Eimeria tenella contains a pyrophosphate-dependent phosphofructokinase and a pyruvate kinase with unusual allosteric regulators

To determine whether the satiating effects of fiber are due to delaying gastric emptying or slowing absorption of meals, 3% guar gum was added to high- and low-fat soups and gastric emptying rate, hunger, and satiety were measured in eight male volunteers. Guar gum delayed the emptying of the low-fat soup but the small delays in the return of hunger and decline of fullness were significantly correlated with the gastric emptying, suggesting mediation by gastric mechanoreceptors. The high-fat soup also emptied more slowly but this had no effect on the return of hunger or the decline in fullness. The delays in the return of hunger and decline of fullness were far greater when guar gum was added to the fatty soup; these delays were not correlated with the small additional delay in gastric emptying. This is more compatible with slowed absorption and prolonged contact of nutrients with intestinal chemoreceptors.     

Characterization of pyruvate kinase from the liver of a patient with aberrant erythrocyte pyruvate kinase, PK Nagasaki

The characterization of the L-type PK were made of PK extracted from the liver of a patient with congenital hemolytic anemia associated with an erythrocyte PK variant, PK Nagasaki. The L-type PK of PK Nagasaki showed the following parameters: slow migration on electrophoresis, high Km for PEP without F-1,6-P2, less activation by F-1,6-P2, normal Km for ADP, high utilization of UDP, acidic pH optimum, and instability to urea and heat. These tests served to differentiate this L-type PK variant from the other variants previously reported. At the same time, both the Km for PEP with F-1,6-P2 saturation and the electrophoretic mobility of L-type PK were found to be different from those of the erythrocyte PK and PK Nagasaki. Though the liver cell, with regard to L-type PK, has only the less functional and less stable mutant L-type PK there is no evidence of liver dysfunction or damage, although there is chronic hemolytic anemia.     

Continuous neurogenesis in the olfactory brain of adult shore crabs, Carcinus maenas

To scrutinize the common belief that the number of neurons in the CNS of adult decapod crustaceans stays constant, in spite of their dramatic postlarval increase in size, I counted olfactory projection neurons (OPNs) in the brains of differently-sized postlarval shore crabs, Carcinus maenas, and performed in vivo labeling of proliferating cells with 5-bromo-2'-deoxyuridine (BrdU) on brains of adults. The number of OPNs increases continuously throughout the postlarval life of shore crabs and approximately doubles from the very young to the oldest animals. Brain sections from adult crabs labeled with BrdU revealed ongoing proliferation of cells in the lateral soma cluster, which consists of OPN cell bodies, and in the cluster of somata of hemiellipsoid body local interneurons, which are the targets of the OPNs. Post-injection survival times from 5.5 to 120 h revealed a small but relatively constant number of labeled nuclei with neuronal morphology in both soma clusters of all specimens (31.3 +/- 9.5 S.D. nuclei per lateral cluster, n = 29; 20.1 +/- 4.5 S.D. nuclei per hemiellipsoid body cluster, n = 10). The labeled nuclei were located in a distinct proliferative zone in each cluster. There were significantly more labeled nuclei in both soma clusters after a prolonged post-injection survival time of 1 month (71.3 +/- 7.8 S.D. nuclei per lateral cluster, n = 4; 38.2 +/- 7.1 nuclei per hemiellipsoid body cluster, n = 6). In both soma clusters the labeled nuclei formed a compact group that was dislocated from the proliferation zone towards the outer edge of the cluster. In the proliferation zone of the lateral cluster histological stainings revealed cell bodies of typical neuronal shape that are slightly smaller and more intensely stained than the surrounding OPN somata. Some of these cell bodies were captured in various stages of mitosis. Collectively, these data indicate that continuous neurogenesis occurs in the central olfactory pathway of the brain of shore crabs throughout their entire adult life. This unexpected structural plasticity may enable long-lived decapod crustaceans to adapt to ever-changing olfactory environments.     

The binding of Cd(II) to the hemocyanin of the Mediterranean crab Carcinus maenas

In idiopathic dilated cardiomyopathy (IDC), the relation between the signal-averaged electrocardiogram and ventricular tachycardia (VT) remains unclear. In this study, conventional time domain and frequency domain analyses (2-dimensional, spectral temporal mapping and spectral turbulence analysis) of the signal-averaged electrocardiogram were performed in 64 patients with IDC. Eight patients had a history of symptomatic sustained VT and an additional 24 had nonsustained VT recorded during ambulatory electrocardiography. Conventional time domain analysis, using the 25 and 40 Hz filter, and spectral temporal mapping, detected late potentials within the terminal QRS in 8 (13%), 14 (22%) and 18 (28%) patients, respectively. Late potentials were seen more often in patients with than without VT, and in patients with sustained versus nonsustained VT, but these differences were not significant. The predictive accuracy of these techniques in detecting either form of VT were: sensitivity, 22, 25 and 31%; specificity, 97, 81 and 75%; and overall predictive value, 59, 53 and 50%, respectively. Two-dimensional frequency domain analysis of the signal-averaged electrocardiogram revealed a higher energy and area ratio in patients with than without VT (entire QRS), and in patients with sustained versus nonsustained VT (entire QRS and terminal QRS). Spectral turbulence analysis was abnormal in 24 patients (39%), but no differences were observed between patients with and without VT. During follow-up (mean duration 18 +/- 14 months), 5 patients had arrhythmic events (3 died suddenly, 1 had aborted sudden death and 1 developed sustained VT).(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro superoxide production by hyaline cells of the shore crab Carcinus maenas (L.)

Routine clinical follow-up for distant metastatic disease among women with early stage breast cancer is of uncertain clinical benefit. In order to evaluate current practice patterns, we administered a mailed survey to a stratified random sample of physician members of the American Society of Clinical Oncology (ASCO) (N = 435). The survey assessed the frequency and motivation for ordering follow-up medical tests in asymptomatic postmenopausal women with stage I or II breast cancer. The response rate was 55%, distributed as 39% radiation oncologists, 32% medical oncologists, and 29% surgeons. In the first year after treatment, physicians performed, on average, one physical examination every 3 months, one blood panel (CBC, alkaline phosphatase and liver function tests) every 4 months, and one chest radiograph every 9 months. In addition, 38% of the respondents ordered CEA and 21% ordered CA 15-3 levels, 28% ordered bone scans, and less than 4% ordered CT scans, bone surveys, or bone marrow biopsies in the first year after treatment. A logistic regression analysis controlling for physician age, gender, sub-specialty, practice type, years of experience, number of breast cancer patients treated annually, geographic region, and community size, showed that surgeons were less likely to order blood tests (p < 0.001) and tumor markers (p < 0.0029) than medical oncologists in years 3 and 5 of follow-up. Compared to physicians practicing in the northeast, those from the midwest were less likely to order chest radiographs in year 3 of follow-up (p = 0.0028). Other provider characteristics had no significant effect on test ordering behavior. The results of this survey suggest that relatively uniform practice patterns in regard to the follow-up of postmenopausal women with early stage breast cancer exist among ASCO physicians.     

Interaction of M1 and M2 isozymes pyruvate kinase from human tissues with phospholipids

This study aimed to measure energy intake (EI) and total energy expenditure (TEE) of asthmatic males and to validate diet history as a method of estimating their energy requirements. EI was assessed by dietary history and TEE by the heart-rate monitoring method in a group of asthmatic and nonasthmatic males. Resting energy expenditure (REE) adjusted for fat-free mass was higher in asthmatic than in nonasthmatic males (5,037 versus 4,839 kJ x day(-1), p<0.05). TEE (93+/-1.8 versus 8.4+/-1.4 MJ x day(-1), respectively; p=NS) and EI (9.2+/-15 versus 8.8+/-15 MJ x day(-1), respectively, p=NS) were not statistically different in asthmatic and nonasthmatic male. EI was not statistically different from TEE in both groups of males. Asthmatic males showed an acceptable agreement between TEE and EI at the individual level (range of agreement: -3.2 to 2.9 MJ x day(-1)), and a good agreement at the group level (95% confidence interval for the bias, - 1.1 to 0.8 MJ x day(-1)). Males with mild-to-moderate asthma have a higher metabolic activity per unit fat-free mass than nonasthmatic males. This increased requirement is apparently well compensated by an adequate energy intake. Diet history is a suitable method for estimating energy requirements in males with mild-to-moderate asthma.     

Ligand interactions with the acetylcholine receptor from Torpedo californica. Extensions of the allosteric model for cooperativity to half-of-site activity

The solubilized acetylcholine receptor from Torpedo californica showed positive cooperativity in acetylcholine binding with a dissociation constant of 1.2 X 10(-8) M. Blockade of acetylcholine binding by nicotine was competitive; blockade by d-tubocurarine appeared to result from an allosteric interaction that altered half of the acetylcholine binding sites to a lower affinity form; decamethonium blockade displayed properties of competitive and allosteric inhibition suggesting less specificity for decamethonium binding than seen with either nicotine or d-tubocurarine. The d-tubocurarine inhibition data were evaluated by several possible models involving either differential competitive inhibition or allosteric inhibiton. The data were best described by the allosteric model.     

The pyruvate dehydrogenase complex from thermophilic organisms: thermal stability and re-association from the enzyme components

Parabuthus transvaalicus, P. granulatus, and P. villosus are three medically important scorpion species occurring in southern Africa which can cause severe envenoming among people. In contrast to many other genera, no data is available on the venom composition of scorpions belonging to the genus Parabuthus. Here we have investigated the components which may contribute to the venomous potential. The constancy of venom composition within each of the three species and between the three species was investigated by means of gel filtration chromatography. The venoms of the three species each were characterized by a constant and typical elution pattern, resulting in a 'gel filtration fingerprint' which allows distinction between each species. It appears that certain components in the venoms are common to either all three species, or to two of the three species. This points to a clear interspecies relationship within the genus. We also describe the isolation and characterization of some of the polypeptide toxins present in the venoms of P. villosus, P. transvaalicus and P. granulatus by means of reversed phase chromatography and screening of the toxic components on voltage-activated potassium and sodium channels. Our results confirm that toxins which inhibit potassium channels and alter sodium channel gating are present in the venoms studied.     

Infestation of an introduced host, the European green crab, Carcinus maenas, by a symbiotic nemertean egg predator, Carcinonemertes epialti

The recent introduction of the European green crab, Carcinus maenas, to the west coast of the U.S. has provided an opportunity for host transfer of the symbiotic nemertean egg predator, Carcinonemertes epialti, from its native shore crab host, Hemigrapsus oregonensis to the exotic C. maenas. Two surveys of C. maenas in Bodega Harbor, California, revealed that, in March 1995 prevalence of C. epialti on C. maenas was significantly lower than on H. oregonensis (11% versus 74%), but in November 1995 there was no significant difference between the 2 species (79% versus 98%). Only juvenile C. epialti were recovered from C. maenas in March 1995. However, in November 1995, ovigerous C. maenas were harboring actively feeding adult worms. Prevalence in both crab species significantly differed from March to November. Laboratory studies revealed that C. epialti fed and reproduced on eggs of C. maenas. The feeding rate of C. epialti on C. maenas eggs (2.5 eggs/trial) was not significantly different from that on H. oregonensis eggs (3.6 eggs/trial). Our findings suggest that this nemertean may have less host specificity than was previously thought. If C. epialti causes brood mortality of C. maenas in nature, it could potentially impact populations of this exotic crab.     

Purification and properties of the pyruvate kinase isoenzymes type L and M2 from chicken liver

A procedure for the simultaneous purification of both isoenzymes of pyruvate kinase (type M2 and L) from chicken liver has been worked out. Each isoenzyme produces a single band in dodecylsulfate gel electrophoresis. Each has a molecular weight of 190 000 and contains four apparently identical subunits of Mr = 50 000. The isoenzymes differ in their isoelectric points (type L: 6.3; type M2: 8.3) and their kinetic behaviour. Pyruvate kinase type L had an S-shaped phosphoenolpyruvate saturation curve (K 0.5=0.79 mM) which was transformed into an hyperbola in the presence of fructose 1,6-bisphosphate, while type M2 had a phosphoenolpyruvate saturation curve of the Michaelis-Menten type (K0.5=0.2mM). Antibodies against pyruvate kinase type L from chicken liver inactivated type L from rat and partially inactivated type M2 from chicken and rat; but the antibodies against type L did not react with type M1 from chicken breast muscle. It is therefore concluded that, contrary to a previous report (Strandholm, J.J. et al. (1975) Biochemistry 14, 2242-2246), avian liver contains pyruvate kinases type M2 and L as the mammalian liver does.     

Recombinant pyruvate kinase type I from Escherichia coli: overproduction and revised C-terminus of the polypeptide

Exposure to continuous darkness and chronic treatment with melatonin, for six weeks, stimulated reproduction in the male Indian desert gerbil as evidenced by morphometric data. Exposure to continuous light, for same duration, on the other hand inhibited reproduction. The results are opposite to those reported from similar studies on temperate zone species. Surface areas of abdominal scent glands increased following both, exposures to continuous darkness and the treatment with melatonin. Exposure to continuous light decreased the scent gland surface area. Assessment of scent gland activity could be useful in evaluation of reproductive function as they are dependent on sex steroids. Melatonin that mediates photoperiodic influence on reproduction is not always inhibitory to gonads.     

A dehydroalanyl residue can capture the 5''-deoxyadenosyl radical generated from S-adenosylmethionine by pyruvate formate-lyase-activating enzyme

OBJECTIVE: To analyze the effects of carotid endarterectomy on the retrobulbar circulation of patients with severe occlusive carotid artery disease (OCAD) by means of color Doppler imaging (CDI). DESIGN: Prospective. PARTICIPANTS: A total of 17 consecutive patients with severe OCAD and neurologic symptoms (with a history of transitory ischemic attack or cerebral vascular accident) participated. INTERVENTION: All 17 patients underwent carotid endarterectomy. The CDI of both orbits was performed by one masked investigator before surgery and at 1 week and 1 month after surgery. MAIN OUTCOME MEASURES: Peak systolic velocity, end diastolic velocity, and resistive index of the ophthalmic, central retinal, and temporal short posterior ciliary arteries were measured. The authors compared the hemodynamic parameters at all intervals. RESULTS: Peak systolic and end diastolic velocities in the ophthalmic, central retinal, and temporal short posterior ciliary arteries increased significantly 1 week and 1 month after carotid endarterectomy (P < 0.05). After surgery, the resistive indices in the central retinal and temporal short posterior ciliary arteries decreased significantly at both intervals (P < 0.05). The six patients who had reversed ophthalmic artery flow before surgery showed forward ophthalmic artery flow after carotid endarterectomy. The contralateral orbits showed no significant hemodynamic change after endarterectomy (P < 0.05). CONCLUSIONS: Hemodynamic changes in patients with severe OCAD undergoing carotid endarterectomy suggest improvement in the ipsilateral retrobulbar blood flow.     

An observational investigation of sibling interactions in married and divorced families.

Examined differences in sibling relations in divorced and married families and the extent to which sibling relationship quality is associated with the quality of spousal/ex-spousal, mother–child, and father–child relations. Observed 96 sibling dyads, 48 from mother-custody divorced families and 48 from biological two-parent families while playing a popular board game. Mothers completed a battery of questionnaires assessing the quality of other dyadic relationships within the family and family functioning. Results revealed that dyads from divorced families containing an older male were more negative, more resistant, and less compliant than older female dyads from divorced families or older male dyads from married families, with the older children more negative than their younger siblings, especially when interacting with their younger sisters. Quality of sibling interactions was found to be related to the quality of other dyadic relations within the family and overall family functioning. Quality of spousal/ex-spousal relationships contributed more than marital status of the parents in predicting whether siblings were negative. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Aspartate transcarbamylase from the hyperthermophilic eubacterium Thermotoga maritima: fused catalytic and regulatory polypeptides form an allosteric enzyme

In the allosteric aspartate transcarbamylase (ATCase) from the hyperthermophilic eubacterium Thermotoga maritima, the catalytic and regulatory functions, which in class B ATCases are carried out by specialized polypeptides, are combined on a single type of polypeptide assembled in trimers. The ATCases from T. maritima and Treponema denticola present intriguing similarities, suggesting horizontal gene transfer.     

Creatine kinase. Modification of the working enzyme

The action of amiodarone (1.5 X 10(-5) M) on sinus node activity of spontaneously beating isolated right atria of rabbit at 30 degrees C was investigated using a microelectrode technique. The drug significantly increased the action potential duration and decreased the slope of diastolic depolarization, both effects leading to a reduction of the sinus rate. In contrast to beta-blocking agents, amiodarone reduced but did not completely abolish the adrenergic effects on the sinus node activity. It is concluded that the amiodarone-induced bradycarida observed in clinical trials might be due to a direct effect of the drug on the sinus node.     

Requirements for the adaptor protein role of dihydrolipoyl acetyltransferase in the up-regulated function of the pyruvate dehydrogenase kinase and pyruvate dehydrogenase phosphatase

The dihydrolipoyl acetyltransferase (E2 component) is a 60-mer assembled via its COOH-terminal domain with exterior E1-binding domain and two lipoyl domains (L2 then L1) sequentially connected by mobile linker regions. E2 facilitates markedly enhanced function of the pyruvate dehydrogenase kinase (PDK) and pyruvate dehydrogenase phosphatase (PDP). Human E2 structures were prepared with only one lipoyl domain (L1 or L2) or with alanines substituted at the sites of lipoylation (Lys-46 in L1 or Lys-173 in L2). The L2 domain and its lipoyl group were shown to be essential for markedly enhanced PDP function and were required for greatly up-regulated PDK function. The complete absence of the L1 domain reduced the enhancements of both of these activities but not the maximal effector-stimulated PDK activity through acetylation of L2. With nonlipoylated L2 present, lipoylated L1 supported a lesser enhancement in PDK function with significant stimulation upon acetylation of L1. Prevention of L1 lipoylation in K46AE2 removed this competitive L1 role and enhanced L2-facilitated PDK activity beyond that of native E2 when PDK activity was measured in the absence or in the presence of stimulatory effectors. Thus, the E2-L2 domain has a paramount role in facilitating enhanced PDK and PDP function but inclusion of E2-L1 domain, even in a noninteracting (nonlipoylated) form, contributes to the marked elevation of these activities.     

Structural and catalytic properties of CMP kinase from Bacillus subtilis: a comparative analysis with the homologous enzyme from Escherichia coli

CMP kinases from Bacillus subtilis and from Escherichia coli are encoded by the cmk gene (formerly known as jofC in B. subtilis and as mssA in E. coli). Similar in their primary structure (43% identity and 67% similarity in amino acid sequence), the two proteins exhibit significant differences in nucleotide binding and catalysis. ATP, dATP, and GTP are equally effective as phosphate donors with E. coli CMP kinase whereas GTP is a poor substrate with B. subtilis CMP kinase. While CMP and dCMP are the best phosphate acceptors of both CMP kinases, the specific activity with these substrates and ATP as donor are 7- to 10-fold higher in the E. coli enzyme; the relative Vm values with UMP and CMP are 0.1 for the B. subtilis CMP kinase and 0.01 for the E. coli enzyme. CMP increased the affinity of E. coli CMP kinase for ATP or for the fluorescent analog 3'-anthraniloyl dATP by one order of magnitude but had no effect on the B. subtilis enzyme. The differences in the catalytic properties of B. subtilis and E. coli CMP kinases might be reflected in the structure of the two proteins as inferred from infrared spectroscopy. Whereas the spectrum of B. subtilis CMP kinase is dominated by a band at 1633 cm-1 (representing beta type structures), the spectrum of the E. coli enzyme is dominated by two bands at 1653 and 1642 cm-1 associated with alpha-helical and unordered structures, respectively. CMP induced similar spectral changes in both proteins with a rearrangement of some of the beta-structures. ATP increases the denaturation temperature of B. subtilis CMP kinase by 9.3 degrees C, whereas in the case of the E. coli enzyme, binding of ATP has only a minor effect.