Variability in the anterior extralaryngeal branch of the recurrent laryngeal nerve: clinical implications.

BACKGROUND: This study aimed to identify the anterior and posterior extralaryngeal branches (AELB, PELB) of the recurrent laryngeal nerve (RLN), measure these branches when present, and determine relationships between gender, sidedness and neck length. MATERIALS AND METHODS: Dissection was completed to level of the thyroid on 45 cadavers. The course of the RLN was then traced superiorly from its entry into the neck. Careful reflection of the thyroid and dissection of the lateral thyroid ligament permitted visualisation of the full course of the nerve. If extralaryngeal branching (ELB) was present, measurements were taken from the point of bifurcation of the RLN to the point of laryngeal entry through the cricothyroid membrane. Neck measurements, from the spinous process of C7 to the superior nuchal line, were taken. Gender of the specimen was noted. Data was analysed in SPSS. RESULTS: Extralaryngeal branching was found in 77.78% of our sample, 77.14% on the left and 54.29% on the right. A significant difference was found between AELB length on the left and right, indicating that the left branch will be longer than the right when present. A significant difference in neck length between those with and without ELB was also found, indicating that people with longer necks more often display ELB. Neither neck length and AELB length, nor gender and AELB length were strongly correlated in this sample. CONCLUSIONS: Extralaryngeal branching can occur in all populations, but there are definite trends in its incidence and length. Surgeons should be aware of these trends before operating on patients.

Safety considerations for sample analysis using a near-infrared (785 nm) Raman laser source.

Raman spectroscopy is often considered a nondestructive analytical technique; however, this is not always the case. The 300-mW 785-nm near-infrared (NIR) laser source used with many commercially available instruments has sufficient power to burn samples. This destructive potential is of special concern if the sample is irreplaceable (e.g., fine art, forensic evidence, or for in vivo medical diagnostics) or a hazardous energetic material (explosive or pyrophoric samples). This study quantifies the heat resulting from illuminating an extensive color array with a 785-nm NIR laser and relates these values to the hazards associated with Raman analysis. In general, darker colors were found to be more problematic. Since visible colors are not ideally correlated with absorptive characteristics at 785 nm, predictions based on thermography are not perfect; however, this approximation gives a useful method for predicting the thermal response of unknown samples to NIR exposure. Additionally, experimental studies evaluated the analysis of flammable organic solvents, propellants, military explosives, mixtures containing military explosives, shock-sensitive explosives, and gunpowders (i.e., smokeless, black, and Pyrodex powders). Safety guidelines for analysis are presented.

Selective stationary phase for solid-phase microextraction analysis of sarin (GB).

A number of critical field applications require monitoring air samples for trace levels of chemical warfare agents. Solid-phase microextraction (SPME) is a convenient format to conduct these analyses. Measurements could be significantly improved if a SPME phase selective for nerve agents were substituted for non-selective polymers typically used (e.g., polydimethylsiloxane). This paper evaluates a novel stationary phase, previously developed for methylphosphonate sensor applications, for use with SPME sampling. The phenol-based polymer, BSP3, was found to offer far higher selectivity toward sarin (GB) than polydimethylsiloxane due to a pronounced affinity toward the target analyte and a lower affinity toward hydrocarbons.

Blind field test evaluation of Raman spectroscopy as a forensic tool.

Analytical instrumentation for Raman spectroscopy has advanced rapidly in recent years to the point where commercial field-portable instruments are available. Raman analysis with portable instrumentation is a new capability that can provide emergency response teams with on-site evaluation of hazardous materials. Before Raman analysis is accepted and implemented in the field, realistic studies applied to unknown samples need to be performed to define the reliability of this technique. Studies described herein provide a rigorous blind field test that utilizes two instruments and two operators to analyze a matrix that consists of 58 unknown samples. Samples were searched against a custom hazardous materials reference library (Hazardous Material Response Unit (HMRU) Spectral Library Database). Experimental design included a number of intentionally difficult situations including binary solvent mixtures and a variety of compounds that yield medium-quality spectra that were not contained in the HMRU library. Results showed that over 97% of the samples were correctly identified with no occurrences of false positive identifications (compounds that were not in the library were never identified as library constituents). Statistical analysis indicated equivalent performance for both the operators and instruments. These results indicate a high level of performance that should extrapolate to actual field situations. Implementation of Raman techniques to emergency field situations should proceed with a corresponding level of confidence.

Identification of carbohydrate-binding domains in the attachment proteins of type 1 and type 3 reoviruses.

The reovirus attachment protein, sigma1, is responsible for strain-specific patterns of viral tropism in the murine central nervous system and receptor binding on cultured cells. The sigma1 protein consists of a fibrous tail domain proximal to the virion surface and a virion-distal globular head domain. To better understand mechanisms of reovirus attachment to cells, we conducted studies to identify the region of sigma1 that binds cell surface carbohydrate. Chimeric and truncated sigma1 proteins derived from prototype reovirus strains type 1 Lang (T1L) and type 3 Dearing (T3D) were expressed in insect cells by using a baculovirus vector. Assessment of expressed protein susceptibility to proteolytic cleavage, binding to anti-sigma1 antibodies, and oligomerization indicates that the chimeric and truncated sigma1 proteins are properly folded. To assess carbohydrate binding, recombinant sigma1 proteins were tested for the capacity to agglutinate mammalian erythrocytes and to bind sialic acid presented on glycophorin, the cell surface molecule bound by type 3 reovirus on human erythrocytes. Using a panel of two wild-type and ten chimeric and truncated sigma1 proteins, the sialic acid-binding domain of type 3 sigma1 was mapped to a region of sequence proposed to form the more amino terminal of two predicted beta-sheet structures in the tail. This unit corresponds to morphologic region T(iii) observed in computer-processed electron micrographs of sigma1 protein purified from virions. In contrast, the homologous region of T1L sigma1 sequence was not implicated in carbohydrate binding; rather, sequences in the distal portion of the tail known as the neck were required. Results of these studies demonstrate that a functional receptor-binding domain, which uses sialic acid as its ligand, is contained within morphologic region T(iii) of the type 3 sigma1 tail. Furthermore, our findings indicate that T1L and T3D sigma1 proteins contain different arrangements of receptor-binding domains.

Sample introduction and separation in capillary electrophoresis, and combination with mass spectrometric detection.

Capillary-electrophoresis methods are attracting interest owing to the ability to yield rapid high-resolution separations, but many aspects, such as sample injection, separation conditions and detection, need further development. Effects related to sample injection and buffer composition have been investigated. Automated methods for electromigration injection of nl-size sample volumes are shown to give a precision of approximately +/-1%. Problems encountered with manual injection procedures have been examined by an electric field reversal technique. The effect of buffer pH on capillary zone-electrophoresis (CZE) separations can be attributed to changes in electro-osmotic flow velocities and to changes in the isoelectric points of analytes. The interfacing of capillary electrophoresis with mass spectrometry is described and demonstrated for a range of conditions, with a quaternary phosphonium salt mixture. Separations obtained by CZE and capillary isotachophoresis are compared and the relative advantages of the two techniques discussed.

Rapid and high resolution capillary supercritical fluid chromatography (SFC) and SFC/MS of trichothecene mycotoxins.

The potential application of capillary column supercritical fluid chromatography (SFC) and SFC/mass spectrometry (SFC/MS) for the separation and analysis of mycotoxins of the trichothecene group was examined. Trichothecenes present significant analytical problems for both gas and liquid chromatography with a major difficulty for the latter being the lack of sufficiently sensitive and selective detectors. Supercritical carbon dioxide mobile phases at temperatures up to 100 degrees C were used with deactivated fused silica columns coated with crosslinked stationary phases. Separations were obtained under pressure ramped conditions using long (15 m) 50-micron i.d. columns for several trichothecenes (diacetoxyscirpenol, deoxynivalenol, and T-2 toxin) and related higher molecular weight macrocyclic (roridin and verrucarin) trichothecenes. In addition, new rapid pressure programming techniques with short (less than 2m) 25- to 50-micron i.d. capillary columns were used to obtain fast separations in as little as 1 min. SFC/MS with ammonia chemical ionization provided high selectivity and sensitive detection (with approximately 1-pg detection limits) for trichothecene mixtures. The extension to complex sample matrices is discussed and the application of selective MS/MS detection is demonstrated.

Three rat monoclonal antibodies to human C3.

Three monoclonal antibodies to human C3 have been obtained from a fusion of the rat myeloma line Y3 Ag 1.2.3. with spleen cells from rats immunized against C3. One, from clone 4, reacts with an antigenic determinant in C3c showing the expected reactivity of the 'C' antigen of C3. The specificity of the other two monoclonal antibodies correspond less clearly with known C3 antigens. By agglutination analysis of complement coated cells the determinant reacting with clone 3 is present in C3d while that for clone 9 appears as a neoantigen on C3bi. In both cases the co-precipitation results are anomalous and more direct studies are needed to define the exact specificity. The possibility that internal sequence duplications in C3 may explain some anomalies is discussed. None of the monoclonal antibodies significantly inhibit C3 functions. The monoclonal antibodies have been found to have unusual properties in co-precipitation assays being able to diffuse through a precipitation line with which they react to react with a further line. One antibody is also able to react strongly with the anodal half of what appears as a single line with a polyclonal antiserum.

Health care planning: acts, agencies, and activities of the federal government.

Healthh care planning and provision are societal responsibilities supported by the federal government since 1798. To maximize benefits of money spent on health care during the past two decades, the federal government created in succession Regional Medical Programs, Medicare and Medicaid, area-wide Comprehensive Health Planning Agencies, and Professional Standards Review Organizations, and enacted the National Health Planning and Resources Development Act of 1974. Health care planning consists of three phases: assessment, planning, and implementation. Intrinsic to these activities are decisions concerning the extent to which society is willing to devote its resources, the types of health care to be given, and to whom services will be rendered.

Lipoxygenic micromethod for specific determination of lipase activity in serum and duodenal fluid.

We propose a rapid enzymatic micromethod for the specific determination of lipase (EC 3.1.1.3) activity in serum and duodenal fluid. Free linoleic acid produced during 10-min incubation of 10 mul of sample with 1 ml of substrate (trillinolein emulsion) at 30 degrees C is converted by lipoxygenase (EC 1.99.2.1), in a coupled reaction, to its hydroperoxide, which is measured photometrically after solubilizing the reaction mixture in ethanol. Lipase activity is calculated from the rate of hydroperoxide formation, with linoleic acid as primary standard. The velocity of the reaction is greatest at pH 8.8, 35-37 degrees C, and a deoxycholate concentration of 3.6 mmol/liter. The energy of activation is 6.7 kcal/mol. The differing "apparent" Km values obtained for lipase in undiluted serum (4 X 10(-5) mol/liter) and in albumin-based diluents (1 X 10(-5) mol/liter) indicate the presence of a competitive inhibitor in the serum matrix. We detected no lipase activity in urine. Results by the proposed method correlate well with those by a copper soap extraction method (r = 0.95), but values are significantly higher for pancreatitis patients' sera (slope 1.6). The linear dynamic range extends to 1000 U/liter. Hemolysis, lipemia, and hyperbilirubinemia do not interfere. The normal range is 40-60 U/liter. Lipase activity of pancreatitis patients generally exceed 1000 U/liter during the acute phase and 250 U/liter for as long as 10 days after it.

Measurements of starch breakdown as estimates of glycolysis during thermogenesis by the spadix of Arum maculatum L.

The rates of starch breakdown at thermogenesis by clubs of the spadices of Arum maculatum L. were measured in individual clubs still attached to the plants. The values obtained were used as estimates of the rate of glycolysis at thermogenesis. Such rates were shown to exceed the maximum catalytic activities of phosphofructokinase (E.C. 2.7.1.11.), aldolase (E.C. 4.1.2.7.), and glyceraldehydephosphate dehydrogenase (E.C. 1.2.1.12.) in developing clubs. The marked increases in the activities of the above enzymes that occur during the development of the club have been shown to be a prerequisite for the attainment of the high rate of glycolysis found at thermogenesis, and thus to be an example of coarse control of glycolysis.

Pathways of carbohydrate oxidation during thermogenesis by the spadix of Arum maculatum.

1. The aims of this work were to discover the pathways of carbohydrate oxidation prior to and during thermogenesis by the club of the spadix of Arum maculatum, and whether there was coarse control of these pathways. 2. 14C02 production from [1-14C]-, [3,4-14C]-, and [6-14C]glucose, the detailed distribution of 14C from [1-14C]- and [6-14C]glucose, and the maximum catalytic activities of phosphofructokinase, fructose-1,6-diphosphate aldolase, glucose-6-phosphate dehydrogenase, and phosphogluconate dehydrogenase were determined at different stages in the development of the spadix. The results indicate that in the early stages carbohydrate is oxidized via both the pentose phosphate pathway and glycolysis, and that a shift to glycolysis occurs during development so that just before and during thermogenesis glycolysis predominates almost exclusively. 3. During development the activities of phosphofructokinase and glucose-6-phosphate dehydrogenase per club increased 100- ans during spadix development, and indicated that the onset of rapid glycolysis at thermogenesis is regulated by fine control or availability of substrate.

New, simple maltogenic assay for mechanized determination of alpha-amylase activity in serum and urine.

We report a new method for the mechanized determination of serum and urinary alpha-amylase by use of a continuous-flow system, based on the measurement of maltose formed by incubating the sample with amylodextrin at pH 7 and 40 degrees C. After dialysis, maltose is converted enzymatically to glucose, which is measured by Trinder's glucose oxidase-peroxidase method [J. Clin. Pathol. 22, 246 (1969)]. The reaction is linear for amylase activities up to 1400 Somogyi units/dl (2560 U/liter) and for maltose concentrations through 1500 mg/dl. No blank assay is required; consequently precision is improved and the automated system is simplified. Calibration with primary maltose standards increases accuracy and reliability. Common reducing substances in serum and urine do not interfere at their normal concentrations. There is a linear correlation between the results of this method and those of chromogenic and iodometric methods for normal and pathologic sera and urines. The chromogenic method yields significantly higher results and the iodometric method significantly lower results than this maltogenic method for elevated amylase activities. The normal range is 40-140 Somogyi units/dl (73-256 U/liter).