Connective tissue activation. VII. Evidence supporting a role for prostaglandins and cyclic nucleotides.

Prostaglandins added to synovial cultures stimulated hyaluronic acid (HA) synthesis and glycolysis. The order of potency of the prostaglandins was: PGE1 greater than PGE2 greater than PGF2alpha greater than PGF1alpha, PGE1 and PGE2, 1.0 mug per milliliter, stimulated synovial cells, whereas F-series prostaglandins required 5 mug per milliliter for stimulation. Connective tissue-activating peptide (CTAP) activation of synovial cells was markedly potentiated by all four prostaglandins, and by PGE1 in concentrations as low as 0.01 mug per milliliter. Exogenous prostaglandins caused a prompt and marked increment in synovial cell cyclic-AMP, while CTAP caused a delayed peak of cyclic-AMP of lesser magnitude. Treatment of synovial cultures with cortisol (1.0 mug per milliliter), cycloheximide (10 mug per milliliter), or indomethacin (15.0 mug per milliliter) failed to block stimulation by PGE1, 7-OXA-13-Prostynoic acid, a prostaglandin antagonist, substantially inhibited the action of PGE1 and suppressed the effect of CTAP on synovial cells. It is possible that both exogenous and endogenous (synovial prostaglandins are involved in the connective tissue activation sequence.     

School-age outcome of children with prenatal cocaine exposure following early case management

The impact of prenatal cocaine exposure on neurodevelopmental outcome in childhood continues to be in question. Objectives of this study were to determine if there are differences in growth, behavioral, cognitive or verbal functioning or caregiver-child dyad interactions for inner-city, school-aged children with and without prenatal cocaine exposure, and if these measures would be affected by early family case management. From an initial sample of 118 cocaine-exposed children who were evaluated to 36 months of age, 47 were able to be contacted and 39 (83%) completed a follow-up evaluation at 84 +/- 11 months of age. Subjects had been assigned at birth to early case management (CM, 24) and routine care (RC, 15). Eighteen children from the initial non-cocaine-exposed comparison sample of 41 children were able to be contacted and 12 (67%) were evaluated at 85 +/- 16 months of age. Evaluation included growth measurements, Stanford-Binet IQ, receptive and expressive language quotients, parent-reported Child Behavior Checklists (CBCL), and assessment of caregiver-child interactions by scored videotaped free-play sessions. No differences were found in growth, mean IQ, language quotients, CBCL scores or videotaped caregiver-child play interactions as a function of cocaine exposure. Within the cocaine-exposed group, there were no outcome differences between CM and RC children. However, during the play sessions, CM caregivers had more positive affect in interactions with the child than RC. Within the RC dyads, birthmothers had significantly more positive interactions than non-birthmothers. In summary, no differences in growth, behavioral, cognitive, or language function were evident for inner-city, school-aged children related to prenatal cocaine exposure or early case management. CM caregivers demonstrated more positive interactions during a play session than RC; within the RC group, non birthmothers had the least positive affective interactions. Further study of influences on caregiver-child interactions may lead to effective intervention strategies for drug-exposed infants.     

Rapid Detection of Human Cytomegalovirus UL97 and UL54 Mutations Directly from Patient Samples

Human cytomegalovirus (CMV) is a significant contributor to morbidity and mortality in immunocompromised patients, particularly in the transplant setting. The availability of anti-CMV drugs has improved treatment, but drug resistance is an emerging problem. Here, we describe an improved, rapid, sequencing-based assay for the two genes in CMV where drug resistance occurs, the UL97 and UL54 genes. This assay is performed in 96-well format with a single master mix and provides clinical results within 2 days. It sequences codons 440 to 645 in the UL97 gene and codons 255 to 1028 in the UL54 gene with a limit of detection of 240 IU/ml. With this assay, we tested 43 specimens that had previously been tested for UL97 drug resistance and identified 3 with UL54 mutations. One of these patients had no concurrent UL97 mutation, pointing toward the need for an assay that facilitates dual UL97/UL54 gene testing for complete resistance profiling.     

Connective tissue activation

Alpha- and beta-adrenergic blocking agents and imipramine inhibit the increased hyaluronate synthesis that may be induced in human synovial cultures by connective tissue activating peptide (CTAP). Considerations of drug concentration requirements, actions of analogues, and time studies all indicate that the adrenergic blockers do not act in this circumstance as conventional blockers of alpha or beta receptor sites. It is suggested that the membrane-stabilizing properties of these agents may be the important determinant for their limited “antiactivation” effect. Ethacrynic acid, a potent and more complete inhibitor of connective tissue activation, appears to act via a different mechanism.     

Using the Elaboration Likelihood Model to Address Drunkorexia among College Students

Background: The many consequences related to alcohol consumption among college students are well documented. Drunkorexia, a relatively new term and area of research, is characterized by skipping meals to reduce caloric intake and/or exercising excessively in attempt to compensate for calories associated with high volume drinking. Objective: The objective of this study was to use the Elaboration Likelihood Model to compare the impact of central and peripheral prevention messages on alcohol consumption and drunkorexic behavior. Methods: Researchers employed a quasi-experimental design, collecting pre- or post-test data from 172 college students living in residence halls at a large Midwestern university, to assess the impact of the prevention messages. Participants in the treatment groups received the message in person (flyer), through email, and via a text message in weekly increments. Results: Results showed that participants exposed to the peripherally framed message decreased the frequency of their alcohol consumption over a 30-day period (p =.003), the number of drinks they consumed the last time they drank (p =.029), the frequency they had more than five drinks over a 30-day period (p =.019), as well as the maximum number of drinks they had on any occasion in the past 30 days (p =.014). Conclusions/Importance: While more research is needed in this area, the findings from this study indicate that researchers and practitioners should design peripheral (short and succinct), rather than central (complex and detailed), messages to prevent drunkorexia and its associated behaviors.     

Assessment of fetal pulmonic stenosis by ultrasonography

This study was designed to determine (1) the value of Doppler echocardiography in depicting the presence of a fetal pulmonary stenosis, (2) its reliability in the assessment of the severity of the lesion, and (3) the usefulness of additional markers from the left side of the heart as criteria of severity. Fourteen pregnant ewes were included in this study (gestational age, 90 to 120 days). Banding of the fetal main pulmonary artery created mild (n = 3), moderate (n = 3), and severe (n = 5) stenosis. Three lambs were sham operated. Intrauterine fetal Doppler echocardiographic data obtained 15 days after surgery were compared with preoperative values. Peak velocities recorded through the band increased linearly from baseline in the groups with mild and moderate stenosis but did not show any further increase in the group with severe stenosis. Compared with the sham-operated group, right ventricular output in the group with stenosis was either similar or reduced significantly. The increase in right ventricular free wall thickness was significantly greater in the groups with stenosis compared with that of the sham-operated group; the correlation with the degree of severity was r = 0.65 and p < 0.05. A stronger positive correlation was found between the severity of stenosis and aortic valve diameters; r = 0.82 and p < 0.01. The strongest correlation was found for right ventricular/left ventricular outputs (r = 0.92; p < 0.001). Thus Doppler peak velocities through the obstruction can help detect pulmonic stenosis but are not reliable for the assessment of its severity during fetal life. Other ultrasound measurements such as the size of the aortic anulus and especially the ratio of right ventricular/left ventricular output could be used as sensitive markers of the severity of stenosis.     

Structural and biological characteristics of connective tissue activating peptide (CTAP-III), a major human platelet-derived growth factor.

Connective tissue activating peptides (CTAPs) extracted from leukocytes and platelets stimulate glycolysis and synthesis of glycosaminoglycan and DNA in cultured human connective tissue cells. CTAP-III, isolated from fresh or outdated human platelets, is a low molecular weight single-chain protein with an isoelectric point of 8.5 that markedly stimulates DNA synthesis and multiple aspects of glycosaminoglycan and proteoglycan metabolism. This report presents a definitive comparison of CTAP-III prepared by two methods [one designated (A), alternative] with similar platelet proteins described by others, beta-thromboglobulin (beta-TG) and low-affinity platelet factor 4 (LA-PF-4). CTAP-III, CTAP-III(A), LA-PF-4, and beta-TG have common antigenic determinants documented by immunoprecipitation and radioimmunoassay. CTAP-III, CTAP-III(A), and LA-PF-4 are biologically active in that they stimulate DNA and glycosaminoglycan synthesis by human synovial cells; beta-TG is inactive. Carboxyl-terminal digestion gave identical terminal sequences for CTAP-III, CTAP-III(A), and beta-TG. Amino-terminal sequence data indicate that CTAP-III and CTAP-III(A) (also LA-PF-4) are identical and differ from beta-TG only by an additional amino-terminal tetrapeptide (Asn-Leu-Ala-Lys-). The biologically active molecule, CTAP-III, may be proteolytically converted to its inactive degradation product (beta-TG) in the course of platelet aging, platelet storage, release from the platelets, or initiation of biological activity.