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PubMed2001

Multisite phosphorylation of a CDK inhibitor sets a threshold for the onset of DNA replication.

SCF ubiquitin ligases target phosphorylated substrates for ubiquitin-dependent proteolysis by means of adapter subunits called F-box proteins. The F-box protein Cdc4 captures phosphorylated forms of the cyclin-dependent kinase inhibitor Sic1 for ubiquitination in late G1 phase, an event necessary for the onset of DNA replication. The WD40 repeat domain of Cdc4 binds with high affinity to a consensus phosphopeptide motif (the Cdc4 phospho-degron, CPD), yet Sic1 itself has many sub-optimal CPD motifs that act in concert to mediate Cdc4 binding. The weak CPD sites in Sic1 establish a phosphorylation threshold that delays degradation in vivo, and thereby establishes a minimal G1 phase period needed to ensure proper DNA replication. Multisite phosphorylation may be a more general mechanism to set thresholds in regulated protein-protein interactions.

Binding Sites↗
PubMed1991

Renal ammoniagenesis in humans with chronic potassium depletion.

Renal ammonia production and distribution and ammonia precursor utilization were evaluated in eight patients with chronic potassium depletion (CPD) and aldosterone-producing adenoma and in 20 controls. In CPD, urinary ammonia excretion and ammonia added to renal venous blood were about twofold higher than in controls; thus, total ammonia production was significantly augmented (88.0 +/- 10.3 mumol/min.1.73 m2 vs. 45.0 +/- 2.6 in controls). Total ammonia production was inversely correlated with serum potassium and directly correlated with urine flow. Stepwise multiple regression analysis showed that both factors, mainly serum potassium, significantly influence ammonia production and account for 61.4% of variations in ammonia production. Renal extraction of glutamine was significantly increased (56.6 +/- 5.9 mumol/min.1.73 m2 vs. 34.6 +/- 3.1 in controls), and this could account for ammonia production. The ratio of urinary ammonia excretion to total ammonia production, an index of the intrarenal ammonia distribution, was similar in patients and controls, and was significantly correlated with urine pH and true renal blood flow (RBF). Stepwise multiple regression analysis showed that RBF, urine pH and urine flow also significantly affected ammonia distribution. However, these factors accounted for only 41.7% of variations in intrarenal ammonia partition, urine pH having a minor role. We conclude that in patients with CPD other factors besides urine pH, urine flow and RBF intervene in the ammonia partition between urine and blood.

Acid-Base Equilibrium↗
PubMed1996

Left ventricular abnormalities in children, adolescents and young adults with renal disease.

The cardiac abnormalities that complicate chronic renal failure and renal replacement therapy are not well characterized in young people. These abnormalities are becoming more important because successful renal transplantation has resulted in children with end-stage renal failure living longer. Echocardiographic abnormalities of cardiac function and structure were studied in children and young adults (< 27 years old) with chronic renal failure (CRF, N = 32), end-stage renal failure treated with chronic peritoneal dialysis (CPD, N = 10) or renal transplantation (N = 30) or controls (N = 60). Left ventricular mass indexed for height (LVM/Ht and LVM/Ht2.7) and body surface area (LVM/SA), fractional shortening, measurement of left ventricular diastolic function (peak E and A wave velocities and the EA ratio) and structural (such as valvular) abnormalities were determined by echocardiography. The median (and range) of LVM/Ht in the groups were control 51.8 (23.1 to 119.8), CRF 60.2 (22.2 to 135.8), CPD 80.2 (14.5 to 100.9) and transplant group 97.8 (51.2 to 182.1) g/m. The increases in LVM/Ht, LVM/Ht2.7 and LVM/SA in the transplant group were significant (P < 0.01). The CRF group had significantly increased LVM/Ht2.7 and LVM/SA (P < 0.01). Systolic function was not significantly different between the groups. A significant correlation between creatinine and LVM indexed for height was found in the CRF group. Systolic or diastolic blood pressure could not be correlated with LVM indices in the transplant group. Changes in diastolic function were found (increased peak A wave velocity and decreased E/A ratios in the CRF and CPD groups, and increased peak E wave velocity in the transplant group). The study demonstrated that left ventricular hypertrophy is a frequent and often severe finding in children with chronic renal failure and those treated with renal replacement therapy. Factors other than hypertension and anaemia are important, and evidence was found for a link between serum creatinine and increased left ventricular mass prior to end-stage renal failure.

Adolescent↗
PubMed2004

Nucleotide insertion opposite a cis-syn thymine dimer by a replicative DNA polymerase from bacteriophage T7.

Ultraviolet-induced DNA damage poses a lethal block to replication. To understand the structural basis for this, we determined crystal structures of a replicative DNA polymerase from bacteriophage T7 in complex with nucleotide substrates and a DNA template containing a cis-syn cyclobutane pyrimidine dimer (CPD). When the 3' thymine is the templating base, the CPD is rotated out of the polymerase active site and the fingers subdomain adopts an open orientation. When the 5' thymine is the templating base, the CPD lies within the polymerase active site where it base-pairs with the incoming nucleotide and the 3' base of the primer, while the fingers are in a closed conformation. These structures reveal the basis for the strong block of DNA replication that is caused by this photolesion.

Bacteriophage T7↗
PubMed2000

A role for clinical audit and peer review in the identification of continuing professional development needs for general dental practitioners: a discussion.

The purpose of this paper is to discuss how the role of peer review and clinical audit may be used in the identification of the continuing professional development (CPD) needs of general dental practitioners (GDPs). Clinical audit and peer review are intrinsically valuable in terms of the continued professional development of GDPs. Collaborative clinical audit, in particular, can provide a framework for short course input and there are particular benefits to this combination of activities which might usefully be more widely encouraged. If open to analysis in a way which retains individual anonymity, peer review and clinical audit resumes, these could be used to inform the provision of CPD and, linked to the knowledge of audit facilitators, short courses might more closely match the CPD needs of local dentists.

Dental Audit↗
PubMed2006

A comparison of the perceived effects on Scottish general dental practitioners of peer review and other continuing professional development.

AIM: To produce evidence to inform future provision of continuing professional development (CPD). METHOD: A postal questionnaire survey of 268 dentists who had participated in the pilot peer review scheme in Scotland. RESULTS: Courses, both Section 63 and privately run, were both seen as highly effective in changing both knowledge and practising behaviour. Most of the free form comments invited were positive about the effects and acceptability of peer review as a mode of CPD. Courses and reading journals were felt to be better at changing knowledge while all the other modes (vide intra) were felt to be better at changing practice, except computer assisted learning (CAL). CONCLUSIONS: Peer review in Scotland is perceived as an acceptable and effective CPD mode for changing knowledge and practice. It is not felt to be superior to the established classical modes of courses and reading of journals.

Adult↗
PubMed2006

Psychosocial effects on caregivers for children on chronic peritoneal dialysis.

The study was designed to explore the psychosocial effects on caretakers of children in Taiwan on chronic peritoneal dialysis (CPD). This is a case-control study, performed with subjects drawn from eight medical centers. The study group consisted of caretakers of 32 children with renal failure being treated with CPD. For comparison, a control group of caretakers of 64 healthy children as well as the regional Taiwanese studies were used. Two instruments were used to explore the presence of probable depression and quality of life (QOL) of the caretakers: the Taiwanese Depression Questionnaire, and the World Health Organization QOL BRIEF-Taiwan Version. In the study group, only 25% of caregivers had full-time jobs, and 66% of families had an annual income of less than US dollar 15,000. Of the 32 families in the study group, 16% had only a single parent. The prevalence of probable depression was significantly more common in the study group compared with control and referent group (28% vs 5% and 9.44%; P = 0.001). QOL scores in four domains were also significantly lower in the study group. In conclusion, even with the advances of peritoneal dialysis techniques, caring for children on CPD in Taiwan has significant adverse psychosocial effects on the primary caregivers. Attention should be paid to the psycho-social status of the caregivers.

Adolescent↗
PubMed2006

Tidal PD: its role in the current practice of peritoneal dialysis.

The role of tidal peritoneal dialysis (TPD) has been the subject of several studies over the past 30 years. The use of the newest generation of cyclers combined with the increasing number of chronic peritoneal dialysis (CPD) patients being maintained on cycler therapy has stimulated a reexamination of the role of TPD in the care of CPD patients. Several studies over the past decade have examined solute clearances with TPD in patients. These studies suggest that TPD does not result in an increase in clearances when compared to conventional intermittent peritoneal dialysis (IPD). TPD is now primarily used for comfort in patients who experience pain at the start of inflow and/or at the end of outflow. In TPD, the presence of at least some fluid in the abdomen during the exchanges generally eliminates these episodes of pain. It has recently been suggested that accurate assessment of drain and fill phases during automated PD may be helpful in redefining a role for TPD in CPD patients. If the 'slow' drainage time can be kept to a minimum, then it is possible that the efficiency of PD could be enhanced. Defining the critical volume and then optimizing the TPD regimen could perhaps increase the clearances noted with TPD.

Dialysis Solutions↗
PubMed1997

Sequence of human carboxypeptidase D reveals it to be a member of the regulatory carboxypeptidase family with three tandem active site domains.

We have cloned the cDNA for human carboxypeptidase D (CPD), a new B-type metallocarboxypeptidase that is membrane bound and has an acidic pH optimum. The 5.8 kb of cDNA sequenced contains an open reading frame of 4131 bp encoding 1377 amino acid residues. The sequence is similar (75% identity) to duck gp180, a protein that was isolated, cloned and sequenced as a hepatitis B virus-binding protein but not characterized as a carboxypeptidase. Hydropathic analysis revealed a hydrophobic region at the N-terminus, representing the signal peptide, and one near the C-terminus that probably represents the transmembrane anchor. The most striking feature is the presence of three tandem carboxypeptidase homology domains that have sequence similarity to the regulatory B-type carboxypeptidase family, typified by carboxypeptidases M, E and N. Because of the three repeats, CPD is about three times larger (175-180 kDa) than other members of this family (approx. 50-62 kDa). Domain 2 is most closely related to carboxypeptidases M, E and N (45-48% identity), followed by domain 1 (37-38%) and domain 3 (20-27%). There is a much higher sequence identity in regions containing putative active site residues, and all catalytically important residues are strictly conserved in domains 1 and 2. In domain 3, however, only 1 of 8 active site residues is conserved, indicating that this portion might not be catalytically active. Northern blotting of mRNA from human tissues and cells showed high levels of CPD mRNA in placenta, pancreas and Hep G2 hepatoma cells, and smaller amounts in skeletal muscle, heart and HT-29 colon carcinoma and melanoma cell lines.

Amino Acid Sequence↗
PubMed2001

An alternative technique for evaluating the effectiveness of continuing professional development courses for health care professionals: a pilot study with practice nurses.

BACKGROUND: Increasing recognition of the need for continuing professional development (CPD) for health professionals has created an inevitable expansion in course provision. One criterion of a successful course demands that CPD must meet local health service needs, as well as the personal and professional requirements of the participants. At a more general level, a key pedagogical requirement of any educational course is that the content and process must meet the stipulated aims and objectives, and this requires appropriate evaluation procedures. Within health care CPD, very little course evaluation routinely takes place, possibly because the area is relatively new, and secondly because relevant evaluation may be difficult to operationalize. AIMS: The current study reports a small exploratory pilot investigation into the use of a valid and reliable training needs analysis questionnaire as a means of evaluating a nurse practitioner (NP) training scheme for practice nurses (PN). METHODS: Using a before/after design, the instrument involved the participants in a pre and post course self-assessment on a range of tasks. These assessments were made according to three criteria: how critical each task is considered to be to the effective delivery of the respondent's current PN role; how critical each task is considered to be to the effective delivery of the role of the NP; and how well each task is currently performed. Various comparisons of the ratings provide a considerable amount of information that may be of use to the nurse manager in planning NP programmes. Of particular relevance to the current paper, however, are the comparisons between salience of tasks for the NP role and performance on these, before and after the course. This provides an assessment of the extent to which the course has met educational requirements. RESULTS: The present study demonstrated that overall, the course reduced skill deficits in all but seven of the tasks. Of these, six related to research and audit and the seventh to clinical examination of patients. CONCLUSION: The study is discussed in terms of the implications of the technique for course development and planning to take account of local needs, as well the instrument's reliability and validity, for use in this way.

Competency-Based Education↗
PubMed1996

The value of maternal height as a risk factor of dystocia: a meta-analysis.

Ten publications and studies on the relation between maternal height and the risk of dystocia due to cephalopelvic disproportion (CPD) are analysed. The rate of Caesarean sections was chosen as the CPD indicator. When maternal height is presented in percentiles, curves can be superimposed, and sensitivities and specificities of the various studies may be analysed together. One biased study was excluded; the remaining 9 were pooled and regression lines calculated for sensitivity (Se) and specificity (Sp) of the entire set of points. The resulting model, i.e. Se = 10.9 + 1.99 Y and Sp = 99.9 - 0.99 Y, permits easy calculation of the expected sensitivity and specificity for each percentile Y. When the frequency of Caesarean section due to CPD is known, positive and negative predictive values can also be calculated. The proposed formulas can also be used to determine confidence intervals. The findings in terms of the sensitivity and specificity of low maternal height as a risk factor for dystocia indicate that 1 out of 5 pregnant women would have to be referred for further investigation to identify half of the cases of mechanical dystocia necessitating Caesarean section. The predictive value for a Caesarean rate of 2% (a value often seen in developing countries) for this 20th percentile would be only 5%. Practical ways of choosing a reference criterion are suggested. A two-track strategy (antenatal check-ups and community monitoring) is proposed.

Body Height↗
PubMedDate not supplied

Preferential decrease in thymus dependent lymphocytes during storage at 4 C in anticoagulant.

Human lymphocytes stored at 4 C either as leukocyte concentrates (LCs) in citrate-phosphate-dextrose (CPD) or as whole blood anticoagulated with CPD show a rapid and marked decrease in the relative and absolute numbers of thymus derived (T) lymphocytes. Determinations were made on cells recoverable on a Ficoll-Hypaque (F-H) gradient. In evacuated LCs, the relative percentage of T cells dropped to less than 10 per cent within 72 hours with a concomitant increase in the relative percentage of bone marrow derived (B) cells to 80 per cent or more. LCs opened to the air and subsequently stored at 4 C displayed an even more precipitous decline in the relative percentage of T cells, reaching a 10 per cent level within 72 hours. The relative percentage of T cells in CPD-anticoagulated whole blood samples stored at 4 C displayed similar decreases, reaching 20 per cent levels within 24 hours. The change in the relative percentage of T cells at the Ficoll-Hypaque interface was shown to reflect a decrease in the total numbers of T cells placed on the F-H gradient with time, since determinations of T and B cell numbers in NH4Cl-treated whole blood showed a 65 to 80 per cent decrease in the numbers of T cells within 24 hours in anticoagulated whole blood held at 4 C. Thus, it may be inferred that the T cell decrease is mediated via some interaction of anticoagulant, storage time, and some component(s) present in both LCs and whole blood.

Anticoagulants↗
PubMedDate not supplied

Blood preservation XXVII. Fructose and mannose maintain ATP and 2,3-DPG.

Mannose and fructose as well as glucose have been shown to be effective for maintaining ATP and thus viability of stored red blood cells. Normal 2,3-DPG levels are desirable in stored red blood cells to provide the needed oxygen transport upon transfusion. ATP levels in sotred concentrated red blood cells in the new preservative, CPD-adenine (citrate-phosphate-dextrose-adenine) become critically low in the 5th week. In this study two hexoses and two pentoses are compared with dextrose in their ability to maintain ATP and 2,3-DPG. ATP levels were best maintained by fructose, then dextrose and mannose. ATP levels had fallen to critically low levels by four weeks with ribose and xylose. Red blood cell 2,3-DPG concentrations were also maintained by hexoses, with mannose being best, dextrose and fructose being similar. When ribose was used in addition to dextrose in CPD-adenine, ATP maintenance was improved and under the same conditions xylose improved 2,3-DPG maintenance. Fructose and mannose may be as useful as dextrose in citrate-phosphate preservatives for maintaining ATP and 2,3-DPG levels. Also, ribose and xylose may help the maintenance of ATP and 2,3-DPG, respectively, in CPD-adenine.

Acid-Base Equilibrium↗
PubMedDate not supplied

Blood preservation. XXXIV. DHA maintains 2,3-DPG and its adverse effect on ATP is reversed with extra phosphate.

We have found that the addition of 10 mM inorganic phosphate to DHA in CPD-adenine maintains ATP levels at normal or higher than normal values for six weeks of storage. 2,3-DPG values are slightly lowered by the extra phosphate, but are still maintained at approximately half normal for four weeks by the DHA. The addition of a higher phosphate concentration, 20 mM, to DHA produced lower levels of ATP and 2,3-DPG than those observed with 10 mM phosphate, although both levels were better than in the CPD-adenine control. pH values in this experiment were lowest in the three preservatives containing DHA, probably indicating increased lactate production due to metabolism of this triose sugar, in addition to dextrose present in CPD.

Adenine↗
PubMedDate not supplied

In vitro evaluation of platelets stored in CDP-adenine formulations.

Little information is available about the effect of adenine and added glucose on stored platelets. Two new formulations, CPDA-2 and CPDA-3, contain 34 mg adenine per 63 ml preservative and extra glucose (1.75 and 2.0 times the glucose in standard CPD). We have studied the in vitro integrity of platelet concentrates stored in CPD, CPDA-1, CPDA-2, and CPDA-3 at 22 C for 72 hours. Morphology score, pH, platelet size, population distribution parameters, and electron microscopic ultrastructure did not show any adverse effects which could be ascribed to the presence of adenine or extra glucose or both. No differences in platelet adenosine triphosphate (ATP) concentration or plasma glucose utilization during storage were found between CPD and CPDA-1 platelets. The results suggest that adenine and added glucose in these preservatives are not detrimental to platelets in vitro by the measures employed.

Adenine↗
PubMedDate not supplied

Storage-induced changes in human newborn red cells.

Fetal red cells are well suited for intrauterine life; however, little is known about their response to postnatal environments. The purpose of this work was to investigate the metabolic and membrane changes affecting newborn red cells during their exposure to storage in citrate-phosphate-dextrose (CPD) and citrate-phosphate-dextrose-adenine (CPDA-1). The findings suggest that newborn red cells are affected more by storage than are adult cells. These accelerated storage changes in the red cells of newborns may be related indirectly to the rapid adenosine triphosphate (ATP) decline. As is the case with adult red cells, fetal cells withstand storage in CPDA-1 better than in CPD. The storage lesion in these cells was partly reversible, as in adult cells, by incubation with adenosine. It was therefore concluded that newborn red cells obtained from placentas and stored for several weeks in CPD or CPDA-1 media or other media that improve the metabolic profile of these cells may be acceptable for transfusion.

Adenosine↗
PubMed2001

The effect of the transfusion of stored RBCs on intestinal microvascular oxygenation in the rat.

BACKGROUND: Although it is known that the transfusion of stored RBCs does not always improve tissue O(2) consumption under conditions of limited tissue oxygenation, the efficiency of O(2) delivery to the microcirculation by stored RBCs has never been determined. STUDY DESIGN AND METHODS: In a rat hemorrhagic shock model, the effects of resuscitation with fresh or 28-day-old RBCs stored in CPD plasma, saline-adenine-glucose-mannitol, and CPDA-1 plasma were investigated. Systemic hemodynamic and intestinal oxygenation measures were monitored. Intestinal microvascular PO(2) was determined with the O(2)-dependent quenching of palladium-porphyrin phosphorescence, and the RBC deformability was measured with a Laser-assisted optic rotational cell analyzer. RESULTS: Hemodynamic and oxygenation measures were significantly decreased during hemorrhagic shock. Intestinal oxygen consumption and mesenteric venous pO(2) were restored with the transfusion of both fresh and stored RBCs, except for CPD-stored RBCs. The intestinal microvascular pO(2) improved only with the transfusion of fresh RBCs. Deformability of the stored RBCs was significantly decreased. CONCLUSION: In contrast to that of fresh RBCs, the transfusion of stored RBCs did not restore the microcirculatory oxygenation, possibly because of impaired O(2) unloading, but, except for CPD-stored RBCs, the storage-induced changes were not enough to impair intestinal VO(2) and mesenteric venous pO(2).

Animals↗
PubMed2002

Comparison of two micronutrient supplements in children with chronic renal failure.

OBJECTIVE: To compare a newly formulated renal-specific micronutrient (RSM) supplement (vitamins C, E, K, and B complex, copper, zinc) with Ketovite (Paines & Byrnes, Middlesex, UK) tablets (vitamins C, E, K, and B complex) in children with chronic renal failure (glomerular filtration rate [GFR] < 25 mL/min/1.73 m(2)) or on chronic peritoneal dialysis (CPD) and hemodialysis (HD). DESIGN: Children currently prescribed Ketovite tablets were changed to RSM for 3 months. Questionnaires on palatability, acceptability, and ease of administration were assessed while on Ketovite and after 3 months on RSM along with plasma levels of zinc, copper, folate (serum, red cell), vitamin B(12), and homocysteine. SETTING: Regional pediatric nephrology unit. PATIENTS: Fifteen children (10 male) with a mean age of 10.4 years (range, 1.1 to 16 y) were recruited (11 had GFR < 25 mL/min/1.73 m(2), 2 CPD, 2 HD). Two children received overnight gastrostomy button feeding. MAIN OUTCOME MEASURES: Fourteen children (1 child refused RSM after 1 week) and their families completed questionnaires using a Likert scale: 1 (liked) to 7 (disliked). Plasma levels were analyzed at baseline on Ketovite and after 3 months on RSM. RESULTS: Children disliked the smell of Ketovite compared with RSM (P =.004). The size of Ketovite was preferable to the size of the RSM (P =.015) and was believed to be easier to administer (P =.046). There were no differences in patient/parent rating of appearance, texture, or taste, but 8 of 15 patients (53%) expressed a preference for the RSM if available. Plasma values of copper, zinc, folate, and vitamin B(12) were within the normal reference range on Ketovite and showed no significant change on RSM. Mean plasma homocysteine levels were above normal reference ranges in all groups (GFR < 25 mL/min/1.73 m(2), 7.6 [SD, 3.1]; CPD, 11.5 [SD, 1.6]; HD, 12 [SD, 8.7]) on Ketovite and were unchanged after 3 months on RSM. CONCLUSIONS: This pilot study confirms the acceptability of the RSM, particularly in relation to its smell and chewability, but the current size may preclude its widespread use. The incorporation of zinc and copper will be beneficial for many children, but reduction in homocysteine levels may necessitate an increase in folic acid content.

Adolescent↗