Application to the study of mitochondrial proteins.

1. The double-isotope concept [Arias, Doyle & Schimke (1969) J. Biol. Chem. 224, 3303--3315] for the measurement of protein turnover was used to estimate the turnover of proteins in subcellular and submitochondrial fractions prepared from rat liver. 2. Double-isotope experiments with [3H]leucine as first precursor and [14C]leucine as second precursor were used to measure the turnover rates of proteins in subcellular and submitochondrial fractions. Solvent extraction procedures designed to remove lipids and nucleic acids from trichloroacetic acid precipitates only changed the isotope ratio of the microsomal fraction. It was not possible to measure turnover of proteins in mitochondrial and submitochondrial fractions with these precursors. 3. Double-isotope experiments were designed to minimize first-precursor reutilization by employing NaH14CO3. [3H]Arginine was used as second precursor. The turnover rates of protein in subcellular and submitochondrial fractions was measured. Solvent extraction procedures designed to remove lipids and nucleic acids showed changes in the isotope ratio for all subcellular fractions, especially in microsomal and detergent-soluble mitochondrial fractions. Isotope ratios of precipitates after solvent extraction indicate that, whereas considerable heterogeneity exists for the average rates of protein turnover in subcellular fractions, little heterogeneity is observed in the average rates of protein turnover in submitochondrial fractions.

Relative rates of turnover of subunits of mitochondrial proteins.

1. The double-isotope concept [Arias, Doyle & Schimke (1969) J. Biol. Chem. 244, 3303--3315] for the measurement of protein turnover was used to estimate the turnover rates of protein subunits from rat liver submitochondrial fractions resolved by means of polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. NaH14CO3 and [5-3H]arginine were used as first and second precursors respectively. 2. Marked heterogeneity of protein subunit turnover rates is seen for protein subunits from water-soluble, salt-soluble and Tween 20-soluble mitochondrial proteins. 3. Much lower heterogeneity is seen in the turnover of protein subunits in Triton X-100-soluble material not binding to DEAE-cellulose at low ionic strength. The relative rates of turnover of proteins in this fraction are lower than for proteins in any other submitochondrial fraction. This fraction contains the integral membrane proteins. 4. Incorporation of [3H]arginine into subunits of the cytochrome oxidase complex is greatest for subunits with molecular weights in excess of 20000. 5. No correlation is seen between protein subunit size and the rate of turnover of the protein subunits in any of the submitochondrial fractions.