Is conservative organ-sparing treatment of penile carcinoma justified?

PURPOSE: The results of different conservative organ-sparing methods - radiotherapy, chemotherapy and radiochemotherapy - in the treatment of penile carcinoma were studied. MATERIALS AND METHODS: Conservatively treated 223 patients with penile carcinoma seen in three hospitals between 1959 and 1996 years were studied retrospectively. Among them 155 received radiotherapy, 33 chemotherapy, and 35 radiochemotherapy. RESULTS: The local control was achieved in 135 (60.5%) of 223 patients. The difference in local control rate among these three groups of patients has not reached statistical significance. The efficacy of conservative treatment was highly associated with three factors: tumor size, grade and patient's age. Overall, 24 of 135 patients (17.7%) had local recurrence. Ten patients (4.5%) developed regional metastases. The recurrence rate did not correlate with tumor size and grade and was similar for all treatment modalities. Long-term results of each method were approximatley equal and 5-year survival varied from 78 to 88%. CONCLUSION: Conservative organ-sparing treatment of early-stage penile carcinomas is justified. Failure should be corrected by surgery without compromising survival.

A liquid-stable reagent for lactic acid levels. Application to the Hitachi 911 and Beckman CX7.

We evaluated the use of a new lactate oxidase-based reagent for the determination of serum and plasma lactic acid levels with the Hitachi 911 (Roche Diagnostics, Indianapolis, IN) and the Beckman CX7 (Beckman Instruments, Brea, CA). Evaluation studies demonstrated on-board stability of at least 3 months and a calibration stability of more than 5 months. Within- and between-day imprecision of this reagent was less than 2% for both applications. The reagent is free of the deleterious effects of triglyceride up to levels of 1,400 mg/dL (15.8 mmol/L), bilirubin to concentrations of 24.6 mg/dL (420 mumol/L), and hemoglobin, from lysed erythrocytes, to levels of more than 0.3 g/dL (3.0 g/L). When used on the Hitachi 911 for the determination of plasma lactate concentrations, the reagent correlates with the Dade aca III (Dade International, Deerfield, IL). When applied to the Beckman CX7 for the determination of serum lactate levels, the method correlates with the Beckman method.

Application of a sensitive and specific reagent for the determination of serum iron to the Bayer DAX48.

We describe a modification of a previously described serum iron procedure applied to the Bayer DAX48 (Bayer Diagnostics, Tarrytown, NY) automated chemistry analyzer. The iron-ligand used in this assay, 2-(5-nitro-2-pyridylazo)-5-(N-propyl-N-sulfopropylamine) phenol (nitro-PAPS), has a molar absorptivity of 94,000 L mol(-1) cm(-1), which is three to four times more sensitive than the more commonly used ligands. The increased sensitivity of the iron-ligand complex facilitates modification of a Ferene S method that requires a smaller sample volume while it maintains the precision of the assay. Because the reagent does not contain ascorbate, the "onboard" stability has been increased to more than 4 weeks. The reagent seems to be quite insensitive to icterus and hemolysis. Furthermore, the interference of turbidity caused by triglycerides, abnormal proteins, or fibrinogen, present in samples from patients undergoing anticoagulant therapy, seems to have been eliminated.

Calcium determination in serum with stable alkaline Arsenazo III and triglyceride clearing.

We describe an analytical procedure for determining serum calcium, using the ligand Arsenazo III in an aqueous alkaline medium. The choice of pH for the proposed technique differs from current procedures, which are for the most part carried out in a slightly acidic medium. An acidic medium avoids interference from magnesium, but is spectrophotometrically suboptimal for this pH-dependent reaction: the molar absorptivity of the Arsenazo III complex with calcium at acid pH is 13,787 L mol-1 cm-1, about one-half of that at a more optimal alkaline pH (26,574 L mol-1 cm-1). We have included a clearing technique in the reagent to avoid spectral aberrations from hypertriglyceridemic samples. alpha-Cyclodextrin absorbs the nonesterified fatty acids liberated from triglycerides by a microbial lipase. This modification may also be helpful for binding nonesterified fatty acids, which are known to interfere with calcium procedures by forming calcium soaps and thus preventing the reaction with intended ligands. The use of 8-hydroxyquinoline sulfonate as the magnesium-masking agent facilitates the use of alkaline pH. The less-water-soluble alternative, 8-hydroxyquinoline, commonly used as a precipitating agent for several methods, is difficult to solubilize in the alkaline reagent and tends to precipitate when complexed to magnesium. Finally, the use of alkaline pH results in a prolonged (> or = 6 weeks) shelf life for the reagent.

Fluorometric determination of phosphatidylcholine as a measure of phospholipid methylation.

The successive methylation of phosphatidylethanolamine to phosphatidylcholine (phospholipid methylation) has been measured by the incorporation of S-[methyl-3H]adenosylmethionine or colorimetric assay of phosphatidylcholine extracted from adipocyte plasma membranes. A fluorometric assay for phosphatidylcholine was developed to measure phospholipid methylation. This assay is 10 times more sensitive than the colorimetric assay and demonstrates no significant interference with other methylated phospholipids. The fluorometric assay was used to determine a biphasic insulin dose response in adipocyte plasma membranes. This fluorometric assay for phosphatidylcholine represents an alternative method for monitoring phospholipid methylation, especially when increased sensitivity is required.

Phosphatidylglycerol in amniotic fluid. Comparison of an "ultrasensitive" immunologic assay with TLC and enzymatic assay.

Phosphatidylglycerol (PG) in amniotic fluid is recognized as a good indicator of fetal lung maturity and is unaffected by moderate amounts of blood or meconium contamination. A rapid immunologic agglutination assay, Ultrasensitive AmnioStat-FLM (FLM), was compared with two-dimensional thin-layer chromatography (TLC) and an enzymic, colorimetric procedure (E-PG). Eighty amniotic fluid specimens were analyzed. FLM results were reported as high (H), intermediate (I), or low positive (L). TLC was compared with FLM:H (n = 27), mean 0.14 (fraction of total phospholipids); I (n = 7), mean 0.11; L (n = 9), mean 0.03; negative results had no detectable PG by TLC. In 33 cases E-PG was compared with FLM:H (n = 9), mean 7.0 mumol/L; I (n = 5), mean 8.1 mumol/L; L (n = 3), mean 3.0 mumol/L; negative (n = 16), mean 3.2 mumol/L. Records were reviewed in 70 cases. Thirty cases were excluded: sample to delivery time was greater than 72 hours; steroids were given or sepsis was documented. Fetal lung immaturity was clinically present in six cases: respiratory distress syndrome in three cases and transient tachypnea of the newborn (TTN) in three cases. One false positive result was identified (TTN, FLM:H). FLM sensitivity for fetal lung maturity was 85.3%, specificity was 83.3%, and the positive predictive value for fetal lung maturity was 96.7%. FLM is a fast, reliable indicator of fetal lung maturity.

A method for the sequential colorimetric determination of serum triglycerides and cholesterol.

A simple spectrophotometric method for the sequential determination of triglycerides and cholesterol from a single serum sample was developed. In this two-stage procedure, the triglycerides and cholesterol esters are first hydrolysed to glycerol and free cholesterol respectively, with simultaneous scavenging of the liberated free fatty acids, a technique that ensures clarity of the sample. The glycerol is subsequently reacted to result in an intense red chromogen with a peak absorption maximum at 510 nm following a series of enzymic reactions. In the second stage, addition of cholesterol oxidase leads to oxidation of free cholesterol generated from the cholesterol esters in the first stage and the free cholesterol normally present in the sample, yielding in a similar fashion the identical red chromogen whose absorbance is also measured at 510 nm. Results obtained with the proposed method demonstrate good correlation with established individual procedures for triglycerides and cholesterol.

Problems with measurements caused by high concentrations of serum solids.

There have been numerous reports of spectrophotometric and volume problems caused by elevated levels of lipids in blood. The offending lipids, primarily triglycerides, not only cause turbidity leading to optical aberrations when added to analytical reagents, but also result in short-sampling errors leading to the measurement of inaccurate volumes of sample. Numerous methods have been developed to clear the lipemia, including ultracentrifugation organic solvent extraction, chemical precipitation and, most recently, enzymic hydrolysis. Although the latter procedures eliminate the optical problems, they do not deal with the volume dilution error created by the triglycerides. In turn, corrective mathematics have been developed to compensate for the inaccurate pipetting caused by the elevated lipids in a sample; however, these empirical calculations are not truly accurate at high concentrations of total lipids. This monograph will describe the problems caused by the presence of elevated lipids and the means available for treating them.

Potential problems in serum protein electrophoresis.

Potential problems are described that one could encounter in carrying out an electrophoretic procedure including its ancillary phases of visualization (staining) and quantification (densitometry). Endpoint-like measurements for separated isoenzymes may provide artifactual kinetic values as well, because stain measurement is fixed at a single time whereas reagent blanking in the electrophoretic medium is substituted for the conventional serum initial absorbance readings of test-tube determinations. Truncation of separated electrophoretic zones or opacity of an electrophoretic anti-convection medium such as uncleared cellulose acetate may also interfere with absolute quantification procedures.

The clinical use of alkaline phosphatase enzymes.

The enzyme alkaline phosphatase is an important serum analyte and its elevation in serum is correlated with the pressure of bone, liver, and other diseases. The analysis of the isoenzymes of alkaline phosphatase is an aid in diagnosing liver and/or bone disease, especially the high molecular weight isoenzymes that appear in cholestatic liver disease.

Enzymic clearing of lipaemic serum following total parenteral nutrition: determination of neonatal bilirubin as a model.

Bilirubin determinations in the newborn infant are one of the many analytical tests that can yield misleading results when the specimen is either iatrogenically or naturally lipaemic. Incorporation of a recently reported enzymic clarification system into a commercially available test kit enables one to conveniently and accurately quantify both total and direct bilirubin within the present procedural characteristics of the Bilirubin Stat Analyser Photometer. This instrument measures the former directly with bichromatic spectrophotometry and the latter with a conventional diazo-type reaction. The proposed modification of existing reagents allows one to apply the assay to samples with as much as, and possibly more than 16 g/l of triglycerides which has been introduced into the vascular circulation as intravenous total parenteral nutrition.

Risk of nosocomial infection with human T-cell lymphotropic virus type III/lymphadenopathy-associated virus in a large cohort of intensively exposed health care workers.

To assess the risk of nosocomial transmission of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV), we prospectively evaluated a cohort of 531 health care workers. One hundred fifty of these employees reported percutaneous or mucous membrane exposures to blood or body fluids from a patient with the acquired immunodeficiency syndrome (AIDS) during the treatment of 238 such patients since 1981. None of these 150 employees had serologic evidence of HTLV-III/LAV infection on follow-up from 6 to 46 months after exposure. Of the 150, 46 were studied immunologically and 29 had lymphocytes cultured for HTLV-III/LAV. Results of all studies were normal. Of the 531 employees, 3 (0.56%) had serologic evidence of HTLV-III/LAV infection. All were seropositive at the time of study entry; none reported adverse nosocomial exposures. All acknowledged membership in one or more established risk groups for AIDS. This study provides strong evidence that the risk of nosocomial transmission of HTLV-III/LAV is extremely low.

A kinetic colorimetric procedure for quantifying magnesium in serum.

We have developed a kinetic colorimetric procedure for determination of magnesium in serum. The magnesium-dependent enzyme glycerol kinase is used to phosphorylate glycerol to glycerol 3-phosphate, the latter being oxidized to dihydroxyacetone phosphate and hydrogen peroxide by glycerophosphate oxidase. The generated hydrogen peroxide is then reduced by peroxidase with the simultaneous oxidative coupling of 4-aminoantipyrine and 2-hydroxy-3,5-dichlorobenzenesulfonate, producing a red reaction product with an absorption maximum at 510 nm. The rate of color production is proportional to the concentration of the Mg X ATP complex, which is, in turn, proportional to the magnesium concentration in serum. This method is rapid and precise, avoids the use of expensive instrumentation, is easily automated, and results compare well with those by the Du Pont aca and manual Magon sulfonate methods.

On-line clarification for the measurement of serum glucose in hyperlipidemic specimens.

The optical aberrations due to the presence of the turbidity caused by hyperlipidemia has been eliminated in two serum glucose procedures. This has been accomplished by incorporating lipase and alpha-cyclodextrin into the two glucose reagents. The hydrolytic action of the lipase generates water soluble glycerol and insoluble fatty acids. By including the chemical scavenger alpha-cyclodextrin into the reagent the fatty acids are solubilized and thus the production of a second turbidity is avoided. Because the clearing reagents are incorporated into the glucose reagents, this is an on-line process and no additional labor is required to clear the sample-reagent mixture. Furthermore, no additional time is required as the clearing occurs in the same period of time that it takes for the indicator reactions to reach equilibrium.

Enzyme reagents for measurement of phospholipids of amniotic fluid.

Enzymic procedures have been developed for the specific determination of three phospholipids in an effort to make the estimation of the phospholipids of amniotic fluid more sound analytically. Apart from the actual determination of these analytes (lecithin, sphingomyelin, and phosphatidylglycerol), these enzymic procedures facilitate the evaluation of a number of the basic premises and procedural steps involved with the traditional procedures for the evaluation of fetal lung maturity. To this end, the changes are reported in sphingomyelin concentrations with gestational age. Although lacking in sufficient clinical data to assign "cut-off" values as yet, the enzymic procedures seem to correlate well with the existing procedures and are analytically accurate and precise.

Peroxidase-coupled method for kinetic colorimetry of total creatine kinase activity in serum.

We describe a peroxidase-coupled method involving a colorimetric indicator reaction for determining the total activity of creatine kinase (EC 2.7.3.2) in serum. The kinetically favorable reverse reaction is exploited to generate adenosine 5'-triphosphate, which is used in the glycerol kinase-catalyzed phosphorylation of glycerol. The glycerol 3-phosphate so generated is oxidized in the presence of alpha-glycerophosphate oxidase to produce hydrogen peroxide, which is reduced in the presence of peroxidase with the simultaneous oxidation and coupling of 4-aminoantipyrene and 2-hydroxy-3,5-dichlorobenzenesulfonate to produce an intensely colored red chromogen. Results of the proposed method (y) correlate well with those of the Boehringer-Mannheim "CK-NAC UV" method as applied to the Hitachi 705 chemistry analyzer (y = 1.025 chi - 18.1, r = 0.9985, n = 100, range = 19-4531 U/L). The sensitivity of the method, based on molar absorptivities, is nearly fourfold that of procedures involving the reduction of NADP+.

The variable reagent blank: protein determination as a model.

Three total protein assays were analyzed to determine the extent of deviation encountered when a constant measured reagent blank is compared to a continuously decreasing true reagent blank. This blank effect owes its regressive nature to the consumption of the active reagent ingredient by the protein reactive species, variably and sometimes, with certain reactants, nonlinearly in the presence of increasing protein concentrations. However, the blank effect of interest here is apparent only when the reagent and the reagent-protein complex present overlapping spectra and therefore absorb at the wavelength of measurement. Thus it was found that while the biuret and the Coomassie brilliant blue assays produced pronounced differences in the variable true reagent blanks, the Folin-Ciocalteau reaction did not develop a deviation from the true blank since the reagent blank does not absorb to any extent at the assay wavelength. In this manner, the latter procedure could serve as a marker against which the former two blank reactions can be shown to display relatively excessive deviations.