Characterization of RAFTK, a novel focal adhesion kinase, and its integrin-dependent phosphorylation and activation in megakaryocytes

We have recently isolated a cDNA encoding a novel human intracellular tyrosine kinase, termed RAFTK (for a related adhesion focal tyrosine kinase). The RAFTK cDNA, which encodes a polypeptide of 1,009 amino acids, shares 65% homology to the focal adhesion kinase (FAK), including several consensus motifs. In this report, we describe the biochemical characterization and functional analysis of the RAFTK protein. Coexpression of RAFTK and FAK proteins in megakaryocytic cells and blood platelets was observed. Using a specific antibody to RAFTK and the monoclonal antibody 2A7 to FAK, FAK and RAFTK could be distinguished antigenically. RAFTK had intrinsic tyrosine kinase and autokinase activities. It was phosphorylated on tyrosine in growing cultures of COS cells transfected with the pCDNAIII/flag-RAFTK expression vector containing the RAFTK cDNA ligated with the 8 amino acid flag peptide sequence. Similar to FAK, dephosphorylation of RAFTK was observed when adherent transfected COS cells were detached. Phosphorylation was regained upon replating of these cells on the fibronectincoated dishes. Analysis of tyrosine-phosphorylated RAFTK from adherent transfected COS cells showed that the Src homology 2 (SH2) domains of the Src and Fyn protein kinases as well as the Grb2 adaptor protein were able to specifically associate with RAFTK. Tyrosine phosphorylation of endogenous RAFTK was observed upon fibronectin-induced activation of human megakaryocytic cells. Furthermore, colocalization of RAFTK protein with vinculin, a focal adhesion protein, was observed by confocal microscopy in focal adhesion- like structures in adherent CMK cells and in transfected pCDNAIII/flag- RAFTK COS cells upon fibronectin activation. These data suggest that RAFTK is a novel member of the FAK family, that it localizes to focal adhesion-like structures in CMK megakaryocytic cells, that it participates in integrinmediated signaling pathways in megakaryocytes, and that it is able to associate with the tyrosine kinases Src and Fyn as well as the adaptor protein Grb2 via SH2-phosphotyrosine interactions.     

Anesthetic action of opiates: correlations of lipid solubility and spectral edge

The ability of opiates to be a complete anesthetic has been assessed in animals. These studies have investigated the serum levels of opiate required to produce a decrease in anesthetic requirement for a concomitantly administered inhalation anesthetic. A linear dose-response relation has been observed between opiate serum level and reduction in anesthetic requirement up to the level of 50% reduction in minimum alveolar anesthetic concentration (MAC). These studies have not demonstrated the production of one MAC anesthesia by the opiates. Recent EEG studies have provided another means of comparing the central nervous system effects of opiates and inhalation anesthetics. The serum levels of several opiates associated with a 50% reduction (IC50 or 50% inhibitory concentration) in maximal spectral edge frequency (SEF) have been reported. The free, unionized serum levels of each opiate at IC50 in humans or 50% MAC reduction in animals are remarkably similar. We calculated brain lipid opiate content at these serum levels using available physiochemical data. The calculated nanogram and molar brain lipid contents of the drugs fell within a 10-fold range while serum levels varied by 5000-fold. This similarity in membrane lipid content in association with EEG and anesthetic effects suggests that opiate "anesthesia" may involve a membrane effect in addition to the well established receptor interaction.     

Feasibility analysis for medical services.

Feasibility analysis is an important aspect of business planning. A feasibility study is a study of the impacts of revenues, cost, and returns on a decision. It helps answer the questions of "What share of total sales can be attained?" "What revenues will be generated?" "What costs will be incurred?" "What will be the return on investment?" The discipline of going through a systematic approach to analyze market demand, cost requirements, and return on investment will improve business decisions by eliminating risk and increasing the possibility of successful ventures.     

Attitudes toward prevention of cardiovascular diseases among first-year students at eight American medical schools, 1983-1985

First-year medical students at eight U.S. medical schools were surveyed by written questionnaire in 1983-1985 to determine their attitudes toward cardiovascular diseases prevention at medical school entry. An overall response rate of 92% was achieved (2,654 questionnaires), and 97% of responders provided complete and analyzable survey data. Response rates at five of eight medical schools were 98-100%, and one school each had rates of 67, 84, or 90%. Differences in mean attitude responses from school to school were small, as were differences between men and women or between blacks and whites. This survey found that entering medical students have generally positive attitudes toward the effectiveness of preventive cardiology practice as well as toward the importance of research efforts in cardiovascular disease prevention. Students frequently indicated, however, that it is "extremely difficult" to change patients' unhealthful habits and that "physician encouragement" may not be sufficient to help patients achieve more healthful behaviors. These findings could be helpful in directing educational efforts for medical students. The data suggest that major emphasis should be placed on conveying facts regarding the physicians' efficacy in clinical preventive cardiology and on teaching the skills of preventive cardiology practice. Less emphasis appears to be necessary on encouraging positive attitudes about the importance of prevention since current students' attitudes appear to be already positive in this dimension.     

An update on digoxin

This review deals briefly with recent developments in the therapeutic drug monitoring of digoxin. Strategies for decreasing the interference by digoxin metabolites, digoxin-like factors, and spironolactone metabolites in immunoassays of digoxin are discussed. Other issues addressed include the development of alternative methods of analysis, such as receptor assays and "high-pressure" liquid chromatography; digoxin-like factors in hypertension; drug-drug interactions; redistribution of digoxin stores in the body; and forensic considerations.     

Effects of lung hyperinflation on mean arterial pressure and postsuctioning hypoxemia

Our purpose was to determine the effect of five different lung hyperinflation volumes (tidal volume, 12 cc/kg, 14 cc/kg, 16 cc/kg, and 18 cc/kg lean body weight) on mean arterial pressure and postsuctioning hypoxemia (arterial blood gases). Subjects received three consecutive lung hyperinflations at one of the five randomly ordered volumes in 15 seconds via a ventilator "sigh" control at a fraction of inspired oxygen of 1.0. The three lung hyperinflations were followed by 10 seconds of continuous suction (flow rate 16 L/min). The procedure was repeated three times. The sample consisted of eight men and women 4 hours after coronary artery bypass graft surgery. Data indicated a statistically significant (by analysis of variance, p = 0.000) mean increase of 15 mm Hg in mean arterial pressure over the three lung hyperinflation sequences that was not volume dependent. There was a significant increase (p = 0.0001) in arterial oxygen pressure at 0 seconds after suctioning that increased with each increasing lung hyperinflation volume.     

Individual psychotherapy with victims of incest

In treating victims of incest, therapists need to be aware of the gamut of symptoms, personality traits, interpersonal difficulties, and clinical syndromes that occur with special frequency in this population. A list of these features is provided. Vignettes with typical therapist-patient dialogue are offered, demonstrating techniques for handling the initial consultation and opening phase of treatment. During later phases of treatment the focus will usually shift to the discussion of the pertinent symptoms (such as suicidal feelings, substance abuse, dissociative phenomena, sexual disturbances, and distorted self-image). Further on, therapy will focus on abnormal personality traits and attitudes that the incest experience may have set in motion. Examples include mistrustfulness, jealousy, seductiveness, hostility, impulsivity, defiance and emotional volatility. Many incest victims develop borderline personality disorder. Highly polarized attitudes ("splitting") toward sexual partners are characteristic of this group. Therapy relies upon supportive, re-educative, and interpretive work designed to help the victim develop more realistic and integrated views of self and other.     

Some factors affecting the strength of the cement-metal interface

A manufacturing technique to increase the bonding between bone cement and metal prostheses has been assessed in the laboratory by "push-out" tests, and the effects of contamination of the cement and of the prosthesis with blood and intramedullary contents have been studied. The technique, known as pre-coating, increases bond strength; this increase is preserved after contamination of the cement which does, however, cause some lowering of interface shear strength. The implications for clinical practice are discussed.