Penetration of natural prostaglandins and their ester prodrugs and analogs across human ocular tissues in vitro

The objective of this study was to assess the corneal and scleral permeabilities of natural prostaglandins as well as their prodrugs and analogs through human cornea and sclera in vitro. The "apparent permeability coefficients" (Papp) of natural prostaglandins (PGF2alpha, PGD2 and PGE2), ester prodrugs of PGF2alpha (1-isopropyl PGF2alpha, 11-pivaloyl PGF2alpha and 11,15-dipivaloyl PGF2alpha) and four analogs (16-m-chlorophenoxy tetranor PGF2alpha, 17-phenyl trinor PGF2alpha, 17-phenyl trinor PGE2 and AH 13205) were measured using modified Ussing perfusion chambers and quantitative high performance liquid chromatography. Our results indicate that the corneal penetration of natural prostaglandins (PGs) is poor (the Papp values ranged from 1.65 x 10(-6) to 2.38 x 10(-6) cm/sec), while the PGF2alpha prodrugs showed higher corneal penetration than PGF2alpha. The 11-pivaloyl ester of PGF2alpha penetrated the cornea faster than both 1-isopropyl ester and the lipophilic 11,15-dipivaloyl ester. The PG analogs also showed poor corneal penetration (Papp values ranged from 0.696 x 10(-6) to 1.49 x 10(-6) cm/sec) except for AH 13205. All compounds tested showed good scleral penetration (Papp values ranged from 6.90 x 10(-6) to 17.1 x 10(-6) cm/sec) except PGF2alpha 11,15-dipivaloyl (Papp = 1.22 x 10(-6) cm/sec). The penetration profiles correlated well with tissue uptake ratios (ratio of final tissue concentration to initial dose) for all compounds except 11,15-dipivalate PGF2alpha. All ester prodrugs (but not the PGs and analogs) underwent corneal first-pass metabolism. The study results demonstrate that transcleral absorption may play a significant role in the ocular absorption of these compounds.     

Protein chemical shift analysis: a practical guide

Proper protein chemical shift analysis requires careful experimental measurements and the implementation of standardized referencing procedures. In this article we outline the steps necessary to ensure proper chemical shift referencing and the selection criteria for choosing appropriate "random coil" amino acid chemical shift values for predicting, comparing, and assigning 1H, 13C, and 15N resonances in proteins. By making use of these standardized conditions we demonstrate how several recently developed methods, namely homologous assignment techniques and empirical chemical shift contour maps (or hypersurfaces), can significantly improve the accuracy of chemical shift prediction for 1H, 13C, and 15N nuclei. In addition to illustrating the potential utility of chemical shift prediction, we also outline procedures for identifying secondary structure elements through heteronuclear chemical shift analysis and further demonstrate how empirical shift contour maps can actually be used to refine, and more importantly generate, reasonably good three-dimensional protein structures.     

Obstetric postanesthesia care unit stays: reevaluation of discharge criteria after regional anesthesia

BACKGROUND: Obstetric patients may have long postanesthesia care unit (OB-PACU) stays after surgery because of residual regional block or other conditions. This study evaluated whether modified discharge criteria might allow for earlier discharge without compromising patient safety. METHODS: Data were prospectively collected for 6 months for all patients (N=358) who underwent cesarean section or tubal ligation and recovered in the OB-PACU. Regional anesthesia was used in 94% of patients. The duration of anesthesia and PACU stays, the presence and treatment of events in the PACU, and the regression of neural blockade were recorded. Discharge from the OB-PACU required a 60-min minimum stay, stable vital signs, adequate analgesia, and ability to flex the knees. After completion of prospective data collection, events that kept patients in the PACU after 60 min were reevaluated as to whether patients needed to stay in the PACU for medical reasons. "Needed to stay" events included bleeding, cardiorespiratory problems, sedation, dizziness, and pain. "Safe to leave" conditions included pruritus, nausea, and residual neural blockade. The cumulative duration of OB-PACU stays not clearly justifiable for medical reasons was calculated. RESULTS: Residual block and spinal opioid side effects accounted for the majority of "unnecessary" stays. Annually, 429 h of PACU time could have been saved using the revised criteria. Complications did not develop subsequently in any patient deemed "safe to leave." CONCLUSIONS: In many obstetric patients, the duration of PACU stays could safely be shortened by continuing observation in a lower-acuity setting. This may result in greater flexibility and more efficient use of nursing personnel.     

Health surveillance using an occupational medical database

This pilot study sought associations between liver function tests (LFTs) and membership in homogeneous exposure groups (HEGs) at a target plant as pre-clinical indications of possible future occupational health problems. A large company database yielded linear models for each of six LFTs (total bilirubin, alkaline phosphatase, gammaglutamyl transferase, lactate dehydrogenase, aspartate transaminase, and alanine transaminase) in terms of sex, body mass index, age, race (white/non-white), alcohol and cigarette consumption, and production/non-production (P/NP) job, permitting control for these in analyses of LFTs vs HEGs at the plant. These analyses, with HEG substituted for P/NP in the large group model, resulted in loosely "suspect" associations significant at P < 0.10. Collapsed HEG variable (containing "suspects" separately and all other non-significant HEG levels pooled) yielded "confirmed suspects" at P < 0.05 in the analysis of an independent LFT set taken at the plant approximately one year later.     

Deblurring visual evoked potentials using commercially available software

Visual evoked potentials are useful clinical tools to study visual pathways of the brain. Although the temporal resolution is unsurpassed by other brain imaging technologies, the spatial resolution is diminished or blurred by the low conductance of the electrical signals through the skull. Methods have been proposed to improve the spatial resolution by downwardly projecting the electrical signals measured on the scalp to the surface of the cerebral cortex through the inverse solution of the equations governing static current flow. We describe the adaptation and combination of commercially available engineering software programs to solve this inverse problem and report the results of a sample run of the system. Before deblurring, the visual evoked potentials appeared to be diffusely localized over the posterior scalp. After deblurring, the visual evoked potentials were only found at the electrodes closest to the visual cortex, as would be predicted by our current knowledge of neuroanatomy.     

Why Johnny can't program [computing professionals]

Innumerate, illiterate, and overwhelmed, today's computing professionals ('Johnny') are torn between system design and program coding. Computing professionals must know vastly more now than they needed to know 40 years ago (1960s), but even then the field often distinguished between programmers and systems analysts. Meanwhile, Johnny must know too much to be at the same time a skilled programmer and a skilled system engineer. That is why he's overwhelmed. But Johnny's problem goes deeper. If he wants to be a professional programmer, he must learn on the job. Programmers have few trade courses to select from, many fewer than those available to carpenters, plumbers, and electricians. Further, very few professional courses will assist Johnny if he wants to become a straightforward, generalist, humanist, system engineer. Specialists such as computer system engineers and software engineers appear to dominate the field. No wonder Johnny has a problem. Let's move to give him the proper training to become whichever he wants to be: programmer or system engineer.     

Managing Customer Expectations

Users who have high expectations for a new system too often become disappointed once they discover the system does not live up to their dreams. Some IS managers circumvent this problem by purposely setting expectations too low. However, high expectations do not have to be a guarantee for disappointment and dissatisfaction.     

The theoretical limits of DNA sequence discrimination by linked polyamides

Linked polyamides bind in the minor groove of double-stranded DNA in a partially sequence-specific manner. This report analyzes the theoretical limits of DNA sequence discrimination by linked polyamides composed of two to four different types of heterocyclic rings, determining (i) the optimal choice of base-binding specificity for each ring and (ii) the optimal design for a polyamide composed of these rings to target a given DNA sequence and designed to maximize the fraction of the total polyamide binding to the specified target sequence relative to all other sequences. The results show that, fortuitously, polyamides composed of pyrrole, a naturally occurring G-excluding element, and imidazole, a rationally designed G-favoring element, have features similar to the theoretical optimum design for polyamides composed of two different rings. The results also show that, in polyamides composed of two or three types of heterocyclic rings, choosing a nonspecific "placeholder" ring, which binds equally strongly to each of the four bases, along with one or two base-specific rings will often enhance sequence specificity over a polyamide composed entirely of base-specific rings.     

Short-term treatment of a Central American torture survivor

The short-term treatment of a Honduran torture survivor is recounted. Torture--the "counter-therapy of the State" (Ritterman 1987, p. 43)--involves intentional physical and psychological destruction of human beings. The socio-political context in which the traumatic events occurred is described, as are the theoretical and ethical assumptions underlying the work. Treatment of victims of organized violence is a formidable challenge. Obstacles and advantages of short-term psychotherapy and use of the therapist's emotional reactions to understand survivors' experiences are emphasized.     

Cloning and mapping of a cDNA for methionine synthase reductase, a flavoprotein defective in patients with homocystinuria

Methionine synthase catalyzes the remethylation of homocysteine to methionine via a reaction in which methylcobalamin serves as an intermediate methyl carrier. Over time, the cob(I)alamin cofactor of methionine synthase becomes oxidized to cob(II)alamin rendering the enzyme inactive. Regeneration of functional enzyme requires reductive methylation via a reaction in which S-adenosylmethionine is utilized as a methyl donor. Patients of the cblE complementation group of disorders of folate/cobalamin metabolism who are defective in reductive activation of methionine synthase exhibit megaloblastic anemia, developmental delay, hyperhomocysteinemia, and hypomethioninemia. Using consensus sequences to predicted binding sites for FMN, FAD, and NADPH, we have cloned a cDNA corresponding to the "methionine synthase reductase" reducing system required for maintenance of the methionine synthase in a functional state. The gene MTRR has been localized to chromosome 5p15.2-15.3. A predominant mRNA of 3.6 kb is detected by Northern blot analysis. The deduced protein is a novel member of the FNR family of electron transferases, containing 698 amino acids with a predicted molecular mass of 77,700. It shares 38% identity with human cytochrome P450 reductase and 43% with the C. elegans putative methionine synthase reductase. The authenticity of the cDNA sequence was confirmed by identification of mutations in cblE patients, including a 4-bp frameshift in two affected siblings and a 3-bp deletion in a third patient. The cloning of the cDNA will permit the diagnostic characterization of cblE patients and investigation of the potential role of polymorphisms of this enzyme as a risk factor in hyperhomocysteinemia-linked vascular disease.