Lactocin AL705 as quorum sensing inhibitor to control Listeria monocytogenes biofilm formation.

AIMS: The control of Listeria monocytogenes biofilm formation using lactocin AL705 bacteriocin at sub-minimum inhibitory concentrations (MICs) through an antiquorum sensing strategy, was preliminarily investigated. METHODS AND RESULTS: The screening for biofilm formation of different Listeria species at 10°C allowed selecting L. monocytogenes FBUNT for its use as biofilm producer. MIC and minimum bactericidal concentration of lactocin AL705 purified extract against the pathogen was determined. Bacteriocin sub-MICs were used to evaluate biofilm reduction. Concentrations between 2·5-20 AU ml-1 of lactocin AL705 produced significant decreases in biofilm formation without affecting the growth of the pathogen after 3 days of incubation. When bacteriocin concentrations (5-20 arbitrary units per millilitre (AU ml-1 )) were investigated as quorum sensing (QS) inhibitors using Vibrio harveyi as reporter strain, a significant reduction in luminescence by lactocin AL705 (20 AU ml-1 ) was observed. Even when L. monocytogenes produced AI-2 like molecules as recognized by the reporter strain, bacteriocins did not interfere with this compound. CONCLUSION: Antilisterial lactocin AL705 used to disrupt QS through a signal molecule inactivation was able to control L. monocytogenes FBUNT biofilm formation. Other molecule(s) different from the AI-2 involved during biofilm formation could be acting as target of the bacteriocin. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of bacteriocins derived from food-grade micro-organisms as a QS inhibition represents an effective strategy to control pathogens as well as an environmentally friendly sanitation method to mitigate postprocessing food contamination.

Lactobacillus spp. impair the ability of Listeria monocytogenes FBUNT to adhere to and invade Caco-2 cells.

OBJECTIVES: The objective of this study was to evaluate the ability of Lactobacillus curvatus CRL705, CRL1532, and CRL1533 and Lactobacillus sakei CRL1613 to survive under simulated gastrointestinal conditions. Moreover, a microencapsulation approach was proposed to improve gastrointestinal survival. Finally, experiments were performed to demonstrate that Lactobacillus spp. can modulate the ability of Listeria monocytogenes FBUNT to adhere to and invade Caco-2 cells. RESULTS: Lactobacillus strains were encapsulated in alginate beads to enhance the survival of bacteria under in vitro gastrointestinal conditions. All strains hydrolyzed bile salts using chenodeoxycholic acid as a substrate and adhered to Caco-2 cells. Cell-free supernatants (CFSs) showed antimicrobial activity against L. monocytogenes as demonstrated by agar diffusion assays. The average percentages of L. monocytogenes adhesion decreased from 67.74 to 41.75 and 38.7% in the presence of 50 and 90% (v/v), respectively, for all CFSs tested. The highest concentrations of CFSs completely inhibited the L. monocytogenes invasion of Caco-2 cells. CONCLUSIONS: The studied Lactobacillus strains have protective effects against the adhesion and invasion of L. monocytogenes FBUNT. Alginate encapsulation of these bacteria improved gastrointestinal tolerance such that they could be further studied as potential probiotics against intestinal pathogenic bacteria.

Lactobacillus sakei CRL1862 improves safety and protein hydrolysis in meat systems.

AIMS: The capacity of Lactobacillus sakei CRL1862 to prevent the growth of pathogens and its ability to degrade sarcoplasmic and myofibrillar proteins in pork meat systems was evaluated. In addition, basic safety aspects of Lact. sakei CRL1862 such as production of biogenic amines and antibiotic susceptibility were addressed. METHODS AND RESULTS: The bacteriocin-producing Lact. sakei CRL1862 showed respectively bactericide and bacteriostatic effect against Listeria monocytogenes and Staphylococcus aureus in beaker sausage assay during 9 days of storage at 22 °C. The hydrolytic effect of Lact. sakei CRL1862 on protein extracts was evaluated by SDS-PAGE and reverse phase HPLC. A more pronounced proteolysis was evidenced in inoculated sarcoplasmic proteins compared with myofibrillar extracts with the generation of predominantly hydrophilic peptides and increase of total free amino acids concentration. Lactobacillus sakei CRL1862 produced neither histamine nor tyrosine and exhibited no resistance to the antibiotics assayed. CONCLUSIONS: Lactobacillus sakei CRL1862 effectively controlled the growth of L. monocytogenes and Staph. aureus; moreover, it was able to hydrolyse pork meat extracts generating peptides and amino acids, which may improve hygienic and sensorial attributes of fermented meat products. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of an integrated approach to evaluate the major traits of Lact. sakei CRL1862 showed it can be applied as an autochthonous functional starter in meat fermentation.

Cetuximab, a chimeric human mouse anti-epidermal growth factor receptor monoclonal antibody, in the treatment of human colorectal cancer.

The recent successful development of monoclonal antibodies that target key components of biological pathways has expanded the armamentarium of treatment options for patients with colorectal cancer (CRC). In particular, the epidermal growth factor receptor (EGFR), a tyrosine kinase growth factor receptor involved in CRC development and progression, is exploited by the newest monoclonal antibody that is available for use in CRC patients. Cetuximab, the first chimeric monoclonal antibody, which has been generated against the EGFR, is currently registered in USA, Europe and worldwide, in combination with irinotecan in the treatment of metastatic CRC patients who have progressed on irinotecan containing chemotherapy. Cetuximab is well tolerated and does not exacerbate the toxicity of concomitant chemotherapy. Furthermore, a series of phase III clinical trials are currently evaluating the combination of cetuximab with standard chemotherapy regimens in the first-line treatment chemotherapy-naïve patients with metastatic CRC.

A review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina.

Several lactic acid bacteria (LAB) associated with meat products are important natural bacteriocin producers. Bacteriocins are proteinaceous antagonistic substances that are important in the control of spoilage and pathogenic microorganisms. The use of LAB as bioprotective cultures to extend the shelf life of fresh meat can improve microbial stability and safety in commercial meat preservation. Lactobacillus curvatus CRL705 used as a protective culture in fresh beef is effective in inhibiting Listeria innocua and Brochothrix thermosphacta as well as the indigenous contaminant LAB, retaining its inhibitory effect at low temperatures and having a negligible effect on meat pH. In addition to the hurdle represented by low temperature and vacuum-packaging, the use of live cells of Lb. curvatus CRL705 seems more feasible from an economic point of view - and without legal restrictions - compared to the addition of purified bacteriocins. A description of meat-borne bacteriocins and their application in meat to extend shelf life is discussed.

[Study on the interaction between the skin detoxifying enzyme glutathione S-transferase and the substances listed in the CE/39/2000 rules with the "skin" annotation, finalized to the biological monitoring of exposed subjects]. / Studio dell'interazione fra l'enzima detossificante cutaneo glutatione S-trasferasi e le sostanze elencate nella normativa CEE/39/2000 con notazione 'skin' finalizzato al monitoraggio biologico di soggetti esposti.

The interaction among chemicals listed in the Directive CE/39/2000 with skin notation and glutathione S-transferase (GSTP1-1) was studied by following two different experimental approaches. The compounds were incubated with the purified GST isoenzyme GSTP1-1 as well as with the human keratinocytes (PR5) selectively expressing GSTP1-1. Some of the molecules affected the enzymatic activity of both the purified and the intracellular GSTP1-1. In particular, 1,2-dichlorobenzene (DCB), ethylbenzene (ETB), cumene, Sulphotep and 2-eptanone (2-EPT) behaved as inhibitors of the purified GSTP1-1 enzyme, with different inhibition properties according to molecular structure. With the exception of Sulphotep showing a Ki value of 0.2 mM, all compounds reported above were characterized by high Ki values (between 2 and 16 mM) and therefore by low affinity towards GSTP1-1. These results make unlikely the use of a biosensor, based on immobilized GSTP1-1, for the detection of these molecules. On the contrary, Sulphotep can be the object of future investigations. It has to be stressed that the above listed compounds were effective on human keratinocytes, at concentrations two order of magnitude lower than that effective on purified GSTP1-1. In particular, cumene and DCB triggered a clear increase of the intracellular GSTP1-1 activity at concentrations lower than 0.1mM. These interesting results let to hypothesize the use of GSTP1-1 present in the keratinocytes as a marker for biological monitoring of workers exposed to these compounds as well as to evaluate the skin permeability of toxic compounds, not yet identified with a skin notation.

DNA ploidy as a significant prognostic factor after radical resection for large bowel carcinoma: a prospective study.

It is controversial whether cell DNA measurement is a reliable method to predict prognosis in radically resected large bowel carcinomas. A study using strict rules was undertaken on 54 consecutive patients to clarify the usefulness of DNA ploidy by image cytometric analysis as a prognostic indicator. The tumors were 39% diploid and 61% aneuploid. DNA ploidy was not related to more advanced stages and it, with histological grade and Dukes' stage, was an independent covariate and influenced both disease-free and overall survival. Aneuploid tumors showed the worse prognosis in all Dukes' stages. This prospective study shows that DNA ploidy is an important independent prognostic indicator of clinical outcome and recurrence rate, and it should be used to both select high-risk patients and guide postoperative treatments, particularly in early-stage large bowel carcinomas.

In vitro sodium salicylate release from a sucrose polyester matrix.

Sucrogel H-70, a recently synthesized polyester of sucrose, was investigated as a potential drug delivery matrix. In this study, the release of sodium salicylate was examined from a reservoir-controlled release device. It is shown that sodium salicylate release follows zero-order kinetics in the three media studied (i.e., water and simulated gastric and intestinal fluids).

Intraotic administration of gentamicin: a new method to study ototoxicity in the crista ampullaris of the bullfrog.

A new method of local gentamicin administration was tested in the bullfrog inner ear to achieve ototoxic-induced hair cell destruction. Gelfoam pledgets soaked with known amounts of gentamicin were inserted into the perilymphatic cisterna of the bullfrog through a ventral surgical approach. A dose of 1.20 mg gentamicin, consistent with a perilymphatic concentration of 65 microg/ml, resulted in the desired ototoxic-induced hair cell damage, that is, complete hair cell destruction with minimal disruption of other components of the sensory epithelium. This study demonstrates that this is a useful and simple method to investigate the process of vestibular ototoxicity and hair cell regeneration, including aspects of hair cell destruction and repair.

Hemodynamic and pulmonary changes during open, carbon dioxide pneumoperitoneum and abdominal wall-lifting cholecystectomy. A prospective, randomized study.

BACKGROUND: Carbon dioxide (CO2) pneumoperitoneum effects are still controversial. The aim of this study was to investigate cardiopulmonary changes in patients subjected to different surgical procedures for cholecystectomy. METHODS: In this study, 15 patients were assigned randomly to three groups according to the surgical procedure to be used: open cholecystectomy (OC), CO2 pneumoperitoneum cholecystectomy (PP), and laparoscopic gasless cholecystectomy (abdominal wall lifting [AWL]), respectively. A pulmonary artery catheter was used for hemodynamic monitoring in all patients. A subcutaneous multiplanar device (Laparo Tenser) was used for abdominal wall lifting. To avoid misinterpretation of results, conventional anesthesia was performed with all parameters, and the position of the patients held fixed throughout surgery. The following parameters were analyzed: mean arterial pressure (MAP), heart rate (HR), cardiac output (CO), cardiac index (CI), stroke volume index (SVI), central venous pressure (CVP), systemic vascular resistances index (SVRI), mean pulmonary arterial pressure (MPAP), pulmonary capillary wedge pressure (PCWP), pulmonary vascular resistances index (PVRI), peak inspiratory pressure (PIP), end-tidal CO2 pressure (ETCO)2, CO2 arterial pressure (PaCO2), and arterial pH. RESULTS: All the operations were completed successfully. The Laparo Tenser allowed good exposition of the surgical field. A slight impairment of the cardiopulmonary functions, with reduction of SVRI, MAP, and CI and elevation of pulmonary pressures and vascular resistance, followed induction of anesthesia. However, these effects tended to normalize in the OC and AWL groups over time. In contrast, CO2 insufflation produced a complex hemodynamic and pulmonary syndrome resulting in increased right- and left side filling pressures, significant cardiac index reduction, derangement of the respiratory mechanics, and respiratory acidosis. All of these effects normalized after desufflation. CONCLUSIONS: Cardiopulmonary adverse effects of general anesthesia were significant but transitory and normalized during surgery. Carbon dioxide pneumoperitoneum caused a significant impairment in cardiopulmonary functions. In high-risk patients, gasless laparoscopy may be preferred for reliability and absence of cardiopulmonary alterations.

Protonation and silver(I) complex-formation equilibria of some amino-alcohols.

Formation constants of the silver(I) complexes with some amino-alcohols have been determined at 25 degrees C in 0.5 M KNO(3) by means of two independent potentiometric measurements employing glass and silver electrode. The ligands considered are: sec-butylamine, 2-amino-1-propanol, 2-amino-1-methoxy-propane, 2-amino-2-methyl-1-propanol, 2-amino-1-butanol, 2-amino-1-pentanol, 2-amino-1-hexanol, 2-amino-1,3-propanediol, 2-amino-1,3-hexandiol, 2-amino-2-methyl-1,3-propanediol and Tris(hydroxymethyl)-aminomethane. Protonation constants of the selected ligands have also been determined. Calculations were made using HYPERQUAD computer program. The influence exerted by the introduction of hydroxy groups and by the presence of alkyl residuals in the ligand structure on the formation equilibria, is discussed.

Complex formation equilibria of some beta-amino-alcohols with lead(II) and cadmium(II) in aqueous solution.

A study of complex formation equilibria of some beta-amino-alcohols with lead(II) and cadmium(II) ions at 25 degrees C and in 0.5 M KNO(3) is reported. The amino-alcohols considered are 2-amino-1-propanol, 2-amino-1-butanol, 2-amino-1-pentanol and 2-amino-1,3-propanediol. sec-Buthylamine and 2-amino-1-methoxy-propane have been also considered for comparison. The results are discussed in terms of ligand structure, paying attention to the number of hydroxyl groups and to the length of the alkyl residual. A weak contribution of the alcoholic oxygen in the coordination of cadmium(II) and the presence of a mixed hydroxyl species in lead(II) containing systems are hypothesized.

A spectrophotometric-partial least squares (PLS-1) method for the simultaneous determination of furosemide and amiloride hydrochloride in pharmaceutical formulations.

A numerical method, based on the use of spectrophotometric data coupled to PLS-1 multivariate calibration, is reported for the simultaneous determination of furosemide and amiloride hydrochloride in synthetic samples and commercial tablets. The method was applied in the concentration ranges of 8.0-13.0 mg l(-1) for furosemide and 1.0-1.6 mg l(-1) for amiloride hydrochloride. Its accuracy and precision were determined, and it was validated by the analysis of synthetic mixtures of both drugs. The method was successfully applied to the quantitation of furosemide and amiloride hydrochloride in three different pharmaceutical formulations, providing results in agreement with those obtained by HPLC. It allowed the rapid, accurate and precise simultaneous estimation of the concentration of both analytes of interest in spite of their important spectral overlap, high concentration relationship and the presence of small amounts of different, unmodelled, absorbing excipients.

Granular cell tumors of the vulva.

Granular cell tumor (GCT), although nearly ubiquitous, is seen infrequently in the vulva. A review of the surgical pathology files from Grady Memorial Hospital, Atlanta, Georgia, from 1983 through 1987 identified eight cases of vulvar GCT. Five of the eight patients had more than one skin and soft tissue lesion. Two of the five had biopsy-proven multicentric GCT with a unique clinical course. One of the patients was a 32-year-old woman with multiple vulvar, lingual, laryngeal, bronchial and pulmonary GCT, necessitating multiple excisions and ultimately pneumonectomy. The second patient had multiple GCTs in the vulva and inguinal area and finally in both lungs, resulting in her death at age 39. No dependable microscopic features could be identified to distinguish benign GCT from its more aggressive variant. However, Feulgen DNA histomorphometry demonstrated aneuploidy in the patient with apparent lung metastases, whereas the tumors from patients with a benign course as well as from the patient with multiorgan involvement were diploid. In three of five patients who could be interviewed there was a history of soft tissue tumors in members of the family. The multifocal nature and possible familial component of GCT need to be explored further.

Retroperitoneal abscess after retained stones during laparoscopic cholecystectomy.

Laparoscopic cholecystectomy is associated with a significant risk of gallbladder perforations with bile and stone spillage. The retrieval of dropped stones is sometimes impossible, and intraperitoneally retained stones can be the source of serious complications, such as inflammatory masses or abscesses. The authors describe a patient in whom a large retroperitoneal abscess developed as a result of missed stone fragments during cholecystectomy. Although several cases of intraperitoneal abscess have been reported in the literature, retroperitoneal collection is very uncommon. Crushed and infected stones seem to be the essential prerequisite for abscess formation. Therefore, every attempt should be made to avoid stone spillage and intra-abdominally retained stones. Conversion to open surgery has to be considered in the presence of adverse factors, such as primary acute cholecystitis or cholecystitis induced by previous sphincterotomy, or bilirubinate stones believed to be infected. Abdominal abscess treatment requires removal of missed stones by surgical incision or new laparoscopic procedure because simple percutaneous drainage is usually unsuccessful.

[Transforming growth factor beta (TGF-beta) in Gaucher's disease. Preliminary results in a group of patients and their carrier and non-carrier relatives]. / Transforming growth factor beta (TGF-beta) en la enfermedad de Gaucher. Resultados preliminares en un grupo de enfermos y familiares portadores y no portadores.

BACKGROUND: The poor phenotype/genotype correlation in Gaucher's disease makes difficult therapy-decision-making and prevention of complications. Gaucher's cells and tissue fibrosis are the earliest findings of the disease. Transforming growth factor ss (TGF-beta1) is the key cytokine involved in the regulation of tissular scarring and fibrosis. The aim of the study was to ascertain if there are differences in plasma TGF-beta1 between Gaucher's disease patients, carriers and non-carriers healthy people and whether there is any correlation between plasma TGF-beta1 and clinical phenotype among patients. PATIENTS AND METHOD: Plasma TGF-beta1 was measured in 11 patients with Gaucher's disease, 12 carriers and 10 healthy people. Patients were further evaluated to know their liver and spleen size, bone involvement, hemoglobin, leukocyte and platelet count and the Zimran's severity score index (SSI). Plasma concentration of TGF-beta1 was determined by RIA phenotypic sandwich antibodies assay and quantified by a colorimetric procedure. Sensitivity was 25 pg/ml and specificity (cross reactivity) < 5% with beta2-TGF and beta3-TGF. STATISTICS: ANOVA and T-test were applied for mean comparisons and subgroup analyses. RESULTS: Plasma TGF-beta1 values were increased in Gaucher's disease patients (98.4 [91.4] pg/ml) over carriers (47.2 [21,7] pg/ml; p = 0.04) and healthy relatives (40.8 [9.8] pg/ml; p = 0.02). No differences in patients subgroups, with regard to SSI or bone involvement, were observed. CONCLUSIONS: Plasma TGF-beta1 levels are increased in this group of patients with Gaucher's disease. Since there is no correlation between the plasma values and the phenotypic expression, TGF-beta1 could merely be a marker of macrophage activation.

[Hereditary colonic cancer. Analysis of 3 families at risk]. / Il cancro del colon eredo-familiare. Analisi di tre famiglie a rischio.

Hereditary colorectal cancer is a clearly identified entity today; it is transmitted as a dominant autosomal heritage, and represents about 25% of large bowel malignancies. The poor prognosis of this tumor, one of the most frequent in Western Countries, despite the improvements in therapeutic field, is due to a delated diagnosis that affects an advanced stage of disease in which the results of treatment are significantly worse, whereas cumulative 5-year survival rate is 35% vs 95% in early stage tumors. Existence of an inherited cancer, even in the absence, at present, of specific biomarkers, allows to identify a high risk population in which early diagnosis of colorectal cancer is possible, on the basis of anamnestic and clinical features. Only in this way, with the extension of the results to general population, prognosis of colorectal cancer will be improved.

[Multidisciplinary treatment of hepatocarcinoma]. / Il trattamento multidisciplinare dell'epatocarcinoma.

Radical treatment of the hepatocellular carcinoma (HCC) is complete surgical removal; it may be done by resection or total hepatectomy. Although multicentric carcinogenesis predicts that liver transplantation is likely adequate to treat both the hepatoma and the underlying cirrhosis, it doesn't seem justified in the advanced stages or in absence of end-stage liver disease and therefore liver resection remains the treatment of choice for radical cure of HCC. However, low resectability and high recurrence rate make surgery alone ineffective. Unresectable HCC may be converted to resectable by multimodality radiation/chemotherapy, and embolization of portal branch feeding tumour, improving the function of the nonembolized liver, can extend the surgical indications for HCC. Adjuvant chemoembolization has already shown to reduce recurrence rate after radical resection and it should be widely applied. In unresectable or not converted HCCs as well as in postoperative recurrence, alternative therapies, particularly as multimodality treatment, can improve survival rate. To date, multidisciplinary treatment of hepatocellular carcinoma, waiting for further studies on newer modalities (prevention and gene therapy, especially), represents the best way to improve long-term results.

[Survey of carriers of Neisseria meningitidis in the health area of Gran Canary]. / Encuesta de portadores de Neisseria Meningitidis en el Area de Salud de Gran Canaria.

BACKGROUND: A) To ascertain the rate of carriers and the Types of Neisseria Meningitidis circulating in the population resident in the health jurisdiction of Gran Canaria. B) to ascertain the pattern of distribution of such carriers. METHODS: A descriptive transversal design was made, with a random sampling in multiple stages and by conglomerates. A minimum sample size was determined at 707 individuals for an expected prevalence of 8.6%, with a rate of reliability of 95.6% and a precision of 0.02. Assuming that 15% if the individuals would not be willing to co-operate, the sample size was increased to 831 individuals, distributed in each conglomerate in proportion to the existing population. This size was distributed in turn into four groups by age and sex, in proportion to their significance in each basic health care zone selected at random. The individuals in the sample were identified from among those who attended the blood extraction units, and after they had passed the criteria of exclusion, their co-operation was requested as volunteers in the study. If they accepted, a questionnaire was filled out with a number of variables of epidemiological interest and a pharyngeal smear was taken. Since the Primary Care units were selected on a simple random basis, and the same method was used to select the individuals within the units, the estimate of the prevalence was made by means of an unbiased estimator. RESULTS: A total of 828 samples were obtained, that is, 99.6% of the number foreseen. With the exception of three, all of the individuals selected participated voluntarily in the study, a circumstance rendering it highly representative. All of the strains obtained corresponded to N. Meningitidis Serogroup B, except for one identified as N. Meningitidis Serogroup C Sero/Subtype 4:P1.2,5. The strains of N. Meningitidis serogroup B identified corresponded to 25 different sero-subtypes. The prevalence determined after having studied the sample was 6.45%, the variance = 0.0275 and the standard error = 1.66. We can conclude with a 95% degree of reliability that the prevalence of carriers of N. Meningitidis in the Gran Canaria health jurisdiction is estimated to be between 3.2% and 9.7%. CONCLUSIONS: A clear predominance of N. Meningitidis serogroup B strains among carriers has been verified. There are no statistically significant differences in the prevalence observed among the different age groups nor between sexes.

[Aging of the population and incidence of primary acquired hemopathies in the Autonomous Community of Aragón]. / Envejecimiento poblacional e incidencia de hemopatías primarias adquiridas en un área de la Comunidad Autónoma de Aragón.

BACKGROUND: Aging probably comprises one of the major factors contributing to the onset of acquired primary blood diseases (APBD's) most of which are of a chronic type. The purpose of this study is to analyze the rate of occurrence (RO) of HPA in a population of 522,621 inhabitants (Males: 252,721; Females: 269,900) showing a negative vegetative growth (-1.4/10(5) inhabitants/year), said occurrence being dealt with separately for the population under age 60 and the population over age 60. METHODS: In January-December, 1994, a estimate was made of the HPA rate of occurrence and rate of analyses among the patients from the area in question, dealing separately with those under age 60 and over age 60. The diagnostic criteria applied were Monoclonal Gammopathies of Undetermined Significance (MGUS's) in keeping with Kyle's criteria. Multiple Myeloma (MM) and Chronic Lymphatic Leukemia (CLL) in keeping with the Myeloma Task Force criteria. Non-Hodgkin's Lymphoma (NHL) and Hodgkin's Disease (HD) in keeping with the REAL classification, Myelodisplasic Syndromes (MS's) and Acute Leukemia (AL) in keeping with the FAB classification, Chronic Myeloproliferative Syndromes (CMS's) in keeping with the PVSG. For calculating the rates of occurrence, descriptive epidemiological methods were used. RESULTS: The highest rates of blood analyses as the result of suspected APH's fell within the over 60 age group (p < 0.0001). During the length of time analyzed, a total of 302 APH's (< 60/> or = years: 100/202, p < 0.0001) were diagnosis, being worthy of special mention: 84 MGUS's; 21 MM's; 57 NHL's; 26 CLL's; 33 CMS's; 11 AL's and 14 HD's. The spread by gender was: Males: 177; Females 125. Average age: 63.54 years (age range 19-92). The rates of occurrence (cases/10(5) inhabitants/year) were (< age 60/> or = age 60): overall: 31.31/178.86; MGUS: 7.37/52.87; MM: 1.84/13.21; NHL: 5.53/34.36; CLL: 1.53/18.50; MS: 0.62/27.31; CMS: 5.52/16.74; AL: 1.53/5.29; HD: 3.68/1.76. CONCLUSIONS: The highest rate of analyses as the result of suspected APH was found among those over 60 years of age. The overall occurrence of APH is significantly higher in those over age 60, as well as for each type of APH taken into account, except for AL and HD. The highest rate of occurrence is that of MGUS, NHL and MS's, especially in males.