[Comparative study of the detection of E. coli after time in seawater with partially modified standard media]. Fecal coliform bacteria (E. coli) are the most important parameter when monitoring the hygienic quality of bathing water according to EC guidelines. Among other things the number of E. coli is clearly dependent on the length of exposure of the bacteria to marine water as well as the media used to establish the number. In a seven-day salt water stress experiment the best rate of recultivation was achieved on average with DEV lactose peptone broth. The Brila-MUG medium produced a slightly lower rate of recultivation and the fluorocult-lauryl-sulfate-broth the lowest rate of recultivation. After a shorter exposure of bacteria to marine water conditions (up to 24 h) the DEV-lactose-broth produced slightly lower rates of recultivation. An alteration of the standard components of the media did not lead to an improvement in the rate of recultivation.


Faecal micro-organisms on the hands of carriers: Escherichia coli as model for Salmonella. To evaluate the public health hazard caused by Salmonella carriers as food handlers, a study was carried out to gather more quantitative data about faecal contamination of hands after stools. Faecal E. coli was used as a model bacterium for Salmonella. In total 92 subjects cooperated in this study. The hand was sampled before toilet use, and also after stools, with or without washing of the hands. Besides E. coli the number of Enterobacteriaceae was also determined. It appeared that hands may be contaminated with Enterobacteriaceae, regardless of toilet use, for a well before as after stools about 60% of the sampled hands carried a detectable number of Enterobacteriaceae. That is why the presence of Enterobacteriaceae on hands is not a good indicator for toilet hygiene. In 4% of the samples before stools and in 25% of the samples taken after stools E. coli could be detected (> 20 CFU/sample). The average 10log CFU of E. coli in the positive samples taken before and after stools were about the same: 2.30 per sample. Hand washing after stools reduced the numbers of Enterobacteriaceae and E. coli on the hands. It was concluded that symptomless Salmonella excretors in the period starting two weeks after infection form only a low risk in carrying over Salmonella by their hands to food. Especially if normal hygiene is practiced like washing hands after stools, the number of contaminated hands will be very low and furthermore the number of faecal micro-organisms will be very small.(ABSTRACT TRUNCATED AT 250 WORDS)


[The determination of the discrepancy between the mathematically ascertained and experimentally provable efficiency of UV facilities for water disinfection]. Using three UV-plants of different technical designs for water disinfection, we studied the conformity between experimental germ reduction using standard test organisms and calculated UV-doses under various water flow conditions. Taking into consideration the style of construction of the UV-plants, the irradiation area and the layer thickness were used as constant parameters for dose calculations. This was also employed for the irradiation intensity, since the experiments were performed for a relatively short period compared of the life span of the UV-irradiators. Both exposure time and water transmission were employed as variable parameters in the dose calculations and experimental procedures respectively. The calculated UV-dose and experimentally obtained germ reduction values were comparatively the same for two of the three UV-plants studied. However, no correlation was observed between the reduction of E. coli and the corresponding calculated UV-dose values. Therefore, the calculated UV-dose values for any given UV-plant should be considered to be relative and by no means absolute values. We are of the opinion that within a certain range of water flow rate and transmission, antimicrobial effectiveness of different UV-plants should be demonstrated independent of dose values, technical and other construction characteristics. The applicability of the UV-plants studied is discussed.


[RoTrac capillary pore membranes for laboratory filtration. II. Bacteria-free filtration]. Because of their special characteristics Capillary pore membranes (CPM) are now applied in several branches of separation techniques and analytics. Besides applications in particle analytics and microfiltration of different media capillary pore membranes can be used in microorganism separation. It was shown that RoTrac CPM can be used for bacteria free (or so called sterile) filtration. Acceptable fluxes were reached in separation of Pseudomonas diminuta (test species ATCC 19146). Membranes with pore diameters of 0.2 micron and smaller always assure a bacteria free filtrate even for a very high bacteria count of about 10(7)-10(8) bacteria/ml. In filtration of Mycoplasma arginini no sterile filtrate was obtained for a pore diameter of 0.08 micron and a high bacteria count of 3 * 10(7) bacteria/ml. The bacteria rejection by a factor of 10(5) was however remarkable. Only for 0.05 and 0.08 micron with reduced bacteria load the filtrate was bacteria free.


[Case study of a Legionella epidemic in a rehabilitation clinic]. A series of nosocomial Legionella infections in a rehabilitation center is reported. In a three months period a total of 10 pneumonias with 3 deaths occurred (8 patients, 1 companion, 1 staff member). Serologic analysis proved additional Legionella infections within the nursing staff. The warm-water system was proved to be the source of infection by isolating Legionella pneumophila serogroup 1 subtype Pontiac both in warm-water and patients samples. The air conditioning system could not be ruled out as another (secondary) route of exposure because of shortcomings in construction. Conclusions about prevention and the course of the disease are discussed and standards for warm-water and air conditioning systems are proposed.


[The development of an enzyme immunoassay for the detection of the herbicide metolachlor in water samples]. Since December 1990, strict limits for pesticide content in drinking water have been in effect in the FRG. These limits have to be enforced and controlled regularly. Because the classical analytical methods, like GC/MS and HPLC are relatively expensive and complicated, concentration and clean up of the samples are necessary. Therefore, the significance of Enzyme Immunosorbent Assays has increased in environmental analysis. The topics of our research are synthesis of suitable Metolachlor conjugates for immunization and as coating antigen, as well as development of an Enzyme Immunosorbent Assay. The chloroacetanilide Metolachlor was coupled with an AMSA-Spacer on Sheep IgG and with AHT-Spacer on BSA. The epitope density was measured photometrically with TNB-derivate. Polyclonal antibodies were produced in white New Zealand rabbits. Metolachlor-AMSA-Sheep IgG conjugate was served as immunogen. The test is indirect and competitive. Microtiter plates are coated with the coating antigen (Metolachlor-AHT-BSA conjugate). In the next step the Metolachlor sample competes with the coating antigen for the antibodies. Detection of bounded antibodies is indirect with a peroxidase-labelled secondary antibody. The chromogenic substrates ABTS, TMB and OPD were tested. TMB showed the best qualities, fast turnover, as well as good and steady colour. Titer of serum and crossreactivity of other chloroacetanilide pesticides and their metabolites were tested. With the exception of Alachlor and Dimethachlor, no pesticide showed a high interaction with serum. Quantitative interpretation was done with the logit/log transformation, which allows comparison of different sera and test series. The antibodies are specific but the affinity is not good enough to detect the limits assigned by the "deutsche Trinkwasserverordnung". Raising the concentration, as is done in classical analytical methods, would solve the problems so that the test can be used as a screening test.


[Emission measurements of fibrous dust in the Federal Republic of Germany. Fibrous emissions from brake linings of motor vehicles]. The brake and clutch linings of automobiles contain 10 to 70% of asbestos, mainly chrysotile. They are therefore considered to be a significant source of carcinogenic asbestos fibers in ambient air. This could be confirmed by measurements done in urban ambient air in different parts of West Germany. Asbestos fiber concentrations till 1000 fibers/m3 (for fibers longer than 5 microns) were found. It could be also shown, that materials used in the brake lining production as well as dust emitted during car braking contained asbestos fibers with high carcinogenic potency.


The effect of progestins on prolactin receptor gene transcription in human breast cancer cells. The sex steroid hormone progesterone modulates the developmental and lactogenic activity of prolactin in the mammary gland. Regulation of the level of prolactin receptor (PRLR) provides one possible mechanism by which this may occur, prompting this investigation of the molecular mechanisms involved in progestin regulation of prolactin receptor levels. Treatment of T-47D and MCF-7 human breast cancer cells with 10 nM of the synthetic progestin ORG 2058 for 24 hr resulted in an increase in all four PRLR mRNA transcripts detected. The effect of ORG 2058 was shown in T-47D cells to be time- and concentration-dependent and resulted in an approximate two-fold increase in PRLR mRNA after 24 hr of treatment with 10 nM or 100 nM ORG 2058. Nuclear run-on assays indicated that ORG 2058 increased the rate of T-47D PRLR gene transcription at all times between 1 hr and 28 hr of treatment. The protein synthesis inhibitors cycloheximide and puromycin abrogated the induction of PRLR gene transcription at 1 hr and 2 hr, which demonstrated that on-going protein synthesis was required for the ORG 2058 effect and suggested that progestins may exert some transcriptional effects via the induction of an intermediary protein. These experiments demonstrated that progestin induced a transcriptionally based increase in PRLR gene expression and provided a mechanism by which progesterone may modulate the mitogenic activity of prolactin during mammary gland development.


Complete human NF1 cDNA sequence: two alternatively spliced mRNAs and absence of expression in a neuroblastoma line. Neurofibromatosis type 1 (NF1) is caused by mutations in a large gene on chromosome 17q11.2. Previously described partial cDNAs for this gene predicted a protein related to yeast IRA1/IRA2 and the mammalian RAS GTPase activator protein GAP. To initiate a detailed study of the role of this gene in NF1, we have characterized a set of overlapping cDNAs that represent its complete coding sequence. Our results show that two differentially expressed human NF1 mRNAs differ by a 63-bp insertion in the GAP-related domain. These mRNAs predict two 2,818- and 2,839-amino acid proteins with calculated molecular masses of approximately 317 and 319 kD. Extensive similarity to IRA proteins is evident in a 1,450-amino-acid central segment, roughly between amino acids 900 and 2,350. However, the remainder of the NF1 protein is not significantly similar to other proteins. Interestingly, the SK-N-SH human neuroblastoma line expresses no detectable NF1 mRNA, indicating that expression of NF1 is not essential for viability of this neural crest-derived tumor cell line.


Genomic organization of the mouse OSF-1 gene. The mouse OSF-1 protein (also known as pleiotrophin, HB-GAM, HBGF-8, or HBNF) gene was isolated from a mouse genomic library and sequenced. OSF-1 is a 15-kD secreted protein specifically expressed in bone and brain, and is believed to play a role in brain development and osteogenesis. The mouse OSF-1 gene consists of at least 5 exons and 4 introns and spans > 32 kb. Computer analysis of approximately 4 kb of 5'-flanking sequence of the OSF-1 gene revealed two candidate promoter regions. One candidate promoter contains a thyroid hormone/retinoic acid-responsive element and the other contains two glucocorticoid-responsive elements. DNA sequence analysis of novel OSF-1 cDNA clones indicates that two promoters can be utilized in MC3T3-E1 osteoblastic cells. The overall organization of the mouse OSF-1 gene is similar and the locations of the three exon-intron junctions within the coding region are identical to the mouse gene encoding the differentiation-related factor midkine (MK). Based on this similarity and on the high degree of nucleotide sequence homology (approximately 55%) of mouse OSF-1 and mouse MK, we conclude that OSF-1 and MK are generated from a common ancestral gene and are members of a family of structurally and probably functionally related proteins.


The HXY box regulatory element modulates expression of the murine IA antigen-associated invariant chain in L fibroblasts. The murine invariant chain (Ii) gene has been shown to be interferon-gamma (IFN-gamma)-inducible in a number of nonlymphoid cell types. In mouse L cells, steady-state levels of Ii mRNA are barely detectable in untreated cells but increase sharply upon IFN-gamma treatment. In IFN-gamma treated L cells, transcription starts 23, 28, 38, and 40 bases downstream of the TATA box. To identify cis-acting elements regulating expression of the Ii gene, reporter plasmids containing deletions of the Ii promoter have been constructed and transfected into mouse L cells. Deletion of the H box results in a 50-100% increase in basal expression. Deletion of both the H and X boxes increases basal expression by 200-300% above that seen in constructs containing all three elements. A 25% decrease in basal level expression is seen for constructs that lack the Y-box element when compared to constructs containing the Y-box element but not the H- and X-box elements. DNase I footprinting analysis demonstrates protection of the H, X, and Y boxes as well as a nonconserved region between the H and X boxes. Mobility-shift experiments detect a factor specifically interacting with the Y box. Although the H-, X-, and Y-box elements interact with nuclear protein and are regulatory elements in L cells, these elements do not appear to play a role in IFN-gamma induction suggesting that other regulatory mechanisms must account for IFN-gamma's induction of the Ii in L cells.


Characterization of liver-enriched proteins binding to a developmentally demethylated site flanking the avian apoVLDLII gene. Although the avian apoVLDLII gene is normally expressed exclusively in the liver of the laying hen, the gene can be activated by estrogen in birds of either sex beginning between days 7-9 of embryogenesis. Developmentally programmed demethylation of sites in the 5'- and 3'-flanking regions of the gene have been shown to occur during this period of embryogenesis, suggesting that they may reflect changes in protein-DNA interactions that are involved in the acquisition of competence to activate the apoVLDLII gene. We have detected specific protein interactions at one location approximately 2.6 kb upstream from the apoVLDLII gene, that includes an Msp I site whose methylation status changes between days 7 and 9 of embryogenesis. The sequence of this region bears significant similarity to binding sites of members of the bZIP family of liver-enriched or -specific factors such as C/EBP, DBP, and LAP, that are characteristically produced relatively late during liver development. In the studies described here, we demonstrate that proteins binding to the upstream apoVLDLII site do not correspond to previously identified liver-enriched or -specific factors. They also display a pattern of activity during development and in human and avian hepatoma cell lines indicating that their expression is increased in proliferating cells. Southwestern blotting and UV cross-linking studies indicate that two proteins of approximately 60 kD are capable of binding to the site and we describe the purification of these factors from crude nuclear protein extracts obtained from rooster liver.


Identification and characterization of additional members of the cytochrome P450 multigene family CYP52 of Candida tropicalis. Using different DNA probes from the first two previously described alkane-inducible cytochrome P450 genes of the Candida tropicalis CYP52 gene family, we isolated five independent additional members by screening a genomic library under low-stringency conditions. These genes are not allelic variants and, when taken gogether, constitute the largest gene family known in this organism. The seven members of this gene family are located on four different chromosomes and four of them are tandemly arranged on the C. tropicalis genome. The products of the seven genes, alk1 to alk7, were compared to each other and revealed a high degree of divergence: the two most diverged proteins exhibit a sequence identity of only 32%. Six of the seven genes were shown to be induced by a variety of different aliphatic carbon sources but repressed when the organism was grown on glucose. Three of the five additional CYP52 genes could be successfully expressed in Saccharomyces cerevisiae and display different substrate specificities in in vitro assays with model substrates: alk2 and alk3 exhibit a strong preference for hexadecane, while alk4 and alk5 preferentially hydroxylate lauric acid.


The first human genes for tRNA(ArgICG), tRNA(GlyUCC), and tRNA(ThrIGU) and more tRNA(Val) pseudogenes: expression and pre-tRNA maturation in HeLa cell-free extracts. A functional tRNA(Val) gene, which codes for the major tRNA(ValIAC) isoacceptor species, and three new tRNA(Val) pseudogenes have been isolated from human genomic DNA. Two tRNA(Val) pseudogenes and a tRNA(Val) variant gene were found to be associated with tRNA genes encoding tRNA(ArgICG), tRNA(GlyUCC), and tRNA(ThrIGU), respectively, on distinct DNA fragments. All tRNA genes, including the pseudogenes, are actively transcribed in HeLa nuclear extract. Pre-tRNAs of tRNA(Val), tRNA(Arg), tRNA(Thr), and tRNA(Gly) genes are correctly processed to mature-sized tRNAs, whereas the three tRNA(Val) pseudogenes yield stable pre-tRNAs in vitro. These findings reveal that, together with the three known pseudogenes, half of the members of the human tRNA(Val) gene family are pseudogenes, all of which are active in homologous nuclear extracts in vitro and presumably also in vivo.


Rapid isolation of flanking genomic DNA using biotin-RAGE, a variation of single-sided polymerase chain reaction. A method is described for quickly and reproducibly isolating genomic DNA contiguous with known DNA sequence by means of the polymerase chain reaction (PCR). Flanking genomic DNA is isolated using a biotinylated sequence-specific primer in combination with a generic hybrid primer that binds to a deoxyoligonucleotide sequence artificially added to the ends of the genomic DNA. Amplified sequences that include the biotinylated primer are purified from nonbiotinylated amplification products by binding to a solid-phase streptavidin matrix. The biotinylated amplification product(s) are subjected to a further round of amplification, after which they can be subcloned and analyzed. This technique was applied to the isolation of three intron-exon junctions. Verification of the identify of these junction sequences was accomplished by designing primers based on the intron sequences isolated by Biotin-RAGE, amplifying across the exon using these intron primers, and sequencing the PCR-generated product.


Immobilization of aminoacylase from Aspergillus oryzae on synthetic modified polyacrylamides. A series of acrylamide-bisacrylamide copolymers modified by the Mannich Reaction was prepared. The immobilization of aminoacylase from Aspergillus oryzae on the copolymers was studied. All the polymers adsorbed the enzyme and the activity of the immobilized enzyme dependent on the amine used, viz. secondary amine, diamine, or aniline derivative. However, the activity was also influenced by the degree of crosslinking of the polymer. The surface morphology of the dimethylamine-modified polymer, with varying degrees of crosslinking, was analyzed by scanning electron microscope; the polymers having the largest pore diameter possessed the highest enzyme activity. One of the best polymers (DMA-A9B8) was used for immobilization of aminoacylase and its properties were studied. It had high enzymatic activity and good operational stability, i.e., retaining 90% of its original activity after being used for 42 days. The use of these copolymers for the preparation of immobilized enzymes is discussed.


Preparative in vitro synthesis of bioactive human interleukin-2 in a continuous flow translation system. Recombinant human interleukin-2 has been synthesized in vitro in a continuous flow translation system based on the wheat germ extract. In the course of translation of mRNA the interleukin-2 becomes aggregated due to the adsorption of this protein onto the ribonucleoprotein complex. This process correlates with the cessation of translation that is usually observed in 25-30 min. This can be prevented by the use of a flow system that allows continuous removal of the synthesized protein and maintains a steady concentration of all the necessary components. This approach permitted a yield of 1,500 protein molecules per mRNA molecule. The interleukin obtained promotes the proliferation of the interleukin-2-dependent CTLL-2 cell line. The biological activity of interleukin-2 not subjected to oxidative refolding was 10(5) units per milligram of protein.


Interactions of bioactive lipopeptides, iturin A and surfactin from Bacillus subtilis. The antifungal activity of iturin A and its interaction with erythrocyte membranes were enhanced in the presence of surfactin. The modification of the properties of iturin A was explained by the formation of mixed iturin A-surfactin micelles. Such mixed micelles were easily generated when both lipopeptides were in aqueous solutions in the absence of mineral salts but the formation of these micelles did not occur when the solutions contained a high molarity of mineral cations.


Use of helper-free retroviral vector to direct a high expression of porcine growth hormone in mouse fibroblast cells. A retroviral vector has been employed to express the cDNA coding for porcine growth hormone (pGH) in the mouse fibroblast cell NIH 3T3 in large quantity. In this study, a single gene vector which contained no selectable marker was used. We have coinfected NIH 3T3 cells with pGH retrovirus and Neo(r) retrovirus to obtain a stable, high-expression clone. Using a superinfection strategy, we further increased the copy number of proviral DNA in the host chromosome, thus increasing the pGH secretion from 22 to 55 micrograms/10(6) cells/24 h. The recombinant pGH produced from mouse fibroblast cells was heterogeneous at the N-terminus, which mimicked the situation with bovine growth hormone either from natural sources or from recombinant products derived from mouse fibroblasts. This technology is useful for many biologically important genes to be stably transduced by retroviral vector into mammalian cells and highly expressed.


Red blood cells as an antigen-delivery system. The use of adjuvants is usually required to induce strong immunological responses to protein antigens. However, in many cases these adjuvants cannot be extensively applied in human and veterinary vaccinations because of associated inflammatory reactions or granuloma formation. We show here that protein antigens (bovine serum albumin, hog liver uricase, and yeast hexokinase), coupled to autologous red blood cells by way of a biotin-avidin-biotin bridge, elicit an immunological response in mice similar to or higher than that obtained by the use of Freund's adjuvant. Quantities as low as 0.5 micrograms/mouse are high enough to generate these immunological responses. Furthermore, splenocytes of mice immunized by red blood cell-coupled antigens can be used to generate hybridomas secreting monoclonal antibodies. Thus, the delivery of antigens by autologous red blood cells is an effective way to avoid the use of adjuvants for producing anti-peptide antibodies and possibly to generate peptide vaccines.


Biochemical and rheodynamic properties of red blood cells crosslinked with glutaraldehyde. New data on the properties of red blood cells (RBC) cross-linked with glutaraldehyde are presented (see also Biochem. Biophys. Res. Commun., 1988, 156, 970-977). Equilibrium and kinetic data show that by carrying out the fixation procedure in the absence of oxygen but in the presence of allosteric effectors (e.g., stabilizing the low-affinity (T) quaternary state of hemoglobin), it is possible to maintain, at least in part, the biochemical functions of the crosslinked hemoglobin inside the cell. Moreover, we show that the oxygen affinity of fixed red blood cells (RBC) is still modulated, even though to a smaller degree, by the allosteric effector bezafibrate (BZF), which is able to cross the fixed RBC membrane. Membrane filtration experiments indicate that the higher rigidity of fixed RBC alters significantly their rheodynamic properties and show that in order to exploit "engineered" RBC as "blood substitutes," more flexible cross-linking reagents may offer significant advantages.


The effect of early enucleation on the orbit in animals and humans. Facial asymmetry and cosmetic deformity can occur following enucleation. The effect of enucleation on the growth of the orbit can be demonstrated in the rabbit and cat by early enucleation, X-ray study and dry skull measurements. In 42 human patients the anophthalmic orbital changes are determined by roentgenograms. The influencing factors of age and the use of implants are discussed as to changes in the rim, orbital walls and volume, and optic foramina measurements. Growth retardation due to roentgen ray therapy and other clinical aspects are considered which would influence the appropriate clinical management.


Temporalis muscle transfer in the treatment of the severely contracted socket. The treatment of severely contracted sockets represents one of the most difficult situations in ophthalmic plastic and reconstructive surgery. In particular, the treatment of the postirradiated socket has been considered "hopeless surgery." In recent years a modified approach to this problem has been tried, in which the use of the temporalis muscle provided a well-vascularized bed for delayed reconstructions.


Dermis-fat orbital implantation and complex socket deformities. Autogenous dermis-fat grafts implanted within the orbit survive best, with little loss of volume, when they are placed within Tenon's capsule immediately following the removal of the globe; when the rectus muscles (and anterior ciliary arteries) are anastomosed to the dermal edge of the graft; when no cautery has been applied to the recipient bed; and when the anterior diameter of the graft is no larger than 22 mm. Primary grafting in patients without systemic vascular disease is more effective than secondary procedures, performed on patients with fibrotic, compromised recipient beds and without direct apposition of the rectus muscles to the graft.


The role of flaps in the management of contracted eye sockets. Based on the pathology of the eye socket and periorbital deficiencies, three distinct classes of patients can be recognized: I. Those who solely have eye socket deficiency with normal orbital and periorbital tissue. The suggested surgical treatment for this class of patient would be a skin or mucosa graft. II. Patients who have inadequate lining, as well as orbital volume deficiency. The preferred reconstructive approach includes cartilage (rib or ear) with or without fat graft, and skin or mucosa grafts for eye socket expansion. III. For failed reconstructions of classes I and II or for patients with severe orbital and periorbital deficiencies, the choice is one of three flaps: If the superficial temporal vessels and the postauricular skin is intact, the ideal flap is postauricular fasciocutaneous. If the postauricular skin has previously been used yet the superficial vasculature is intact, a secondary flap is the better choice. In cases where both postauricular skin and superficial temporal vessels have been sacrificed the recommended flap is a free flap with microvascular anastomosis.


Indications and surgical techniques for orbital exenteration. Indications for the mutilating operation of exenteration are enumerated. They usually involve a malignant neoplasm of the orbital contents, primary, direct extension, or adnexal tissue that cannot be controlled by simple excision or irradiation. Surgically, subtotal exenteration with partial preservation of lids and even conjunctiva may be achieved occasionally. However, total exenteration may be lifesaving. Techniques and precautions are discussed. Advantages and disadvantages of skin grafting that influence the post-operative care are noted.


Radical orbital resections.

Radical orbital resections. The authors describe the techniques for subtotal, total and radical orbital exenteration. The aspects of primary and late reconstructive surgery are also discussed, with special reference to Frezzotti's personal technique for temporalis muscle transplantation after subtotal exenteration.


Frontalis muscle transfer in the reconstruction of the exenterated orbit. A technique using the frontalis muscle to reconstruct the exenterated orbit is described. The technique is simple and does not leave a depression in the temporalis fossa.


The history and development of facial prostheses. This paper includes the historical development of the modern-day facial prosthesis, the various materials used from the early days to the present, and the historical input of various people and their contributions towards the development of the qualities in fabricating a lifelike facial prosthesis.


Three dimensional imaging and computer-designed prostheses in the evaluation and management of orbitocranial deformities. Three dimensional images reconstructed from two dimensional CT scans allow improved analysis of complex orbitocranial bony deformities. This evaluation may be useful in patients with defects resulting from trauma, tumor, congenital abnormalities, or developmental disorders. Diagnosis, surgical management, and long-term follow-up evaluation may be aided by improved understanding of bony contour and volume analysis. Computer designed prostheses can be fabricated to precisely match bony defects and may be used as an alloplastic implant or as a model to aid intraoperative contouring of an autogenous bone graft. The limitations of three dimensional imaging include artifacts in the reconstructed images, increased radiation exposure, and increased cost. The technology is still evolving and the indications and benefits remain undefined at the present time.


Impressions for oculofacial prosthetics. Primary to the development of an acceptable and functional facial prosthesis is the impression procedure. The fabrication of a facial prosthesis combines the art and science of anatomy, cosmesis, and function, to develop a nonliving substitute to replace altered, missing, or defective regions of the head and neck area. Success depends on patient's cooperation, motivation, and commitment to treatment as well as the technical and artistic scope of the facial prosthetic service.


Impression making, sculpting, and coloring of orbital prostheses. The replacement of any anatomic structure by artificial means remains a challenge. This is particularly true in the facial area. The replacement must be one which blends with the adjacent tissues as well as replaces the missing structures. Careful planning and meticulous attention to detail in fabrication of a prosthesis can enable the maxillofacial prosthodontist to make a major contribution in the rehabilitation of the patient with an orbital defect.


The ocularists' management of congenital microphthalmos and anophthalmos. Early socket stimulation is crucial for management of congenital anophthalmos and microphthalmos among infants. Progressive sized hard conformers and lid expansion devices can expand the small socket in these patients. The ocularists' management of these two conditions is discussed and techniques are introduced.


Hydrophilic expanders for the congenital anophthalmic socket. The congenitally contracted socket often poses a challenging management problem; early surgery may be necessary in spite of the attendant difficulties. A series of patients is presented, with particular emphasis upon the use of a new hydrophilic expander in the severely contracted socket, to illustrate our current management protocol. Factors pertinent to orbital development and growth are discussed. Two types of contracted sockets are identified that appear to correlate with the presence or absence of an eye or ocular remnant. Use of a hydrophilic expander in the severely contracted anophthalmic socket with marked lid phimosis has enabled subsequent fitting and retention of a hard conformer in many of these cases, thus obviating early interventional surgery.


An expansion prosthesis for the microphthalmic socket. An expansion prosthesis has been developed for the anophthalmic or microphthalmic socket. Orthodontic wire covered with silicone tubing is fused onto a conventional methyl-methacrylate ocular prosthesis. The use of this device is illustrated in a case report.


Orbito-palpebral reconstruction in anophthalmos and severe congenital microphthalmos. In patients with congenital anophthalmos and severe microphthalmos, a tiny orbit and socket exist with little eyelids, frequently preventing retention of a standard conformer or prosthesis. Socket expansion is sometimes impossible with microorbitism; the retention of a prosthesis is also difficult when malformations of the eyelids exist. The treatment of these difficult cases includes three stages. The first stage is orbital expansion that depends on the cephalometric studies of the patient: transverse osteotomy on the maxilla and the zygomatic bone with lateral bar by extracranial route, vertical osteotomy on the roof of the orbit by intracranial route. In some cases, the osteotomy includes expansion in the transverse and vertical diameter with bone grafts in the defects and on the lateral and superior rims. Simultaneously, socket expansion is performed by incision of the conjunctival sac circumferentially, with mucosal or split skin grafts on a conformer. The second stage includes eyelid reconstruction by different flaps. A third stage is frequently needed for correction of eyelid malposition on the prosthesis: ptosis, entropion surgery. Two cases of congenital anophthalmos are reported. Methods and indications of treatment are discussed.


Orbito-palpebral reconstruction in two cases of incomplete cryptophthalmos. Two cases of congenital symblepharon (variant of cryptophthalmos) are reported. Cryptophthalmos is a very rare congenital defect, with incomplete or complete failure in the development of one or both eyelids with skin recovering the anterior segment. Surgical treatment is described including expansion of the conjunctival fornix with eyeball conservation if possible. At the same time or later, the upper eyelid is reconstructed by inferior eyelid flap. The ophthalmic features of cryptophthalmos and its systemic associations are reviewed.


The myofibroblast and the anophthalmic socket. One of the greatest advances in the understanding of wound healing was the identification and characterization of the myofibroblast by Gabbiani in 1971. Since that time this contractile cell has been found in the early stages of wound healing and in many pathologic states. In a recent study, the myofibroblast was found in healing and contracting anophthalmic sockets.


Structure of 2,3-naphthalenedicarboxylic acid. C12H8O4, M(r) = 216.20, monoclinic, C2/c, a = 5.087 (2), b = 19.222 (3), c = 9.552 (2) A, beta = 93.81 (3) degrees, V = 932.0 (5) A3, Z = 4, Dx = 1.54 g cm-3, lambda = 0.71073 A, mu = 1.09 cm-1, F(000) = 488, T = 295 K, R = 0.037 for 687 unique reflections having I > 3 sigma (I). In the structure reported here, a twofold axis lies in the plane of the molecule and bisects the three central ring bonds. The average C--C bond distance in the naphthalene core is 1.401 (27) A and the average interior angle 120.0 (14) degrees. The average deviation of the C atoms comprising the naphthalene rings from the best least-squares plane describing the rings is 0.021 (8) A. The plane of the single unique carboxylic acid group makes a dihedral angle of 33.1 (1) degrees with the least-squares plane of the naphthalene core. The carboxylic acid group, in which the observed O--H distance is 1.00 (3) A, forms a cyclic-dimer hydrogen bond across a center of inversion with the H...O(acceptor) distance 1.68 (3) A, the O...O-(acceptor) distance 2.676 (2) A and the O--H...O-(acceptor) angle 174 (3) degrees. The C--O bond lengths indicate no disorder in the carboxylic acid dimers. The molecules are arranged in stacks with the naphthalene cores parallel to the (102) plane and with the hydrogen bonds of the acid groups linking adjacent stacks. This structure of 2,3-naphthalenedicarboxylic acid is essentially the same as that of o-phthalic acid, but with the b cell edge correspondingly larger to accommodate the additional aromatic ring.


Structure of 9-epiquinine hydrochloride dihydrate versus antimalarial activity. 9-Epiquinine hydrochloride dihydrate [(9S)-6'-methoxycinchonan-9-ol hydrochloride dihydrate], C20H25N2O2+.Cl-.2H2O, M(r) = 396.9, orthorhombic, P212121, a = 8.059 (2), b = 11.537 (3), c = 22.311 (6) A, V = 2074.1 (9) A3, Z = 4, Dx = 1.271 g cm-3, Cu K alpha, lambda = 1.54178 A, mu = 18.58 cm-1, F(000) = 848, room temperature, final R = 6.56% for 1344 reflections with magnitude of Fo > 3 sigma (F). 9-Epiquinine crystallized as a hydrated tertiary amine hydrochloride salt. The intramolecular N(1)+ ...O distance is 2.816 A. All H atoms attached to O or N atoms form intermolecular hydrogen bonds. The Cl ion is involved in four hydrogen bonds including one with the hydroxyl group of 9-epiquinine. The N(1)+-H moiety hydrogen bonds to a water molecule. The O(12)-C(9)...N(1)+-H(1) torsion angle was equal to -0.2 (3.8) degrees in comparison to 97.0 degrees for quinidine sulfate [Karle & Karle (1981). Proc. Natl Acad. Sci. USA, 78, 5938-5941]. Two theories have been proposed in the literature to explain the low antimalarial activity of 9-epiquinine. The crystal structure of 9-epiquinine hydrochloride is not consistent with the hypothesis that 9-epiquinine prefers to form intramolecular rather than intermolecular hydrogen bonds, but is consistent with the hypothesis that N(1) and the hydroxyl group of 9-epiquinine are in an orientation which is unfavorable towards exerting antimalarial activity.


Ethyl (2R,3R,SR)-3-(ethoxycarbonyloxy)-2-[1-(p-tolylsulfinyl) cyclopropyl]-1-pyrrolidinecarboxylate. The title compound, derived in four steps beginning with the cyclopropanation reaction of the anion of the alpha-sulfinyl ketimine (4R,SS)-4-(tert-butyldimethylsilyloxy)-3,4-dihydro-5-(p-tolysul  finyl)methyl)-2H-pyrrole with 2-chloroethyl trifluoromethanesulfonate, contains a shorter than expected bond (C(7)--C(8) 1.48(l) A) and a larger than expected angle (S(1)--C(6)--C(2) 123.2(6) in the cyclopropane framework.


Racial differences in maximal vasodilatory capacity of forearm resistance vessels in normotensive young adults. This study was performed to determine whether alterations in vascular structure exist in a biracial population of young (age 22.3 +/- 0.6 yrs [mean + SE]) normotensive men. We examined maximal vasodilatory capacity in 21 blacks and 20 whites (average blood pressure = 122/75 and 118/72 mm Hg, respectively). Forearm blood flow was determined at rest and after 10 min of ischemic handgrip exercise using venous occlusion plethysmography. Forearm vascular resistance was computed from blood flow and mean arterial blood pressure determined by auscultation. Minimum forearm vascular resistance was 23% higher in blacks (2.60 +/- 0.60) than in whites (2.11 +/- 0.41) (P = .005), and was unrelated to parental history of hypertension. The regression equation for minimum forearm vascular resistance (Y) and casual blood pressure (X) for blacks was Y = -1.782 + 0.0487X (r = 0.522); for whites it was Y = -1.165 + 0.0367X (r = 0.418). When the data were covaried on resting mean arterial blood pressure, blacks still had a higher minimum forearm vascular resistance (P = .014). The results suggest a racial difference in the vascular structure of the forearm resistance vessels.


The relation of culturally influenced lay models of hypertension to compliance with treatment. Problems in compliance with treatment and illness management have frequently been traced to differences between patients' explanatory models of illness and the biomedical model. We investigated the relationship of cultural beliefs about hypertension to compliance with treatment. Using semistructured interviews, we elicited the explanatory models of hypertension held by 60 black hypertensive women being treated at Charity Hospital in New Orleans. The patient sample was followed for 2 months to obtain data on compliance with antihypertensive treatment and blood pressure control. As described, 53% of the patient sample recognized two basic folk illness models: "high blood" and "high-pertension." Patients' illness models were significantly related to compliance with treatment at the P = .01 and .001 levels. Compliance was related to blood pressure control at the P = .05 level. These results indicate that culturally influenced health beliefs are an important influence on compliance and, in turn, blood pressure control. Improved understanding of these problems by physicians may improve the management of their patients' illness.


Relationship of the renin-angiotensin system and systemic arterial pressure to sodium excretion during atrial natriuretic peptide infusion in men. The goal of this study was to evaluate the relative contribution of the renin-angiotensin system and mean arterial pressure to sodium excretion and urine flow rate during an infusion of atrial natriuretic peptide (ANP) at physiologically relevant doses in humans. Eight normal volunteers were studied during five periods: (1) baseline in the supine position; (2) during an infusion of ANP at physiologic doses (0.01 micrograms/kg/min) in the supine position; (3) during ANP infusion and 60 degrees head-up tilt; (4) during ANP infusion, head-up tilt, and interruption of the renin-angiotensin axis with the angiotensin converting enzyme inhibitor (ACEI) enalaprilat; and (5) in the supine position during ANP infusion and ACEI. Infusion of ANP in the supine posture significantly increased urine flow rate and sodium excretion compared to baseline while mean arterial pressure and plasma renin activity were unchanged. During head-up tilt and ANP infusion, urine flow rate and sodium excretion were no longer significantly elevated over baseline while mean arterial pressure decreased and plasma angiotensin II levels increased. Addition of ACEI caused a marked diminution of urine flow rate and sodium excretion compared to baseline levels despite continued ANP infusion. Although mean arterial pressure after ACEI administration was lower than baseline, it was not significantly different from the non-ACEI head-up tilt state. Placing subjects in the supine position during ANP infusion and ACEI administration increased mean arterial pressure to levels that were no longer different from baseline, but urine flow rate and sodium excretion remained significantly depressed to the same degree as during head-up tilt with ACEI.(ABSTRACT TRUNCATED AT 250 WORDS)


Angiotensin II regulates cytochrome P-450 steroid hydroxylase enzyme expression in human adrenal glomerulosa cells. We have examined the regulation of cytochrome P-450 side chain cleavage enzyme (P-450SCC) and P-45011 beta (18) hydroxylase (P-450(11) beta) enzyme expression by angiotensin II (AII), the major regulator of aldosterone biosynthesis, in cultured human adrenal glomerulosa cells. We also examined the effect of lipoxygenase products of arachidonic acid on the expression of these enzymes since it has been previously shown that the 12-lipoxygenase pathway plays a key role in AII-induced aldosterone synthesis in rat and human adrenal glomerulosa cells. AII (10(-7) mol/L) induced a 3-fold stimulation of P-450SCC and over a 2-fold increase in P-450(11) beta protein expression. The 12-lipoxygenase product, 12-hydroxyeicosatetraenoic acid (12-HETE) caused a 2-fold increase in P-450(11) beta levels without altering P-450SCC levels. These results show for the first time that AII can directly increase the levels of P-450SCC and P-450(11) beta enzymes in glomerulosa cells. The results also suggest that 12-HETE may mediate long term effects of AII action by stimulating P-450(11) beta levels.


Ganglion atrial natriuretic peptide in NaCl sensitive spontaneously hypertensive rats. Reports from other laboratories have shown that atrial natriuretic peptide (ANP) stores in sympathetic ganglia are increased during dietary NaCl supplementation in normotensive rats. We have previously demonstrated that dietary NaCl supplementation in NaCl sensitive spontaneously hypertensive rats (SHR-S) exacerbates hypertension and enhances peripheral sympathetic nervous system activity, while NaCl resistant Wistar-Kyoto (WKY) rats show neither response. Since endogenous ANP may inhibit ganglion transmission, an inability of SHR-S to increase ganglion ANP appropriately in response to high NaCl feeding could contribute to the NaCl induced increase in sympathetic nervous system activity and blood pressure in this model, while an increase in ganglion ANP in NaCl supplemented WKY would tend to prevent sympathetic activity and blood pressure from rising. The current study tested the hypothesis that ganglion ANP levels increase in WKY but not in SHR-S during dietary NaCl supplementation. Male SHR-S and WKY rats were placed on 1% or 8% NaCl diets at 7 weeks of age. The rats were decapitated without prior anesthesia 3 weeks later, and the superior cervical and celiac ganglia were removed for the measurement of ANP by radioimmunoassay. Dietary NaCl supplementation produced significant increases in blood pressure in SHR-S, but not in WKY rats; the high NaCl diet was associated with significant increases in the ANP content of superior cervical and celiac ganglia in WKY rats, but not in SHR-S.(ABSTRACT TRUNCATED AT 250 WORDS)


Usefulness of atrial natriuretic peptide assay in primary aldosteronism. Atrial natriuretic peptide (ANP) can be elevated in conditions which are characterized by increased atrial pressure and or expanded plasma volume. We and others have previously shown a significant increase of ANP plasma levels in a small number of patients with primary aldosteronism. In this study we have extended the assay of plasma ANP to a larger number of patients. We studied ANP plasma levels before and after upright posture and acute sodium load in 16 patients with aldosteronoma (APA) and 13 with idiopathic aldosteronism (IHA). The study was repeated also after the removal of aldosteronoma. In patients with primary aldosteronism, the mean supine ANP plasma level was significantly higher than in the age matched normal subject group; supine ANP was significantly higher in the APA than in the IHA group. The decrease of ANP levels after upright posture was significant in both groups. The ANP increase after acute saline load was similar in APA and in IHA. After the removal of aldosteronoma ANP values returned to normal. In conclusion, it is confirmed that plasma ANP levels are elevated in primary aldosteronism and could reflect a greater volume expansion in patients with APA. Despite this difference, ANP still responds to physiological stimuli in both groups. Finally, ANP measurement can provide an additional tool in the differential diagnosis between APA and IHA.


Effects of endothelin-1 and vasopressin on resistance arteries of spontaneously hypertensive rats. Previous studies have shown an impairment of responsiveness of resistance arteries to endothelin-1 in experimental hypertensive rats. This study was undertaken to demonstrate whether responses were also blunted in adult 20-week-old spontaneously hypertensive rats (SHR). Resistance arteries from the mesenteric vascular bed exhibited normal active tension responses to endothelin-1 and arginine vasopressin, and exaggerated responses to norepinephrine. Since the media was thicker in SHR, media stress responses to endothelin-1 and arginine vasopressin were significantly impaired, while norepinephrine media stress responses were similar in SHR and Wistar-Kyoto rats (WKY). Active pressure responses to endothelin-1, arginine vasopressin, and norepinephrine were significantly amplified by the narrowed lumen of blood vessels in SHR. Additionally, media cross-sectional area was similar in SHR and WKY, but was greater in SHR when normalized for the smaller body weight of the hypertensive rats. These results demonstrate the presence of remodeling in resistance arteries of 20-week-old SHR, and show that the altered morphology of these blood vessels may significantly amplify impaired wall stress responses to endothelin-1 and arginine vasopressin, which may contribute to elevation of blood pressure.


Lack of effect of parathyroid hormone on calcium homeostasis in rat platelets. The in vitro effects of parathyroid hormone on Ca2+ handling by fura-2 loaded rat platelets were studied. The incubation of these platelets with rat parathyroid hormone (1-34) for 10 min had no effect on intracellular fura-2 metabolism or [Ca2+]i in the resting state. The [Ca2+]i response to 0.1 U/mL thrombin was unaffected by preincubation with parathyroid hormone in the presence or absence of extracellular Ca2+. Furthermore, the recovery rate of [Ca2+]i after the thrombin-induced peak in Ca(2+)-depleted media was not altered with parathyroid hormone. These data indicate that parathyroid hormone may not have a significant effect on Ca2+ homeostasis in rat platelets in unstimulated and stimulated conditions.


Glucose, insulin, and lipid metabolism in doxazosin-treated patients with hypertension. The metabolic changes associated with doxazosin treatment of hypertension were evaluated in ten patients with mild hypertension (mean +/- SEM = 150 +/- 3/100 +/- 1 mm Hg) and a plasma triglyceride (TG) concentration > 1.50 mmol/L. The blood pressure was lower after 4 to 6 months of doxazosin treatment (mean +/- SEM = 134 +/- 4/87 +/- 1 mm Hg), which was also associated with a significantly lower plasma insulin response to a 75 g oral glucose load, and lower plasma TG and cholesterol concentrations. In addition, insulin-mediated glucose uptake was significantly greater after doxazosin treatment. These data suggest that doxazosin treatment of patients with mild hypertension is associated with changes in insulin and lipid metabolism that should decrease the risk of coronary heart disease.


In vivo measurement of atrial natriuretic peptide receptors using nuclear imaging. We have successfully visualized atrial natriuretic peptide (ANP) receptors in vivo using nuclear imaging. 123I-Labelled ANP, injected in green vervet monkeys, was rapidly bound to ANP receptors in the kidneys and lungs. That the observed uptake was receptor mediated was demonstrated with competition studies using simultaneous injection of unlabelled ANP 99-126. It was possible to distinguish between the ANP receptor subtypes by the use of selective antagonists. Thus coinjection of ANP 102-121-des[Gln, Ser, Gly, Leu, Gly] (C-ANP), an ANP analog that selectively binds to the ANP C-receptor, decreased uptake in the kidneys by 50% but increased relative uptake in the lungs and soft tissues. This method permits for the first time, the dynamic in vivo analysis of ANP receptors and their interaction with endogenous ligand. Differences and changes in local ANP receptor concentrations and occupancy could be detected. Since ANP receptor density and affinity are influenced by various physiological and pathological conditions, clinical and diagnostic applications seem possible.


Effects of angiotensin converting enzyme inhibitors and calcium antagonists on atrial natriuretic peptide release and action and on albumin excretion rate in hypertensive insulin-dependent diabetic patients. Abnormalities in sodium homeostasis and in atrial natriuretic peptide (ANP) behavior could play a role in determining and accelerating the development of glomerular hypertension, hypertension, and microalbuminuria in insulin-dependent diabetes. The aim of the present study was to investigate in 32 hypertensive insulin-dependent diabetic patients (HD) with an altered albumin excretion rate the natriuretic response and ANP release to saline load (2 mmol/kg 90 min, and the effects angiotensin converting enzyme inhibitor therapy 2.5 to 5.0 mg cilazapril, once daily), and calcium antagonists (sustained release verapamil: 120 to 240 mg Isoptin Press, once daily, and long acting nifedipine: 20 to 40 mg Adalat AR, twice daily) on sodium homeostasis and albumin excretion rate. Eight normal subjects matched for sex, age, and weight served as controls. The 32 HD patients showed a blunted response in ANP release and sodium excretion during saline infusion in comparison with controls. The cilazapril and verapamil treatments were tested in 16 of the 32 HD patients and were both effective in ameliorating natriuretic and ANP response to saline load and in decreasing albumin excretion rate. The combined cilazapril and verapamil treatment further improved both these parameters in these patients, although blood pressure levels were comparable. The other 16 HD patients underwent sequential verapamil and nifedipine treatment. Verapamil was more effective than nifedipine in improving natriuresis and ANP release to saline load and in lowering the albumin excretion rate. The results of the present study demonstrate that sodium homeostasis and ANP release are altered in hypertensive nephropathic patients, and both cilazapril and verapamil are more effective than nifedipine in ameliorating natriuresis, ANP release, and albumin excretion rate.


Utilization of biosensors and chemical sensors for space applications. There will be a need for a wide array of chemical sensors for biomedical experimentation and for the monitoring of water and air recycling processes on Space Station Freedom. The infrequent logistics flights of the Space Shuttle will necessitate onboard analysis. The advantages of biosensors and chemical sensors over conventional analysis onboard spacecraft are manifold. They require less crew time, space, and power. Sample treatment is not needed. Real time or near-real time monitoring is possible, in some cases on a continuous basis. Sensor signals in digitized form can be transmitted to the ground. Types and requirements for chemical sensors to be used in biomedical experimentation and monitoring of water recycling during long-term space missions are discussed.


Binding kinetics of antigen by immobilized antibody: influence of reaction order and external diffusional limitations. The influence of reaction order and external mass transfer limitations on the binding kinetics of antigen in solution to antibody covalently or non-covalently attached to a cylindrical fiber-optic biosensor is presented. Both single-step and dual-step binding of antigen to antibody is considered. The rate of attachment of antigen to antibody is linear for all reaction orders in the time frame (100 min) considered. The rate of attainment of saturation levels of antigen in solution close to the surface is very rapid (within 20 min). An increase in the presence of mass transfer (denoted by a Damkohler number increase) decreases the saturation level of the antigen close to the surface, and the rate of antigen attachment to the antibody covalently or non-covalently bound on the surface for one-half, one, one and a half, and second-order reaction. As intuitively expected, an increase in the initial antigen concentration in solution increases the saturation level of the antigen close to the surface, and the rate of antigen attachment to the antibody covalently bound on the surface for all the reaction orders considered. Non-dimensional plots presented in the analysis help extend the analysis to different antigen-antibody systems. A decrease in the external diffusional limitations has an effect of decreasing the effect of reaction order on the saturation levels of antigen close to the surface and the rate of attachment of the antigen in solution to the antibody on the surface.


Calibration in dogs of a subcutaneous miniaturized glucose sensor using a glucose meter for blood glucose determination. The feasibility of calibrating a glucose sensor by using a wearable glucose meter for blood glucose determination and moderate variations of blood glucose concentration was assessed. Six miniaturized glucose sensors were implanted in the subcutaneous tissue of conscious dogs, and the parameters used for the in vivo calibration of the sensor (sensitivity coefficient and extrapolated current in the absence of glucose) were determined from values of blood glucose and sensor response obtained during glucose infusion. (1) Venous plasma glucose level and venous total blood glucose level were measured simultaneously on the same sample, using a Beckman analyser and a Glucometer II, respectively. The regression between plasma glucose (x) and whole blood glucose (y) was y = 1.12x-0.08 mM (n = 114 values, r = 0.96, p = 0.0001). The error grid analysis indicated that the use of a Glucometer II for blood glucose determination was appropriate in dogs. (2) The in vivo sensitivity coefficients were 0.57 +/- 0.11 nA mM-1 when determined from plasma glucose, and 0.51 +/- 0.07 nA mM-1 when determined from whole blood glucose (t = 1.53, p = 0.18, n.s.). The background currents were 0.88 +/- 0.57 nA when determined from plasma glucose, and 0.63 +/- 0.77 nA when determined from whole blood glucose (t = 0.82, p = 0.45, n.s.). (3) The regression equation of the estimation of the subcutaneous glucose level obtained from the two methods was y = 1.04x + 0.56 mM (n = 171 values, r = 0.98, p = 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)


Amperometric glucose biosensor with extended concentration range utilizing complexation effect of borate. Borate buffer strongly decreases amperometric response of a glucose oxidase linked pO2 or H2O2 sensing electrode, extending substantially its linear calibration range. With increasing pH and concentration of the buffer the upper limit for glucose can be varied between 1 and 30 mmol l-1 glucose. The effect of borate ion is explained by the rapid complexation of glucose decreasing the equilibrium concentration of free beta-anomer, the specific substrate of glucose oxidase. The high loading of cross-linked enzyme inside the sensor membrane is necessary for the measurement to ensure an almost constant response factor (delta i per 1 mmol l-1) between pH 5 and 10. Analysis in stirred solution and in a flow-through system has been employed for the measurement of elevated glucose levels in heparinized human blood or plasma samples.


Structural and ultrastructural pattern of the embryonic and early foetal human liver. With the aim of stressing the main steps which characterize the early development of the human foetal liver, fragments of hepatic buds of 7-12 weeks of pregnancy have been processed for light and scanning electron microscopy; Vial and Porter technique for isolated cells has been utilized too. The obtained results can be summarized as follows. At the 7th-8th weeks the hepatocytes show a globose shape, their surface is furnished with scattered and irregular evaginations and they are arranged in loose and narrow ribbons, separated by vascular spaces; the hepatocytes are tightly connected with haemopoietic cells, usually furnished with hyperchromatic nuclei. On the other hand, the hepatic parenchyma shows a compact pattern at the 11th-12th weeks. The hepatocytes have acquired a polyhydric shape and their faces are usually interlocked with the plasmatic membrane of the adjoining elements than in three fields: (a) the face projecting to the vascular walls with which they are going to form the Disse spaces; (b) the opposite side in which the membranes of neighbouring cells often appear spaced to line the primitive biliary canaliculi; and (c) the areas in which hemispheric and deep hollows of the hepatocyte surface hold haemopoietic elements. The obtained results demonstrate that the developing hepatic buds undergo deep structural changes, at level of the parenchyma and vascular system, in the studied period.


Borderline hypertension: relationship between job and psychophysiological profile. To identify relationships among hypertension, job and cardiovascular reactivity we studied 81 borderline hypertensives divided into labourers (L), white collars (W) and managers (M). After behavioral analysis, they underwent 4 tests: arithmetic, Sacks, acoustic, electric. Along the entire sitting, muscular contraction, skin conductance (SCL), peripheric temperature (THP), SBP, DBP and HR were taken, every 30". Depression, obsessive-compulsive, anxiety and neurotic traits were found in W. SBP, DBP and HR were not significantly different. Failed recovery curves of SCL were identified in M and W, but the presence of abnormal response profile, of both, SCL and THP, only in W. This autonomic dysreactivity, previously recognized as a possible characteristic of the prehypertensive condition, could uncover the role of certain work stressful condition to increase the sympathetic drive underlying hypertension.


Cells from human bone giant cell tumors show a [Ca2+]o-sensing. Giant cells from a human giant cell tumor of bone, showing several osteoclast features were tested for their capability of detecting the [Ca2+]o by a receptor like [Ca2+]o sensing. We found that cultured cells responded to elevation of [Ca2+]o, obtained adding 4 mM Ca2+ to the 2 mM Ca2+ containing buffer, by a transient increase of [Ca2+]i. Proliferative cells induced to differentiate by treatment with 10(-8) M 1,25 dihydroxyvitamin D3, were upregulated in their capability of responding to elevated [Ca2+]o. In fact, in this circumstance, the peak of [Ca2+]o-induced [Ca2+]i rise was increased compared to untreated cells. This suggests that 1,25 dihydroxyvitamin D3 induces a more efficient regulation of osteoclast activity.


[Use of a protein-free medium for the in vitro development of sheep embryos]. The employment of protein-free medium for the culture of ovine embryos collected at the 1-2 cell stage from superovulated ewes was investigated. For this purpose sheep zygotes were randomly allocated in four treatment groups: T1) CZB medium + bovine serum albumin (BSA) on sheep oviductal monolayer (SOM), T2) CZB + polyvinyl alcohol (PVA) + SOM, T3) CZB + PVA + SOM supplemented with inositol (I) and serine (S), T4) TCM 199 + 10% fetal calf serum + SOM. Standard culture conditions were 2 ml of medium in 35 mm Petri dishes, under 5% CO2 in air at 39 degrees C. The percentages of morulae and blastocysts were recorded after 4 and 7 days of culture. After 4 days of culture there was no significant difference (P > 0.05) in the percentage of morulae between embryos cultured in T1 (86%), T2 (85%), T3 (88.8%), and T4 (87.5%). After 7 days the percentages of blastocysts were T1 (70%), T2 (50%), T3 (55.5%) and T4 (46.8%). These data suggest that a protein-free medium, CZB + PVA and CZB + PVA + I + S, can support ovine preimplantation embryo development in vitro; however CZB medium supplemented with BSA enhances development to blastocyst.


[Microencapsulation in Na-alginate and in vitro development of sheep blastomeres]. Embryos at 4 cell stage obtained from Sarda ewes superovulated with FSHp (Sigma) were micromanipulated in order to obtain single blastomeres (1/4 E). The 1/4 E have been located randomly in two groups. In the first (Group A n. 30) the 1/4 E have been put back in empty zonae pellucidae; in the second (Group B n. 21) they have been microencapsulated in sodium alginate (1.1%) by dropping cell-alginate solution in a 1.5% CaCl2. Each capsule (1 mm diameter) contained four 1/4 E. The blastomeres have been co-cultured for 5 days in CZB medium on oviductal cell monolayer in a humidified incubator (5% CO2, 95% air, 38.5 degrees C). No differences were found between the groups reaching blastocyst stage after the end of the culture period (A 50%-B 47%).


[Muscle reinnervation in rats treated with vitamin E]. During muscle reinnervation, a transitory phase of polyinnervation occurs. In reinnervated muscles of vitamin E deficient rats, sprouting and polyinnervation are increased with respect to reinnervated controls. In this work, polyinnervation was observed in reinnervated extensor digitorum longus (edl) muscle of rats treated with pharmacological doses of vitamin E. Sciatic nerve was crushed and edl muscle was examined electrophysiologically at 30, 40 and 60 days after denervation. The percentage of polyinnervated cells in controls peaked at 30 days and thus it decreased. In muscles of vitamin E treated rats, the time course of percentage of polyinnervated muscle cells was qualitatively the same, but it was decreased at all times.


[Different behavior of phagocytes in young and old subjects]. Ageing is a dynamic phenomenon in which there is a physiological decay in all the functions of the individual. The consequence is an increased susceptibility to infections, autoimmune diseases and cancer. Phagocytic cells as polymorphonuclear leukocytes (PMNL) and monocytes (Mo) are of prime importance in the defence against invasive agents, PMNL and Mo seek out and destroy invading micro-organisms. Chemotaxis and phagocytosis are two mechanisms that are activated by these cells for this purpose. In this study, using "in vitro" techniques, we have verified if, at the level of such functions of cell defense, there could be variations in elderly subjects with respect to younger subjects. Our results show a chemotactic activity of PMNL in the elderly that is higher and a phagocytic activity that is lower. As regards Mo, there is a lower chemotactic activity in the elderly and only a slight difference in phagocytic activity with respect to the younger subjects. These results are in agreement with those found at the clinical level showing the elderly less protected from infection with respect to younger subjects.


[Modifications of the Portoscuso ecosystem. Distribution, accumulation,and dynamics of pollutants. I. Observations on animal tissues]. In order to assess the extent of environmental pollution by Pb, Cd and Zn in the industrial area of Portoscuso (Southwestern Sardinia-Italy), anatomohistopathological, histochemical and chemical tests were carried out on the heart, liver, kidneys and bones of sheep slaughtered in the local abattoir. On microscopic examination, degenerative lesions were severe in liver and kidneys and slight in the heart; histochemical stainings clearly showed a notable presence of Zn, while spectrophotometric examination confirmed, besides Zn pollution, very high tissue concentrations of Pb and Cd. The Authors conclude that other trial observations over larger areas are necessary.


Effect of one booster dose in antirabies vaccination. The antibody response obtained after one booster dose of rabies vaccine prepared in suckling mice brains at Serviço de Saúde Pública da Cidade do Rio de Janeiro is described. Four prime vaccinal groups were used: group I, persons who had received 16 doses 10 years before this investigation; group II, persons who had received 5 doses of the vaccine 10 years before; group III, persons who had received 9 doses 5 years before; and group IV, persons who had received 16 doses 5 years before. One booster dose of the vaccine was administered to all persons involved in the study. Blood samples were collected before vaccination (day 0), then 7 and 30 days after vaccination. Antibody titres were determined by seroneutralization test in mice (SWM). The results demonstrated that all persons who had been treated with 5, 9 or 16 doses of the vaccine 5 to 10 years before had their antibody titres increased on the 7th and 30th days after one booster dose.


Purification process monitoring in monoclonal antibody preparation: contamination with viruses, DNA and peptide growth factors. Administration in vivo of monoclonal antibodies to humans is challenged by considerations regarding their safety. Contamination with viruses, potentially oncogenic nucleic acids and biologically active components like growth factors and hormones forms a serious point of concern in this respect. We have investigated the potential risk of viral contamination by measuring the reduction of 12 different viruses (after spiking) in the standard downstream purification process of ascitic fluid. Depending on the type of virus added and the purification step employed, the reduction of infectious virus particles varies considerably. The overall reduction ranges from about 10(3), observed for a member of the family of Papovaviridae, to more than 10(12) for members of the families of Herpesviridae and Orthomyxoviridae. Using hybridization analysis with a mouse (genomic) DNA probe, we show that the amount of residual DNA in ascitic fluids may also vary considerably, ranging from 75 ng/ml to 1 microgram/ml. In crude preparations produced in cell culture, much lower DNA concentrations are found (0.3 ng/ml). When standard downstream purification procedures are applied to ascitic fluid, a significant reduction of residual DNA levels is observed in the purified monoclonal antibody preparations and in intermediate fractions. The overall reduction factors vary from about 10(3) to 10(4), which is also confirmed by spiking experiments with either purified DNA or crude chromatin-like DNA. Using in-vitro cellular assays, we further show that peptide growth factors like PDGF and TGF beta are present in considerable amounts in ascitic fluids. The observed biological activities, however, are completely eliminated during the purification steps applied.


The use of bovine fibrin-streptokinase films for the determination of recombinant human plasminogen. Plasminogen is a key component of the haemostatic system in man and the plasma-derived protein molecule has been actively investigated. Within the last few years cDNA and the gene encoding plasminogen have been cloned and the protein has been expressed in a number of eukaryotic or prokaryotic systems. Yields of expressed plasminogen are frequently low. Currently available assays for plasminogen generally rely on the determination of antigen or utilize tripeptide substrates for measuring functional activity, and they have certain limitations. Assays employing relevant protein substrates offer an alternative way to measure function and overcome the drawbacks associated with the other tests. The use of fibrin films for the assay of low levels of recombinant plasminogen has not been described fully before. The two fibrin film-based assays described in this paper are significant additions to the array of assays available for plasminogen molecules.


High growth reassortant influenza vaccine viruses: new approaches to their control. When a new strain of an influenza virus is required to be incorporated into influenza vaccine, attempts are made to recombine such strains with laboratory adapted viruses, which will grow to high titre in order to improve the yield of the vaccine strain. It is important that such high growth reassortant vaccine strains are not contaminated with genes coding for the antigenic determinants of the high growth laboratory strain. We describe the characterization of two recent high growth reassortants and the application of the polymerase chain reaction to ensure their genetic identity and purity.


Analysis of therapeutic growth hormone preparations: report of an interlaboratory collaborative study on growth hormone assay methodologies. Recombinant DNA-derived human growth hormone (somatotropin) is widely used to treat growth hormone-deficient children. The potency of this product is determined by in-vivo bioassay in hypophysectomized rats, which is imprecise, costly and invasive, and there have been suggestions that it could safely be replaced with in-vitro or physico-chemical alternatives. In this report we present the results of a collaborative study designed to test this proposal. Somatotropin was modified by mild or severe proteolysis, mild or severe oxidation or treatment at high pH, and compared in a multi-centre collaborative study with unmodified somatotropin or with dimerized somatotropin. Participating laboratories included manufacturers and national control laboratories, and pharmacopoeial bioassays were compared with in-house in-vitro and physico-chemical bioassays. Although performing adequately with untreated somatotropin, for degraded samples the in-vivo bioassays were relatively unresponsive to changes in the growth hormone molecule. In contrast, the physico-chemical assays, in particular the reverse-phase HPLC, performed with a high degree of selectivity. We conclude that in the case of somatotropin, the in-vivo bioassay can be removed from the routine product specification with an acceptable degree of security. This however does not obviate the requirement rigorously to demonstrate biological activity in-vivo during product development, nor may the conclusions of this study be applied to other therapeutic recombinant proteins without similar collaborative investigations.


A collaborative study to assess the proficiency of laboratory estimates of potency of live measles vaccines. A collaborative study was undertaken to assess the variability in estimates of the potency of measles vaccines. Overall a median variation of 2.0 log10 between estimates was observed. This was reduced to a median of 1.0 log10 when the potencies were expressed relative to a reference vaccine. A difference in the sensitivity between plaque assays and TCID50 assays was also reduced when relative potencies were used. The benefit of including a common reference preparation in vaccine assays was therefore demonstrated. For the vaccines assayed in this study, it was not necessary to use a measles reference of the same strain as the vaccines tested. We therefore recommend that measles vaccines be assayed against a single international reference preparation.


Effect of low fresh gas flow rates on inspired gas composition in a circle absorber system. To determine the effects of fresh gas flow on inspired gas composition during low-flow anesthesia. Randomized trial with 2-hour observation periods in patients assigned to one of three groups. Inpatient surgery clinic at a medical center. Thirty-six patients undergoing abdominal surgery with low-flow anesthesia. Fresh gas flow was given at a starting rate of 5 L/min for 6 minutes. Thereafter, the fresh gas flow setting was nitrous oxide (N2O) 1 L/min and oxygen (O2) 0.6 L/min (Group 1), N2O 0.5 L/min and O2 0.5 L/min (Group 2), and with a moderate surplus of N2O and O2 with respect to the patient's O2 consumption (Group 3). The inspired O2 concentration (FIO2) was measured using a paramagnetic technique, and N2O levels were measured with infrared sensors; the inspired nitrogen concentration (FIN2) was calculated by the following formula: FIN2 = 1-FIO2-FIN2O, where FIN2O is the inspired N2O concentration. After 1 hour of anesthesia, FIO2 was significantly lower in Group 1 than in Groups 2 and 3 (p < 0.01), and FIN2 was significantly higher in Groups 2 and 3 than in Group 1 (p < 0.01). After 2 hours of anesthesia, FIN2 returned to normal in Group 2 but continued to increase in Group 3. FIN2O was close to 0.7% only in Group 1. The same initial period of denitrogenation is not adequate to denitrogenate the circle system in all cases. The lower the fresh gas flow, the longer the initial period of denitrogenation should be. Various levels of fresh gas flow for low-flow anesthesia have been suggested, but none guarantees adequate control of inspired gas composition unless flowmeters are continuously adjusted.


Inguinal herniorrhaphy in young infants: perianesthetic complications and associated preanesthetic risk factors. (1) To prospectively observe and tabulate all perianesthetic complications in young infants undergoing herniorrhaphy with general anesthesia and (2) to identify all major postnatal complications and determine which, if any, might be significant risk factors for perianesthetic complications. Prospective case control study. Columbus, Ohio, Children's Hospital, a teaching and tertiary referral center. One hundred two consecutive infants 60 weeks postconceptual age (PCA) or younger undergoing herniorrhaphy with general anesthesia. None All perianesthetic complications occurring during anesthesia, in the postanesthesia care unit (PACU), during the remaining hospital stay, and within 30 days of anesthesia were recorded, and a detailed postnatal history was compiled. Fifty-five percent of 60 preterm infants [37 weeks gestational age (GA) or younger] and 50% of 42 term infants (older than 37 weeks GA) experienced at least one perianesthetic complication. Following discharge from the PACU, in-house complications were confined to the preterm group. Significant risk factors included a history of apnea, bradycardia, and ventilatory support for at least 24 hours after birth, mainly for respiratory distress syndrome. In a teaching hospital, prospectively observed perianesthetic complications can occur in more than 50% of infants 60 weeks PCA or younger undergoing herniorrhaphy with inhalation anesthesia. Infants younger than 49 weeks PCA with a significant preanesthetic risk factor should be monitored overnight for apnea and bradycardia.


Performance evaluation of a draw-over vaporizer with a nonbreathing circuit during simulated adverse conditions. Performance evaluation of a draw-over vaporizer with a nonrebreathing circuit during simulated adverse conditions. Open laboratory study. University hospital laboratory. The output (%) of a temperature-compensated Ohmeda Cyprane PAC (portable anesthesia complete) draw-over vaporizer (Ohmeda, Madison, WI) using isoflurane attached to a nonrebreathing circuit was tested in the laboratory during manual ventilation under normal and simulated adverse conditions. The adverse conditions tested were high ambient temperature and static and dynamic positional variation. The results show that the output bias of the PAC vaporizer has a generally consistent profile as tested, except when placed in the 180 degrees full-inversion or 90 degrees anterior-tilt position. We also conducted trials with the Penlon ventilator (Ohmeda, Abingdon, Oxford, U.K.) attached to the circuit at each of its input ports. Ventilation at one input port produced dangerously high airway pressures (within the circuit), thus supporting the manufacturer's recommendation against the use of this port for positive-pressure ventilation. Using the recommended port, the test lung was seen to inflate and deflate appropriately, but, surprisingly, no vapor output was detected by the agent monitor at any vaporizer setting when using the ventilator at the recommended port. Anesthetists should be aware of the pitfalls and possible problems that may be associated with this type of anesthetic delivery system.


Desflurane potentiates atracurium in humans: a comparative study with isoflurane. (1) To evaluate the neuromuscular effects of desflurane and its interactions with atracurium and (2) to compare desflurane and isoflurane in these effects. Sequential entry of informed and consenting patients randomly assigned to receive desflurane (n = 25) or isoflurane (n = 25). Operating suite of a county-university medical center. Fifty adults, ASA physical status I, undergoing elective orthopedic surgery. Following establishment of steady desflurane or isoflurane anesthesia, at 1.25 minimum alveolar concentration (MAC) exhaled for 15 minutes, a randomly predetermined dose of atracurium (0.05, 0.1, or 0.15 mg/kg) was injected intravenously (IV). At the end of surgery, neostigmine 0.04 mg/kg IV was given to reverse the residual block. The neuromuscular effects of desflurane or isoflurane alone, and the dose-response relationship, time course, and reversibility of the neuromuscular effects of atracurium with either anesthetic, were examined in detail and compared using electromyographic quantification of the response of the first dorsal interosseous muscle to train-of-four (TOF) stimulation of the ulnar nerve. TOF fade and depression of the first response (T1) of the TOF were measured in response to desflurane or isoflurane, atracurium, and neostigmine. Desflurane caused more TOF fade than isoflurane prior to atracurium administration. The TOF ratios were 0.91 +/- 0.02 and 0.98 +/- 0.01, respectively (p < 0.05). For other measured neuromuscular parameters, atracurium-induced depression tended to be greater in the presence of desflurane than in the presence of isoflurane, but none of the measured differences reached the statistical significance level of p < 0.05. The ED50, ED95, and 25-75% recovery index of atracurium were 0.038 mg/kg (95% confidence level; range 0.030 to 0.047 mg/kg), 0.11 mg/kg (0.095 to 0.14 mg/kg), and 31 +/- 4 minutes (means +/- SEM) with desflurane anesthesia, versus 0.043 mg/kg (0.035 to 0.052 mg/kg), 0.13 mg/kg (0.11 to 0.16 mg/kg), and 23 +/- 4 minutes with isoflurane anesthesia (p = 0.1-0.2). Continuation of either anesthetic at 1.25 MAC prevented complete recovery of neuromuscular functions spontaneously or following neostigmine 0.04 mg/kg. In ASA physical status I adults, 9% desflurane has neuromuscular effects equal to or slightly in excess of those of 1.6% isoflurane.


Pharmacodynamic effects of three doses of ORG 9426 used for endotracheal intubation in humans. To determine the pharmacodynamic characteristics of three incremental doses of ORG 9426 used for endotracheal intubation in patients. Double-blind, randomized administration of one of three doses of intravenous ORG 9426. Inpatients requiring surgery at Georgetown University Medical Center. Thirty-six patients, ages 18 to 65, ASA physical status I, II, and III, scheduled for general surgery. After Georgetown University Institutional Review Board approval and patient consent, patients were premedicated with midazolam or droperidol. Anesthesia was induced with thiopental sodium and fentanyl. Anesthesia was maintained with 60% nitrous oxide in oxygen. The ulnar nerve was stimulated supramaximally with a 2 Hz train-of-four (TOF) every 20 seconds. Thumb contractions were measured with a force transducer. When TOF and anesthesia were stable, 2, 2.5, or 3 times the ED95 of ORG 9426 (570 micrograms/kg, 710 micrograms/kg, or 850 micrograms/kg) was administered randomly. Tracheal intubation was attempted at maximal depression of the first TOF response (T1). The following parameters were measured: time interval from the injection of ORG 9426 to 90% depression of T1 (T1 90% block), maximal T1 depression (onset time), intubating conditions, clinical duration (time for return of T1 to 25% of control), heart rate (HR), blood pressure (BP), and any adverse clinical experience. ORG 9426 provided adequate intubating conditions in all patients but two, independent of the dose used. Its onset time was rapid, but increasing the dose did not shorten the onset. T1 90% block was achieved rapidly (75 +/- 25 seconds to 78 +/- 18 seconds, means +/- SD). The clinical duration of ORG 9426 was relatively short and lengthened with increasing doses (from 36 +/- 18 minutes at 570 micrograms/kg to 42 +/- 10 minutes at 850 micrograms/kg. Spontaneous twitch recovery from 10% to 25% was similar in all dosage groups (5 +/- 1 minutes to 6 +/- 4 minutes). No clinically significant changes in HR and BP and no adverse clinical experiences were noted in any group. These findings warrant further clinical evaluation of ORG 9426 for induction and maintenance of muscle relaxation in humans.


The effect of respiratory alkalosis on oxygen consumption in anesthetized patients. To investigate whether hyperventilation significantly altered oxygen consumption in anesthetized and paralyzed patients undergoing surgery. Open crossover trial with 1 hour of hyperventilation preceded and followed by 1 hour of normoventilation. University medical center. Eight patients (five men and three women) undergoing lengthy orthopedic surgery with general anesthesia and muscle paralysis. After baseline normoventilation for 1 hour (Period 1), the anesthetized patients were hyperventilated to an arterial carbon dioxide tension (PaCO2) of 20 to 25 mmHg for 1 hour (Period 2). Patients then experienced another hour of normoventilation (Period 3). Hemodynamic variables, electrocardiography, temperature, end-tidal partial pressure of CO2 (PETCO2), oxygen consumption (VO2), carbon dioxide production, and minute ventilation were continuously followed throughout the study, and arterial blood gases were drawn at the beginning and end of each study period. During the period of hyperventilation, pH was significantly higher and P.ETCO2 and PaCO2 significantly lower compared with the periods of normoventilation. VO2 was significantly increased during hyperventilation compared with the periods of normoventilation. Hemodynamic variables and temperature were similar in the three study periods. In anesthetized paralyzed patients, there is an increase in whole-body VO2 with hypocapnic alkalosis.


Ketorolac or fentanyl to supplement local anesthesia? To evaluate the usefulness of ketorolac in the treatment of intraoperative pain refractory to the administration of local anesthetic alone. Intraoperative acute-pain treatment model consisting of awake, nonsedated patients who randomly received either an opioid or a study drug in a double-blind fashion. University medical center. Eighty patients who underwent breast biopsy, lumpectomy, or central venous catheter placement. Patients received either ketorolac 1 mg/kg intravenously (IV) up to a total dose of 60 mg or fentanyl 3 micrograms/kg IV up to a total dose of 250 micrograms to supplement the local anesthetic. Verbal pain evaluation and the visual analog scale (VAS) were used for perioperative measurement of pain. Heart rate (HR), blood pressure, and respiratory rate (RR) were recorded before and after analgesic drug injections at 10-minute intervals, both intraoperatively and while the patient was in the postanesthesia care unit (PACU). Speed of recovery was quantified by p-deletion and digit substitution tests on admission to the PACU and at 30-minute intervals until discharge. The frequency of nausea, vomiting, and pruritus were recorded. There were no differences between the groups in perioperative verbal pain evaluation, VAS scores, HR, systolic blood pressure, diastolic blood pressure, or RR. Patients who received ketorolac exhibited a significantly lower frequency of intraoperative and postoperative medication administration intraoperatively, than those who received fentanyl. No additional pain medication was required by patients in the PACU in either group. Ketorolac is a useful alternative to fentanyl for the treatment of intraoperative pain refractory to the administration of local anesthetic alone during monitored anesthesia care. A decided advantage of ketorolac over fentanyl is the absence of nausea and vomiting in the intraoperative and postoperative periods.


Anesthetic considerations of an infant with Beckwith-Wiedemann syndrome. The case of a 3-day-old infant with Beckwith-Wiedemann syndrome who required anesthetic care during closure of an abdominal wall defect is presented. Beckwith-Wiedemann syndrome comprises a constellation of clinical features, including macroglossia, macrosomia, omphalocele, visceromegaly, mild microcephaly, facial nevus flammeus, horizontal earlobe creases, and renal medullary dysplasia. Due to the high rate of omphalocele in this syndrome, anesthetic care is frequently required during the neonatal period. Many of these infants (greater than 50%) are born prematurely. Therefore, their anesthetic care may be further complicated by associated diseases of prematurity, such as hyaline membrane disease. Additional anesthetic implications of this syndrome relate to the occurrence and management of hypoglycemia and polycythemia. Careful intraoperative management of glucose homeostasis is particularly important, since eventual neurologic outcome and intelligence will be normal provided prolonged neonatal hypoglycemia is avoided. Preoperative evaluation of the cardiac and genitourinary system, including echocardiography and renal ultrasound, are recommended because of the frequent occurrence of associated anomalies with omphalocele.


Patient-controlled drug administration during local anesthesia: a comparison of midazolam, propofol, and alfentanil. To evaluate the perioperative effects of alfentanil, midazolam, and propofol when administered using a patient-controlled analgesia (PCA) device during local anesthesia. Randomized, single-blind comparative study. Outpatient surgery center at a university teaching hospital. Ninety outpatients undergoing minor elective surgical procedures with local anesthetic infiltration were assigned to one of three treatment groups. After premedication with midazolam 1 mg intravenously (IV) and fentanyl 50 micrograms IV, patients were allowed to self-administer 2 ml bolus doses of either alfentanil 250 micrograms/ml, midazolam 0.4 mg/ml, or propofol 10 mg/ml at minimal intervals of 3 minutes to supplement a basal infusion rate of 5 ml/hr. The total intraoperative dosages of alfentanil, midazolam, and propofol were 2.7 +/- 1.1 mg, 4.7 +/- 2.7 mg, and 114 +/- 42 mg, respectively, for procedures lasting 48 +/- 28 minutes to 51 +/- 19 minutes (means +/- SD). Propofol produced more pain on injection (39% vs. 4% and 6% in the alfentanil and midazolam groups, respectively). Episodes of arterial oxygen saturation less than 90% were more frequent with alfentanil (28%) than with midazolam (3%) or propofol (13%). Using the visual analog scale, patients reported comparable levels of discomfort, anxiety, and sedation during the operation in all three treatment groups. Postoperative picture recall was significantly decreased with midazolam versus alfentanil and propofol. Finally, postoperative nausea was reported more frequently in the alfentanil group (29%) than in the midazolam (10%) or propofol (18%) groups, contributing to a significant prolongation of the discharge time in the alfentanil-treated patients. When self-administered as adjuvants during local anesthesia using a PCA delivery system, alfentanil, midazolam, and propofol were equally acceptable to patients. However, propofol and midazolam were associated with fewer perioperative complications than was alfentanil.


Intravenous nitroglycerin for uterine inversion. Uterine inversion following vaginal delivery or during Cesarean section is rare. Cardiovascular instability resulting from blood loss is possible. This article describes the use of intravenous nitroglycerin as an alternative to the induction of general anesthesia and administration of volatile anesthetics to provide uterine relaxation.


Leukemia inhibitory factor--a puzzling polyfunctional regulator. LIF seems likely to have important functions in the early developing embryo and in adult life can influence platelet formation, osteoblast and neuronal function, calcium and lipid metabolism and the production of acute-phase proteins. LIF appears usually to be produced and to function locally in various tissues, an arrangement that would minimize unwanted actions of this polyfunctional regulator. Nevertheless it remains puzzling what purpose is achieved by use of a regulator with potent actions on such a wide range of apparently unrelated tissues.


To define a specialty: a brief history of the American Board of Anesthesiology's first written examination. The initial written examination of the American Board of Anesthesiology, a division of the American Board of Surgery, was given on March 28, 1939. For all anesthesiologists, this date has double significance. First, what was meant by anesthesiology as a medical specialty was defined through the questions posed on the first examination. Second, the physicians being tested that day were among the first physician-anesthetists to exploit the newly created path to recognition as specialists in the science and art of anesthesia by the American medical hierarchy. Gaining the support of organized medicine was an involved and arduous struggle that consumed most of the 1930s. A triumvirate of visionaries, Paul Wood, John Lundy, and Ralph Waters, was necessary to crystalize the goal of specialty recognition of physician-anesthetists. The first written examination was the consummation of this dream of equal status for anesthesia. The examination would not become repetitious, and within the first decade of testing, the style would change from an essay format to multiple-choice questions similar to the current form.


Regulation of amphiregulin mRNA by TGF-beta in the human lung adenocarcinoma cell line A549. Transforming growth factor beta is a strong growth inhibitor for many types of normal and transformed cells, although little is known on the mechanism of this anti-proliferative effect. The human lung adenocarcinoma cell line A549 is growth arrested by TGF-beta 1 and serves as a model for studying this effect. We describe that, concurrent with the inhibition of A549 cell growth, TGF-beta 1 treatment causes a dramatic reduction in the level of expression of the amphiregulin (AR) gene, a recently identified member of the EGF/TGF alpha family. Similar results were also observed with TGF-beta 2. Peak inhibition occurred at 24 hr of treatment and was reversible upon removal of TGF-beta 1. The level of AR protein secreted by A549 cells was also decreased by TGF-beta 1. In contrast, TGF-alpha mRNA was not detected in these cells regardless of TGF-beta 1 treatment. Another TGF-beta inhibited cell line, PC-3 (human prostatic adenocarcinoma) also exhibited a decrease in AR message levels following exposure to TGF-beta 1. The growth inhibitory effects of TGF-beta may in part be mediated by modulation of AR expression.


Cell cycle-dependent localization of immunoreactive basic fibroblast growth factor to cytoplasm and nucleus of isolated ovine fetal growth plate chondrocytes. Basic fibroblast growth factor (basic FGF) is a potent mitogen for chondrocytes in vitro and is present in developing cartilage in vivo. Studies of intracellular basic FGF localization in other cell types have revealed a transient nuclear presence. We have examined ovine fetal growth plate chondrocytes for the presence of intracellular basic FGF by immunocytochemistry. Chondrocytes were isolated from the proximal tibial growth plate of lamb fetuses between 75 and 80 days' gestation using collagenase, and were cultured in monolayer before use between passages 3 and 6. In non-synchronized cell cultures 58 +/- 6% of cells (mean +/- s.d., n = 3) demonstrated cytoplasmic staining for immunoreactive basic FGF. Of these cells, 18 +/- 3% also exhibited strong nuclear staining. Chondrocytes were growth-restricted and restarted into the cell cycle with 2% (vol/vol) fetal calf serum. The timing of S phase was followed by nuclear labelling of nuclei with [3H] thymidine followed by autoradiography, or by the incorporation of [3H] thymidine into trichloroacetic acid-precipitable DNA in parallel cultures. A cytoplasmic presence of immunoreactive basic FGF did not appear, following immunocytochemistry, until the second half of G1 with 97% of cells immunopositive 2 hr prior to S phase. Nuclear staining for basic FGF appeared 2 hr before peak nuclear labelling index, and 56% of cell nuclei were immunopositive. Following entry into S phase cytoplasmic and nuclear basic FGF immunostaining rapidly disappeared. When these experiments were repeated with or without the presence of anti-basic FGF antibody or heparin, the presence of the antibody significantly reduced peak [3H] thymidine incorporation into DNA during S phase while exposure to heparin increased this. However, the proportion of cells demonstrating cytoplasmic or nuclear staining for immunoreactive basic FGF, and the time of onset of staining, were unaltered. Incubation of cells with suramin blocked subsequent DNA synthesis and no intracellular basic FGF was visualized. Cell-conditioned culture medium, extracellular matrix and cytoplasm from synchronized cultures of chondrocytes were taken at time points throughout the cell cycle and assessed for basic FGF content by radioimmunoassay. Basic FGF was detectable in each compartment and steadily rose throughout the second half of G1 to reach maximum values around the S phase. The accumulation of basic FGF in medium, matrix and cytoplasm was blocked by the presence of cycloheximide.(ABSTRACT TRUNCATED AT 400 WORDS)


Identification of bone morphogenetic protein-2 in early Xenopus laevis embryos. Polyclonal antibodies capable of reacting with amphibian bone morphogenetic protein (BMP-2 and -4) were raised in rabbits by immunization with a synthetic 21 amino acid peptide which corresponds to a sequence residing in the mature protein of Xenopus BMP-2 (xBMP-2). The antibodies recognized an embryonic BMP as well as mammalian and bacteria expressed recombinant xBMPs. The antibodies detected, under reducing conditions, a 30 kDa protein in the extract of oocytes and embryos during early development. Interestingly, acidification of the extract from each developmental stage yielded a protein band of smaller molecular weight of 18 kDa, which is similar in size to reduced form of mature BMPs purified from mammalian species. Two-dimensional electrophoresis employed to examine the molecular weight of unreduced forms using the antibody, revealed that both molecular forms are monomeric in the embryos. The result suggests that at least BMP-2 mRNA previously detected in early embryos, is translated into peptide but the dimerization may be incomplete or strictly limited in these embryos.