Cycling and early pregnant endometrium as a site of regulated expression of the vitamin D system.

In addition to its calciotropic function, the secosteroid 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), has potent anti-proliferative/immunomodulatory effects on various tissues. Consistently, the enzyme that catalyzes the synthesis of 1,25(OH)(2)D(3), 1alpha-hydroxylase (1alpha-OHase) and the vitamin D receptor have a widespread tissue distribution. Among site-specific functions, the hormone has been suggested to be involved in uterine physiology. However, molecular analysis of the vitamin D system in normal endometrium throughout the menstrual cycle as well as its regulation in the context of endometrial physiological and pathological events have received very limited attention. Thus, we have studied expression, localization and regulation of 1alpha-OHase in human cycling and early pregnant endometrium. The capacity for 1alpha-hydroxylation and the presence of vitamin D receptor in endometrial cells have also been evaluated. The functional significance of these findings has been tested by evaluating gene expression of the catabolic enzyme, vitamin D 24-hydroxylase, and of the adhesion protein, osteopontin. Finally, to verify any potential dysfunction of the vitamin D system in endometriosis, a reproductive disease characterized by immune-mediated anomalies, we have analyzed expression of 1alpha-OHase in both eutopic and ectopic endometrium of affected patients. Results obtained showed that the active form of the 1alpha-OHase gene was expressed in human endometrial stromal cells independent of the cycle phase but with a significant increase in early pregnant decidua. A similar profile was observed for the protein, which was abundantly expressed in the cytoplasm of both endometrial stroma and epithelial glands. Both cycling and early pregnant endometrial cells also expressed the vitamin D receptor. In the same cells, 1alpha-OHase mRNA levels were significantly stimulated by the pro-inflammatory cytokine interleukin (IL)-1beta (50 and 500 pg/ml) while addition of the active form of the hormone could modulate both CYP24 and osteopontin gene expression. The 1alpha-OHase gene was also expressed in ectopic endometrium and its levels were increased in proliferative phase cultures derived from patients with endometriosis. Human cycling endometrium may be included among the extrarenal sites able to synthesize vitamin D. The IL-1beta-mediated induction of 1alpha-OHase gene and the hormonal modulation of osteopontin support a role for the hormone in the immunological mechanisms underlying uterine function. Abnormalities of this system are present in endometriosis.

Creatinine measurement proficiency testing: assignment of matrix-adjusted ID GC-MS target values.

The results of an external quality-assessment experiment for serum creatinine measurement are described. Fifty-one laboratories performed quintuplicate analyses during three different analytical runs on six lyophilized sera and two frozen human serum pools. Isotope dilution gas chromatography-mass spectrometry (ID GC-MS) target values were assigned to all the materials. Intralaboratory within- and between-run imprecision results were very similar for all the materials tested (CV < or = 2.20% and < or = 4.70%, respectively). The overall imprecision obtained was high (CV 6.5-20.0%) because of increased interlaboratory-intermethod variability. A significant positive bias (+ 9.2-+43.7%) was found for all the materials at lower creatinine concentration. By using two human sera at different concentrations, we could calculate the constant and the proportional calibration bias displayed by each peer group. The majority of the lyophilized materials showed a behavior divergent from the frozen pools, indicating matrix-related problems. We propose a new algorithm for calculating matrix bias correction factor instrument-reagent specific for each material.

Automated fluorimetric assay procedure for glucohydrolases using a routine centrifugal analyser assay of enzymes of lysosomal origin in plasma, II.

The manual fluorimetric procedure, considered as a reference method for the determination of N-acetyl-beta-D-glucosaminidase, beta-D-glucuronidase and beta-D-galactosidase in human plasma, was automated as a routine method, using the IL Monarch centrifugal analyser. Using a liquid standard with a known enzyme content, the automated assay correlated fairly well with the reference manual method (r values very close to 1). Its analytical imprecision was much lower than that of the manual method. The automated assay of N-acetyl-beta-D-glucosaminidase, beta-D-glucuronidase and beta-D-galactosidase gave coefficients of variation of 5.7-6.9, 3.6-5.0 and 3.8-4.2%, respectively, detection limits of 4, 2 and 1 mU/l plasma respectively, and linear responses of up to 73, 8.4 and 0.9 U/l of plasma respectively. Furthermore, the method required only small volumes of undiluted plasma (4-10 microliters). This method appears to be reliable, sensitive, simple enough for routine analyses and as cost effective as the most common routine serum enzyme assays.

Assessment of skeletal muscle viability by PET.

UNLABELLED: We investigated the use of [18F]fluoro-2-deoxyglucose (FDG) PET scanning for assessment of skeletal muscle viability in patients with peripheral vascular disease and in patients following free-flap skeletal muscle transfer for closure of open wounds. METHODS: We obtained 32 FDG-PET scans from 30 patients, either at the time of admission for peripheral vascular disease (n = 16) or between 1 and 15 days after surgery for skeletal muscle transfer (n = 16). Ratios between injured and contralateral limb FDG tracer activity uptake were correlated with clinical outcome at 1 mo to 3 yr follow-up. RESULTS: Viable muscle uptake ratios ranged from 0.47 to 7.88 (mean: 2.26 +/- 1.81; n = 26), while nonviable muscle uptake ratios ranged from 0.12 to 0.46 (mean: 0.27 +/- 0.12; n = 6; p < 0.02). After skeletal muscle transfer, two patients with viable tissue, as documented by PET, required amputation due to osteomyelitis, and one patient with peripheral vascular disease who showed viable tissue by PET required amputation 3 mo after the PET scan because of recurrent ulcers. CONCLUSION: FDG-PET scanning can determine skeletal muscle viability in patients with peripheral vascular disease and in patients following free-flap transfer.

[Immediate post-operative enteral nutrition]. / La nutrizione enterale post-operatoria immediata.

Patients with cancer of the upper digestive tract may present with malnutrition, which may cause immunodepression and an increased rate of postoperative complications. In this study we describe the rationale and the feasibility of immediate postoperative enteral nutrition (NEPI) and evaluate its effectiveness for the nutritional support of patients undergoing surgery of the upper digestive tract. We studied 46 patients undergoing the following procedures: total gastrectomy (n = 22); oesophageal resection (n = 12); duodenocephalopancreasectomy (n = 12). NEPI was started on postoperative day 0 with a polymeric diet (calories: 53% as CHO, 22% as proteins, 25% as lipids) aiming at a calorie intake of 25 Kcal/Kg/day by postoperative day 4. It was possible to administer 24 Kcal/kg/day with the enteral diet from the 4th to the 10th postoperative day. Oral intake was resumed on average on postoperative day 13, and the mean hospital stay was 27 +/- 17 days. Tolerance of NEPI was good in most patients: only 5 patients (11%) interrupted the enteral nutrition. The mean postoperative weight loss was 3.7%. The rate of septic complications was 27%; mortality was nil. The surgical procedures caused a transient and reversible acute-phase decrease of nutritional and immunological parameters in the early postoperative period. However the NEPI preserved the nutritional status postoperatively as shown by the lack of significant changes in the nutritional indices at 10-14 days after surgery, as compared with baseline. In summary, we documented that NEPI can be started from postoperative day 0 with good intestinal tolerance, allowing adequate nutritional support, after extensive surgical procedures on the upper digestive tract.

Preparation of a stable liquid material for calibration and quality control for lysosomal enzymes in plasma. Assay of enzymes of lysosomal origin in plasma, I.

Several lysosomal enzymes present in human plasma (N-acetyl-beta-glucosaminidase, beta-glucuronidase, beta-galactosidase, alpha-galactosidase, alpha-L-fucosidase, alpha-mannosidase, beta-glucosidase) were maintained in a fully active state for at least 8 months by the addition of ethylene glycol (300 milligrams final concentration) to freshly prepared plasma and storage at -20 degrees C. Pools of human plasma from healthy humans, stabilized and stored as above, and containing a low, medium or high content of the above enzymes, were used to establish the analytical imprecision (within-run, day-to-day and total imprecision) of the fluorimetric assay. Ten replicates in ten different analytical series, covering a period of two months, were performed. The total imprecision (expressed as coefficient of variation) was in general lower than 10%; in a few cases, particularly plasma samples with a low enzyme content, the total imprecision was 18%. The isozymes A, B, I1, and I2 of N-acetyl-beta-glucosaminidase displayed the same stability upon storage as the unfractionated enzyme. It is concluded that pools of human plasma containing known amounts of lysosomal enzymes, stabilized by the addition of 300 micrograms ethylene glycol and stored at -20 degrees C, are suitable liquid materials for calibration and quality control for the assay of the same enzymes.

Beta-blockade during whole-body hyperthermia: a toxicity study in the dog.

A radiant heat device (RHD) for whole-body hyperthermia (WBH) has been safely and effectively used with a dog model. The cardiovascular changes which occur in the dog during WBH--including heart rate, blood pressure, cardiac output, stroke volume and ejection fraction--agree qualitatively and quantitatively with changes observed in previous WBH-RHD studies done in pigs and humans. We elected to study the effect of propanolol in dogs during WBH in order to evaluate this drug's potential use in human cancer patients who are ineligible for WBH because of coronary artery disease. This report details cardiovascular changes which occur with beta-blockade during 42 degrees C WBH in the dog. Our results show that the level of beta-blockade needed to control heart rate during WBH produces acute cardiovascular decompensation.

Impairment of left ventricular function during maximal isometric dead lifting.

High-intensity short-duration lifting is frequently performed by athletes and laborers. Little is known about the magnitude and pattern of blood pressure response and resultant effects on left ventricular (LV) function during this form of intense isometric exercise. We monitored brachial intra-arterial pressure and LV ejection fraction (LVEF) during upright isometric dead lifting performed on a force platform. Fourteen healthy male subjects (age 27 yr) maintained maximal sustained isometric dead lift (140 +/- 34 kg) for 32 s. LVEF was measured by 99mTc first-pass radionuclide ventriculography. Mean arterial pressure increased from 107 +/- 15 mmHg at rest to a peak of 174 +/- 28 mmHg and fell precipitously to 88 +/- 13 mmHg within 10 s after release of the dead lift. LVEF decreased from 63 +/- 8 to 51 +/- 14% (P less than 0.02) in seven subjects with technically acceptable ventriculograms. We conclude that maximal upright isometric dead-lift exercise produces a marked increase in arterial pressure and corresponding LV afterload that is associated with a transient reduction in LVEF in normal men.

[Agranulocytosis caused by dipyrone. Case reports]. / Agranulocitosi da dipirone: contributo casistico.

Two cases of dipyrone-induced agranulocytosis are described. In the first one, the disease was due to a single administration, in the second one, to prolonged therapy. The possible pathogenic mechanism is discussed, which can be immunologic and/or toxic. The effectiveness of some of the drugs and general supportive measures applied is discussed. Health education of the patient, in conjunction with careful attention by the physician, may play an important role in the prevention of this pathological condition.

Hematological values from quantitative buffy coat analysis (QBC II) system: evaluation in a hematological/oncological outpatient section.

The quantitative analysis of the buffy coat (QBC II system) of centrifuged whole blood samples allows one to obtain hematocrit (Ht), platelet (PLT) and white blood cell (WBC) values. We have tested the QBC II system in a hematological/oncological Outpatient Section, where it is necessary to know patient's hematological condition immediately before administering chemotherapy. One hundred ten blood samples from 110 patients with solid or remission hematological tumors were measured in duplicate with the QBC II system and with the counter instrumentation (Sysmex TOA 800) of the Central Laboratory, where samples were sent for emergency determination. Reliable values for Ht and WBC were obtained in all samples, and for PLT in 101 samples. The QBC II System failed to furnish very low and very high PLT counts. The time needed for the QBC II system analysis was shorter than that required to obtain the Central Laboratory data. The QBC II system analysis is a useful means for rapidly evaluating the basis hematological condition in most chemotherapy patients.

Blood flow in the carotid arteries: quantification by using phase-sensitive MR imaging.

The feasibility of MR phase-sensitive imaging for the quantification of blood flow in the carotid arteries was studied in two normal volunteers and six patients with carotid artery and/or cerebrovascular disease. The technique consists of sensitizing the phase of the MR signal to blood flow velocity gated to different times in the cardiac cycle. Flow velocities and volumes were measured by using transverse planes in the common, internal, and external carotid arteries, and flow curves were generated. Measurements made by using flow phantoms correlated well with calculated results. The MR measurements yielded values between 250 and 580 ml/min for the total flow through each of the common carotid arteries in the two normal volunteers. Markedly reduced flow (about 50% below normal) was detected in a patient whose arteriogram showed severe occlusion of the internal carotid artery. In a second patient, who had a large frontal intracranial arteriovenous malformation noted by arteriography, the MR-quantified flow was abnormally high (about 1 liter/min). In the remaining four patients, the findings on phase-sensitive quantification were consistent with those expected from clinical and other laboratory studies (including arteriography and sonography). These preliminary findings suggest that MR phase mapping may be a feasible tool for the noninvasive quantification of carotid blood flow.

Comparison of seven serum assays on four automatic analysers.

Serum enzyme assays used on four different analysers (Hitachi 737, Hitachi 705, Cobas-bio and RA-2X) were compared by determining the activity of seven different enzymes (AST, ALT, LD, ALP, GGT, CK and AMS). Performance checks (quality control procedure) and replications (the study of the total analytic imprecision and of its components) were conducted and the methods were compared by linear regression analysis with statistical inference on the curves following the protocols of the National Committee for Clinical Laboratory Standards (NCCLS: PSEP- 2, PSEP-3, PSEP-4). The correlation coefficients between the methods (r = 0.991-0.999), together with the other statistical parameters, indicated that the methods are well correlated on all the instruments. The total imprecision was good for all analytes, except ALT. Among the instruments tested, the RA-2X gave more variable results, although the total imprecision was acceptable. There was no relevant carry-over effect. The evaluation of performance claims indicated that the expected error did not substantially affect the results at the level of clinical decisions.

[Radiotherapy in the treatment of carcinomas of the base of the tongue. Analysis of the literature and personal experience]. / La radioterapia nel trattamento dei carcinomi della base della lingua. Analisi della letteratura ed esperienza personale.

Carcinoma of the base of the tongue is usually treated with radiation therapy. After a review of the literature, the findings are reported of 129 patients with squamous cell carcinoma of the base of the tongue treated in 1979-1983, with a minimum of a 3-year follow-up: 8 T1, 40 T2, 60 T3 and 21 T4 were found; only 48 cases were N0. Advanced stages (stage III and stage IV) were 83% of the total. External photon irradiation (Cobalt-60) was used in all cases; 33 patients underwent chemotherapy or surgery as additional treatment. The overall local control rate was 45.7%. Local control decreased as T stage advanced: T1 62.5%, T2 55%, T3 41.7% and T4 33.3%. The overall regional control rate for lymph nodes clinically palpable was 51.8%. The overall actuarial 5-year survival rate was 29.1%. The 5-year survival rate according to the N-staging varied from 37% for N0-stage to 17.4% for N3-stage. The majority of failures (92.8%) occurred within 2 years since the end of the therapy. Treatment complications, secondary carcinomas and causes of death are also discussed. Radiation therapy has proven to be effective for early-stage carcinoma of the base of the tongue; in more extensive lesions results are poor. Improved results could be obtained by optimal application of radiotherapeutic techniques. Knowledge of the various prognostic factors is essential to administer the therapeutic regimen for a given patient by the characteristics of his particular tumor.

[Automatic tracing of conversion scales from conventional units to the SI system of units]. / Tracciamento automatico delle scale per la conversione delle unità convenzionali in unità del sistema SI.

American medical journals, as the Journal of the American Medical Association (JAMA), and the American Journal of Clinical Pathology (AJCP), the Journal of the American Society of Clinical Pathologists (ASCP), are shifting to selected SI (Système International d'Unités) units for reporting measurements. Further discussion by the AMA, the ASCP and other organizations is required before consensus in the US medical community can be reached as to the extent of and time frame for conversion to SI for reporting clinical laboratory measurements: however this decision will certainly greatly speed up the process of conversion in European countries too. Transition to SI units will require the use of different reference ranges, and there will be a potential for serious misinterpretation of laboratory data unless well-planned educational programs are instituted before the change. A simple program written in Microsoft Basic for automatically tracing on one's personal computer (PC) monitor a dual scale, in the conventional and in the SI system of units, is presented here. The program may be easily implemented and run on every PC operating under MS-DOS, equipped with a CGA or an AT&T6300 graphic card: through the operating system the scales may also be printed on a dot-matrix graphic printer. We believe that this, and other tools of this kind, will be useful in the thorough educational process of those reading the reports, and will be an important factor in the success of conversion to SI reporting.

Oblique magnetic resonance imaging of the bronchi.

The trachea and main bronchi of a supine patient in a magnetic resonance (MR) scanner are not contained in a single standard coronal plane, but instead intersect this coronal plane at some angle, usually 20 degrees - 35 degrees. We have developed a new MR imaging protocol to determine the oblique imaging plane which best contains the trachea and main bronchi. The resulting oblique images simplify anatomical identification, and allow the user to select additional oblique planes which cut any desired portion of main bronchus in true cross section. Accurate lumen shapes and areas may then be extracted from these cross-sectional images. The method does not require the patient to be moved or rotated, and does not require hardware modification. We demonstrate the clinical application of the protocol with both a normal volunteer and a patient with an endobronchial tumor. The use of gradient echo pulse sequences together with this protocol to distinguish between vessels and bronchi is presented. We provide phantom verification to demonstrate the quantitative accuracy of the method to provide lumen areas.

[Results of radiotherapy in the treatment of 218 carcinomas of the tonsillar area]. / Analisi dei risultati della radioterapia nel trattamento di 218 carcinomi della regione tonsillare.

After a brief review of the literature, the results of the treatment by radiation therapy in 218 patients with tonsillar region carcinoma are reported. The patients were treated at the Turin University Radiotherapy Department (131 cases) and at the Radiotherapy Department of the Ospedale Maggiore, Novara (87 cases) from 1976 through 1984. The series included 29 T1, 88 T2, 64 T3 and 37 T4; 86 patients (40%) were N0. Only 49 patients were stages I and II (1/4 of the total). The local control (T) rate at the end of therapy was 63.5% for the overall series (139/218) ranging from 100% in T1, 72.6% T2, 54.6% T3 to 30% T4; the regional control (N) for patients with cervical lymphadenopathy was 66% (88/132). The overall 5-year survival was 38%; the 5-year disease-free survival was 35%. The 5-year survival according to T-stage was 52% for limited cases (T1-T2) in comparison to 22% for advanced ones (T3-T4). The 5-year survival for N0 stage was 55%; when neck nodes were present (N # 0) the survival was 29% after 5 years. Complications included 6 cases of trismus and 2 osteonecroses of the mandible. Second primary malignant diseases were seen in 17 cases (7.8%). The most common cause of death was failure in primary tumour control (80% of cases). In this series the results of radiotherapy of early stage tonsillar carcinomas are quite good; patients with large tumour masses fare less satisfactorily. Better results should be obtained in the future by identifying prognostic factors, monitoring potential causes of treatment failure and optimizing the radiotherapeutic treatment plan and technique.

[Determination of urine density: comparison of refractometry and measurement of the period of oscillation using the protocols of the National Committee for Clinical Laboratory Standards]. / Determinazione della densità delle urine: confronto tra rifrattometria e misura del periodo di oscillazione, con l'impiego dei protocolli del National Committee for Clinical Laboratory Standards (NCCLS).

Two methods for the determination of urine density (relative density or specific mass) have been evaluated and compared using NCCLS protocols: the refractometric method and a new method based on the measure of shifts in harmonic oscillation (SHO). Within the range investigated (from 1,000 up to 1,034) the methods were linear. Total imprecision, including the carry-over, was the same for both: the 95% confidence limits, at the level of 1,003, 1,013 and 1,030, were 1,001-1,005, 1,011-1,015 and 1,028-1,032 respectively. In analysing urines having known density, the SHO method was always accurate, while the refractometric method was affected by a significant bias at protein and haemoglobin concentrations greater than 4 and 2 g/L respectively. A direct comparison of the results obtained by the two methods was made on 300 untimed specimens of urine with density ranging from 1,002 to 1,034; the equation of regression line (refractometric method on the x axis, SHO on the y axis), calculated with a non-parametric statistical method, was SHO = refractometric method + 0,001. Despite some urine specimens (24, i.e. 8%) showed density differences (absolute value) of more than 0,003 (between 0,004 and 0,007 relative density units), the good mean agreement observed allows to conclude that both methods give clinically useful analytical results.

Stability constants of haemoglobin cyanide and azide measured by two-wavelength spectrophotometric method.

A two-wavelength spectrophotometric procedure for the simultaneous determination of haemoglobin and haemoglobin cyanide (HiCN) (or of haemoglobin and haemoglobin azide (HiN3)) concentrations in mixtures has been developed and applied to the determination of the stability constants of HiCN and HiN3. The analytically reliable procedure allowed stability constants to be estimated with about 10% (relative standard deviation, coefficient of variation) uncertainty. Values of 1.9 X 10(6) and 2.0 X 10(5) 1 X mol-1 were obtained for HiCN and HiN3, respectively. These results are discussed in relation to the optimal composition of the reagents for blood haemoglobin assay by the two methods.

The accumulation of a platelet protein, thrombospondin, at the site of arterial thrombus formation: preliminary report.

There are many methods available for the detection of thrombosis, none of which are noninvasive, rapid and accurate. Thrombospondin is a platelet protein that is present in the developing thrombus and may be an effective substance to use for imaging thrombosis. Vascular stenosis and thrombosis were produced in coronary, carotid and femoral arteries in eleven adult mongrel dogs. 131I labeled thrombospondin was administered to each animal to determine whether the radiotracer accumulated at the site of thrombus formation. The radioactivity per gram of vessels with thrombi was significantly different from the control vessels or whole blood (p = 0.0037 and p = 0.0015, respectively, paired t-test). This preliminary work suggests that iodinated thrombospondin accumulates at the site of thrombus formation. Labeled thrombospondin may be a rapid, safe and accurate method of detecting arterial thrombosis.

Pulse sequence generated oblique magnetic resonance imaging: applications to cardiac imaging.

A pulse sequence procedure for producing oblique magnetic resonance images is described. Using this procedure we present a new, accurate method to obtain true short-axis views and true long-axis views (both parallel and perpendicular to the septal plane) of the heart. The method is accurate regardless of the orientation of patient's heart. The method does not require the patient to be rotated, nor otherwise moved, and does not require any additional hardware. The method is experimentally verified with both human and phantom studies. The phantom study indicates accuracy of approximately 1 degree with a commercial scanner that reports angular measurements to a precision of 1 degree. Application of the short-axis views to measurement of left ventricular volume, and possible advantages of Gauss-Legendre integration for this measurement are discussed. Finally, multiphase oblique cardiac images are presented.