Quantitative proteomic analysis in symptomatic and asymptomatic apical periodontitis.

AIM: To quantitatively and qualitatively compare the host proteomic profile in samples of symptomatic and asymptomatic apical periodontitis (AP) using nano-liquid chromatography-electron spray tandem mass spectrometry. METHODOLOGY: Samples were obtained from 18 patients with radiographically evident AP, divided into symptomatic and asymptomatic groups (nine per group) according to clinical characteristics. After sample collection, protein extraction, purification and quantification of the samples were performed, which were analysed by reverse-phase liquid chromatography coupled to mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in expression of proteins between the groups were calculated using the Monte Carlo algorithm, considering P < 0.05 for down-regulated proteins and 1 - P > 0.95 for up-regulated proteins. Proteins were identified with the embedded ion accounting algorithm in the software and a search of the Homo sapiens UniProt database. RESULTS: A total of 853 individual human proteins were identified. In the quantitative analysis, common proteins to both groups accounted for 143 proteins. Differences in expression between groups resulted in 51 up-regulated proteins (1 - P > 0.95) in the symptomatic group, including alpha-1-antitrypsin, protein S100-A8, myeloperoxidase, peroxiredoxin and lactotransferrin. This group also had 43 down-regulated proteins (P < 0.05), comprising immunoglobulin, neutrophil defensin, pyruvate kinase and alpha-enolase. The qualitative analysis considered only the exclusive proteins of each group. For the symptomatic group, 318 complete proteins and 29 fragments were identified, such as dedicator of cytokinesis protein, intersectin, prostaglandin, phospholipase DDHD2 and superoxide dismutase. For the asymptomatic group, 326 complete proteins and 37 fragments were identified, including azurocidin, C-reactive protein, collagen alpha, cathepsin, heat shock and laminin. CONCLUSIONS: Quantitative differences in the expression of common proteins in cases of symptomatic and asymptomatic AP were found, which were mostly related to host immune response in both groups. Exclusive proteins in the symptomatic group were mainly related to the host response to the presence of viruses in endodontic infections, oxidative stress and proteolytic enzymes. The results provide a basis for a better understanding of cellular and molecular pathways involved in AP, establishing specific proteomic profiles for symptomatic and asymptomatic conditions.

Concurrence of lower jaw skeletal anomalies in triploid Atlantic salmon (Salmo salar L.) and the effect on growth in freshwater.

Triploid Atlantic salmon populations are associated with higher prevalence of lower jaw skeletal anomalies affecting fish performance, welfare and value deleteriously. Anomalous lower jaw can be curved downward (LJD), shortened (SJ) or misaligned (MA). Two separate groups of triploid Atlantic salmon (~12 g) with either normal lower jaw (NOR) or SJ were visually assessed four times over three months for presence and concurrence of jaw anomalies (with severity classified) and opercular shortening to understand the relatedness of these anomalous developmental processes. The prevalence of jaw anomalies increased in both groups over time (NOR group - SJ, LJD and MA combined 0-24.5%; SJ group - LJD and MA combined 17-31%). SJ and LJD occurred both independently and concurrently whereas MA exclusively concurred with them. All three anomalies could be concurrent. Severity of both LJD and SJ increased in the SJ group only. Opercular shortening recovery was observed in both groups but at a slower rate in the SJ group. The SJ group specific growth rate (SGR) was significantly (P < 0.05) lower than the NOR group. This study demonstrated the concurrence of SJ, LJD and MA and showed possible deleterious consequences deriving from the conditions.

Skeletal anomaly assessment in diploid and triploid juvenile Atlantic salmon (Salmo salar L.) and the effect of temperature in freshwater.

Triploid Atlantic salmon tend to develop a higher prevalence of skeletal anomalies. This tendency may be exacerbated by an inadequate rearing temperature. Early juvenile all-female diploid and triploid Atlantic salmon were screened for skeletal anomalies in consecutive experiments to include two size ranges: the first tested the effect of ploidy (0.2-8 g) and the second the effect of ploidy, temperature (14 °C and 18 °C) and their interaction (8-60 g). The first experiment showed that ploidy had no effect on skeletal anomaly prevalence. A high prevalence of opercular shortening was observed (average prevalence in both ploidies 85.8%) and short lower jaws were common (highest prevalence observed 11.3%). In the second experiment, ploidy, but not temperature, affected the prevalence of short lower jaw (diploids > triploids) and lower jaw deformity (triploids > diploids, highest prevalence observed 11.1% triploids and 2.7% diploids) with a trend indicating a possible developmental link between the two jaw anomalies in triploids. A radiological assessment (n = 240 individuals) showed that at both temperatures triploids had a significantly (P < 0.05) lower number of vertebrae and higher prevalence of deformed individuals. These findings (second experiment) suggest ploidy was more influential than temperature in this study.

Identification of components of health complexity on internal medicine units by means of the INTERMED method.

BACKGROUND: The INTERMED was developed for the early identification of biological, psychological, social and health system factors considered interacting in health complexity. This is defined as the interference with the achievement of expected or desired health and service use outcomes when patients are exposed to standard care. OBJECTIVE: The aim of this study was to test the INTERMED's ability to identify 'case' and 'care' complexity, identifying patients that would especially benefit from integrated care. METHODS: Observational longitudinal study of Internal medicine in patients in two National Health System hospitals in Spain using the INTERMED (patients scoring ≥ 21 were considered to be 'complex'); the Cumulative Illness Rating Scale (CIRS), a severity of illness assessment; and standard clinical variables. RESULTS: Six hundred and fifteen consecutives were included, and the prevalence of health complexity was 27.6%. The greatest differences between patients with and without health complexity were observed in the non-biological domains. Eighty-five per cent of patients with health complexity had non-biological items considered to require timely (immediately or soon) assistance or intervention compared to 30% of those without, nearly a threefold difference. Complex patients had a significantly higher number of medical diagnoses (p = 0.002) and number of psychiatric referrals (p = 0.041), but there were no differences in CIRS scores or lengths of stay. CONCLUSION: The INTERMED has the potential to identify a considerable subset of complex internal medicine inpatients for which timely corrective action related to non-biological risk factors not typically uncovered during standard medical evaluations would be considered beneficial.

Isolation and characterization of a female-specific DNA marker in the giant freshwater prawn Macrobrachium rosenbergii.

In this study, a female-specific DNA marker in the freshwater prawn Macrobrachium rosenbergii was identified through amplified fragment length polymorphism (AFLP). The AFLP-derived sequence-characterized amplified region (SCAR) marker was tested in over 200 individuals, giving reproducible sex identification. Further molecular characterization of the sex-marker's genomic region (∼ 3 kb long) revealed the presence of tandem and inverted repeats. The ∼ 3-kb sequence was identified both in male and female prawns, but with subtle differences: a deletion of 3 bp (present in female prawn but absent in male prawn) identified upstream of the SCAR marker sequence and two female-specific single-nucleotide polymorphisms, both indicating that male prawns are homozygous, whereas female prawns are heterozygous in this locus. Fluorescent in situ hybridization showed the ∼ 3-kb sequence to be unique: to the best of our knowledge, this is the first report of a unique sex-specific sequence observed in situ in crustaceans. The sex-specific marker identified in M. rosenbergii may have considerable applied merit for crustacean culture in that it will enable the determination of genetic sex at early developmental stages when phenotypic differences are not identifiable.

Incidence and lifetime risk of dementia and Alzheimer's disease in a Southern European population.

OBJECTIVE: To calculate both the incidence rates and the lifetime risk (LTR) of dementia and Alzheimer's disease (AD). METHODS: A two-phase case-finding procedure was implemented in a cohort of 4057 cognitively intact individuals 55+ years of age living in Zaragoza, Spain, and followed-up at 2.5 and 4.5 years. Age- and sex-specific incidence rates were calculated. A mortality-adjusted, multivariate model was used to document LTRs. RESULTS: The incidence rate of dementia continued to rise after the age of 90 years, but was slightly lower than in North and West European studies. Only a tendency for an increased LTR with age was observed. Thus, LTR was 19.7% for a 65-year-old woman and 20.4% at the age of 85 years, the corresponding figures for AD being 16.7% and 17.6%. The LTR of AD was higher in women and was about twice as high among illiterate individuals when compared with individuals with higher educational levels. CONCLUSIONS: The incidence rate of dementia in this Southern European city was slightly lower than in previous studies in North-West Europe. LTR of dementia and AD seems to be slightly increased with age. The association of illiteracy with higher LTR of AD is intriguing.

Development and Interrogation of a Transcriptomic Resource for the Giant Triton Snail (Charonia tritonis).

Gastropod molluscs are among the most abundant species that inhabit coral reef ecosystems. Many are specialist predators, along with the giant triton snail Charonia tritonis (Linnaeus, 1758) whose diet consists of Acanthaster planci (crown-of-thorns starfish), a corallivore known to consume enormous quantities of reef-building coral. C. tritonis are considered vulnerable due to overexploitation, and a decline in their populations is believed to have contributed to recurring A. planci population outbreaks. Aquaculture is considered one approach that could help restore natural populations of C. tritonis and mitigate coral loss; however, numerous questions remain unanswered regarding their life cycle, including the molecular factors that regulate their reproduction and development. In this study, we have established a reference C. tritonis transcriptome derived from developmental stages (embryo and veliger) and adult tissues. This was used to identify genes associated with cell signalling, such as neuropeptides and G protein-coupled receptors (GPCRs), involved in endocrine and olfactory signalling. A comparison of developmental stages showed that several neuropeptide precursors are exclusively expressed in post-hatch veligers and functional analysis found that FFamide stimulated a significant (20.3%) increase in larval heart rate. GPCRs unique to veligers, and a diversity of rhodopsin-like GPCRs located within adult cephalic tentacles, all represent candidate olfactory receptors. In addition, the cytochrome P450 superfamily, which participates in the biosynthesis and degradation of steroid hormones and lipids, was also found to be expanded with at least 91 genes annotated, mostly in gill tissue. These findings further progress our understanding of C. tritonis with possible application in developing aquaculture methods.

Prevalence and implications of psychopathological non-cognitive symptoms in dementia.

OBJECTIVE: Clinical experience and recent population studies suggest that psychopathological, non-cognitive symptoms are both frequent and relevant in dementia. METHOD: A representative community sample (n = 4,803 individuals, 55 + years) was interviewed in a two-phase design. The Geriatric Mental Sate (GMS) was used for assessment and cases were diagnosed according to DSM-IV-TR criteria. RESULTS: The prevalence of non-cognitive symptoms (1 + symptoms) in cases of dementia (n = 223) was 90.1%, and negative-type symptoms were most frequently found. A GMS 'apathy-related symptom cluster' (anergia, restriction of activities and anhedonia) was significantly more frequent in the demented (55.6%) than in non-cases (0.7%; specificity = 99.2%). In both dementia of Alzheimer's type and vascular dementia, number of symptoms tended to be inversely related to severity of dementia, but psychopathological profiles differed. CONCLUSION: Non-cognitive, negative-type symptoms are very frequent in cases of dementia living in the community. They have powerful specificity in the distinction with non-cases, and might change current concepts of dementia.

Testosterone measured from amniotic fluid and maternal plasma shows no significant association with directional asymmetry in newborn digit ratio (2D:4D).

Foetal sex hormones can have powerful and far-reaching effects on later phenotype. However, obtaining accurate measurements is difficult for ethical reasons, and researchers often employ proxy variables to examine their effects. The relative length of the second and fourth fingers (digit ratio or 2D:4D) is frequently used for this purpose, as it is hypothesized to index variance in prenatal androgen and oestrogen exposure. Most studies employing this method examine digit ratio for the right hand (R2D:4D) and/or left hand (L2D:4D), though the mean value (M2D:4D) (i.e., the average of R2D:4D and L2D:4D) and directional asymmetry (D[R-L]) (i.e., R2D:4D minus L2D:4D) are also commonly used. As no published studies have examined M2D:4D or D[R-L] in relation to testosterone measured from amniotic fluid, we conducted a secondary analysis of data published by Ventura et al. The sample comprises 106 mothers from Portugal who underwent amniocentesis during the second trimester and their neonates. Newborn M2D:4D was negatively correlated with amniotic testosterone in females (P<0.05) but not in males; no significant association was observed between amniotic testosterone and D[R-L] in either sex. In addition, we examined testosterone measured from maternal circulation during the second trimester, and found that it was not a significant predictor of M2D:4D or D[R-L] in male or female infants. Further research should aim to measure the ratio of testosterone to oestradiol present in amniotic fluid and maternal plasma, to examine whether either is a predictor of digit ratio variables at different stages of postnatal development.

Neurogenic bladder in the settings of congenital Zika syndrome: a confirmed and unknown condition for urologists.

INTRODUCTION: Congenital Zika syndrome (CZS) is a recently discovered condition that affects central nervous system structures that control the lower urinary tract. The first cases of neurogenic bladder (NB) were recently reported as a sequalae of CZS in neurologically impaired children. OBJECTIVE: Our goal is to further evaluate NB in the setting of CZS, identifying urological risk indicators in hopes that early diagnosis will mitigate the impact of the disease. STUDY DESIGN: Urological assessment was performed in all patients with CZS and neurological impairment who were referred to our urodynamic clinic between June 2016 and May 2018. Neurogenic bladder was confirmed by urodynamic evaluation, and urological risk was based on urodynamic results. RESULTS: Sixty-nine patients with CZS were tested. The majority (63 patients, 91.3%) presented with overactive bladder with increased pressures and reduced capacity for age (table 1). Different urodynamic patterns were observed, and the association of reduced bladder capacity for age, high bladder-filling pressure, and increased postvoid residual were frequently observed. DISCUSSION: NB continues to be consistently diagnosed in our cohort of CZS, mostly with high-risk indicators for renal impairment. When not intervened upon in a timely manner, NB can cause progressive damage to the urinary tract, but the lack of knowledge that CZS causes NB delays investigation and treatment. Parents and health professionals will need to be sensitized to the risks that ZIKV can pose to the urinary tract so that appropriate therapies are initiated to prevent irreversible renal damage. CONCLUSION: NB is a common condition among our patients with CZS and microcephaly. This is a new cause of NB, unknown to urologists. While further investigation is necessary to understand long-term disease behavior and therapeutic response, increased knowledge among urologists may help to reduce morbidity related to untreated NB and to mitigate the disease burden for patients and families.

Identification and Characterization of an Insulin-Like Receptor Involved in Crustacean Reproduction.

Sexual differentiation and maintenance of masculinity in crustaceans has been suggested as being regulated by a single androgenic gland (AG) insulin-like peptide (IAG). However, downstream elements involved in the signaling cascade remain unknown. Here we identified and characterized a gene encoding an insulin-like receptor in the prawn Macrobrachium rosenbergii (Mr-IR), the first such gene detected in a decapod crustacean. In mining for IRs and other insulin signaling-related genes, we constructed a comprehensive M. rosenbergii transcriptomic library from multiple sources. In parallel we sequenced the complete Mr-IR cDNA, confirmed in the wide transcriptomic library. Mr-IR expression was detected in most tissues in both males and females, including the AG and gonads. To study Mr-IR function, we performed long-term RNA interference (RNAi) silencing in young male prawns. Although having no effect on growth, Mr-IR silencing advanced the appearance of a male-specific secondary trait. The most noted effects of Mr-IR silencing were hypertrophy of the AG and the associated increased production of Mr-IAG, with an unusual abundance of immature sperm cells being seen in the distal sperm duct. A ligand blot assay using de novo recombinant Mr-IAG confirmed the existence of a ligand-receptor interaction. Whereas these results suggest a role for Mr-IR in the regulation of the AG, we did not see any sexual shift after silencing of Mr-IR, as occurred when the ligand-encoding Mr-IAG gene was silenced. This suggests that sexual differentiation in crustaceans involve more than a single Mr-IAG receptor, emphasizing the complexity of sexual differentiation and maintenance.

Degree of cognitive impairment and mortality: a 17-year follow-up in a community study.

BACKGROUND: To test the hypothesis that cognitive impairment in older adults is associated with all-cause mortality risk and the risk increases when the degree of cognitive impairment augments; and then, if this association is confirmed, to report the population-attributable fraction (PAF) of mortality due to cognitive impairment. METHOD: A representative random community sample of individuals aged over 55 was interviewed, and 4557 subjects remaining alive at the end of the first year of follow-up were included in the analysis. Instruments used in the assessment included the Mini-Mental Status Examination (MMSE), the History and Aetiology Schedule (HAS) and the Geriatric Mental State (GMS)-AGECAT. For the standardised degree of cognitive impairment Perneczky et al's MMSE criteria were applied. Mortality information was obtained from the official population registry. Multivariate Cox proportional hazard models were used to test the association between MMSE degrees of cognitive impairment and mortality risk. We also estimated the PAF of mortality due to specific MMSE stages. RESULTS: Cognitive impairment was associated with mortality risk, the risk increasing in parallel with the degree of cognitive impairment (Hazard ratio, HR: 1.18 in the 'mild' degree of impairment; HR: 1.29 in the 'moderate' degree; and HR: 2.08 in the 'severe' degree). The PAF of mortality due to severe cognitive impairment was 3.49%. CONCLUSIONS: A gradient of increased mortality-risk associated with severity of cognitive impairment was observed. The results support the claim that routine assessment of cognitive function in older adults should be considered in clinical practice.

Increased enterocyte apoptosis and Fas-Fas ligand system in celiac disease.

Our aim was to evaluate whether increased enterocyte apoptosis was responsible for mucosal flattening in celiac disease (CD), and, since the mechanisms responsible for tissue injury in this condition are unknown, we studied the possibility that the Fas-Fas ligand (FasL) system may be involved. Endoscopic duodenal biopsy specimens from 12 patients with untreated and 12 with treated CD and 12 control subjects were evaluated for enterocyte apoptosis by the terminal deoxynucleotidyl transferase-mediated digoxigenin-deoxyuridine triphosphate nick-end labeling assay and for Fas and FasL expression by immunohistochemistry. A coculture of isolated enterocytes (targets) and purified lamina propria mononuclear cells (LPMCs) (effectors) was performed in the absence or presence of an antagonistic ZB4 anti-Fas antibody. We found a significant correlation between the degree of villous atrophy, morphometrically evaluated, and the level of enterocyte apoptosis, suggesting that mucosal flattening is a consequence of exaggerated epithelial cell death. Most celiac enterocytes express Fas, and LPMCs express FasL. The abolishment of enterocyte apoptosis observed in the presence of ZB4 antibody suggests that enterocytes are potential targets of lymphocyte infiltrate. These results directly demonstrate that FasL-mediated apoptosis is a major mechanism responsible for enterocyte death in CD.

[Sclerotherapy of thyroid cysts with tetracycline hydrochloride]. / Sclerosi delle cisti tiroidee mediante tetraciclina cloridrato.

In order to verify sclerotherapy efficiency in solitary thyroid cysts, needle aspiration of the cyst followed by instillation of tetracycline hydrochloride into the cyst cavity was performed in 110 patients; in thirty of these the aspirations was ultrasonically guided. A tight dressing on the neck was applied for 48 hours in order to maintain pressure on the cyst cavity. All patients were treated with L-T4 in TSH-suppressive doses. In a few patients (< 155), the tetracycline instillation caused local pain, lasting less than two hours. In no case, did cytological examination of the sediment of the aspirated fluid reveal any evidence of malignant cells. In 82 patients (84.54% of the 97 controlled patients), the last ultrasonic follow-up (performed at 3, 6, 9, 12 or 24 months) showed a decrease in lesion size. In particular, 16 patients (16.50%) had a complete cyst disappearance, 41 patients (42.17%) had a volume reduction of the cyst between 85 and 65%, and in 25 patients (25.77%) the reduction was more than 25%. In the remaining 15 patients (15.46%) a second treatment was performed, 3 of them also required a third treatment but in all these cases, the results seemed to be less satisfactory. The distinction in cyst size seemed to indicate that in the case of a large cyst (> 36 mm diameter), the treatment gives better results (92% success rate against 82% of the other patients). In conclusion, we are able to affirm that needle aspiration of the thyroid cyst with instillation of tetracycline hydrochloride is a simple, well tolerated and low cost technique.(ABSTRACT TRUNCATED AT 250 WORDS)

Surgical anatomy of the papilla of Vater and biliopancreatic ducts.

The present research was aimed at defining the surgical anatomy of the biliopancreatic ducts and of Oddi's sphincter. The numerous anatomic variations of the papilla, the millimetric distribution of its muscle fibres, and any morphological detail of clinical significance have been investigated. An integrated analysis of radiographic, tridimensional (casts), and histologic findings has been carried out in 49 of 64 autoptic bilio-duodenopancreatic specimens. Exact limits of the choledocus and Wirsung sphincters were defined. A consistent accumulation of circular muscle fibres could be seen, on the choledocus duct side, up to a mean distance of 13.6 mm from the papillary pore. However, more rarefied fibres were present up to 20.5 mm. Muscle fibres were seen to stop roughly on the pancreatic duct side at 7.3 mm from the papillary pore. The beginning of the sphincter was observed 2-3 mm above the papillary pore. There was no evidence suggesting the presence of upper, middle, and lower biliary sphincters. Five anatomic diversities of the Wirsung-choledocus confluence were found. The Y type was the most frequent (61.2%), followed by the U type (22.4%), V (14.3%), and II (2.1%). Santorini's duct with a normal papilla was present in 16 per cent of the cases. These data along with other interesting observations on antireflux mechanisms (Santorini's valves) and on the ductal space orientation appear to be useful guidelines for a physiopathological understanding of bilio-pancreatic diseases and for any therapeutic procedure on these structures.

Lack of prognostic significance in Goseki grading of gastric carcinoma.

The Goseki grading system, based on intracellular mucin content and tubular differentiation, was originally created to study the influence of histologic type on the mode of extension of gastric carcinoma. The prognostic value of this grading system was subsequently proposed and even recently supported, but controversies still remain about this topic. We applied the Goseki system on 114 cases of node-negative primary gastric cancer and compared Goseki groups with the other clinicopathologic features of the patients. Statistical analysis showed a significant correlation between Goseki grading and Laurèn classification, but failed to reveal any prognostic significance for this grading system. We believe that Goseki classification should not be routinely used for prognostic purposes.

Müllerian adenosarcoma of the uterus.

A case of adenosarcoma of the uterus in a 59-year-old woman is here reported. Adenosarcoma is a low malignant potential tumor with a benign glandular and a malignant stromal component. The treatment is usually hysterectomy with bilateral salpingo-oophorectomy. Debated is the usefulness of adjuvant chemotherapy, while radiation treatment is not beneficial. Long term follow-up is necessary for these patients because of high recurrence risk, mostly in cases with myometrial invasion.

Histopathological study of 110 cystectomy specimens for bladder cancer by an original mapping method.

The management of cystectomy specimens represents the first and most important step in the study of bladder cancer and related lesions. We carried out a study on 110 patients, applying an original mapping protocol which allowed to determine the exact topography of lesions, recognizing even the smallest ones and putting in evidence some rare histotype. A prevalence of high-grade, high-stage tumors was noted, as well as a remarkable frequency of precancerous lesions, mostly found in Brunn's nests. This latter finding could mean that in many cases a flat carcinoma becomes invasive within a Brunn's nest rather than in surface urothelium. We were also able to accurately evaluate prostatic pathology, finding incidental malignant lesions of this gland in 24.2% of the cystectomized males. The apparently worst prognosis of the patients who underwent chemotherapy depends on the fact that they had grades and stages higher than the untreated subjects. In conclusion, we believe that a more extensive sampling of cystectomy specimens gives highly reliable prognostic data and represents an unreplaceable tool in understanding bladder neoplasms.

Association between chronic lymphocytic leukaemia and secondary tumours: unusual occurrence of a neuroendocrine (Merkell cell) carcinoma.

The authors describe a case of cutaneous neuroendocrine (Merkell cell) carcinoma in a patient previously treated for Chronic Lymphocytic Leukaemia (CLL). The authors discuss some of the mechanisms concerning the increased frequency of a second tumour in patients with CLL and focus attention on the high incidence in CLL of secondary tumours, especially skin tumours, of which Merkell cell carcinoma is a rare example. The authors consider the possible therapeutic approaches, reported in the literature, for the treatment of this particular skin carcinoma, which on account of its aggressiveness and the preexistence of a leukaemic process requires a particularly careful therapeutic approach.

Establishment and characterization of a human prostatic carcinoma cell strain obtained from exfoliated tumor cells after prostatic massage.

During a control campaign connected to our main program on early diagnosis of prostatic carcinoma through tissue culture of prostatic fluid samples obtained after prostatic massage (M. Bologna et al., Eur. Urol., 14, 474-476, 1988), we isolated and characterized a human prostatic carcinoma cell strain from a 58-year-old patient with a grade III prostatic carcinoma. The epithelial cell strain, named PMU-23, has been passaged in vitro for 31 subculture cycles during a period of approximately 8 months, after which cell proliferation slowed down irreversibly. The isolation of this cell strain constitutes a renewed confirmation of the validity of our method for the early diagnosis of prostatic carcinoma and demonstrates some intermediate features in the progression of prostatic tumors. In addition, the study of limited-lifespan tumor cell strains in culture may extend the knowledge on prostatic cell biology, particularly toward the identification of intermediate steps of tumor progression, for a better approach of tumor therapy and prevention of metastatic spread.