Different expression patterns of LGALS1 and LGALS3 in polycythemia vera, essential thrombocythemia and primary myelofibrosis.

Despite all the knowledge, the cellular and molecular mechanisms involved in myeloproliferative neoplasm (MPN) pathophysiology remain unclear. Authors have shown galectin-1 (Gal-1) and 3 playing roles in tumour angiogenesis and fibrosis, which were correlated with poor prognosis in patients with MPN. In the present study LGALS1 and LGALS3 were differently expressed between polycythemia vera, essential thrombocythemia (ET) and primary myelofibrosis (PMF) diseases. Increased LGALS3 expression was associated with a negative JAK2 V617F status mutation in leucocytes from PMF but not in patients with ET without this mutation. However, a positive Janus kinase 2 (JAK2) V617F cell line established from patients with ET (SET-2 cells) when treated with JAK inhibitor presented high levels of LGALS3. Additionally, high LGALS1 expression was found in CD34(+) cells but not in leucocytes from patients with PMF, in absence of JAK2 V617F mutation, and also in SET-2 cells treated with JAK inhibitor. Thus, our findings indicate that differential expression of LGALS1 and/or LGALS3 in patients with MPN is linked with JAK2 V617F status mutation in these diseases and state of cell differentiation.

Abnormal US response of main pancreatic duct after secretin stimulation in patients with acute pancreatitis of different etiology.

To assess changes in caliber of the main pancreatic duct, we performed abdominal ultrasonography after maximal stimulation with secretin (US-S test) in 14 patients with idiopathic recurrent acute pancreatitis, in six with recurrent acute pancreatitis secondary to pancreas divisum, in 14 recovered from a single attack of acute pancreatitis, and in 21 control subjects. In five patients, the test was repeated 10 days after endoscopic sphincterotomy. We repeated the test 48 h later in nine subjects to evaluate its reliability. We evaluated changes in lipase serum values in some of these subjects. In the acute pancreatitis patients, the main pancreatic duct diameter was significantly increased over baseline and control values throughout the observation period. In the patients undergoing sphincterotomy, the poststimulation diameter of the main duct was substantially reduced after the operation. The reliability of the test ranged from 77 to 91.5%. In the acute pancreatitis patients, serum enzymes after secretin stimulation showed a persistent increase over controls. These results suggest that pancreatic outlet obstruction, mainly at the sphincter of Oddi level, may be an important pathogenetic factor in the course of the disease and that, if this condition is present after an attack of acute pancreatitis, endoscopic sphincterotomy may be in order. The simplicity and satisfactory reproducibility of the US-S test suggest a strong case for its routine clinical use.

Glucose counterregulatory response to acute hypoglycemia in hyperthyroid human subjects.

To evaluate the impact of hyperthyroidism on the counterregulatory response to hypoglycemia, eight hyperthyroid and eight sex-, age-, and body mass index-matched healthy women were given an iv insulin bolus (0.1 U/kg BW), and blood was drawn from 0-120 min for glucose, epinephrine, norepinephrine, glucagon, GH, ACTH, and cortisol measurements. In the basal state plasma glucose, GH, and cortisol levels were similar in the two groups, whereas plasma glucagon and ACTH were increased (135 +/- 17 vs. 80 +/- 10 ng/L and 6.4 +/- 1.5 vs. 2.6 +/- 0.4 pmol/L, respectively; both P < 0.025), and plasma catecholamines were reduced [epinephrine, 142 +/- 25 vs. 371 +/- 71 pmol/L (P < 0.025); norepinephrine, 0.41 +/- 0.07 vs. 1.41 +/- 0.12 nmol/L (P < 0.001)] in hyperthyroid patients. After insulin injection, plasma glucose similarly declined in the two groups (nadir, 1.5 +/- 0.2 vs. 1.6 +/- 0.2 mmol/L). Conversely, recovery from hypoglycemia was significantly faster in the hyperthyroid patients. In this respect, it is noteworthy that the plasma glucagon response had remarkably increased in the latter (peak, 444 +/- 56 vs. 198 +/- 17 ng/L; P < 0.005). On the other hand, the epinephrine responses were similar in the two groups, whereas norepinephrine levels remained consistently lower (peak, 0.97 +/- 0.20 vs. 2.61 +/- 0.24 nmol/L; P < 0.001), and the GH increase was severely impaired (peak, 10.6 +/- 1.9 vs. 29.6 +/- 6.2 micrograms/L; P < 0.01) in hyperthyroid patients. Plasma ACTH remained slightly higher in hyperthyroid subjects, but there were no substantial differences in the cortisol response between the two groups. In conclusion, hyperthyroidism affects plasma levels of several counterregulatory hormones, either in the fasting state or after insulin-induced hypoglycemia, with increased efficiency of plasma glucose recovery from hypoglycemia.

Systemic and renal effects of a new angiotensin converting enzyme inhibitor, benazepril, in essential hypertension.

Seventeen essential hypertensive patients with normal renal function were treated with a new non-sulphydryl orally active angiotensin converting enzyme (ACE) inhibitor, benazepril, 10 mg given once or twice daily, according to diastolic blood pressure levels, for 6 weeks. In all patients, changes in blood pressure, systemic and renal hemodynamics, plasma renin activity and urinary aldosterone and albumin excretions were assessed at the end of a 2-week placebo run-in period and at the end of the study. Benazepril monotherapy controlled blood pressure well. No changes in cardiac output, heart rate or stroke volume were observed, while peripheral vascular resistance was significantly decreased (-11%, P less than 0.05). Plasma volume was unaltered. The glomerular filtration rate was stable, but effective renal plasma flow was increased because of the marked reduction in renal vascular resistance (-35%) and, therefore, the filtration fraction was decreased. Urinary albumin excretion remained unchanged. A significant increase in plasma renin activity (P less than 0.001) and a decrease in urinary aldosterone excretion were seen. No side effects were observed during the treatment period. In conclusion, our results suggest that benazepril alone is an effective antihypertensive agent in patients with essential hypertension. The blood pressure lowering effect is due mainly to systemic vasodilation and is observed up to 24 h after administration of the drug. The vasodilation appears to be more consistent in the renal than in the systemic circulation.

Development of a screening system for cystic fibrosis: meconium or blood spot trypsin assay or both?

High blood trypsin levels during early days of life are found in newborns subsequently diagnosed to be affected by cystic fibrosis. The authors compared the validity of the traditional meconium test with the blood immunoreactive trypsin (IRT) assay, carried out in parallel on 113,302 neonates from three regions of North-eastern Italy. The meconium test showed a sensitivity of 57.7%. The sensitivity of the IRT test was higher (96.1%). It was possible to identify by IRT 10 out of 11 false negative CFs at the meconium test. A shortcoming of neonatal IRT, however, is its low specificity; 1.6% of the newborns had to be retested. A new screening policy was therefore proposed and carried out on 69,640 newborns: the Lactase test (LACT) on meconium was introduced as a complementary assay in IRT positive newborns. If LACT exceeded 2 U/g dry meconium, a confirmatory sweat test was immediately requested; if LACT test was negative and IRT exceeded 85 micrograms/l, IRT was repeated. Postneonatal retesting values above 25 micrograms/l required a sweat test. As a result, the estimated prevalence of CF was 1:4,352, the sensitivity was 93.3%; the specificity turned out to be 99.6%, considering all false positive newborns investigated with retesting and/or direct sweat test.

Plasma concentrations of glucagon during hyperglycemic clamp with or without somatostatin infusion in obese subjects.

The aim of the present study was to evaluate whether the inhibitory effect on pancreatic A-cell exerted by hyperglycemic hyperinsulinemia and/or by somatostatin administration is impaired in human obesity. For this purpose plasma glucagon concentrations were measured in 8 obese and 8 nonobese nondiabetic subjects during a 4-h hyperglycemic clamp. Synthetic cyclic somatostatin-14 was infused at the rate of 2.5 nmol/min during the third hour of the study. Fasting plasma glucagon was higher in obese than in nonobese subjects (242 +/- 32 vs 163 +/- 15 pg/ml, p less than 0.05) (mean +/- SEM). In the last 20 min of the glucose infusion period preceding somatostatin administration (100-120 min of the study) plasma glucagon averaged 195 +/- 26 pg/ml in obese and 122 +/- 13 pg/ml in nonobese subjects (p less than 0.05), with a reduction of 19 +/- 3% in the former and 28 +/- 4% in the latter (p = n.s.). In both groups somatostatin infusion did not result in a further decrease in plasma glucagon, which averaged 192 +/- 27 pg/ml in obese and 123 +/- 16 pg/ml in nonobese subjects (p less than 0.05) in the 160-180 min period of the study. Also after discontinuing somatostatin infusion plasma glucagon levels did not change. These results suggest that in human obesity hyperglycemic hyperinsulinemia has a normal inhibitory effect on pancreatic A-cell and that somatostatin administration has no additive effect on hyperglycemia and hyperinsulinemia in either obese or nonobese nondiabetic subjects.

An evaluation of an enzyme immunoassay method for immunoreactive trypsin in dried blood spots.

A novel monoclonal antibody based enzyme immunoassay (EIA) method for the measurement of the human cationic trypsinogen (NeoScreen, AGEN Biomedical Ltd., Acacia Ridge, Australia) in dried blood spots for the neonatal screening of cystic fibrosis was evaluated. The calibration standards provided as dried blood spots by AGEN are highly unstable and must be replaced with user prepared materials. Reference values from control individuals were obtained by parametric methods. A preliminary comparison with a polyclonal antibody based RIA method (Trypsik, SORIN Biomedica, Saluggia, Italy) was performed. Regression analysis between the RIA and the EIA methods gave a coefficient of correlation of 0.58 for RIA values less than 40 micrograms/L and of 0.77 for RIA values greater than or equal to 40 micrograms/L. Average CV of the within-run imprecision for the EIA method was 19.6% and for the RIA method 28.8%. CVs of the between-run imprecision at low, intermediate and high values for the EIA method were 23.7%, 15.8%, 15.6% and for the RIA method 20.6%, 14.4%, 11.2%. The diagnostic accuracy analyzed by a Receiver Operating Characteristics (ROC) curve of the RIA method gave a maximum accuracy of 190.9 while that of a simulated ROC curve for the EIA method was 193.0. We found that the precision and the diagnostic accuracy of the EIA method (AGEN) are equal to or better than those of one of the RIA methods.

A clinical evaluation of monoclonal (CA19-9, CA50, CA12-5) and polyclonal (CEA, TPA) antibody-defined antigens for the diagnosis of pancreatic cancer.

We measured in 193 patients, admitted to our wards for symptoms and signs suggestive of pancreatic or digestive malignancy, the serum levels of five tumor-associated antigens (CA 19-9, CA 50, CA 125, TPA, CEA) and we evaluated their diagnostic accuracy both when used alone and in combination. For CA 19-9 and CA 50 a sensitivity for pancreatic cancer as high as 92 and 88%, respectively, and specificity of 91.8% were found. A lower sensitivity vs. pancreatic cancer was found for the other tumor markers, and vs. the other digestive and nondigestive malignancies for all tumor markers (apart for CA 19-9 and CA 50 vs. biliary carcinomas). As for the combined assays, the best figures were found vs. pancreatic cancer for CA 19-9 plus CA 50, CA 50 plus CEA, CA 50 plus CA 125; a sensitivity by far worse vs. the other gastrointestinal cancers was found for all the possible combinations. We conclude that in selected symptomatic patients some tumor-marker determinations can be useful in identifying those with a high probability of harboring a pancreatic cancer, to be further studied or operated upon. The clinical relevance of this in patients already symptomatic is at present unclear.

[Behavior of thyroglobulin after fine needle aspiration of the thyroid]. / Comportamento della tireoglobulina dopo agoaspirazione tiroidea con ago sottile.

The authors report the behaviour of serum concentration of thyroglobulin (TG) after fine needle aspiration biopsy of the thyroid. Compared with the basal level, serum concentration of thyroglobulin increase of 70% three minutes and of 341% sixty minutes after this maneuver. The authors agree with those who believe an interval of 10-15 days between fine needle aspiration biopsy of the thyroid and determination of serum concentration of thyroglobulin is necessary.

New method ("SPAC ET") for free thyroxin in serum evaluated.

We analytically and clinically evaluated a new commercial method ("SPAC ET") for simultaneously determining total and free thyroxin in serum. We found the method to be rapid, simple, and precise. Results significantly correlated with those by "symmetric" dialysis and also agreed with the clinical data for patients with extremely low or high concentrations of thyroxin-binding globulin. On contrast, in those patients the most commonly used thyroid indexes (free thyroxin index or thyroxin:thyroxin-binding globulin ratio) failed to correctly reflect the true functional status of the thyroid.

CA 19-9 and carcinoembryonic antigen in pancreatic cancer diagnosis.

CA 19-9 (Centocor, Malvern, PA) and carcinoembryonic antigen (CEA), two recently developed immunoradiometric assays utilizing monoclonal antibodies, were evaluated in the sera of 139 subjects in order to assay their individual and combined value in pancreatic cancer diagnosis and to assess the influence of jaundice. Sensitivity, specificity, and accuracy in detecting pancreatic cancer were 69%, 85%, and 54% for CA 19-9; and 28%, 78%, and 6% for CEA, respectively. Combined evaluation gave the highest specificity (95%) when both, and the highest sensitivity (79%) when at least one, gave pathologic results. The receiver-operating characteristic curves demonstrated that CA 19-9 is more discriminating than CEA, for any serum value. A correlation between serum bilirubin and CA 19-9 was demonstrated in pancreatic and extrapancreatic disease. CEA determination, performed using monoclonal antibodies, seems to be unsatisfactory as compared to CA 19-9 in pancreatic cancer diagnosis, and combined assessment does not improve the results of CA 19-9 alone. Jaundice may influence serum CA 19-9 in pancreatic and extrapancreatic diseases.

Serum elastase 1 in chronic pancreatic disease.

Elastase 1 and immunoreactive trypsin were assessed by a RIA technique in the sera of 29 control subjects, 24 pancreatic cancer patients, 22 patients with chronic pancreatitis and 31 with extra-pancreatic diseases to ascertain and compare their usefulness in chronic pancreatic disease diagnosis. Increased levels of elastase 1 were detected in 60.9% of pancreatic cancer and in 61.1% of chronic pancreatitis patients; low values were found in only two subjects with pancreatic disease. A close correlation between the two enzymes was found in patients suffering from pancreatic cancer and chronic pancreatitis. These data suggest that serum elastase 1, as well as immunoreactive trypsin, is of limited value in chronic pancreatic disease diagnosis; increased levels of the two enzymes always occur simultaneously; low immunoreactive trypsin values together with normal elastase 1 serum levels are detectable in a number of patients with chronic pancreatitis and severe exocrine insufficiency.

Serum pancreatic enzyme alterations in acute viral hepatitis.

In a group of 167 patients with acute viral hepatitis (AVH), 11 with type A hepatitis, 125 with type B and 31 type non A, non B, the following enzymes were studied: serum amylase (S-AMY) and its isoenzymes (pancreatic and salivary type), urinary amylase (U-AMY), serum lipase (S-TGL) and serum immunoreactive trypsin (i-TRY). In all groups of patients, in the acute phase of illness, a significant increase in S-AMY was observed, in particular in hepatitis type B and non A, non B (p less than 0.001). An increase in U-AMY excretion was recorded in patients with type A hepatitis. S-TGL levels were significantly higher in all groups, especially in patients with type A hepatitis. i-TRY was only slightly higher in patients with hepatitis A and non A, non B. S-AMY isoenzymes showed a peculiar pattern: the pancreatic type (2) of isoamylase was found to be prevalent in 66% of patients with AVH while in controls the salivary type (1) was prevalent in most cases. Pancreatic enzyme alterations correlated neither with laboratory hepatic function tests nor with the clinical syndrome. These results suggest that a pancreatic injury is not uncommon in AVH, although it is seldom severe.

Actual role of pepsinogen group I in the study of upper gastrointestinal diseases.

The role of serum PG I in screening patients with chronic atrophic gastritis and gastric cancer, and in detecting peptic ulcer patients with high relapse risk, was ascertained in 276 subjects. Although not diagnostic per se, PG I was found to be under 20 micrograms/L in patients with chronic atrophic gastritis and in some gastric cancer or partially gastrectomized patients. In patients presenting with relapsing duodenal ulcer, PG I values were significantly higher than in the non-relapsing ones, but a satisfactory identification of all the duodenal ulcer patients with high relapse risk was not possible on this basis. Even the correlation between PG I and MAO was not accurate in every subject considered. These results suggest that the value of PG I is limited to assessing patients with upper gastrointestinal diseases in which a reduction of peptic secretion, and therefore of PG I in serum, is present.